Magazine abstract

The Changes in Expression of p35, p25 and Cdk5 Kinase Activity in Hippocampal Neuron Irradiated with X-ray

Mon, 27 Jun 2011 12:26:04 +0000

【Background】The malignant tumor incidence rate roars up globally these years with the speeding up of life tempo and global aging.Now it has become \'the second killer\'of human following the cardiovascular diseases which damage people\'s health seriously as all know.And it endangers human health seriously.Radiotherapy is one of the major therapeutic tools on malignant tumor and the radical cure tools on partial of them.According to WHO statistics in 1998,malignant tumor control rate of all over the body is 45%,operation occupies 22%and radiotherapy 18%.Over 70%malignant tumor patients had radiotherapeutic experience.However,X-ray can result in normal tissues injury at the same time of killing tumor cells.Radiation injury of brain is a radioactive syndrome caused by various kinds of reason and one of the most serious long-term complications after pate malignant tumors radiotherapy.Following oncotherapy level improvement and life span unceasing prolongation,the incidence of radiation brain injury goes up year by year.Radiation brain injury impacts living quality,life span and prognosis of patients seriously.But the mechanism of radiation brain injury is not clear,so the effective treatment is still lack up to now.Therefore,it is very important to study the radiation brain injury and explore new therapeutic targets.Cyclin-dependent kinase 5(CDK5) is a proline-directed protein kinase that phosphorylates serine and threonine residues.It is a special member of the cyclin-dependent kinase family(CDK).CDK5 phosphorylates a diverse list of substrates including KSP protein motif on MAP1b,KSPXK on tau or KSPXX on neurofilament protein.However,in contrast to other members of the cyclin-dependent kinase family,CDK5 is not directly involved in mediating progression through the cell cycle.Although CDK5 is expressed in most tissues,its activity is found predominantly in post-mitotic neurons.The reason for the localized activity of CDK5 is that its activators,p35 and p39,are expressed almost exclusively in the central nervous system(CNS).CDK5 is critical to CNS development.Furthermore,CDK5 likely plays a role in synaptic plasticity,cellular motility and adhesion,drug addiction and neurodegeneration.As monomeric CDK5 displays on enzymatic activity,an interaction with either p35 or p39 anchored to membrane is necessary for CDK5 activation.The proteolysis of p35 to p25,is likely important in regulating CDK5 activity.The truncated activators,p25,appear to have greater stability than their precursors and therefore,this may result in prolonged CDK5 activity resulting in hyperphosphorylation of available substrates.There is evidence to suggest that overexpression of CDK5 and p25 in neurons results in apoptosis.In addition,the expression and activity of CDK5 have been shown to increase during neuronal apoptosis.CDK5 inhibitors protected against both necrotic and apoptotic neuronal cell death in several different models.Focusing on the links between CDK5 activity and components of the cytoskeletal membrane and adhesion systems has allowed us to postulate a role for CDK5 in directing intracellular traffic in neurons.Microtubule dynamics and axonal transport might be targets for CDK5 through its phosphorylation of the microtubule-associated protein(MAP).Aberrant phosphorylation of the neuonal cytoskeleton by CDK5 could alter celluar trafficking and transport and contribute to neuronal dysfunction and death.So it is hypothesized that p25(a truncated form of p35) cause aberrant activation of CDK5,leading to the breakdown of cytoskeletal proteins and intracellular trafficking,which,in turn,contributes to neurodegeneration.Cleavage of p35 to p25,may be a critical step in this pathological cascade,including neurite retraction,microtubule collaose and apoptosis.But the exact mechanisms by which CDK5 can facilitate cell death processes have not been fully elucidated.There is no doubt that CDK5 is critical for the development and functioning of the CNS.In addition,there is increasing evidence that overactivation of CDK5 may contribute to neurodegenerative processes.So the question is whether CDK5 is involved in the pathogenesis of RBI.Thus,the aim of this study is to investigate the relationship between aberrant activation of CDK5 and RBI.On this ground,we exposed the hippocampal neurons at 12days to single x-ray doses of 30Gy,and then we observed the changes in the expression of p35/p25 by western-blot,and finally we observed the effect of inhibition of CDK5 so as to investigate if dysfunction of CDK5 is involved in the pathogenesis of RBI.【Objective】To study the changes in expression of p35,p25 and CDK5 kinase activity in cultured rats hippocampal neurons irradiated with x-ray and provide theoretic basis for prevention and treatment of radiation brain injury.【Methods】(1) The cultured rat hippocampal neurons at 12days were exposed to single x-irradiation with doses of 30Gy.(2) Western-blot was used to detect the levels of protein p35/p25,which correlates with the activity of CDK5.The levels of protein p35/p25 were assayed separately at 3.5h,4h,5h,6h after irradiation.(3) To observe the effect of roscovitine treatment on neuronal apoptosis induced by x-irradiation,the cells were stained with 4\',6-diamidino-2-phenylindole(DAPI) at 24 h after irradiation,the apoptotic changes were measured quantitatively by nuclear pyknosis.(4) All values were expressed as mean±SD.Statistical significance was defined as P≤0.05. 【Results】(1) Western-blot showed that p35(35KD) significantly increased(F(4,15) = 19.338,P=0.000,N=4).Compared with the control group the 3.5 hours group significantly increased to 1.52(P＜0.001),the 4 hours group significantly increased to 1.44(P＜0.001).The result also show that p25(25KD) significantly increased (F(4,15) =6.617,P=0.003,N=4).To compared withthe control group,the 6 hours group significantly increased after x-irradiation(P＜0.001 ).The changes of p35,p25 expression levels suggested that CDK5 may participate in the neuronal death of the hippocampus after x-irradiation.(2) Apoptosis experiment showed that administration roscovitine(10μM) at 15 minutes before x-irradiation can protect the hippocampal neuronal death induced by irradiation(F(2,13.121)= 146.226,P= 0.000,N=9).The percentage of nuclear pyknosis in the group of 30Gy(24.8%±3.97%) was significantly increased over that in the group of 0Gy at 24 h after irradiation(1.82%±1.08%,P＜0.001),while the percentage of nuclear pyknosis in the group of 30Gy & roscovitine(7.74%±2.27%) was significantly different from that in the group of 30Gy at 24 h after irradiation(P＜0.001).There was significant difference between the group of 30Gy & roscovitine and the group of 0Gy(P＜0.001).【Conclusion】(1) x-irradiation may activates CDK5 by enhancing the levels of p35,p25 in hippocampal neurons.(2) Inhibiting CDK5 activity by the CDK5 inhibitor roscovitine can significantly protect hippocampal neuronal cell from death,which suggest that dysfunction of CDK5 may be involved in the pathogenesis of radiation hippocampal neuron injury.

Evaluation of Left Cardiac Function in Healthy Pregnant Women Using Full Volume Three-Dimensional Echocardiography

Mon, 27 Jun 2011 12:26:04 +0000

Objective: Prominent adaptable changes of structure, function and hemodynamics have taken place in normal pregnant women\'s cardiovascular system. Because of its characteristics of convenience, noninvasion, accuratissime, cheap and strong repetitiveness, echocardiography became one of the important means of detecting maternal cardiac function by clinic from the later stage of the seventies. There were a large number of researches evaluating cardiac structure, systolic and diastole function using conventional ME, 2DE and doppler echocardiography. RT-3DE is a great technological break-through in the field of cardiac ultrasonic imaging for the past few years. Full volume imaging is one mode of RT-3DE.But the report of evaluating normal pregnant women\'s cardiac function by full volume imaging is rare now. The study was designed to investigate the value of the early quantitation in healthy pregnant women’s left cardiac function using new technique FV-3DE.Methods: The left heart of 52 healthy pregnant women (15 first trimester, 18 second trimester and 19 third trimester ) and 20 age matching healthy nonpregnant women(control group) were detected by FV-3DE and conventional echocardiography. The left cardiac parameters of healthy pregnant women group and control group were compared and analyzed.They were:left atrial function (LAVmax, LAVmin, LAEF, LASV), left ventricular mass (LVM, LVMI), left ventricular function (LVEDV, LVESV, LVSV, LVEF, LVCO, LVCI, PER, PFR); left cardiac structure and function (LVEDd, IVSd, LVPWd, LAs; E, A and E/A ).Meanwhile, the hemodynamic indexes (MBP,TPR) were analyzed . Results: The imaging satisfactory rates of FV-3DE were 100%(the control group and the first trimester), 88.9%(16/18, the second trimester), 78.9% (15/19, the third trimester), respectively. There was a high-flow,low-resistance hemodynamic state during normal pregnancy. TPR decreased during all three trimesters of pregnancy(P0.05). During gestational period the healthy pregnant women\'s cardiovascular system remodeled because of the growth of circulation volume and the long-term overloading. With the gestational development, the healthy pregnant women\'s left atrial and ventricular diameter and volume (LAs, LAVmax, LAVmin, LVEDd, LVEDV, LVESV), stroke volume (LASV, LVSV), LVCO and LVCI increased gradually, peaked in the last weeks of pregnancy(P0.05).While the left ventricular diastole function decreased a little through gestation(P0.05), showing the compensasive increasing of left ventricular initiative filling ability.Conclusions: Maternal cardiovascular adaptive changes in pregnancy are active preparations. As a new means of reflecting cardiac remodeling and hemodynamic changes in pregnancy ,FV-3DE will be used in evaluating healthy pregnant women cardiac function by clinical routine gradually.

Experimental Study on Effect of Xie Shi Lung Cancer NO.1 in Mice with Lewis Lung Cancer

Mon, 27 Jun 2011 12:26:03 +0000

Objective: Through the establishment of our animal model of lung cancer, the application of the model of lung cancer on the xie shi lung cancer N0.1 side of the treatment intervention, by observing the different treatment groups of mice body weight and tumor weight in mice, tumor cell apoptosis rate, proliferating cell nuclear antigen ( PCNA) data for lung cancer in clinical better I can provide a theoretical basis of the application.Methods: 10d after passage from the source of Lewis lung tumors in mice, under sterile conditions in tumor tissue stripping choice, homogenate, made of tumor cell suspension (count the number of viable cells> 95%). Preparation of cell suspension will be good to 0.2mL / only transplanted in C57BL/6J mice subcutaneous right axillary, reproduce Lewis lung metastatic model mice. After the model 24 hours, the Lewis lung models were made in 40 mice and were randomly divided into four groups: normal group, xie shi lung cancer N0.1group, control group and treatment group. The normal group were given saline by intraperitoneal injection,the control group were given cisplatin 1 mg/kg by intraperitoneal injection, the lung cancer N0.1group were given xie shi lung cancer N0.1 0.6ml by gavage respectively, the treatment group were given xie shi lung cancer N0.1 0.6ml and cisplatin 1mg/kg by intraperitoneal injection every other day. After 15-day`s treatment. The inhibitory tumor rate, proliferating cell nuclear antigen (PCNA) index and apoptotic rate were detected by immuno -histochemistry and flow cytometry.Results: 1, compared with the normal group, the inhibition rate in each group and compared weight were significant (P <0.01), the xie shi lung cancer N0.1group of the inhibition rate, body weight was significantly higher than comparative treatment group, control group and normal group (P <0.05), tips on the xie shi lung cancer N0.1 can significantly inhibit the cancer and increase the mouse weight, improve quality of life in mice. 2, Xie shi lung cancer N0.1 group,treatment group and contral PCNA index group compared with the normal group, with statistical analysis of the significance of a significant (P <0.01). Xie shi lung cancer N0.1 group and control group, treatment group PCNA index, there is significant difference (P <0.05). 3, the apoptosis rate of Xie shi lung cancer N0.1 group was significantly higher than that of treatment group, control group, normal group (P <0.05).Conclusion: lung cancer N0.1 Can significantly induce cell apoptosis,inhibit the proliferation of tumor cells , which may be one of the antineoplastic mechanisms.Lung cancer N0.1 in mice inhibited tumor and improve the quality of life have a significant role.

Study on the Hypolipidemic Effect of Crude Extract from Schisandra in Vitro

Mon, 27 Jun 2011 12:26:03 +0000

Schisandra is a plant from Magnoliaceae,it is a traditional Chinese medicinal plant.In this paper,to study the Schisandra\'s hypolipidemic effect.Schisandra was extracted with 75%ethanol firstly and then with ethylacetate and N-butanol.CHO cell was the cell model. Intracellular cholesterol content was detected through HPLC.The column was Eclipse XDB-C18(5μm,250mm×4.6mm) and the mobile phase consisted of acetonitrile-methanol (55:45) at a flow rate of 1 mL/min.The detection wavelength was 210 nm.Through the preparation of a series of different concentrations of cholesterol standard,the standard curve was established respectively,In this testing conditions,the scope of the peak area was relatively good linear relationship when the cholesterol content was in 0.282～1.410μg,The results showed that Schisandra could reduce the cholesterol content in CHO cell, Schisandra ethylacetate extract was to 25.81 percent,and Schisandra N-butanol extract was to 35.47 percent.The hypolipidemic effect of Schisandra N-butanol extract was better.

Narcissistic Artists

Mon, 27 Jun 2011 12:26:02 +0000

Narcissism which is one of important aspect of modern psychology is worth of being researched for human being。According to the study of modern psychology, narcissism is one of unconsciousness psychology for every person, but for the artists, Narcissism expresses the self-concern and strong behaving desire, the relationship between the artist’narcissism and the artist self-portrait is one kind of painting form related with the artist’narcissism, i.e. the concentrated expression of the artist’Narcissism on and mental visual and artistic creation.Self portrait is an independent drawing method which was popular during the renaissance. As a special drawing method, self portrait became irreplaceable because of its connection with the artist itself. Thus, on one hand, self portraits appear in different forms. On the other hand, they are mirrors and carrier of the artists’personality, sensibility and experience, through which people know the inner-world of the artists. This paper can be divided into two parts according to the role and expression of the narcissism during the self portrait drawing.Part one including the 2nd, 3rd and 4th chapter mainly addressed how“Narcissism”inspires the artists to create the self- portraits and the influence upon the artists. This part firstly summarizes the relevant content of“narcissism”, then analsizes the artists’characters from the mentality of“narcissism”, finally clarifies the relation between“narcissism and painting. Based on the above reasons, the inevitable result is concluded– the emergent of self portrait.Part two i.e. the 5th chapter bases on the mental characters divides the self portrait into two types: Self-Absorbed and Self-Exhibitionism. Self portraits of Self-Absorbed include two typical forms: self-portrait mirror, narrative self-portrait; and symbolic self-portrait for Self-Exhibitionism. The article investigates these three characteristics of self portraits from the portrait works.The discussion of narcissism never goes to an end. And this article which recurs to self-portrait makes more researches how the unconsciousness narcissism influences the creation of self-portrait and artistic individuality. This paper is trying to dig the content of narcissism and to figure out the self-portrait’signification , in order to explore the connection between the mind and the art, thus give some enlightenment to our future art creation.

Study on the Flavonoids in Fruit of the Catalpa Ovata G.Don and Volatile Oil in the Flowers

Mon, 27 Jun 2011 12:26:01 +0000

The flavonoids is a phenolic compounds distributing widely in nature, especially in plant kingdom. Recently, people pay more and more attention to them. Catalpa ovata G.Don belongs to the family of Bignoniaceae. Its plant has many functions, such as clearing-detoxication, and has been used as a traditional Chinese medicine for cirrhosis. But because it lack phlebitis with marked curative effect, it didn’t recorded in pharmacopoeia. The major contents of this research are the flavonoids in the fruit of Catalpa ovata G.Don. and the volatile oil in the flowers. It will provide theory evidence for the development of pharmaceutical industry and cosmetic industry.It was proved that flavonoids was exist in the fruit of Catalpa ovata G. Don by pre-test. The extraction technology of flavonoids from the fruit of Catalpa ovata G. Don used traditional hot organic solvent method. The extraction rate of flavonoids optimized condition(ethanol concentration, extraction temperature, ratio of solid to liquid and extraction time) were determined by orthogonal design. The result showed that the optimized conditions was obtained as: ethanol concentration is 60%; extraction temperature is 70℃; the ratio of solid to liquid is 1:20; extraction time is 2.0h and extract yield is 4.83%. Contrasted with rutin, we studied the total content of flanovoids in different part of Catalpa ovata G. Don. ( fruits, leaves, flowers), by spectrophotometry. The results showed that the total content of flavonoids was highest in fruits.Flavonoid was purified with the AB-8 pocket adsorption resin, the adsorption rate was 93.98% , the desorption rate was 68.60%. Purification flavonoids was mainly contain flavone, flavonol, isoflavone. The result of thin layer chromatographic analysis showed that purification flavonoids may contain rutin.Antioxidan experiment showed that the flavonoids in the fruit of Catalpa ovata G. Don had good oxidation resistance activeness. The antibacterial experiment showed that the purification flavonoids had good inhibition to Bacillus subtilis and Bacillus thuringiensis.The essential oil in the flowers of Catalpa ovata G. Don were extracted by steam distillation and analyzed by using GC-MS, 32 compounds are identified. The compound which content surpass 5% were phenanthrene (27.21%), fluorine (7.71%), tetracotane (6.17%), tetracosane (6.05%).

The Study of Loss of Heterozygosity on Hepatocellular Carcinoma and Nodulars of Altered Hepatocytes

Mon, 27 Jun 2011 12:26:01 +0000

Objective:To study the features of loss of heterozygosity(LOH) on hepatocellular carcinoma(HCC) and focal nodular hyperplasia(FNH), by nest-PCR, denatured polyacrylamide gel electrophoresis and silver staining method,and analysised the frequency of LOH on above loci of HCC and FNH. Furthermore, we analysised their relationship between LOH and clinicopathological characteristics of HCC, and further discussed the relationship between LOH at special locus with occurrence and development of HCC,and it might help to supply new molecular markers for early diagnose and prognosis warning of HCC.Methods:1 Samples collection:44 HCC samples were taken from the paraffin block tissues filed during January of 2006 to October of 2008 by the Department of Pathology of Tangdu Hospital. 42 nodulars of altered hepatocytes (NAH) were collected in FNH samples from the same department as well. Every sample had undergone the examination of histology and had been confirmed by two experienced doctors.2 Microsatellite loci:The sequence of primors were from data Genbank supplied.Eleven microsatellite loci as following:D1S199(1p36.13),D1S507(1p36.1),D1S2843(1p36.1),D6S1008 (6q26),D8S261(8p22-p21.3),D8S277(8p23.3-p22),D8S298 (8p21),D8S1819(8p23.1),D10S215(10q23),D11S1301 (11p13),D16S402(16q24-2-q24-3).3 DNA abstraction :Each paraffin embedded sample was cut into 8 pieces in 15μm to collect tissues with the use of microdissection.Abstracted DNA with equal volume phenol-chloroform method with a final concentration of DNA as 50ng/mL.4 Microsatellite polymorphism analysis :Do nest-PCR on eleven selected microsatellite loci.Total PCR reaction volume should be 50μl and make use of Bio Rad thermal cycler. The mixture of 4μl PCR production and equal volume essay buffer were added to 8% denatured-polyacrylamide gel, and electrophoresised at 120V for 4h.Visulized the gel after silver staining and record data with UVP gel analysing system and optical picturing.Results:LOH was found in 35 of 44 HCCs at least one locus, the total LOH frequency is 79.55% (35/44);top three loci were D11S1301 (64.29%,27/42),D1S2843(62.50%,25/40), and D16S402(54.76%,23/42).Totally, LOH had no apparent correlation with sex, age, serum AFP, HBsAg, the size of tumor, Edmondson grading, cirrhosis of liver, metastasis. But LOH at some particular loci were relevant with certain clinicopathological parameters of HCC.LOH on D8S261 and D1S2843 were significantly higher in the cases with positive serum HBsAg than in those with negative HBsAg(P <0.05 and P <0.01, respectively);LOH on D1S507 and D11S1301 were more frequent in tumors larger than 3 cm in size (P <0.05 and P <0.01, respectively) ;LOH frequency of D11S1301 was significantly higher in poorly or moderately differentiated cases than in well-differentiated cases(P <0.01);LOH on D16S402 was more frequently detected in tumors with intrahepatic metastasis than in those without metastasis(P <0.05).LOH was found in 2 of 42 NAHs at D11S1301, LOH frequency was 4.76%, No LOH was found at any loci else.Conclusion:LOH at some particular loci are associated with occurrence,development and clinicopathological parameters of HCC. This study find out that locus of D11S1301 of the district of 11p13 in chromosome has higher LOH frequency. This discovery implies the potential existance of a new tumor-suppressor at D11S1301 which related to the development of HCC, which supplies a significent foudation and clues to further study the tumor suppressor gene of HCC. It was discovered that LOH occurred at specific locus is associated with such prognosis factors as the occurence of positive HBsAg HCC, differentiated degrees of HCC and the extention or metastasis of it, which enjoys profoundly clinical importance to supply a new theorical foundation to the individualized diagnose, treatment and the prognosis warning of HCC.Moreover, LOH was found in 2 of 42 NAHs at the locus of D11S1301,with a frequency of 4.76%.This result shows that FNH exsists LOH that sharing the same locus with HCC, it points that FNH may be kind of precancerous lesions in the level of molecular genetics,and implies new theorical foundation to the early diagnosis of HCC.

Construction of Prokaryotic Expression Plasmid with Two Subgroups of Human ndrg2 Gene and Expression and Purification and Anti-tumor

Mon, 27 Jun 2011 12:26:00 +0000

Our group discovered the ndrg2 gene(N-myc downstream regulated gene 2)and cloned it from normal human whole brain cDNA library in 1999,which has been adopted in Genbank with the number of AF159092.This gene locates on human chromosome 14q1.11-11.2.There are two subtypes about ndrg2,encoding about 43kDa protein with 357 amino acid and 371 amino acid respectively.The ndrg2 gene is one member of ndrg gene family,which consists of 4 members:ndrg1,ndrg2,ndrg3 and ndrg4.This gene family has high similarity in nucleotide or amino acid sequence from inferior creature to advanced creature.Those indicate that the ndrg gene family plays an important role in life of cells.Our preliminary studies showed that there were higher expression of ndrg2 mRNA in brain,salivary gland,skeletal muscle,lower expression of ndrg2 in bone marrow,testis,peripherial blood and placenta and none expression in bone marrow cells(BMC)of leukaemia patients,colorectal cancer tissue,lung cancer tissue,liver cancer tissue and some tumor cell lines.In addition,the ndrg2 gene could inhibit the proliferation of many tumor cells,such as glioma cell line BT325.All above suggested that gene might be a new candidate tumor suppressor gene(TSG).However,the mechanism on suppressive effect of ndrg2 isn’t clear.First, we constructed the prokaryotic recombinant vectors for two subgroups of human ndrg2 gene, and identified their expression and purification. The coding sequence of two subgroups of human ndrg2 were amplified with specific primers and cloned into the 6×Histidine (6×His)fusion expression vectors pPROEX-HTb. Then the recombinant plasmids were transformed into E.coli BL21 and the expressions of 6×His fusion protein were induced by adding isopropylthiogalactoside (IPTG). Then fusion proteins were purified by Protino Ni-TED Resin. Restriction digestion of the constructed recombinant plasmid revealed the existence of gene segments about 1100bp in length, which were in accordance with what had been expected and the results of sequencing showed that the coding sequence of two subgroup of human ndrg2 gene were correct. Two protein bands with the molecular weight of about 45kd were induced by IPTG in the recombinant plasmids. Western blot assay revealed that the 6×His fusion protein could be specifically recognized by ndrg2 antibody.And by Protino Ni-TED Resin,the fusion proteins were purified successfully.By the test, the coding sequence of two subgroups of human ndrg2 gene have been successfully cloned into the 6×his fusion expression vectors pPROEX-HTb and efficiently expressed and purified, which lay the foundation for study on biological activity and suppressing tumor.The second, we detected the inhibition action of ndrg2-357 protein and ndrg2-371 protein on tumor cell. The fusion protein of ndrg2-357 protein and ndrg2-371 protein were induced to express and purified by Protino Ni-TED Resin.The protein purified were concentrated by ultrafiltration centrifuge tube and the balanced solution were replaced. The inhibition action of ndrg2-357 protein and ndrg2-371 protein on tumor cell of U251,BT325 and A549 were observed by in vitro tests.And the inhibition action of ndrg2-357 protein and ndrg2-371 protein on tumor cell were compared.The tests showed us that the inhibition action of ndrg2-357 protein and ndrg2-371 protein on tumor cells were obvious, but there were no significant difference between the ndrg2-357 protein and ndrg2-371 protein.

Investigation of B7H3 in Benign and Malignant Renal Tumors and Their Angiogenesis

Mon, 27 Jun 2011 12:25:58 +0000

ObjectiveCell surface molecule B7H3 has different expression patterns in a variety of tissues and cancer types;it can also distinguish pathological and physiological angiogenesis.Our purpose was to investigate its expression in benign and malignant renal tumors and their angiogenesis,and to explore its possible values in clinical practice.MethodsFrom July 2006 to August 2008,31 consecutive renal angiomyolipoma(AML) with no history of tuberous sclerosis received surgical treatment in our department.By immunohistochemistry,we compared B7H3 expression patterns in paraffin-embedded specimens,between them and another 31 pathological tumor size paired clear cell renal cell carcinomas(RCC);paired normal renal parenchyma from each of the patients above as well as 4 corpus luteum specimens were also analyzed.Of the selected AML and RCC patients,11 serum samples for each group were extracted from our tissue bank,and the concentration of peripheral blood(PB) soluble B7H3(sB7H3) was detected and compared by enzyme-linked immunosorbent assay(ELISA).Prospectively,twenty PB samples were collected before operation,including 2 from renal cysts,one from incision infections after radical nephrectomy,one from local recurrence of papillary RCC,four from metastatic RCC,as well as one from locally advanced RCC,and the other 11 from renal tumors with difficulty in deciding their malignancy only by radiological study.PB B7H3 mRNA level were analyzed using real-time PCR,and compared with the postoperative pathological results.All tissue and PB samples\' collection and use were in line with the Helsinki Declaration of Human Rights.Results1,Cancer cell expression of B7H3 was found in 22.6%(7/31) of clear cell RCCs,but that of tumor vessels was confirmed in all the 31 cases;focal, moderate and diffuse expression of the tumor vessels in RCCs were 3/31,7/31, and 21/31,respectively;no obvious positive expression was detected in either tumor cells or tumor vessels of renal AML,neither in the 62 paired normal renal parenchyma and their vessels;four cases of luteal blood vessels do not express B7H3,but all the theca folliculin cells were positive for B7H3.2,Serum concentration of sB7H3 from RCC and AML were:16.47ng/ml and 16.31ng/ml,with no significant difference(p＞0.05);sB7H3 had no significant correlation with tumor and blood vessels expression of B7H3(p＞0.05).3,Forty-five cycles of the real-time PCR found four cases with no detectable B7H3 mRNA,of which 2 were simple cysts,and the other 2 AMLs;after normalized,we found minimal level of B7H3 mRNA in PB.The renal AML group had similar B7H3 mRNA level with the normal control(renal cysts and incision infections)(p＞0.05),and significantly lower than the RCC group(p＜0.001); there was no difference between RCCs of different types or tumor size in localized ones(p＞0.05);the level of PB B7H3 mRNA in non-metastatic RCCs was slightly lower than that of metastatic RCCs,although without significant difference(p＞0.05).Conclusion1,A small group of clear cell RCC expressed B7H3 in tumor cells,while tumor cells of renal AML did not express B7H3,and we happened to find theca folliculin cells in the corpus luteum also had expression of B7H3.2,B7H3 selectively expressed in angiogenestic endothelial cells of RCCs,but not in that of AMLs,nor in normal or physiological angiogenestic endothelial cells.3,Serum concentration of sB7H3 from clear cell RCCs and AMLs had no difference in between,and seemed similar to that from normal adult.4,B7H3 mRNA levels in PB could distinguish AML from RCCs,when radiological study confirmed a renal tumor but had difficulties in differential diagnose.Specific expression in malignant blood vessels and some cancer cells made B7H3 a potential role not only in tumor and angiogenesis-targeted therapies,but also in tumor vessel based differential diagnose.

Study on the Application of Electro-neurophysiology Monitoring in the Diagnosis and Treatment of Cubital Tunnel Syndrome

Mon, 27 Jun 2011 12:25:58 +0000

ObjectivesThis study aims at evaluating the application of electroneurophysiology monitoring in the diagnosis/treatment of cubital tunnel syndrome(CTs) and discussing the effect of compressing agent excision and epineurium dissection on the recovery of never function by monitoring change of preoperative,intra operative and postoperative ulnar never conduction velocity as well as analyzing the correlation of severity and prognosis with electroneurophysiology indices to provide scientific proofs for the early diagnosis for the CTs improving the operation method,reducing operation wound and finally,improving the surgical outcome.MethodsThe study was conducted among the sample of 61 in-patients with CTs in orthopedics department of Tianjin Medical University General Hospital to evaluating the application of electroneurophysiology monitoring in the diagnosis/treatment of CTs by using(1)one-sample t test and Spearman correlations analysis to compare the difference of ulnar nerve conduction velocity,latent period and amplitude between patients and normal as well as analyzing the correlation of severity with electroneurophysiology indices;(2)assessing the accuracy of electroneurophysiology position oriented diagnosis by comparing the estimated compressing position(tested by using electroneurophysiology short section movement technique) and actual compressing position.(3) analyzing the change of nerve conduction velocity,latent period and amplitude before-after the compressing agent excision and before-after epineurium dissection by paired-samples t test to discuss the short-term effect of these two processes on the recovery of unlar never function;(4) evaluating the surgical outcome at the 6~(th) month after operation and its correlation with nerve conduction velocity,latent period and amplitude.Results55(90.16%) patients were followed up at the 6th month after operation.(1) the nerve conduction velocity,latent period and amplitude of subjects are 26.16±11.68(m/s)、13.71±3.47(ms) and 0.34±0.20(mv) respectively;comparing to normal value,they have slower nerve conduction velocity and longer latent period.(2) the severity of illness is negative correlation with nerve conduction velocity(r= -0.863,p=0.000) and positive correlation with latent period(r=0.354,p=0.037).(3)the accuracy rate of entrapment position positioning by electroneurophysiology position oriented diagnosis is reached to 85%.(4) nerve conduction velocity is increased,the latent period is shorten with highly statistical significance after compressing agent excision;however,there are no significant change in the electroneurophysiology indices after epineurium dissection.(5)the pathology exam shows that the most of compressing agents(87.26%) are hyaline degeneration of ligaments with hyperplastic adipose tissue and small vessels, 1 of them with calcified cartilage tubercle,3 of them contain skeletal muscle;the microscopic anatomy of other 7 compressing agents are 2 cases of angiolipoma,1 case of osteochondroma,1 case of neurilemmoma and 3 cases of ganglionic cyst.(6)most of subjects(65.46%) have satisfactory therapeutic effect after operation, however,minority of them(12.73%) have poor outcome;the surgical outcome is negative correlation with preoperative nerve conduction velocity(r= -0.862,p=0.000)Conclusion(1) Preoperative electroneurophysiology exam is conducive to the diagnosis, prognosis forecasting and severity judgment of CTs as well as locating the diseased region exactly.(2) Intra operative monitoring can estimate the effects of certain operation procedures on the short-term recovery of never function to provide proofs for the improvement of operation.(3)Nerve function has recovery after compressing agent excised,but no improvement after epineurium dissection according to electroneurophysiology exam,the necessity of epineurium dissection need further study.(4)the surgical therapeutic effect is related to the degree of nerve damage,the more severity damage is,the poorer outcome it would appeared.As a simple, non-invasive and objective exam method,electroneurophysiology exam should be widespread used for the diagnosis and treatment of CTs in the future.

A New Multi-point Puncture Perfusion Technique for Adult Human Hepatocytes Isolation and Detection of Cell Function in Vitro

Mon, 27 Jun 2011 10:37:49 +0000

Background:Artificial liver support system is recognized as the most effective treatment for a variety of acute liver failures and bioartificial liver(Bioartificial Liver,BAL) based liver cell may be a great hope for various acute and chronic liver diseases in the future. The bioartificial liver is mainly made up of the cultivated liver cells with activity in vitro,so bioartificial liver has the same functions of detoxification,secretion, synthesis and transformation as liver and liver cells function as its core part.With the development of modern cell engineering and the latest achievements in cytobiology,tissue culture technique and biology,nowadays hepatocyte has become the hotspot of multiple studies because it can be used as the biological section in the construction of bioartificial liver,the seed cell in cellular transplantation to cure acute and chronic liver failu and the target cell in the drug screening.Thus,the preparation and culture of high-active hepatocytes,in particular the preparation of human liver cells,have become the most basic link among all these studies.There are many kinds of hepatocyte isolation methods,such as mechanical separation,chelation,enzyme digestion,and so on.The mechanical separation began more than half a century ago.In 1956,for the first time,Sorrentino prepared fresh hepatic tissue homogenate using mechanical method and many drug detoxification trials in vivo were performed.Gradually,this method was abandoned because of a small number of hepatocytes with poor motility.In 1967,Howard established collagenase perfusion method,and In 1972,Seglen further developed the method into two-step perfusion,during which the preheated balanced saline without calcium was first infused to remove blood cells within hepatic sinusoid and some nonparenchymal cells,and then balanced saline containing was infused to perform purified in vitro digestion.Following these procedures,collagenase hepatocyte was separated,and obtained.With this method,the number of hepatocyte and its motility were both improved and foreign matters in hepatocyte suspension were reduced.Nowadays,the two-step perfusion has become a common method used in the laboratory.Two-step perfusion method is appropriate in liver separation of small animals such as mice,rats,rabbits and dogs.At present,the main application of two-step perfusion method for human liver which is discarded in transplantation for non-matching.The biggest flaw of this method is the lack of contributive liver in transplant,and the discarded liver for not matching is particularly rare.In this condition human liver cells are completely unable to meet the needs of the basic research,so that many basic researches have to choose animal\'s liver cells to replace human\'s.A lot of scholars have applied small pieces of liver tissue resected from hepatic lobes,through its remnants of the small blood vessels to separate liver cells by two-step perfusion method.This approach requires a larger block of liver tissue, and 3～4 blood vessels but in fact the liver tissue obtained in surgery is usually of irregular shape.So it is difficult to find blood vessels,two-step perfusion can not be implemented.Even if the blood vessels could be found,perfusion can only be done through the remnant irregular-shaped blood vessels,which would lead to insufficient perfusion,and unsatisfying separation result.So it is necessary to set up a practical cell separate technology to separate liver cells from human liver.On the basis of two-step perfusion,this study set up a new method of multi-point puncture perfusion to separate adult liver cells from small pieces of irregular-shaped liver tissue obtained in surgery.In this way,human liver cells will be obtained more simply and easily in many basic researches.This method will provide technical support to the application of human hepatocytes for bioartificial liver,cell transplantation research.This experiment further cultivated human liver parenchymal cells which were isolated by multi-point puncture perfusion in a short-term primary and external way and examined supernatant of the cultured cells for albumin,urea nitrogen,the level of intracellular enzyme,and cell metabolic function.Thus,on one hand,we identified the possibility of separate liver cells with functional activity by multi-point puncture perfusion;on the other hand,we analyzed primary hepatic parenchymal cell function in the course of adult primary liver cells cultured in vitro.Methods1、Human liver tissue samples were obtained from patients with liver cancer or hepatic hemangioma in the department of Hepatobiliary Surgery of Nanfang Hospital. Normal tissues around the lesions were sampled during the surgeries,and each of them weighed about 0.5～5g.2、Multi-point puncture perfusion was adopted to separate adult liver cells.Specific steps include:①multi-point puncture and perfuse with 38℃preheating perfusion fluid by pinhead injector until liver tissues change color from dark red to gray and the outflow of perfusion fluid get clean and bright.The pressure and flow rate during perfusion must be constant and produce no bubbles.This process lasts about 10-15 minutes.②multi-point puncture and perfuse with 38℃preheating collagenase typeⅣsolution with injector until the tissues get loose,lose elasticity, and the surface appears back of turtles.Ⅳcollagenase solution can be recycled.This process takes around 15-20 minutes.③Tear the liver tissues into pieces bluntedly with sterile forceps,remove the residual amicula and fibrous connective tissue, collect the liver cell suspension into sterile bottle,and then continue to digest with shake in collagenase solution for 15 minutes at 37℃.④Make the digest into cell suspension with thick-mouth straw.Filter with 100-mesh stainless steel screen,and collect the liver cell suspension with a centrifuge tube on the ice bath.⑤Centrifuge 5min at 4℃with a speed of 50g,remove the supernatant,add some red cell disruption to the sediments at the bottom,allow it to stand for 3 minutes at room temperature,and then centrifuge at the same speed for 5min again.⑥Collect the bottom sediment,re-suspend the cells with liver cell washing buffer solution containing DNase I,and then filter with 200-mesh stainless steel screen.⑦Centrifuge 5min with 50g at 4℃,remove the supernatant,re-suspend the cells with serum-free DMEM medium,centrifuge again,discard the supernatant,and put the sediment cells on the ice bath for use.3、Collect the liver cells,stain them with trypan blue and then calculate the motility rate and the yield.Culture the liver cells with RPMI-1640(contained 15% fetal bovine serum),and calculate the adherence rate after 24h.4、Observe the liver cells\' shape and structure of the adult primary liver cells when they were freshly isolated and cultured with the inverted phase contrast microscope and the fully automatic biology photomicrographic device.5、Draw a growth curve of adult primary liver cells isolated by multi-point puncture perfusion. 6、Collect cell supernatant of the primary human liver cells that were isolated by multi-point puncture perfusion respectively after cultures for 1 day,3 days,5 days,7 days,9 days.Analyze the concentrations of urea(BUN),alkaline phosphatase(ALP), alanine aminotransferase(ALT),aspartate aminotransferase(AST),alpha-fetoprotein (AFP) etc in cell culture supernatant with fully automatic biochemical analyzer.7、Detect the levels of albumin of adult primary liver cell culture supernatant separated by multi-point separation of puncture perfusion by ELISA.8、Detect AlbmRNA and the expression of cytochrome P450 2E1 mRNA of primary cultured human liver cells by RT-PCR.9、Detect the metabolic function of the primary cultured human hepatocyte by the diazepam transformation test.Results1、The method of multi-point puncture perfusion set up by us was used to separate liver cells from adult small irregular liver tissue resectted in surgery.Survival Rate of liver cells were 87%±7%,a total of(2.2±0.7)×10~7 liver cells were isolated from one gram liver tissue,liver cell adhesion rate was 75%,and there were rare cell debris and other non-parenchymal cell pollution.In addition,the methods are good for isolation of primary liver tumor cells,and the success rate is almost 100%.2、Automatic biochemical analyzer was used to detect the level of biochemical indicators in primary adult liver cell culture supernatant.As a result,the levels of ALT and AST rose with the increased culture days,ALP showed no significant change trend,and AFP was not detected all the time.Although the content of BUN demonstrated a slight downward trend,it maintained at a relatively high level in the short-term culture in vitro.3、ELISA was used to detect albumin secretion of adult liver cells in vitro.The secretion amount reached a peak on the third day and then went down.The cell supernatant concentrations of human albumin excreted by human primary liver cells cultured from 1 day to 7 days were respectively 48.2±4.5 ng/ml、55.2±6.9 ng/ml、63.3±6.4 ng/ml、37.0±10.6 ng/ml、22.5±4.9 ng/ml、14.9±5.3 ng/m、10.0±2.9ng/ml.4、RT-PCR can detect the expression of AlbmRNA and cytochrome P450 2E1 mRNA in the adult primary liver cells obtained from multi-point puncture perfusion. 374bp and 338bp bright bands separately appeared.5、Adult liver cells can be effective in playing its role in drug metabolism in vitro within 7 days via the diazepam transformation test.In vitro on the third day it had the greatest transforming capacity,and then there was a gradual downward trend.Conclusions1、The laboratory operating procedures of multi-point puncture perfusion were established to separate human liver cells,which can reduce the requiment for liver tissue samples.So it is no longer difficult to get primary adult liver cells.The new method was easy,economical and practical.2、Through the functional detection of human primary liver cells obtained from multi-point puncture perfusion,we know that this method can provide liver cells with the biological activity and metabolic function that can be used to do researches on source of human liver cells.3、The dynamic change of functional activity of adult primary liver cells was studied in the process of culture in vitro.

The Study of the Doctrine of Arthromyodynia in Huangdis Classic on Medicine

Mon, 27 Jun 2011 10:37:48 +0000

Huangdi\'s Classic on Medicine is one of the our country\'s earliest medical literature writings existing,It was the foundational classic of traditional Chinese medicine theory and considered as the standard by medical scientists of different dynasty for its abundant record of clinical treatment.The ancient classical learning of heat disease treatment practice was summarized in the heat disease treatment part of Huangdi\'s Classic on Medicine.Informative records on the occurrence,syndrome,outcome and treatment of arthromyodynia formed a preliminary theory and principles.For the benefit of arthromyodynia treatment,this research analyzed comprehensively the achievements of the study on arthromyodynia in Huangdi\'s Classic on Medicine and made it more systemly and orderly.First of all,it studied the name origins and the meaning of disease.According to etiological tactor,location,symptoms characteristics,course of the disease and the season,arthromyodynia was divided into five categories and was analyzed separately.Secondly,the etiological tactor,pathogenesis and treatment of arthromyodynia was analyzed in depth.The paper discussed the etiological tactor of arthromyodynia from the aspects of natural factors and living factors.Qn the influential factors of diseases,the article concludes three factors,including constitution and anti-pathogenic Qi.The progress and transmission of arthromyodynia,discussed in Neijng includes transmission from exterior into interior and transmission among Zang-Fu organs.From the aspects of observasion of complexion and inspect vessels,inspecting the body statue and movements,and touching the skin on the Chi and taking pulse,it taught upon the examinations of arthromyodynia.As to the method of differentiation of syndromes,it discussed "differentiation of syndromes by five Zang-organs principal syndromes"and"Eight-Principal differentiation".What\'s more,for the characters of the treatment of arthromyodynia,the article concluded three characters, including"treatment by differentiation of syndromes","taking precautions against the progression of a disease"and "treatment in accordance with three factorsl".As to therapeutic method,the article discussed acupuncture therapy and drugs therapy.It also talked about prognosis.then,it analyses the value and defect of the doctrine of arthromyodynia,and the influence on later ages.In view of inadequacy of arthromyodynia theories,the paper made study on the standardization of disease named specific measures for improvement,and rich the content of arthromyodynia theories。 The paper elaborated the arthromyodynia theories in Huangdi\'s Classic on Medicine,and made up the shortage of present arthromyodynia theories in this aspect.Study this topic certain to systematize arthromyodynia theories in Huangdi\'s Classic on Medicine, and apply to clinic provide important and worthy data.

The Embedded Systems of Biocheical Analysis of the Parameters

Mon, 27 Jun 2011 10:37:48 +0000

The device of Biochemical analysis of the parameters is one of the necessary equipment in the medical institutions to conduct clinical diagnosis. It is that through analysis a variety of body fluid for determination to obtain chemical and biological parameters of a variety of indicators to help doctors diagnose the disease.On the embedded systems of ARM+Linux platform, The system of Biochemical analysis of the parameters has been studied, including the design of the hardware, building a software platform, the driver and application development., as a result, completed the embedded system of chemical and biological parameters analysis. The hardware design of the system includes the following elements: power, clock and reset circuit, memorizer unit, liquid lane unit, the optical control unit, temperature control units, data acquisition unit, the interactive unit,and detailed in this article on the design process. In the system software design, introduces that how to build the cross-compiler debugging environment, BootLoader (VIVI) of the transplant, Linux kernel of the transplant, the design of device drivers, file system (CramFs) and the design of applications.

Clinical Researches of Retreatment

Mon, 27 Jun 2011 10:37:47 +0000

Various anatomic complications of pulp cavity and canal system, the pathological changes of the root apex before treatment, the differences of medical instrument and drugs, technique level between dentists, all the causes lead to this situation that there are still some failed cases of endodontics treatment though all the clinical operations are done in aseptic condition. The causes for the failure are various, for example, resinifying therapy, pulp mummification, overfilled and underfilled canals, root apical diseases, missing canals, and so on. Sometimes the root canals have clinical symptoms, sometimes the root canals don’t have any clinical symptoms and they just need to have the improved root canal filling before Prosthodontic treatment. The successful key for the endodontic retreatment is analyzing the causes of failure, the former treatments, the situation of diseased teeth and selecting the correct ways of endodontic retreatment. Furthermore, with the modern root canal therapy and instruments are introduced, various special operative microscope and ultrasonic device are used in the endodontic retreatment, the successful rate is increased. Even if most of the hospitals don’t have the operative microscope and ultrasonic device, the chances are also greatly increased if we have the reasonable treatment, cautious clinical operation, right drugs and instruments. So we should try our best to take the conventional endodontic retreatment for the failed cases. If the conventional retreatment fails, apical surgery is recommended.The study, based on clinical cases of endodontic retreatment, includes the reasons and the effective treatments, and we hope the findings and treatment options will be further value in the clinical operations.

The Neuron Loss and the Activation of Neuroglial Cells in Hypoglossal Nucleus after Hypoglossal Nerve Injury

Mon, 27 Jun 2011 10:37:46 +0000

Objective: In this study, a peripheral nerve injury model was established by cutting the left hypoglossal nerve. Using this model the relationship between the neuron loss and the functions of neuroglial cells were studied by observe the changes in the amount and the morphology of astrocytes and microglia in hypoglossal nuclei after hypoglossal nerve injury in adult mouse at different time points.Methods: A total of 30 adult Kunming mice which bodyweight was 30±5g were used in this study. The left hypoglossal nerve of the mouse was cut under 4% chloral hydrate anesthesia, and then the skin was sutured. For Day 7 or day 35 after the injury, Mice were anesthetized with diethylether and were perfused with 4% paraformaldehyde solution (PFA). Brains were removed and stored in the same fixative overnight. Then, the brains were cryoprotected in PBS containing 20% sucrose overnight, embedded in OCT compound, and were cut in 18μm slices for immunohistochemistry(IHC) and thionine staining. The number and morphology of neuroglial cells at different time points were studied by IHC using GFAP and Iba1 antibody which is the marker of astrocytes and microglia,respectively. Neurons were labeled by thionine then be counted. The data statistics was analyzed by using spss11.5 software.Results: On the 7th day after hypoglossal nerve injury, neurons in injured side appeared soma swell, nuclear deviation and nissl body margination, but the amount (8.00±1.35/14400μm2) has no significant difference(P>0.05) compared with that in non-injury side (8.51±1.30/14400μm2). On the 35th day after injury, the shape of neurons in injured side reached normal, however the amount of neurons in injured side (6.70±1.22/14400μm2) was significantly decreased compared with that in non-injury side (8.41±0.61/ 14400μm2) (P 0.05) between the amount of astrocyte in non-injury side (5.33±0.62/14400μm2) and that in injured side (6.67±0.73/ 14400μm2 ). On the 35th day after injury, the amount of astrocytes in injured side (7.85±0.42 /14400μm2) was significantly incresed compared with that in non-injury side (5.41±0.3314400μm2,P<0.05).On the 7th day after injury, activated microglia accumulated in injured side, their thickening and retractive branches wrapped around the neurons, and their amount (9.59±0.80/14400μm2) was significantly increased compared with that in non-injury side (2.52±1.17/14400μm2)(P<0.05)On the 35th day after injury, the morphology of microglia returned to ramified which had long and thin branches. However,the amount of microglia in injured side(5.40±0.95/14400μm2) still significantly more than that in non-injury side (2.18±0.28/14400μm2,P<0.05).Conclusions:1.The neuron loss happened on the 35th day after hypoglossal nerve injury in adult mice.2.Neuroglial cells were in different active state coincide with the neuron loss, indicates neuroglia may be involved in the process of neuron loss.

Study on Correlations of Subclinical Hypothyroidism with Coronary Artery Disease

Mon, 27 Jun 2011 10:37:46 +0000

Objective:To explore the correlation of subclinical hypothyroidism with coronary artery disease(CAD).Method:One hundred and forty-nine patients via coronary angiography were divided into two groups:normal coronary artery group(control group)and coronary artery stenosis group.Coronary artery stenosis group was further divided into two sub-groups,single-vessel coronary artery stenosis group and multi-vessel coronary artery stenosis group,according to the severity of CAD.Serum free triiodothyronine(FT_3),free thyroxine(FT_4) and thyrotropin(thyroid stimulating hormone)(TSH) were tested.Conventional risk factors of coronary artery disease (CAD), clinical characteristics, serum lipid levels, fibrinogen(Fbg) and angiographic results of coronary artery were also examined. Result:1.The prevalence of subclinical hypothyroidism in coronary artery stenosis group (21.7%)was higher than that in the control group(9%) (P<0.05).2.The prevalence of subclinical hypothyroidism in multi-vessel coronary artery stenosis group (28.6%) was higher than that in the control group(9%)(P<0.05).3.The concentration of sTSH in coronary artery stenosis group was higher than that in control group(P<0.05),and the concentration of sTSH in the multi-vessel coronary artery stenosis group was also higher than that in the single-vessel coronary artery stenosis group(P<0.05)and control group(P<0.01).4.Subclinical hypothyroidism was associated with coronary artery disease (OR=2.41 95% CI 0.91-6.38 ) ,and also correlated to multi-vessel coronary atherosclerosis(OR=2.93 95% CI 0.72-12.04).Conclusion:Subclinical hypothyroidism is one of the risk factors for the coronary artery disease .It has positive relationship with the severity of CAD.

Clinical Evaluation of Endoscopic Therapy on Biliary Pancreatitis

Mon, 27 Jun 2011 10:37:45 +0000

Objective: To explore the clinical features of acute biliary pancreatitis and the evaluation of ERCP in treatment of biliary pancreatits.Methods: 76 patients of ABP treated in the First Affiliated Hospital of Dalian Medical University from January 2006 to July 2008 were analyzed on clinieal materials. These cases were divided into endoscopic treatment group (n=20) and non-endoscopic treatment group (n=56) according to their treatment methods. Through clinical observation and follow-up to see wether the average days to be in hospital, the recurrence rate and the time of pain relief between the two groups had any significant importance.Results: Of the 76 with ABP, 57.9% of whom were male (n=44) ,and female 42.1% (n=32) , with mean age of 61.9 years. The main cases of ABP is cholelithiasis and biliary tract infection. 53 cases were combined with hepatic function damaged (69.7%). All the patients got followed up visits for average 20 (ranges4-33) months (97.4%). 20 in 76 cases were accepted ERCP, 0 cases of recurrence, other therapies 56 cases, 11 cases of recurrence, the recurrence rate of ERCP group was significantly lower than other groups (P<0.05); the hospital days of other groups was significantly lower than ERCP group (P<0.01); the time of pain relief of ERCP groups was significantly lower than other groups (P<0.01).Conclusions: ABP remains a frequent disease, especially in the middle-aged and old. The incidence between man and woman is no signi- ficantly different. The most frequent etiologocal factors is cholelithiasiss. The clinical symptom is abdominal pain. With high recurrence and long time of easement of pain if just conservative treatmen, the application of ERCP in the Patients of ABP can prevent its recurrence and shorten the time pain relief.

Chinese Medicine Compound Preparation of the T24 Bladder Tumor Cell Line Proliferation

Mon, 27 Jun 2011 10:37:45 +0000

Objectives.Through orthogonal experimental design study conducted ginsenoside Rg3,curcumin,quercetin three kinds of Chinese herbal compound on cultured human bladder cancer T24 cell line to optimize the role of research,for further research and clinical application to lay the foundation for the interpretation of the Chinese Medicines Board side of the compatibility principle,a scientific group to provide a theoretical basis.Methods.Cytotoxicity assay,respectively,by determination of ginsenoside Rg3, curcumin,quercetin three monomers of Chinese medicine within 24 hours of human bladder cancer cell line T24 of inhibition rate,IC50 calculation and analysis software through three types of Chinese medicine reached within 24 hours of the role of monomer T24 bladder cancer cell line in half the rate of inhibition(IC50).Three monomers in accordance with the IC50 value of traditional Chinese medicine to develop the level of orthogonal test factors.Selected based on factors orthogonal test L9(34) orthogonal test table,the table cytotoxicity test compatibility of each compound preparation of Chinese medicine within 24 hours of the T24 human bladder cancer cell lines inhibited the rate of use of Annexin V-FITC/PI detection wither death,the two tests of the data sequence into the column,the level of pigeon-holing,the use of orthogonal test analysis software Analysis assistant test results(data),using visual analysis,the best dose combination drawn.Obtained for the best compatibility with various doses of medicine Hydroxycamptothecin monomer and comparison,testing division for the control group,traditional Chinese medicine compound preparation group ginsenosides Rg3 group,quercetin group, curcumin group,positive control group(hydroxycamptothecin),the bivariate scatter plot of flow cytometry to observe the traditional Chinese medicine compound effects of pharmaceuticals.Fluorescence microscope 24h role of traditional Chinese medicine compound preparation T24 human bladder cancer cell lines,the cell membrane and the nucleus of the state.Results.Results.Through the cytotoxic test and IC50 calculation and analysis software have been three of the IC50 value of traditional Chinese medicine, ginsenoside Rg3 160μmol/L,curcumin 24μmol/L,quercetin 152μmol/L. According to the IC50 value of the orthogonal test factors in the development of the level of ginsenosides Rg340μmol/L,80μmol/L,160μmol/L,curcumin 6μmol/L, 12μmol/L,24μmol/L,quercetin 38μmol/L,76μmol/L,152μmol/L.Select the L9(34) Form-based test of the orthogonal form,by cytotoxicity assay and flow cytometry detection of apoptosis rate test,the data into the orthogonal form by orthogonal test analysis software assistants,using visual analysis of law,A3,B3,C1 than in other levels,and by the very poor with a primary and secondary order of A＞C＞B,so come to the best dose combination for ginsenoside Rg3 160μmol/L, quercetin 38μmol/L,turmeric Su-24μmol/L.Obtained for the best compatibility with various doses of medicine Hydroxycamptothecin monomer and compared.Flow cytometry with excitation wavelength 488nm,with a wavelength of 515nm filter passband detection of FITC fluorescence,560nm wave grew up in the filter Detection PI,bivariate scatter plot of flow cytometry,the compound preparation of traditional Chinese medicine better efficacy in other groups,its role in human bladder cancer after the tumor cells to apoptosis and death-oriented,and basically no living cells.In the fluorescence microscope can be clearly observed that apoptosis and necrosis of the existence of cells. Conclusions.Based on the results of research both at home and abroad and over the basis of our preliminary work,the subject of the application of orthogonal experimental design combined three kinds of Chinese medicine compound monomer optimized for bladder cancer research,through in vitro to observe their impact on bladder cancer cells induced apoptosis growth inhibition effect and the situation,to exploit their advantages Compound bladder.Confirmed that compound preparation of traditional Chinese medicine and 24h of human bladder cancer cell line T24 cells, have a good ability to induce apoptosis.However,the test method is the use of intuitive analysis of the results of orthogonal test,the best match due to be elected only in relation to the factors and levels,not absolute "best",and thus have a certain one-sidedness.

The Degradation of α-CaSiO_3 and β-CaSiO_3 Bioactive Bioceramics in Vivo and in Vitro

Mon, 27 Jun 2011 10:37:44 +0000

Bone defects resulting from traumas, infections, tumors and other causes are common in the field of orthopedics. Large bone defects are still troublesome in clinical treatment. Common treatments include autograft and allograft of bone. However, both have restricted use. Alternative artificial bones with sound biocompatibility, safety and bone conductivity shall be taken into account. Recent research shows that calcium silicate is a kind of bioactive material with potential and promising applications. But, there are few reports on research of in vitro and in vivo degradability of bioceramics ofα-CS (α-CaSiO3) andβ-CS (β-CaSiO3). In particular, the report on comparison research of the two materials is still not available. Thus, their in vitro and in vivo degradability is of importance. This test is mainly intended to research on in vitro and in vivo degradable features of calcium silicate bioceramics and evaluate their degradability as artificial bones.1. In vitro study of macroporous calcium silicate bioceramics degradation Objective: The degradability of theα-CS andβ-CS bioactive macroporous bioceramic in vitro were comparatively studied, and the results were compared withβ-tricalcium phosphate (β-Ca3(PO4)2,β-TCP) macroporous bioceramics. Methods: The macroporousα-CS,β-CS andβ-TCP bioceramics were obtained after adding pore forming materials and sintering at high temperature. According to the methods as given in GB/T16886.14, prepare Tris-HCl buffer solution. Soak porousα-CS,β-CS andβ-TCP ceramics into the buffer solution separately on shaking table of 37℃for 1, 3 and 7 days. Replace the buffer solution every 24 hours. At every time point, take six samples for every material. Take out samples upon soaking them for certain time. Leach them with deionized water and acetone in sequence, dry them by baking and weigh them. Calculate the percentage of their weight in initial weight, which is represented as their degradable rate. Results: The in vitro experiment result showed that the degradable rates of the three materials are given as belowβ-CS>α-CS>β-TCP.. There is no significant difference in statistics betweenβ-CS andα-CS(p>0.05). There are significant difference betweenβ-TCP and the other two materials at every time point (pα-CS>β-TCP. However statistical analysis does not indicate significant statistical difference (p>0.05). The ossified amounts ofα-CS in 4, 8 and 12 weeks are all greater than that ofβ-TCP of the same period. Conclusion: At 12 weeks, there is not any significant difference between the degradability ofα-CS,β-CS andβ-TCP implanted in vivo. The ossification performance ofα-CS is better than that ofβ-TCP.The said study indicates: although statistical difference of degradability ofα-CS,β-CS andβ-TCP only occurs in vitro test and no significant difference is found in vivo histological observation, qualitative histological observation shows that the degradability ofβ-CS andα-CS is superior toβ-TCP. The whole research indicates that the degradability of porous and active bioceramics ofα-CS andβ-CS is superior to that ofβ-TCP. Bothα-CS andβ-CS are expected to be used for repair of sclerous tissues and as support materials for bone tissue in tissue engineering.

A Mid-term Follow-up of Surgical Ablation for Atrial Fibrillation and a Primary Comparison between Monopole and Bipolar Radiofrequency Ablation

Mon, 27 Jun 2011 10:37:43 +0000

Objective:To review the mid-term effect of maze procedure using monopole radiofrequency ablation in cardiac surgery and do a primary comparison between monopole radiofrequency ablation and bipolar radiofrequency ablation.Methods:237 patients who performed maze procedure using radiofrequency ablationg in 2007 and 2008 were followed up since procedure.Medtronic monopole radiofrequency ablation was used in 210 cases,Medtronic bipolar system was used in 13 cases,Atricure bipolar radiofrequency ablation was use in 14 cases.Results:1.193 cases have been followed up,the follor up rate is 81.43%,171 cases with monopole radiofrequency ablation and 22 cases with bipolar radiofrequency ablation.One patient died in perioperative period,2 died in 3 month after surgery,3 stroke happened and pace-maker installed in 4 cases.2.In the monopole radiofrequency ablation group,the instanct rate of elimination of atrial fibrillation was 99.42%,the rate was 72.78%in 3 month after procedure,while 74.83%in 6 month and 71.30%in one year.3.In the monopole radiofrequency ablation group,the patients with larger left atrium(left atrial diameter＞65mm) has a lower elimination rate of atrial fibrillation.The Left atrial diameter is a risk factor in recurrence of atrial fibrillation(P＜0.05).4.Compared with monopole,bipolar radiofrequency ablation procedure has no better elimination rate of atrial fibrillation and no improvement in cardiac function(P＞0.05).Compared with Atricure bipolar radiofrequency,Medtronic bipolar radiofrequency procedure has no better elimination rate of atrial fibrillation and no improvement in cardiac function(P＞0.05).Conclusion:1.It is safe to perform maze procedure with radiofrequency ablation in cardiac surgery.2.The left atrial diameter was an independent influence factor to the efficacy of monopole radiofrequency ablation maze procedure.3.In a primary comparison,the efficacy of perform maze procedure using bipolar radionfrequency was no better than using monopole radiofrequency,there was no statistical significance.

Effect of Ginsenoside Rg1 on Biobehavior of Human Periodontal Ligament Cells and Expression of Serum Cytokine in Periodontal Tissues in Rats

Mon, 30 May 2011 13:09:08 +0000

Objectives:1.To study the primary culture of human periodontal ligament fibroblasts(HPDLFs) by covering coverslip and to investigate the influences of ginsenoside Rglon the biobehaviors of HPDLFs in primary culture.2.To establish an experimental periodontitis model in Wistar rats by prednisolone acetate injection and ligature of their maxillary molars with steel-wire.Study the effects of ginsenoside Rglon the expression of IL-6 and BGP in periodontal tissues of rat periodontitis model.Methods:1.Morphological and immunocytochemical methods were used to identify the cell origin and the efficiency of the primary culture by covering coverslip.2.A series of experiments were performed to investigate the effects of ginsenoside Rglon the biobehaviors of HPDLFs in vitro.Cell proliferation was determined by MTT assay,the total protein synthesis was measured by Coumassie Bright Blue method and the alkaline phosphatase(ALP) activities were detected by spectrophotometric assay.3.Cell cycle of HPDLFs were examined by cell culture and flow cytometry(FCM).4.The intracellular calcium levels of HPDLFs were measured by the fluorescent Ca~(2+) indicator Fluo-3 and laser scanning confocal microscopy(LSCM).5.One hundred 160-200g Wistar rats were randomized into ten groups.Group A used as the normal group was killed at the beginning of the experiment.Group B-E received the therapy of ginsenoside Rg1 were killed at 1st、2nd、3rd and 4th weeks.Group F-J were experimental periodontitis models and were killed at 0、1st、2nd、3rd and 4th weeks.The level of BGP and IL-6 in the periodontium were measured with immunohistochemistry after the rats were killed.Result:1.Covering coverslip significantly increased the yield of primarily cultured HPDLFs.2.The effect of ginsenoside Rg1 on the proliferation of HPDLCsCompared with the control group,the proliferative abilities of PDLC were increased when treated with 0.005-1μmol/L of ginsenoside Rg1,cells treated with 0.01-0.05μmol/L were significantly higher than other group(P＜0.05).3.The effect of ginsenoside Rg1 on the total amount of protein of HPDLCsCompared with the control group,the total amount of protein of HPDLCs treated with ginsenoside Rg1 in 0.01μmol/L,0.05μmol/L,0.1μmol/L was increased dramatically for 72 hours(P＜0.05).4.The effect of ginsenoside Rg1 on the alkaline phosphatase(ALP)activity of HPDLCsCompared with the control group,ALP activities of PDLCs treated with 0.01μmol/L,0.05μmol/L, 0.1μmol/L of ginsenoside Rg1 for 72h were enhanced significantly(P＜0.05).5.The effect of ginsenoside Rg1 on intracellular free calcium concentration in human periodontal ligament FibroblastsAfter being treated with Ginsenoside Rg1(0.01μmol/L),intracellular calciumin human periodontal ligament fibroblasts decreased.At last,it came to lower level of resting intracellular calcium.6.The effect of ginsenoside Rg1 on cell cycle of HPDLCsAfter 48 hours of ginsenoside Rg1(0.1μmol/L),the FCMs demonstrated that the G1 phase cells reduce (P＜0.05),the S phase cells and(S+G2/M)%significanctly increased(P＜0.05). 7.The successful establishment of rat periodontitis modelThe rats in experimental and control groups showed bad appetite and retardation in movement after the fourth day of injection.These symptoms were accordance with those of "shen xu" in traditional Chinese medicine.The osteoporotic changes in alveolar bone,the absorption of alveolar ridge,the formation of periodontal pocket.None of above pathological changes occurred in periodontal tissues of the normal group.The levels of IL-6 in the periodontitis tissues were significantly higher and BGP much lower than those in the normal groups.(P＜0.05).8.Effects of the ginsenoside Rg1 on the expressions of serum cytokines in periodontal tissues of rats The levels of BGP in the group treated were significantly higher than that in the group periodontitis.The levels of IL-6 in the group treated were much lower than that in the group periodontitis(P＜0.05),and the general physical conditions of rats were improved.Conclusion:Covering coverslip can significantly increase the yield of primarily cultured HPDLCs.Ginsenoside Rgl can promote cell proliferation and the total amount of proteins and ALP activity of HPDLCs.The number of S stage cells and muhiplieation index S+G_2M%markedly increased byGinsenoside Rg1,Calcium participates in the process of Ginsenoside Rg1 message transduction.Experimental periodontitis model may be successfully established in Wistar rats by ligature with steel-wire plus prednisolone injection,and this animal model offers a good means for the study of periodontitis.Ginsenoside Rg1 can down-regulate the expressions of IL-6,up-regulate the expression of BGP in periodontal tissues of rat periodontitis model, promote the regeneration of the periodontal tissues,our studies suggest that ginsenoside Rg1 can promote wound healing and tissue regeneration,and provide scientific evidence for the rational drug administration of periodontal diseases.

Clinical Analysis of Surgical Therapy of Five Clivus Chordoma by Far Lateral Approach

Mon, 30 May 2011 13:09:08 +0000

Objective The chordoma is the remnant tissue of the embryo organizing which is a kind of rarer destructive tumour.The morbidity age is 30～40-year-old between.The male incidence rate is greater than the female.The skull inner chordoma often originate from the center line of the slope,may grow to the vicinity.Owing to the position of the chordoma is very profunda,the demand of the surgical therapy is very strict for the exposure of surgery,the handing of the vertebral artery and nerve root in the skill,can expand the visual fields of surgery by stripping condylus occipitalis and the atlas lateral mass.The merit of the far lateral approach can be sufficient when the tumour is especially bigger,embody.The condylus occipitalis length is 30 +/- 4 millimeters about,when we grind away the condylus occipitalis every 1 millimeter,we can expand the visual fields to abdomen,approximately 2.4°,grind off the condylus occipitalis 1/3,may enhance a visual fields 15.9°,Grind except 1/2 but enhance 19.9°.Because of the the visual fields of the surgery is wild,the scope of the grinding condylus occipitalis can be zoomed out,when the anteroposterior diameter is long、the brain stem be justled to the dorsal.On the contrary, the scope of the grinding condylus occipitalis should be expand.Thereby protect nerve and bilateral vertebral artery in the surgery area much better. To explore a surgical approach to chordoma by farlateral suboccipital approach.Methods Retrospective analysis six patients with chordoma were operated through the far-lateral suboccipital approach from September. 2000 to October.2009.And analysis the merit and demerit of the surgical approach as well as surgical highlight. Results Among the six patients,total removal of tumor was achieved in 3 cases,subtotal removal of tmnor in 1 cases and biopsy in 1 case.No surgical mortality occurred in this series.Conclusion this approach can provide large field of operation and wide surgical exposure to the lesion,and benefit minimalrem～etation of brain stem and main blood vessls or cranial nerves.

Study on Distribution of Sweat Glands and Hair Follicles on the Bodies of Chinese Male Adults

Mon, 30 May 2011 13:09:07 +0000

Objective: To get the message on the distribution of Sweat Glands and Hair Follicles on the Bodies of the Chinese Adults. Accumulate the Anthropology data of the Chinese Male Adults. And give the useful and scientific theory according about the choice of skin flap.Methods: Specimens were obtained with a circular metal punch from 30 body surface areas from 62 cadavers stored in formalin(without any kind of skin disease and scar). The samples were dipped in formalin, embedded in paraffin, sectioned as parallel with the skin surface from epiderm to derm, and then stained with hematoxylin-eosin (HE staining). Get the numbers of sweat glands and hair follicles at the dermoepidermal junction.Results: The sweat glands were found in all of the analyzed 30 areas, and the hair follicles were not found on the palm, sole and dorsum of the distal phalanx. The palm area had the most density of sweat gland and the mammary area had the least. Significant dimorphism for the density of sweat gland was seen between hands and any other area, but no dimorphism was found between upper and lower limbs. The buccal area had the greatest density of hair follicle, while the sparest areas were palm, sole and dorsum of distal phalanx, and the hair follicle doesn’t appear on the dorsum of hand. The order of the hair follicle density was head, limbs, trunk, hands and other areas. Generally speaking, no dimorphism for the hair follicle density was found among upper limbs, lower limbs and trunk. Compared with the densities of sweat glands and hairs reported by K. Huang, the densities of sweat glands and hairs of Chinese Male Adults is less than Korean, but the curves of Korean and Chinese males’density of sweat glands and hair follicles were similar. Therefore we speculate that the densities of hair follicles and sweat glands are similar between Chinese and Korean Male Adults.

Effects of Combined α_2-adrenergic Receptor Agonist and κ-opioid Receptor Agonist on Postresuscitation Cardiac Function in Rabbits

Mon, 30 May 2011 13:09:06 +0000

Objective:Postresuscitation myocardial dysfunction contributes to the high fatality rate following successful resuscitation.The purpose of this animal research to investigate the effects of combinedα_2-adrenergic receptor agonist andκ-opioid receptor agonist on cardiac function in rabbits,and to explore if they can improve postresuscitation myocardial dysfunction.Methods:Ventricular fibrillation(VF) was induced by the external electricity,After setting up cardiopulmonary resuscitation model in rabbits,30 rabbits were randomly divided into 5 groups,namely,epinephrine group(E),vasopressin group(V),U50488H group (U),mivazerol group(M),mivazerol+U50488H group(M+U),medicines were administered respectively during CPR.Hemodynamics、B-type natriuretic peptide and myocardial injury marker cardiac troponin T were examined before VF and the early stage of postresuscitation(30-240min),Animals were killed after experiment ended,The left ventricules of the rabbits were obtained for light microscopic observation.Results:(1)MAP、peak-dp/dt and peak+dp/dt in M+U group were significantly higher than any other groups,and the increase of LVEDP was less than other groups(P＜0.05). (2)The serum concentrations of BNP in M+U group significantly decreased than those in any other groups(P＜0.01 vs E、V、U,P＜0.05vs M).(3)The concentration of cTnT in M+U group significantly decreased than those in any other groups(P＜0.01 vs E、V, P＜0.05 vs M、U).(4) Myocardium damages are shown from heavy to light under light microscopic observation:E、V＞M＞U＞M+U.Conclusion:Combinedα_2-adrenergic receptor agonist(mivazerol) andκ-opioid receptor agonist(U50488H) can improve postresuscitation hemodynamics and reduce the concentration of B-type natriuretic peptide and cardiac troponin T in the early stage of postresuscitation and alleviate myocardial damage in pathological,and improve postresuscitation myocardial dysfunction.

Study on Immaterial Motivation of Medical Staff in Public Hospitals of Chongqing

Mon, 30 May 2011 13:09:06 +0000

With the globalization of trade in services, as well as the reformation of medical institutions, public hosipitals are facing increasingly fierce competition in the market. In the final analysis, the market competition is a talent competition, and the excellent talents play a vital role in the development of hosipitals. How to motivate staff to encourage the staff’s enthusiam and enhance the core competitiveness of hosipital has become an important research topic for hosipital administrators. Immaterial motivation, as an important component of motivation, has enormous meanings to stimulate staff’s work enthusiasm and initiative. At present, most of the domestic documents which study the enterprise refer to the foreign thesis and lack the theory of the public hospital, especially lack the study about the immaterial motivation of quantity study. Thus, this research takes the staff’s needs as the breakthrough point, through discussing the characteristic of medical staff’s needs, the importance of appraisal for immaterial motivation, the percipience to immaterial motivation and the staff’s working activity, proposes the effective countermeasures of immaterial motivation to public hosipital.Through investigation and analysis, this research obtains the main conclusions as follows:①The medical staff’s characteristics of needs in Chongqing public hosipital show that material needs and immaterial needs are both paid much attention to, but immaterial needs are paid more.②Immaterial motivation includes three dimensions: survival motivation, emotional motivation and development motivation. The medical staff of Chongqing public hosipital assert that immaterial motivation is important, of which the most important thing is emotional motivation, followed by survival motivation, and finally the development motivation.③The percipience to immaterial motivation of the medical staff in Chongqing public hospital is in the general level, in which the most perception is emotional motivation, followed by survival motivation, and finally the development motivation.④The working activity for medical staff in Chongqing public hosipital needs to be strengthened.⑤Staff with different sex, age, educational background, rank, position appraise differently in needs, appraisal of importance to immaterial motivation, percipience to immaterial motivation and working activity. The differences in some factors are still analogous to the significance.⑥Creating good hospital culture, providing job-related motivation, creating growth opportunities and other immaterial incentives could improve the medical staff’s working activity effectively.

Research on Germline Mutation of STK11 Gene, STK11 and TGFβ1 Protein Expression in Peutz-Jeghers Syndrome

Mon, 30 May 2011 13:09:05 +0000

BACKGROUNDPeutz-Jeghers syndrome(PJS) is a rare autosomal dominant hereditary disease characterized by gastrointestinal hamartomatous polyps and mucocutaneous melanin deposition.Hemminki and Jenne found STK11(serine threonine kinase 11,also known as LKB1) gene germline mutation in PJS patients and families,proved that STK11 gene is the pathogenic gene of PJS.Germline mutation is related to genetic,which is at the level of blood DNA.The main mutation forms were point mutation and small fragments deletion.Predicted effects are occur premature termination signal,resulting in truncated protein and inactivation of protein kinase. STK11 protein play an important role in chromatin remodeling,cell cycle,cell polarity and regulation of energy metabolism.And play a regulatory role in many signaling pathways such as TGFβ/Smad,NF-κB,PTEN/ phosphatidylinosito13\' kinase,G protein-coupled signaling pathway.STK11 protein inhibit cell proliferation and promote apoptosis,down-regulated of protein expression will lead to other tumor-associated gene mutations in somatic,eventually lead to malignant tumors.TGFβis a disulfide-linked dimer peptide.Its family members in mammals have very high stability in vivo.TGFβcontrols the growth of epithelial cells,indirectly repairs DNA damage,stimulates apoptosis,regulates the function of stem cell.TGFβhas the dual characteristics of promote and inhibit tumor.TGFβ/Smad pathway is the main way of TGFβsignal transduction.TGFβand its receptor composed receptor complex,activate Smad proteins,then transducte signal to the nucleus,performance the function as transcription factors.To different cells,TGFβhas different effects:for fibroblasts,the main effect is to stimulate cell division;for the majority of epithelial cells,its effect is growth inhibition.In recent years,many literatures reported TGFβsignaling pathway disorder is closely related to tumorigenesis,development and metastasis.Human TGFβfamily including TGFβ1,2,3,which located at 19q13, 1q41,14q24.Any component in the pathway may cause signal transduction disorder, then cells escape TGFβ-mediated growth inhibitory effect,leading to tumorigenesis. In cells and tissues,TGFβ1 is the most abundant member of the TGFβ.Its promoter regulation is complex,so TGFβ1 disorder diseases(including tumorigenesis) are most common.TGFβ1 inhibited cell growth with the negative regulation of biological effect,its expression or activation deficiencies lead to cell malignant proliferation.Ki67(Proliferating cell nuclear antigen) is confirmed as a nuclear proliferation marker.It initial expressed in the G1 phase of cell cycle,then gradually increased in S and G2 phase,in M phase reached the peak and quickly disappeared in the division phase,no expression in GO phase.Half-life of Ki67 is short,rapid degradation out of the cell cycle.Ki67 expression can objective reflect tumor proliferative activity.A recent research found TGFβsignaling defect in the stromal of gastrointestinal polyps by PJS animal model(STK11 gene allele deletion mouse),which caused the adjacent mesenchymal epithelial cell proliferation activity.This feature is not found in wild-type mice(non-STK11 deletions).Researchers proposed "STK11 deletion→TGFβsignal down-regulated→excessive glandular epithelium proliferation→PJ polyps formation" hypothesis,speculated the formation of PJ polyps and TGFβsignal abnormal are closely related.This hypothesis provide a new way to explore the formation mechanism of PJ polyps.Objeetives: 1,Detection STK11 gene germline mutation in PJS family,analysis of possible pathogenic mutation and mutation forms,explore the Characteristics of STK11 gene mutation in Chinese PJS families.2,Research on STK11 and TGFβ1 protein expression in PJ polyp tissue, combined with research the proliferation of PJ polyps,preliminary authenticate "STK11 deletion→TGFβsignal down-regulated→excessive glandular epithelium proliferation→PJ polyps formation" hypothesis from tissue protein level.Materials and Methods:1,PJS families(20 people,including 16 patients and 4 family health members) are experimental group.Randomly selected 20 health people who unrelated to PJS families as the control group,peripheral blood were taken from two groups,extract genomic DNA.Design 9 pairs of primers according to all the exons of STK11 gene.Purified PCR amplified products,then detect STK11 germline mutation by DNA direct sequencing method.Initial screening by online BLAST in NCBI,positive samples compared with family health people and control group,analysis the characteristics of STK11 germline mutation and mutation forms in PJS families.2,23 paraffin specimens of PJ polyps(from 11 patients) as the experimental group,10 normal gastrointestinal mucosa specimens as control group. Immunohistochemical(IHC) method detected STK11,TGFβ1 and Ki67 protein expression then calculated the positive rate,detected these three proteins at the same time in order to reduce errors,calculated the positive rate.The positive staining rates were expressed by labeling index(LI).We detected TGFβ1 and Ki67 protein expression of colon adenoma(12 cases) and colon adenocarcinoma(11 cases)also by IHC.3,Using SPSS13.0 statistical software to deal with experimental data.If raw data consistent with parameters test conditions,t test were used to compared the mean of two independent samples;analysis of variance(F test)were used to compared many groups independent samples;bivariate correlation analysis using Pearson correlation analysis.If raw data do not consistent with parameters test,Mannn-Whitney U test were used to compare two independent samples differences;Kruskal-Wallis H test were used to compare many independent samples differences;bivariate correlation analysis using Spearman rank correlation analysis,α=0.05 as a tests standard,P＜0.05 for significant difference or there is a significant correlation.Results:1,In experimental group,we found 15 samples from 20 cases existence STK11 gene mutation,the mutation form is point mutation.The predicted effect is caused abnormal protein coding.No mutation was found in control group.Point mutations were found in exon 4,6 and 8.In family A,3 patients have codon 354(exon 8) heterozygous mutation(GAA→CAA),there is no such mutation in family health person.Meanwhile,no such mutation was found in control group.In addition,some missense mutations were found in several patients in codon 154,155,176 and 270.2,Both epithelium and stromal cells of normal gastrointestinal mucosa expressed STK11 protein:cytoplasmic and nuclear staining.STK11 expression types of PJ polyps were as followed:①reduced expression in epithelial cells,completely loss expression in stromal cells;②completely loss expression in epithelial cells and stromal cells;③similar to normal mucosa expression.Both epithelium and stromal cells of normal gastrointestinal mucosa expressed TGFβ1 protein:cytoplasmic staining,no nuclear staining.Expression of TGFβ1 in PJ polyps:stromal staining, little epithelial cells staining,no nuclear staining.Expression of TGFβ1 in colon adenoma and adenocarcinoma:cytoplasmic,membrane and nucleus staining.Ki67 is specific nuclear staining,Ki67 expressed in PJ polyps and normal tissues are lower than colon adenoma and adenocarcinoma.3,Mann-Whitney U test showed LI average rank of STK11 protein staining in PJ polyps group was 13.5,the normal group was 25.05,u=34.500,P=0.002＜0.05, two groups have significant differences,PJ polyps expressed STK11 protein lower than normal.That of TGFβ1 protein staining in PJ polyps group was 14.61,the normal group was 22.5,u=60.000,P=0.031＜0.05,two groups also have significant differences,PJ polyps expressed TGFβ1 protein lower than normal tissue.Using Kruskal-Wallis H test comparing PJ polyps,normal gastrointestinal tissues,colon adenoma and colon adenocarcinoma of-Ki67 protein expression,the staining LI of average rank were:18.43,15.75,37.79,51.0 as followed.χ~2=39.711,P=0.000＜0.05, four groups were significantly different,according to the average rank,the highest expression of Ki67 is colon adenocarcinoma,followed by colon adenoma group,PJS group,normal group.4,Using Spearman rank correlation analyzed PJ polyps STK11 and TGFβ1 relevance of staining LI,r_s=0.863,P=0.000＜0.05,two have significant positive correlation.TGFβ1 and Ki67 of PJP rank correlation analysis results:r_s=-0.422,P= 0.045＜0.05,two have significantly negatively correlation.TGFβ1,Ki67 in normal gastrointestinal tissues,colon adenoma and colon adenocarcinoma correlation analysis of the LI with Pearson correlation analysis,r=0.216,0.349,0.095,P= 0.549,0.266,0.782,P＞0.05,no significant correlation.Conclusion1,A number of STK11 gene germline mutations in PJS pedigrees,confirmed STK11 gene mutation is closely related with this syndrome.A missense mutations: GAA→CAA had been found in codon 354,this mutation is related with PJS in family A.The missense mutation in codon 154,155,176(exon 4),and codon 270 (exon 6) may also be correspond with pathogenic of those PJS families.2,PJ polyps reduced express STK11 protein suggest that abnormal function of this protein probably play a key role in the formation of PJ polyps.Reduced express TGFβ1 protein suggest the formation of PJ polyps may be related to the up-regulation TGFβsignals.3,In PJ polyps,STK11 protein inactivation may lead to low expression of TGFβ1 protein;low expression of TGFβ1 protein may enhance the activity of epithelial tissue proliferation(Ki67 expression higher).This correlation did not found in normal tissue,colon adenoma and adenocarcinoma,suggested this correlation is probably the characteristics of PJ polyps,the trend of the IHC results, supported the "STK11 deletion→TGFβsignal down-regulated→excessive glandular epithelium proliferation→PJ polyps formation" hypothesis in some extent.

The Purchasing Management and Process Optimization on the Model of F Hospitals Supply Chain

Mon, 30 May 2011 13:09:05 +0000

With the hastened global economic integration and intensified market competition, there will be closer global economic link and more complex business environment. In order to reduce the uncertainty of demand and supply and acquire the advantages in competition, the core has been the competition of supply chain management. The competition among enterprises has been superseded by the competition among supply chains. With the intensive study of supply chain management, the value-added purchasing won the approval. As an important link of demand and supply, purchasing runs through the whole supply chain. The purchasing management in terms of supply chain can reduce the cost and play an important role in the synchronization and integration of the enterprises, which strengthens the competitive power of the enterprises.This paper, with the thesis of purchasing management, analyses the situation of F Hospital which is the largest general hospital of this province. According to the theory of supply chain and the current situation of F Hospital\'s purchasing management, it provides two strategies to improve the purchasing management: to enhance the management and cooperation with the suppliers and to optimize the purchasing process. It makes an exposition of the theory, methods and procedure of the two strategies and provides the optimized process as well. With the increasingly intensive competition, the improvement of management will be the key point of hospital\'s core competitiveness although there is a disparity between public institutions and enterprises. It is hoped that these methods could be useful guidance for those enterprises which bring in purchasing management under supply chain, guaranteeing the smooth operation of supply chain.

Study on Combination Use of Ultra-shortwave and Compound Anisodine in the Treatment of Anterior Ischemic Ophthalmopathy and the Regulation of MAPK Signal Pathways

Mon, 30 May 2011 13:09:04 +0000

Objective:1. High-frequency electromagnetic radiation therapy plays important roles in expanding the blood vessels, promoting local blood circulation and nutrition metabolism in organizations by clinical studies. After clinical treatment of high-frequency electromagnetic waves based on its mechanism, changes of visual function and hemodynamics of ophthalmic artery in anterior ischemic optic neuropathy patients were observed. Compared with treatment of compound anisodine injection, we investigate the clinical characteristics and rules.of high frequency electromagnetic radiation treatment for anterior ischemic optic neuropathy. New clinical methods and theoretical basis for anterior ischemic optic neuropathy were investigated in our study. 2. To study the regulatory effect of p44/42 MAPK pathway on the differentiation of MC3T3-E1 pre-osteoblast. Methods: 1. Forty-two cases (57 eyes) with anterior ischemic optic neuropathy were randomly divided into treatment group and the control group, 20 cases (27 eyes) and 22 cases (30 eyes) respectively. In control group, patients were treated by compound anisodine subcutaneous injection adjacent to superficial temporal artery; high-frequency electromagnetic radiation combining with compound anisodine injection were used in treatment group. Before and after treatment, changes of visual acuity and visual field of all subjects were observed. And changes of hemodynamic parameters, including peak systolic velocity (PSV), end diastolic velocity (EDV) and time average maximum velocity (TAMX), pulsatility index (PI) and resistance index (RI) in ophthalmic artery, central retinal artery and short posterior ciliary arteries were examined by color Doppler ultrasound instrument. 2. The activation of the p44/42 MAPK pathway was continuously suppressed with a specific MEK-1 inhibitor PD98059. The Alkaline Phosphatase activity and calcium deposition of MC3T3-E1 preosteoblasts were examined by histochemical staining. The expression of some osteoblast marker genes was evaluated by Semiquantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR). The transcriptional activity of CBFA1 gene promoter was explored by Luciferase assay.Results: 1. Two weeks after treatment of compound anisodine injection and combination treatment of high-frequency electromagnetic radiation and compound anisodine injection respectively, visual acuity of all subjects were increased in both groups and was significantly higher than the levels before treatment (P <0.01). Compared with the control group, visual acuity of subjects in treatment group was improved significant (P <0.05). Results of visual field examination showed that percentages of visual field defect were decreased significantly after treatment (P <0.05), for all subjects of both groups. Compared with the controls, the percentages of visual field defect of combination treated subjects were reduced significantly (P <0.01).By color Doppler examination, results of ocular artery hemodynamics showed that hemodynamic parameters in central retinal artery had been improved significantly after treatment (P <0.05), both by simple treatment of compound anisodine injection and combination treatment of high-frequency electromagnetic radiation and compound anisodine injection. Compared with the control group, PSV, EDV and TAMX were improved significant (P <0.05) in combination treatment group, but no difference were found in the assay for RI and PI. In control group, PSV and EDV were significantly increased after treatment (P <0.05), but no significant difference were found on TAMX, RI and PI., Except for PI, all the other hemodynamic parameters of ophthalmic artery were improved significantly (P <0.05) in the combination treatment group. Results of PSV, EDV and TAMX examination were higher in combination treatment group than those of control group, and the difference was statistically significant. For hemodynamic parameters of short posterior ciliary artery, PSV and TAMX were increased significantly (P <0.01) after treatment in control group; PSV, EDV and TAMX were also increased significantly (P <0.01) after treatment in combination treatment group and were significant higher than those of control group (P <0.05).2. PD98059 not only increased the Alkaline Phosphatase activity of MC3T3-E1 but also promoted calcium depostition and the expression of some osteoblast marker genes in MC3T3-E1 cells. The treatment of PD98059 significantly increased the Cbfa1 gene promoter activity.Conclusion: 1. Combination treatment of high-frequency electromagnetic radiation and compound anisodine injection can significantly increase visual acuity, improve visual field defect scope in anterior ischemic optic neuropathy patients, levels of improvement are better than that of compound anisodine injection alone. Its treatment mechanisms are increasing peak systolic velocity, end diastolic velocity and average peak flow velocity and improving the ocular blood supply in the ophthalmic artery, central retinal artery and short posterior ciliary artery, for the treatment of anterior ischemic optic neuropathy.2. Continuous suppression of the p44/42 MAPK pathway promotes the differentiation of MC3T3-E1 pre-osteoblasts.

NDRG2 Gene Expression in Colon Carcinoma and Its Significant Function

Mon, 30 May 2011 13:09:03 +0000

The human NDRG2 (N-myc downstream regulated gene 2) cDNA was first cloned and reported from normal human whole brain cDNA library in our lab in 2000. The full length of NDRG2 cDNA is 2,024 bp long. It located on human chromosome 14q11.2. NDRG2 gene consists of 16 exons and 15 introns that encodes a 40 kDa protein with 357 amino acid residues. Preliminary data indicated that NDRG2 was expressed in multiple tissues, with heterogeneous expression in normal brain structures except glioma specimens lacking NDRG2 expression. Until now the reported four members comprise the NDRG family: NDRG1, NDRG2, NDRG3 and NDRG4, with NDRG4 possessing three isoforms: NDRG4-B、NDRG4-Bvar and NDRG4-H. The four NDRG family members share great homology in their composition of amino acid residues with differential expression in diverse tissues. The function of NDRG2 has not been clearly understood .Our results show that NDRG2 is high expressed in normal tissues, while is low or no expressed in cancer tissues and cancer cells. These results indicate that NDRG2 has a close relation with cell differentiation and proliferation of the cancer.To study its function and the relationship of signal transduction has important theoretical meaning. There have been no reports about the precise cellular and molecular function of NDRG2 gene. Our group aimed to establish a colon cancer cell line with NDRG2 gene steady expression and analyze the cellular characteristic, so as to investigate the NDRG2 gene better for future.First, semiquantitative RT-PCR analysis and western-blot were utilized to observe the different expression of NDRG2 gene in SW620, HT29, SW480 after differentiation induced by sodium butyrate. The result indicated that there were many difference in these cancer cell lines. The expression of the NDRG2 gene in SW620, HT29, SW480 could be increased significantly by sodium butyrate.Second,for further investigation on the biological behaviors of HT29 cells, we used flow cytometry, plate clone formation assay and MTT assay to investigate the characteristic of the transfected cells with Ad-NDRG2/HT-29. The cell percentage of G1 phrase was about 57.36%, which is more than each control group. It indicated that the transfection of NDRG2 gene contributed to the inhibition of proliferation and induction of cell cycle arrest in G1 phrase. Compared with control group respectively, the curve of translated cells increased relatively slowly in MTT assay. The plate clone formation assay indicated that plate clone formation efficiency was lower than each control group (p<0.05). These proved that the steady overexpression of NDRG2 could inhibit the proliferation of glioma cells.Conclusion:1. The expression of NDRG2 is significantly increased after induced by sodium butyrate in colon cancer cells, which indicates NDRG2 has a close relation with the differentiation of colon carcinoma cells.2. The propagation of HT29 is significantly suppressed after transfected with Ad-NDRG2 in 72h,with G1 blocking,it indicates that overexpressing NDRG2 can inhibit the growth of colon cancer cells.

Changes and Clinical Significance of vWF and CD62P in the Patients with Chronic Renal Failure Combined Hyperuricemia

Mon, 30 May 2011 13:09:03 +0000

Objective:To investigate the Expression and clinical significance of peripheral blood vWF and CD62P in patients with chronic renal failur combined hyperuricemiaMethods:in our hospital in May 2008～December2008 40Patients with chronic renal failur were divided into two groups.A:group of Chronic renal failure combined hyperuricemia(CRF+HUA).B:simple Cronic renal failur(CRF) group,20 cases of normal served as a control group(CO),In the Morning,took 2ml blood in the elbow vein from the patients who was on empty stomach.The plasma percentage levels of CD62P were measured by flow cytometry(FCM),which made in American.;von Willebrand factor(vWF) were measured by ELISA(Enzyme-linked Immunosorbent Assay).The uric acid(UA)was measured by the uricase method.Triglyceride(TG),total cholesterol(TC)、serum creatinine(Cr)levels were measured by standard clinic biochemistry technique..Then all datas as well as intercommunication were evaluated among three groups.used the spss13.0 statistic software to carry out statistical analysis.Statistic mathods:t test and multiple linear regression analysis.Results:Analysis showed that the serum levels of VWF in patients with CRF combined HUA were higher than those in the health controls (p＜0.01),the percentage of CD62P in patients with CRF combined HUA were higher than that in the health group(p＜0.01),we can draw a conclusion that the levels of CD62P(R=0.902),VWF(R=0.621),Cr (R=0.162)TG(R=0.025),Tch(R=0.135) had a positive correlation with HUA.Conclusions:It was a significant difference between experiment group and the norm neonatal group;the serum levels of VWF and CD62involved in chronic renal failure Hyperuricemia the pathogenesis of cardiovascular complications,high uric acid can damage kidney function in different ways and for reasonable control of serum uric acid levels,and provide a theoretical basis.

Clinical Research of 3-dimensional Conformal Radiotherapy for Stage Ⅲ Non-small Cell Lung Cancer

Mon, 30 May 2011 13:08:56 +0000

Objective: To observe the effects of 3-dimensional conformal ra-diotherapy(3D-CRT) in combined modality therapy for stageⅢnon-s-mall cell lung cancer(NSCLC), estimate the response rate, free surviv-al time and survival rate, analyze adverse reaction.Methods: Between December 2003 and June 2008, 71 cases that have complete followed-up data with stageⅢof NSCLC received ra-diotherapy in the first affiliated hospital of Dalian medical university.The disease was StageIIIA in 39, StageIIIB in 32, squamous-celled c-arcinoma in 40 ,adenocarcinoma in 21, adenosquamous Carcinoma in 7,Large Cell Carcinoma in 2 and carcinoma sarcomatodes in 1.These c-ases were divided into two groups randomly: in group A, 34 patients received chemoradiotherapy, in group B, 37 patients received operationplus chemoradiotherapy. All patients received 3D-CRT with high ener-gy X-ray. Normo-fractionated radiotherapy,1.8~2.0Gy/fraction, 5/fraction/week. The prescribe doses of chest radiation field in group A were60~65Gy. In cases that had excellent conformal the prescribe doses cou-ld be 70 Gy.The course of treatment was 6~7 weeks. The cases with supraclavicular lymph nodes metastasis were given the fields of supra-clavicular metastatic lymphatic region. The field was given irradiationwith X-ray first, the total dose was 40~45Gy, then it received irradia-tion with high-energy electron to a total doses of 60~65Gy. Patients with adjuvant radiotherapy, the prescribe dose of target volume was50Gy, but to the cases whose cutting edge were male,the dose added to55~60Gy. The course of treatment was 5~6 weeks. All patients receiv- ed sequential chemoradiotherapy for 2~6 cycles. Combination chemoth-erapy included TP, GP and NP. Two issues were assessed by usingDo-se volume histogram: dose delivered to Target Volume and normal tis-sues nearby. The Kaplan-Meier method was used to calculate the ove-rall survival rate. Acute and late toxicities were graded according to RTOG/EORTC radiation morbidity scoring criteria.Results: With a median followed-up of 25.5 months to May 2009, the response rate(CR plus PR)of group A was 52.9%. With long-term following up, In group A, the median overall survival time was 16.5 months, the1-,2-,3-year overall survival rates were 68%, 28% and 10% respectively. In group B, the median overall survival time was 34.6 months, the median progression-free survival time(PFS) was 22.3 months, the1-,2-,3-year survival rates were 95%, 81% and 47%. The major toxicity were radioactive esophagitis and pneumonia. The incidence rate of esophagitis was 38%(27/71) and usually occurred in 2~6 weeks after radiotherapy (degree I and II, and no degree III). The incidence rate of acute radioactive pneumonia was 23.9%(17/71) and occurred in 3~6 weeks when radiotherapy proceed (usually degree I and II, degree III in one case). It got better after therapy. By reviewing at 6 months after radiotherapy and following up long-term radioactive pulmonary injury, the rate of occurring iconographic change was 38%(all averaging below Degree II).More than half of patients in group B occurred progression in 2 years. The main reason of progression in group A and B was distant metastasis. The metastatic rate was 76.2%. The main reason of death was nonfunction of multiple organs induced by distant metastasis.Conclusion:1. Combined Modality Therapy should be the main therapy of stageⅢnon-small cell lung cancer(NSCLC).2. 3-Dimensional conformal radiotherapy(3D-CRT) can increase the effective rate of inoperable patients and it is an effective adjuvant therapy. Furthermore, no apparent toxicity is increased.3. The major toxicity of 3-Dimensional conformal radiotherapy were radioactive esophagitis and pneumonia. However, all the toxicity can be tolerated and got better after treatment; 4. 3-Dimensional conformal radiotherapy(3D-CRT) can decrease the dose delivered to normal tissues nearby, while increase the dose in target volume. Survival rate could be raised by 3D-CRT. The development potential of 3D-CRT is excellent.

Effects of ABCG2 Gene Silencing on the Proliferative Ability in Vivo and Morphological Changes in Human Lung Adenocarcinoma Cell GLC-82

Mon, 30 May 2011 13:08:55 +0000

Background and objectivesLung cancer is one of the most common malignant tumors which is a big threat to human\'s health,and its morbidity and mortality rates continue to rise.Molecular, biological and immunological studies show that development of lung cancer is a complicated pathological process with interaction of multiple factors and multiple gene mutations.With evolution of scientific research,the treatment and management of lung cancer has improved a lot,but the 5-year survival rate is still below 15%. Since the detection,treatment and prognosis will not be optimistic without a clear understanding of the underlying mechanisms and effective therapeutic options,it is imperative to explore further the etio-pathogenesis and develop more potent means of prevention and cure.ABCG2 gene is a member of ATP-binding cassette transporter superfamily, participating in multidrug resistance(MDR).Because it was first discovered in MCF-7 breast cancer cell sub-line,the protein it encodes is also referred to as breast cancer resistance protein(BCRP).With more in-depth research,evidence has emerged linking ABCG2 and stem cell,suggesting a role as a potential marker of stem cell.However,research on distribution of ABCG2 in lung cancer tissue and cell lines indicate that it cannot be considered as a single marker of lung cancer and so most research on ABCG2 focuses on it\'s correlation with multidrug resistance and stem cells.Our previous study on "effect of ABCG2 on the biological behavior of lung cancer cell line"showes that ABCG2 gene silencing can induce slow proliferation in vitro,whether it\'s influence on the proliferative ability in vivo is unkown.To reveal the effect of ABCG2 gene silencing on the proliferative ability in vivo,this study was carried out based on subcutaneous tumor of ABCG2 gene silencing in human lung carcinoma cell.Furthermore,we use quantitative computerized image analysis and stereological method to show the morphological and ultrastructural changes in human lung carcinoma cell after ABCG2 gene silencing.That we can gain more insight into the relationship between this transporter and lung cancer,which could provide a more effectual strategy for prevention and cure of this dreaded disease.Materials and MethodsⅠ.Effects of ABCG2 gene silencing on the proliferative ability in vivo of human lung carcinoma cell.(1) Human lung carcinoma cell lines GLC-82,GLC-82/EGFP~+ and GLC-82/EGFP~+/ ABCG2~- were implanted into subcutaneous tissue of nude mice to observe the process of tumorgenesis and compare proliferative ability in vivo.(2) Flow cytometry was used to detect the cell cycle of the heteroploid cells.(3) By immuno-histochemical methods(SP) the expressions of Ki-67,ABCG2, Bcl-2 and p53 was assessed.(4) Quantitative studies and morphometric analysis of AgNORs were conducted on tissues with AgNOR staining technique(Ploton). Ⅱ.Effects of ABCG2 gene silencing on morphological changes in human lung carcinoma cell.(1) Morphological cellular features of GLC-82/EGFP~+/ABCG2~-,GLC-82/EGFP~+, and GLC-82 were observed both in vitro and vivo,with cytoskeleton staining used in vitro cells.(2) Morphological parameters of cells(HE) both in vitro and vivo were examined by quantitative computerized image analysis.The parameters included Cells and nuclear included area(A_c,A_n),Circumference(C_c,C_n),Long axis(d_(maj,c),d_(maj,n)), Short axis(d_(min,c),d_(min,n)),Axis Ratio(Ra_c,Ra_n),Karyoplasmic ratio(Rnp),Form factor PE(PE_c,PE_n),Form factor AR(AR_c,AR_n),Regular form factor(RFF_c,RFF_n),and Form irregular index(FII_c,FII_n).(3) The images of subcutaneous tumor were photographed with electron microscopy and stereological analysis carried out using the Image-Pro-Plus analytic software. The parameters included volume density(V_v),surface density(S_v),ratio of surface to volume(Rsv),numerical density(N_v),mean volume(v),mean surface(s),ratio of nucleus to cytoplasm(Rnp),mean free distance(λ).ResultsⅠ.Effects of ABCG2 gene silencing on the proliferative ability in vivo of human lung carcinoma cell.(1) Compared with tumors from GLC-82 and GLC-82/EGFP~+,the tumorgenesis in GLC-82/EGFP~+/ABCG2~- proceeded slowly with smaller volume tumors.(2) Flow cytometry results showed no difference in the percentage of heteroploid cell numbers in G_2 among the 3 groups.The cell numbers in G_1 increased in GLC-82/EGFP~+/ABCG2~- group while the cell numbers in S decreased.(3) Immunohistochemistry results indicated that ABCG2 protein expressed on the tumors of GLC-82/EGFP~+/ABCG2~- group attenuated.Image analysis demonstrated that the Positive Unity(PU) of ABCG2 protein in GLC-82/EGFP~+/ABCG2~- group was lower than GLC-82/EGFP~+,GLC-82 groups with significant difference.Ki-67 proliferation index of GLC-82/EGFP~+/ABCG2~- cells in vivo was significantly lower than GLC-82/EGFP~+ cells and GLC-82 cells(P＜0.05).There were no significant differences in Bcl-2 and p53 expression among the 3 groups.(4) Image analysis revealed significant differences in AgNORs parameters with lesser number of AgNORs and total AgNORs areas but higher mean AgNORs areas in the cells of GLC-82/EGFP~+/ABCG2~- group.AgNORs in GLC-82/EGFP~+/ ABCG2~- group were most mixed type,while dispersed type were the main pattern in GLC-82/EGFP~+ and GLC-82 group.Ⅱ.Effects of ABCG2 gene silencing on morphological changes in human lung carcinoma cell.(1) GLC-82/EGFP~+/ABCG2~- exhibited visible differences in size and shape appearing mainly spindle shaped,whereas GLC-82/EGFP~+,GLC-82 cells were spherical,oval or polygonal.(2) Cytoskeleton staining results showed the cytoskeleton of GLC-82/EGFP~+/ ABCG2~- to be fine and spread throughout the cytoplasm,while that of GLC-82/ EGFP~+,GLC-82 was coarse and clumped around the nucleus.(3) Image analysis of H&E stained cell slide revealed that when compared with GLC-82/EGFP~+,GLC-82 group,the values of d_(maj,c),C_c,Ra_c,d_(maj,n),Ra_n,Ac,, Acp,FII_c,FII_n in GLC-82/EGFP~+/ABCG2~- cell were the highest,but its PE_c,AR_c, RFF_c,PE_n,AR_n,RFF_n,R_(np) were the lowest.(4) Histologically,the volume of the cells in GLC-82/EGFP~+/ABCG2~- group was increased with clear cytoplasm or presence of vacuoles inside.Negative AB-PAS staining ruled out the presence of mucus inside the cytoplasm.The nuclei of GLC-82/EGFP~+/ABCG2~- cells exhibited marked pleomorphism with numerous bizarre mitoses.Image analysis revealed that the values of PE_n,AR_n,RFFn and R_(np) decreased while FII_n,A_c,Acp increased.(5) Image analysis of HE stained tumor tissue revealed that when compared with GLC-82/EGFP~+,GLC-82 group,the values of FII_n,A_n,Acp,in GLC-82/EGFP~+/ ABCG2~- cell increased,but its PE_n,AR_n,RFF_n,R_(np) decreased(6) Ultrastructural examination showed obvious pleomorphism of nucleus in both GLC-82/EGFP~+/ABCG2~-,GLC-82/EGFP~+ groups with numerous lysosomes.The characteristic changes in GLC-82/EGFP~+/ABCG2~- group were mitochondrial swelling and dilatation of endoplasmic reticulum,with presence of specific "albuminoid material" inside the cytoplasm crushing the nucleus.(7) Three-dimensional quantificational results of the ultrastructure revealed that the V_v,S_v,R_(np) of nucleus decreased withλincreased in GLC-82/EGFP~+/ABCG2~-group;the value of V_v,S_v for mitochondrion was higher andλlower,V_v for endoplasmic reticulum was higher in GLC-82/EGFP~+/ABCG2~- group.Conclusions:1.ABCG2 gene silencing can effectively suppress the proliferative ability of lung carcinoma cells in vivo:the tumorgenesis of GLC-82/EGFP~+/ABCG2~- cells proceeded slowly with smaller volume tumors,indicating that ABCG2 gene is significant to the proliferate of GLC-82 cell in vivo and it could be a curative target to be deeply investigated.2.Ki-67 proliferation index decreased in GLC-82/EGFP~+/ABCG2~- group;Flow cytometry results showed that most percent of cells in GLC-82/EGFP~+/ABCG2~-group were inhibit in G1;number of AgNORs and total AgNORs areas but higher mean AgNORs areas in the cells of GLC-82/EGFP~+/ABCG2~- group.AgNORs in GLC-82/EGFP~+/ABCG2~- group were most mixed type,while dispersed type were the main pattern in GLC-82/EGFP~+ and GLC-82 group. 3.After ABCG2 gene silencing the morphology of GLC-82 cell changed:the lung cancer cells in vitro transform into spindle shaped cells.In vivo,the cell body dilated with nucleus heteromorphism(V_v,S_v,R_(np) decreased andλ,increased) mitochondrial swelling(V_v,S_v increased andλdecreased) and dilatation of endoplasmic reticulum(V_v increased and Rsv decreased) ultrastructurally.Yhe volume of the cells in GLC-82/EGFP~+/ABCG2~- group was increased with clear cytoplasm and nucleus heteromorphism histologically.

The Clinical Observation of Wind Evil Encroach Lung-type Cough Variant Asthma That Treatment of Ke Ping Tang

Mon, 30 May 2011 13:08:55 +0000

Objective:To observe the effect of treating Wind evil encroach lung -type cough variant asthma with ke ping tang united conventional treatment of western medicine and western medicine treatment,comparison of the two effects,With the effects of Cytokines IL-13 and the target of Pulmonary function-the largest peak expiratory flow,Discussion of Chinese medicine treatment of cough variant asthma General advantages,Cytokines involved in the pathogenesis of IL-13 regulation of the advantages and the improving situation Of pulmonary function.Information and methods:1.Selection wind cough disease traditional Chinese medicine belong to Wind evil encroach lung-type,andReference to Western diagnostic criteria of CVA,Select 60 cases of patients,and according to traditional Chinese medicine dialectical standard classification for randomly divided into traditional Chinese and western medicine and western medicine treatment of 30 cases of control group.2.Both groups were given Shu li die(50/100) inhalation treatment,Treatment group at the foundation of western medicine on the use of add Hirayu with the treatment of cough, Compared clinical efficacy of the two groups,Detection of two groups before and after treatment in the largest PEF,Numerical of IL-13,Change of dosage about determination of bronchial provocation test,In the course of 4 weeks after the end of follow-up cases of recurrence of the number of cases cured.3.Statistical treatment:SPSS13.0 statistical software used for the United States the results of statistical treatment,Set for P≤0.05 statistically significant difference.(1) pre-treatment comparability analysis(2) Group matching t test before and after treatment(3) group into a group t test(4) Level of information using chi-square test,Level count data using rank sum test,Comparison of the rate of adoption of the U test.Results:1.Clinical Observation:Comparison of TCM Syndromes points and General efficacy, The two groups before and after treatment within each group of points compared syndrome were significantly statistical significance(P＜0.01),Two groups before and after treatment comparison points TCM Syndrome have decreased significantly.The two groups after treatment of TCM Syndrome margin points and difference between two points were no significant differences(P＞0.05),General effects of the two groups no significant difference (P＞0.05),The efficiency of the two groups after treatment significantly higher than those of comparison group slightly.2.The intervention of ke ping tang on the cytokine IL-13:Compared the two groups before and after the treatment group,IL-13 values,Two groups after treatment and difference after treatment were all significant differences(P＜0.01),Lower levels of IL-13 treatment group was significantly better than the control group.3.The intervention of ke ping tang on PEF:Compared the two groups before and after the PEF treatment group is expected to account for the value of the percentage of value,PEF between the two groups are expected to account for the value of the percentage difference between the value and treatment comparison,Were all significant differences(P＜0.01),PEF improvement in the treatment group was significantly better than the control group.4.The intervention of ke ping tang on airway hyperresponsiveness(Determination of BHR cases dosage Level):The two groups after treatment dosage level of BHR,compared to the situation there is statistically significant difference(P＜0.05),The two groups dosage level of BHR\'s difference before and after treatment is significant differences Treatment group, airway hyperresponsiveness significantly lower than the control group.5.Two cases of recurrence after treatment follow-up compared to the situation:Cured two cases of recurrence after 4 weeks follow-up compared to the situation there is statistically significant difference(P＜0.05),The number of recurrent cases of the treatment group than the control group.6.The course of the treatment group did not find obvious side effects,0 cases of test subjects during the exit.Conclusion:1.Improvement of TCM Syndromes points and General aspects of efficacy to determine,after 4 weeks the treatment of TCM Syndrome General effects of integration The two groups had no significant difference,But follow-up cases of recurrence of the number of cases cured at 4 weeks after the end of treatment,Western medicine treatment of simple cases of the control group recurrence was significantly more than the treatment group,Description that help with Western medicine Wind evil encroach lung-type cough variant asthma treatment effective,And a significant reduction in recurrence rate after cessation of medication.2.A significant reduction in the level of IL-13,improving the level of PEF,With the determination of BHR Change dosage,The treatment group was significantly better than the control group,western medicine alone,Description that ke ping tang to reduce airway responsiveness,inhibition of cytokines involved in the pathogenesis and reduce airway inflammation there is synergy.Maybe are one of the mechanisms of Qufeng Xuanfei,Liyan Yang Jin traditional Chinese medicine.3.ke ping tang has obvious security and effective on Community-acquired pneumonia.

The Interference Effect on HepG2 Human Liver Cancer Through RNA Interference

Mon, 30 May 2011 13:08:54 +0000

Objective:Hepatic carcinoma in China is one of the most common malignant tumor.Its pathogenesis is complex,Which appeare in the tumor cells and tumor-related gene expression is an important mechanism.Midkine (MK) and the tumor is closely related to the occurrence and development. MK can regulate cell growth, survival and differentiation, and participate in tumor pathogenesis.many different types of tumor are high expression including the esophagus, stomach, colorectal cancer, pancreatic cancer, gallbladder, lung, thyroid, breast, bladder, uterus, ovary, prostate and liver cancer, neuroblastoma, malignant gliomas, myeloid leukemia , the positive expression rate of between 45% ~ 90%, MK in the normal tissues did not express or low expression.RNA interference (RNA interference, RNAi) technology is developed in recent years in the field of RNA molecular biology technology,which means making cell-specific mRNA degradation which is mediate by endogenous or exogenous double-stranded RNA (dsRNA) in order to silience target gene expression and generate corresponding function losses.It has greate advatages in a strong inhibitory effect, high stability etc,we preliminary sudy significance on MK in hepatocellular carcinoma cells, provide experimental basis for the treatment of hepatic carcinoma.This experiment, we use Pgenesil-2.1 to construct MK siRNA expression vector,transfect into HepG2.Using flow cytometry, MTT method, RT-PCR, Western-blot,we check the role of MKsiRNA in mRNA and protein expression, the sensitivity of chemotherapeutic drugs as well as the impaction on apoptosis, preliminary study of liver cancer cells at MK expression meaning, in order to open up MK-mediated RNA interference to provide experimental basis for treatments of liver cancer.Method:1.Construct MK siRNA expression vector, indentify inserting sequence’validility.2.Through cationic liposome, MK interference plasmid and general negative interference plasmid (HK) transfected into HepG2 human liver cancer cells, G418 pressure screening, testing the Neo gene expression in cells,and then getting more positive cloning.3.After expanding positive and negative cloning,using AnnexinⅤ-FITC and PI double staining method detect apoptosis in each group of transfected HepG2 cells by flow cytometry.4.Uses MTT assay to check tumor cells sensitivity to chemotherapy drugs.5.Uses RT-PCR to detect expression of MKmRNA in each group.6.Uses Western-blot to detect MK protein expression.Result: .1 HepG2 cells which are transfected MK siRNA interference plasmid, HepG2 cells which are transfected the general negative control plasmid and normal cells change shapes.2.Both HepG2 cells which are transfected MK siRNA interference plasmid and HepG2 cells which are transfected the general negative control plasmid are Neo gene expression.3. HepG2 cells which are transfected MK siRNA interference plasmid have the spontaneous apoptosis, early apoptosis increased by 15.5 times and 9.34 times than the negative control group and blank group.4. By MTT assay,using the cell culture medium which contain different concentrations of cisplatin and 5 fluorouracil culture cells 48 hours, HepG2 cells which is transfected with MK positive interference plasmid have the lowest survival rate in the same concentration of the three groups of cells,which indicate it is sensitive to chemotherapy drugs.5.Protein and mRNA expression of MK in HepG2 cells which are transfected interference plasmid fell respectively 89.3% and 77.3%.Conclusion: HepG2 cells which are transfected MK siRNA interference plasmid can enhance apoptosis of tumor cells and improve the HepG2 cells\'sensitivity to chemotherapy drugs,such as cisplatin and 5-fluorouracil. At the same time MK-mediated RNA interference provide experimental basis for treatments of liver cancer.

Acinetobacter Baumanii as Lower Respiratory Tract Infection: Epidemiology, Drug Resistance and Correlation Factor

Mon, 30 May 2011 13:08:53 +0000

Background and ObjectivesFor the past few years,to control the nosocomial infection caused by multidrug resistant gram-negative bacteria is constantly the important problem in the field of medicine.Although many new antibiotics had come out and been used extensively, effectivelly decreased the rate of nosocomial infections caused by Enterobacteriaceae,they increased the number of Nonfermentative bacteria such as Pseudomonas aeruginosa,Acinetobacter baumannii.In the past,the infections caused by A.baumannii were rare,now because of extensive use of broad-spectrum antibiotics,the isolation rate of A.baumannii is increasing year by year and the second highest,just lower than that P.aeruginosa.A.baumannii has emerged as an important nosocomial pathogen that is mainly responsible for lower respiratory tract infections,also comprising bacteremia,meningitis,soft tissue infection,peritonitis and so on.A.baumannii was usually highly resistant to antibiotics,exhibited multidrug-resistance,especially resistant to carbapenem which was very difficult to treat,so A.baumannii has been nicknamed the "Gram-negative MRSA".Furthermore, A.baumannii resistant to soap can survive for a longer time in hospital environment, and was the most common gram-negative bacteria isolated from the hand of medical workers.In this way,A.baumannii can increase the hazard of cross infection between the patients,easily result in nosocomial outbreak.We should enhance the study of A.baumannii,so as to provide effective scientific basis for its prevention and treatment.Although the study of A.baumannii were considerably reported in the world, because of the difference among environment,sterilized measure,therapeutic tool and risk factors in different regions,so there were significant differences in antimicrobial resistance.For this reason,we should master the epidemiological characteristics and drug resistance in our own hospitals,which is of practical importance to prevent and control nosocomial infection.Therefore,this study had analyzed the data of 198 cases of lower respiratory nosocomial infection caused by A.baumannii,which was discussed as follows:(1)To investigate epidemiological characteristics and drug resistance retrospectively,(2) To study the risk factors and prognosis of hospital acquired pneumonia caused by carbapenem-resistant A.baumannii,(3) To analyze the factors influencing bacterial clearance and treatment strategy.MethodsWe analyzed the clinical data of 198 case of lower respiratory tract infections caused by A.baumannii from January,2005 to December,2007.Meanwhile,a total of 356 A.baumannii isolates were collected to study.1、By descriptive epidemiological methods,the reported epidemic data was analyzed retrospectively.2、Susceptibility of 356 strains of A.baumannii to 14 kinds of antimicrobial agents was detected by standard K-B methods.3、By a case-control study,the risk factors and prognosis of 44 cases of nosocomial infection caused by CRAB were analyzed.4、According to the bacteria therapeutic effect criteria,the patients were categorized as clearance group(n=86) and non-clearance group(n=112).Bacterial eradication rate and the factors influencing bacterial clearance were analyzed respectively.5、All statistical tests were operated with the program SPSS13.0,statistical data was expressed by(?)±SD,Univariatesis(T test and chi-square test) and multivariate logistic regression were used for statistics analysis.All tests were two-tailed,and a value of P≤0.05 was considered significant.Results1、Most of A.baumannii were from lower respiratory tract(79.2%,595/751).The isolation rate of A.baumannii tended to increase year by year(2005:9%, 2006:13.2%,2007:16.3%).The rate of infection was 59.8%(356/595).A. baumannii strains were mainly distributed in the ICUs(70.8%,252/356),ICU of neurology department as the most ones.Most of these patients were diagnosed as stroke in hospitalization.Their mean age was 49.4 years,the ratio of male to female was 2.0:1.There were A.baumannii cases reported in the whole years.Our data of continuous 3 years showed that the rate of monthly infection had 3 epidemic peaks in April,July,September,with the difference less than one month.2、The antibiotic susceptibility test in vitro showed:Drug resistance of A.baumannii was a serious problem,77.2%(275/356)of stains showed multiple resistance. Imipenem(76.7%susceptible) and meropenem(74.2%susceptible) were the most active agents against A.baumannii,followed by cefoperazone-sulbactam(67.1% susceptible),the intermediate rate to cefoperazone-sulbactam(28.1%intermediate) was highest,and other 11 kinds of antimicrobial agents were less than 30%.The resistant rate to cefoperazone-sulbactam was lowest.The resistance rates to cefoperazone-sulbactam,imipenem and meropenem were less than 50%in ICU. Except cefoperazone-sulbactam,the resistance rate in the ICU was(11.1%～31.2%)higher than the others with significant difference(P＜0.05).Less than 11% of carbapenem resistant strains were resistant to cefoperazone-sulbactam,others were over 80%.3、Positive correlation(r=0.896,P=0.000) and linear regression(R~2=0.803)existed between imipenem and meropenem in diameter of inhibition zone,and the regression equation was Y=0.928X+1.241.4、Univariate analysis revealed that five factors associated with the infection caused by CRAB were APACHEⅡscore,chronic pulmonary disease(COPD/bronchiectasis), imipenem/meropenem and fluoroquinolone used 15 days before isolation of CRAB, and early combination therapy of antibiotics.Multivariate logistic regression analysis identified two independent factors:APACHEⅡscore(OR=1.40,95% CI=1.19～1.64) and imipenem/meropenem used 15 days before isolation of CRAB (OR=5.57,95%CI=1.71～18.18).5、Of 44 cases of CRAB infections,14 patients died and 30 patients survived. Univariate analysis revealed that two factors associated with poor prognosis were organ failure,clinical pulmonary infection score(CPIS) rise after three-day treatment. According to multivariate logistic regression analysis,only CPIS rise after three-day treatment(OR=7.01,95%CI 1.23-40.03) was an independent predictive factor.6、The total bacterial eradication rate of lower respiratory infection caused by A.baumannii was 43.4%,but in MDR and Non-MDR A.baumannii was 36.0%, 59.7%respectively(x~2=9.693,P=0.002).The factors influencing bacterial clearance were APACHEⅡscore,MDR,coma,mechanical ventilation,tracheal intubation/incision and CPIS rise after three-day treatment(P＜0.05).7、In clearance group,57 cases were treated according to the results of susceptibility testing and recommended antibiotics,average duration of bacterial clearance was (17.6±9.7) d.Oppositely,29 cases were not treated according to this criteria, average duration of bacterial clearance was(23.8±17.1) d.The factors significantly influencing duration of bacterial clearance were MDR,APACHEⅡscore≥15, mechanical ventilation and tracheal intubation/incision(P＜0.01).8、Different therapeutic strategies(Antimicrobial monotherapy or combination therapy) influencing duration of bacterial clearance were compared,the results showed that In Non-MDR A.baumannii group(n=37),monotherapy vs combination therapy, Z=-0.198,P=0.843;In MDR A.baumannii group(n=20),monotherapy vs combination therapy,Z=-1.941,P=0.052. Conclusion1、The main side of A.baumannii infection is lower respiratory,strains represented severe multiple-resistance.It is important to monitor closely the key departments, vulnerable population and high occurrence time,take effective measures promptly, so as to prevent and control nosocomial infection.2、Carbapenems are still the most active agent against A.baumannii,but the resistant rate to cefoperazone-sulbactam was lowest,and the intermediate rate was highest.So it is possible to improve therapeutic effect by adding doses within safety margin,especially for CRAB.3、Imipenem showed the same sensitivity as meropenem.So it seems to be impossible to alternate medicine between these two drugs.4、APACHEⅡscore and imipenem/meropenem used 15 days before isolation of CRAB were independent risk factors for CRAB infection.CPIS rise after three-day treatment was a predictive factor for the prognosis of CRAB infection.5、The main factors influencing bacterial clearance were APACHEⅡscore,MDR, coma,mechanical ventilation,tracheal intubation/incision and CPIS rise after three day treatment.6、In order to attain satisfactory effect on bacterial clearance,duration of therapy is at least two weeks.It needs prolonged therapy for the patients who have risk factors.It is not significant on duration of bacterial clearance between monotherapy or combination therapy.

Expression of a Subfamily Member of Transient Receptor Potential Gene (TRPV4) in Rats Testis

Mon, 30 May 2011 13:08:53 +0000

BackgroundIn the cell environment,the calcium ion is the key informational molecule and the only way to trigger cell proliferation while the intracellular calcium ion concentration in cytoplasm is regulated by the cell membrane calcium channel.The latest research on the cell membrane calcium channel shows that,it\'s the only way for calcium influx induced by cell membrane activated calcium channel to trigger cell proliferation;non-voltage-dependent calcium channel,Transient Receptor Potential (TRP),has a function to regulate cell survival,growth,death and so on.At many TRP-mediated functions and disease,it at the role of cell proliferation is particularly prominent,which in many tissues and ceils,have been supported by experimental results.For example,the pulmonary artery smooth muscle cells of TRPC1 and TRPC6 expression was significantly higher,and their by-mediated calcium influx stimulate smooth muscle cell proliferation,which eventually led to the occurrence of pulmonary hypertension;source of vascular endothelial growth factor (VEGF) by activating endothelial cells TRP channels,causing calcium influx, stimulating vascular endothelial cell proliferation can lead to tumor angiogenesis; TRPM1 expression and the malignant melanoma is closely related to the degree; These results indicate that,TRP in the regulation of cell proliferation has a role that can not be ignored.At present,many studies have found that,TRP gene family at a number of subtypes have on the expression of the testis,in part or high expression.However, research in this regard is still very small,it is necessary to study.At the above-mentioned study on the testis epididymis are limited to the question whether the expression,since the TRP family in the testis on the expression of such a rich place in the testicular function on it becomes a concern,and on current international research is still There is no relevant research reports.On the current international research shows that:TRPV4 substantial expression in renal distal convoluted tubule of the epithelial tissue,but also a small amount of expression in heart,liver,lung,spleen,testis,and many other organizations.Although functional studies have made considerable achievements,such as:TRPV4 in airway smooth muscle cells may relate to airway hyperresponsiveness,such as asthma; TRPV4 at the expression of Corti may indicate sensorineural hearing damage; TRPV4 with ewes caused by temperature change on the micturition reflex.However, in TRPV4 on testicular function has not yet reported.To study its relationship with the testis will have important practical significance and broad prospects for development.Therefore,the study of TRPV4 and testis,as well as on the distribution function is of great significance.To this end,we experiment on animals through TRPV4 channels and the relationship between testicular tissue for a preliminary study.1 PurposeTo study the expression of TRPV4,a subfamily member of TRP gene in rat\'s testis.To Explore the TRP gene family in testis on the expression for the testicular tissue on TRPV4 function of laying the foundation.2 Method2.1 RT-PCR detection of TRPV4 in the rat testis tissue mRNA expression 2.1.1 TRPV4 gene primer designTRPV4 landing rats genebank,using Primer Premer5.0 software to design primers.2.1.2 Total RNA Extraction and cDNA SynthesisRats by intraperitoneal injection of phenobarbital death,testieular tissues collected and immediately stored into liquid nitrogen.100mg from testieular tissue,in accordance with the common method for molecular cloning TRIzol extracted total cellular RNA;then at 37℃with DNaseI digestion 30min,to remove possible DNA contamination.Nucleic acid/protein quantitative determination of its A260 spectrophotometer and A280.For A260/A280 greater than 18 but less than 20 of the RNA samples,check lug total RNA,in accordance with the instructions synthesis reagents cDNA.2.1.3 RT-PCRThe first chain as a template,amplified by the following conditions:94℃10min, 94℃45s,55℃45s,72℃1min,30cycles,then 94℃1min,60℃1min,72℃10min.PCR products line 0.7%agarose gel eleetrophoresis.Gel imaging Analyzer.2.2 Immunohistochemistry Detect TRPV4 protein in rat testis tissues2.2.1 TRPV4 buy monoclonal antibody and SABC immunohistoehemistry kit TRPV4 monoelonal antibodies.2.2.2 Specimen acquisition and handling:Rats by intraperitoneal injection of phenobarbital death,collected testicular tissue into formalin solution fixed,by the ladder dehydration,xylene and the Baptist transparent wax,the paraffin-embedded tissues.2.2.3 ImmunohistochemistryUsing SABC method,all tissue samples with thickness of 4μm serial sections 3, in accordance with the instructions reagents operation,the use of phosphate buffer solution(PBS) in place of anti-1 for negative control.To the cytoplasm showed brown particles for positive expression.2.3 Western blot detection of the third part of TRPV4 protein in rat testis tissues2.3.1 Specimen acquisition and processingRats by intraperitoneal injection of phenobarbital death,collected testicular tissue,using protein extracted protein extract,using pre-cooling of the PBS wash 2 times,go supematant,Canada on 4×SDS gel sample buffer,denatured in boiling water bath 10min.2.3.2 Western blotPreparation of conventional SDS-PAGE gel,two sets of the same sample points, 120V about 3.5h after electrophoresis,cut one of a group by conventional methods fixed,Coomassie brilliant blue R-250 dyeing and bleaching,as a judge based on Western blot.Another group of samples to cut plastic membrane,nitrocellulose membrane by tailoring the size of gel and do a good job in the beginning and end tags, filter paper on both sides of the buy 3,according to plastic film negative is placed,4℃,18V switch membrane overnight.Remove the membrane with liquid closed closed at least 2h,TBS washing liquid membrane 3 times 10min;add 1:100 diluted anti-1,4℃role 2h;TBS washing liquid membrane 3 times 10min;add 1:5000 diluted goat anti-rabbit IgG secondary antibody at room temperature,the role of 1h; TBS washing liquid membrane 3 times 10min;imaging technology to observe by reflection band.Gel Logia 200 with camera imaging system and analyzed.3 Results3.1 RT-PCR detection of TRPV4 in the rat testis tissue mRNA expressionUV instrument at the next transmission,TRPV4 the lane at the standard nucleic acid molecular weight near a specific amplification bands of approximately 237 bp fragment about the size,image analysis showed that the product of higher purity,with no more than specific interference.3.2 Immunohistochemistry Detect TRPV4 protein in rat testis tissues TRPV4 can be seen under the microscope-positive film,brown many antigen-antibody complex deposition in the testicular tissue in the more mature spermatogenic cells and some Sertoli cell nuclei;TRPV4 negative film at brown no antigen-antibody complex deposition.3.3 Western blot detection of TRPV4 protein in rat testis tissuesWith Gel Logia 200 Imaging System camera,the picture shows:In the test lane, in the 36KDa and 871KDa Department clearly shows that the positive bands,and in the negative control lane,only to see the positive in the Department 36KDa bands can be identified in rat Testicular tissue has TRPV4 protein and GAPDH protein expression,GAPDH protein as a reference.4 ConclusionTRPV4 in the testicular tissue has on the expression,the main expression in more mature spermatogenic cells and supporting cells,showing strong expression.

Behavioral and ERP Study on Heroin Abstainers Attentional Bias

Mon, 30 May 2011 13:08:52 +0000

Drug dependence includes physical dependence and psychological dependence. Relapse is key to cure drug abusers , and psychological dependence is the main reason of relapse. Attentional bias is the main area on cognition study of drug dependence. Attentional bias on drug-related cue has been suggested for two decades. Most learners contend that is a fact. They demonstrate the theory by means of testing reaction time and recording EEG. In China, heroin prevail more extensively than other drugs. At present, there are several literatures about attentional bias on heroin-related cues overseas. No one deploy the emotional Stroop to study the attentional bias in China yet.The present research tried to approach the cognitive mode of heroin abusers . To erect two reference , including neutral and evocative picture, We draw some conclusions. We chose neutral and evocative picture of no emotional motivation from IAPS so as to acquire motivational trait to emotion of heroin-related picture.Some main conclusions were drawn as follows:The main effect was found for cue overall. The RT of heroin-related picture was longer than neutral picture in patients. There was no difference between RT of evocative picture and heroin-related picture in patients. The RT of heroin-related picture in patients was longer than healthy control subjects. These result provide evidence for attentional bias.The reaction mode of patients was different from healthy control subjects. Ragard to amplitude of P300 and SSPW, we draw a conclusion that the amplitude for heroin-related picture was equal to evocative picture and was more than neutral picture in patients, but in healthy control subjects the amplitude for heroin-related picture was equal to neutral picture and less than evocative picture.Someone contend that left frontal sites would be more responsive when subjects were experiencing heroin-related pictures as appetitive. The present research found that amplitude of SSPW was higher at F3 site than right site. We support the standpoint.A significant interaction between cue and group was found for SSPW. This indicate that amplitude induced by heroin-related pictures was higher than neutral pictures and that amplitude of patients was higher than healthy control subjects.Although a significant main effect was found for P300, it would be approached whether it may be regarded as sensitive index.The present research made a meaningful attempt to compare the three cue stimuli. It’s conceivable that human beings will get more systematic understanding about their neurobiological bases and functions with the development of cognitve neuroscience, brain neuroimaging and electrophysiological technology.

Prevention Study of Radiation-induced Lung Injury

Mon, 30 May 2011 13:08:52 +0000

ObjectiveRadiation-induced pulmonary injury, a frequent complication of pulmonary radiation therapy for lung cancer, is usually unavoidable and is associated with a very poor prognosis. Since lesion progression is often irreversible and can lead to fatal respiratory failure Therefore, strategies to prevent and/or ameliorate radiation-induced lung injury could be beneficial both in the clinical setting and in the event of accidental radiation exposure. In this study, we will observed the effect of ulinastatin on radiation-induced lung injury caused by the linear accelerator in SD rats in order to find a novel way to prevent radiation-induced pulmonary interstitial pneumonia and interstitial fibrosis. This study divided into two parts: 1. Establish the model of radiation-induced lung injury on SD rats by Linear accelerator; 2. Effect of pretreatment with high-dose ulinastatin in preventing radiation-induced pulmonary injury in rats; 3. Effect of Recombinant Human Tumor NecrosisFactor-O Receptor (rhTNFR:Fc) on tumor necrosis factor in rats with radiation-induced lung injury. This research is clinical foundation research, which will have great value to benefit both military medicine and civilian medicine.Methods1. Rats were restrained in custom-built apparatuses and were anesthetized by i.p. injection of pentobarbital (40mg/kg, Sigma, USA). Their heads and abdomens were protected from irradiation by lead, and the indicated dose distribution was achieved by placing a 23-mm thick wax block above their chests. Irradiation was performed with a linear accelerator producing 6 MV photons (Siemens, Germany) at a focus-to-surface distance of 100 cm. Radiation was delivered at a single dose of 20 Gy (2.5 Gy/min) with a field size of 4.5 cm×4.5 cm.2. In ulinastatin treatment exprement, rats were randomized into five groups. Group R (negative control; 60 rats) received i.v. intraperitoneal saline injections, in lieu of ulinastatin, pre- and post-irradiation. Groups P1 and P2 (45 rats each) were treated with ulinastatin (Miraclid, Mochida Pharmaceutical, Japan) for 3 d pre- and 4 d post-irradiation at the pre-determined high (400,000 U/kg body weight/d) or low (200,000 U/kg body weight/d) dose, respectively. Groups A1 and A2 (45 rats each) were treated post-irradiation for 7 days with high or low dose ulinastatin, respectively.3. In rhTNFR:Fc treatment erprement, rats were randomized into three groups: group R (negative control) received radiation and i.v. intraperitoneal saline injections, group C (normal control) only received i.v. intraperitoneal saline injections, group Y (treatment control) received radiation and rhTNFR:Fc at a dose of 5mg?kg-1 twice in first week while rats in control group and irradiation group were injected with the same volume of saline. 4. Observed rats and record the mortality of each group rats after radiation; bronchoalveolar lavage fluid was centrifuged at 360×g for 10 min, and the supernatant used for the measurement of TGF-β1 was stored at ?70°C. Two milliliters of blood were obtained by cardiac puncture, collected on ice using EDTA, and centrifuged to test IL-6、TGF-β、LN、Hyp; A piece of lung tissue was and stored at ?70°C until further processing stained with hematoxylin-eosin or Masson’s trichrome to observe the general pathological changes of the lung tissue and collagen fibrils; Immunohistochemical analysis of TGF-βand TNF-α.; Western blot analysis of TGF-βand TNF-α.Results1. Rats in each group suffered different degrees of radiation injury after irradiation. The levels of TGF-β, IL-6 and TNF-αin serum,BLAF and lung tissue were gradually increased which is related to the severity of lung injury, especially the simple radiation group, which suggested that IL-6 and TNF-αand TGF-βplay an important role in radiation-induced pulmonary injury.2. The levels of TGF-β,IL-6 and TNF-αand the severity of radiation-induced lung injury and radiation-induced pulmonary fibrosis in treatment groups were significantly lower than in irradiation group, which means that ulinastatin could prevent radiation-induced lung injury effectively by inhibiting TGF-β, IL-6 and TNF-α.3. After irradiation,pretreatment of high-dose ulinastatin could inhibit TGF-β, IL-6 and TNF-αlevel more effectively than other groups while the lighter severity of radiation-induced lung injury, which was according to the reports that the TGF-β, IL-6 and TNF -αlevel could reflect the degree of radiation-induced pulmonary injury; levels of LN and Hyp in rats of pretreatment of high-dose ulinastatin decreased significantly compared with other treatment groups at 24 weeks after irradiation.4. Expression of TNF-αand IL-6 in rats treated with rhTNFR:Fc decreased significantly compared with rats only accepted radiation.ConclusionsUlinastatin especially pretreatment of high-dose ulinastatin and rhTNFR:Fc can prevent radiation-induced pulmonary injury and fibrosis effectively which provides a novle way to treat radiation-induced pulmonary injury.

Preparation of MRI Contrast Agent Based on Chitosan and Its Relaxation

Mon, 30 May 2011 13:08:51 +0000

As a naturally occurring polysaccharide,chitosan shows many unique properties, such as biocompatibility,biodegradation,low toxicity and biological activity.Many derivatives have been prepared to meet different needs.In this paper,the development of modification of chitosan and the application of its derivatives were introduced. First,we have synthesized several chitosan derivatives with different substituting group,such as AA、DTPA and Ga.Then,these CS derivatives are prepared for chelating Gd~(3+).the Chemical structure and morphology of these CS derivatives were characterized with ~1H NMR,~(13)C CP/MAS NMR spectroscopy,ultraviolet spectroscopy and Transmission electron microscopy.We have studied their relaxation in the pure water.The results show,these CS derivatives presented good water solubility and good relaxation ability.And then evaluated their toxicity in vitro,as a result,CS-Ga-(Gd-DTPA)_n has better biocompatibility than Gd-DTPA which is one of commercial magnetic resonance imaging contrast agent.Therefore,the compound CS-Ga-(Gd-DTPA)_n can be used for further study for macromolecular contrast agents.

Inhibitory Effect on Asthma by Dendritic Cells of Schistosoma Japonicum-infected Mice

Mon, 30 May 2011 13:08:51 +0000

Objective This paper aimed at studying the inhibition of allergic asthma by adoptive transfer of dendritic cells(DCs) from Schistosoma japonicum-infected mice and its mechanisms.Methods DCs from schistosoma japonicum-infected mice and naive mice were respectively isolated and purified using CD11c+ microbeads.Twenty BALB/c mice were divided into four groups randomly.10~6 DCs of Schistosoma japonicum-infected mice were adoptively transferred to each mouse in Group A by tail vein,while 10~6 DCs of naive mice were adoptively transferred to each mouse in Group B in the same way,Group C was asthma group,Group D was control group of naive mice.2 hours later,mice in Group A,B and C were sensitized and challenged with OVA to induce asthma.4 weeks later,all the mice were killed. Sera were collected for measurement of OVA-specific IgE;Bronchoalveolar lavage fluids(BALF) were collected for measurement of various cytokines(IL-4,IL-5, IL-10 and IFN-γ);Spleen cells were collected and cultured,72h later,spleen culture supernatants(SPC) were collected also for determination of IL-4,IL-5,IL-10 and IFN-γ,Lungs were collected for histological analysis.Results Compared with mice in Group B and C,pulmonary inflammation of mice in Group A diminished markedly (P＜0.05).Moreover,the levels of IL-4 in SPC of mice in Group A、B、C and D were(18.22±2.77)pg/ml,(37.88±10.68)pg/ml,(146.61±36.32)pg/ml and (11.28±1.15)pg/ml respectively;The levels of IL-5 were(170.33±21.32)pg/ml, (1021.94±70.71 )pg/ml,(3882.16±493.42)pg/ml and(2.51±1.34)pg/ml respectively: The levels of IFN-γwere(108.59±45.56) pg/ml,(1073.02±450.14)pg/ml, (582.48±207.76)pg/ml and(95.91±5.98)pg/ml respectively.Compared with mice in Group B and C,levels of IL-4、IL-5 and IFN-γin SPC of mice in Group A decreased significantly(P＜0.05).The levels of IL-4 in BALF of mice in Group A、B、C and D were(23.25±1.57 ) pg/ml,(40.70±3.28)pg/ml,(65.23±4.84)pg/ml and (11.28±1.15)pg/ml respectively;The levels of IL-5 were(16.20±4.11) pg/ml, (41.41±7.14)pg/ml,(64.75±16.96)pg/ml and(0.34±0.16)pg/ml respectively;The levels of IFN-γwere(4.48±0.67) pg/ml,(5.90±1.24)pg/ml,(5.97±1.59)pg/ml and (5.77±0.98)pg/ml respectively;The levels of IL-10 were(281.05±38.71)pg/ml, (120.16±37.51)pg/ml,(93.99±29.91)pg/ml and(110.48±22.53)pg/ml respectively. Compared with mice in Group B and C,the levels of IL-4 and IL-5 of mice in Group A decreased significantly(P＜0.05),and the levels of IL-10 were increased(P＜0.05), whereas the levels of IFN-γwere comparable(P＞0.05).The levels of OVA-specific IgE were(283.22±83.67 ) pg/ml,(1180.61±209.02)pg/ml,(1389.26±247.61)pg/ml and (197.83±24.12)pg/ml.Compared with mice in Group B and C,the levels of OVA-specific IgE of mice in Group A decreased significantly(P＜0.05).Conclusion DCs of Schistosoma japonicum-infected mice played a critical role in inhibition of allergic asthma through influence of secretion of Th2 cytokines by receptor mice.

Applied Anatomy of the Endoscopic Endonasal Approach to the Ventral Cranio-cervical Junction

Mon, 30 May 2011 13:08:30 +0000

Craniocervical junction is a complex anatomical structure,Gladstone and Erickson-Powell first described this region in 1815,which includes inferior one-third of clivus,atlas,atlantoaxial and foramen magnum,as well as the important neurovascular structures in the region.Because the region is located adjacent to the skull base,deep brain stem,upper cervical cord and other vital structures,anatomical relationship are so complex that it is difficult to have an operation.Although the traditional treatments of surgical lesions in the region can relieve the lesions on the craniocervical junction at different levels and ease the oppression,but these methods often cause damage to a large extent of bone tissue and treatment of traction of neurovascular.Traumas are too large and have the existence of serious complications. As far as the ventral craniocervical junction lesions are concerned,Kanavel firstly proposed the transoral approach in 1917,which uses the body\'s natural physiological Lacunar,and no essential blood vessels nerves exist in front of craniocervical junction,the approach can directly expose cranio-cervical junction ventral midline lesions from the inferior one-third of clivus to the C_2 vertebral body.Even a considerable number of scholars believe that such a surgical approach is the optimal solution to the lesions of ventral craniocervical.junction.But the traditional transoral approach also have a larger wound because it requires incision of hard palate and soft palate,damaging the teeth and temporomandibular joint,causing tongue edema ischemia even necrosis.In recent years,with economic development,social progress and people\'s living standards improvement,people required surgeons removed their lesions completely, at the same time,also called for as much as possible to reduce the surgical invasion and improve postoperative quality of life.Traditional surgical methods have been replaced by minimally invasive surgery gradually,because of their large trauma,the high incidence of complications,great surgical risks,a long recovery period,patients suffering big disadvantage,and higher medical costs.Since Steven Gans and Bradley Rodgers first diagnosed the disease for infants and children by using laparoscopy and thoracoscopy,minimally invasive surgical techniques has been developed rapidly.Simultaneously with the development of optical fiber,medical laser,micro-electronics,as well as the new biological materials, the minimally invasive surgery which were represented by using a variety of endoscopic have been emerging.In the early 1970s,Messerklinger firstly carried out endoscopic sinus surgery which is known as "Messerklinger technology" has become the cornerstone of surgery endoscopic sinus surgery technology,and it was widely used and expanded endlessly.Alfieri extended the surgical technique to the ventral craniocervical junction area in 2002 and put forward the opinion that treating the craniocervical junction lesions by using the endoscopy.In 2005,Kassam depended on the basic researches which were studied by Alfieri and others,firstly carried out the surgical technique in clinical practice,and achieved the desired results. Compared with transoral approach,the endoscopic endonasal approach to the ventral cranio-cervical junction although has many advantages,but the surgical approach path is longer,relatively narrow space operation,distorted images,two-dimensional image which lack of three-dimensional sense,coupled with the surgical approach to this region adjacenting to important and complex anatomical structure.On the one hand,surgeon should have excellent basic skills of endoscopic sinus surgery,on the other hand,also be familiar with the nasal cavity and paranasal sinuses and nasal skull of the anatomical landmarks.The purpose of this task is through observational study the features of the endoscopic endonasal approach to the ventral region of cranio-cervical junction to establish the structures which is relatively constant at the same time easy to observe as the surgical marking points,and carry out a detailed description of the structures which surround the adjacent of surgical approach.At the same time,we measure the relevant anatomical structures around the important observation measurements,to establish the relative safety range of the operation.And on this basis,using the corpse\'s head to follow the actual process of clinical,simulate to the ventral craniocervical junction surgery,pre-validation studies established the usefulness of surgical marking points,and further proves that the the feasibility and safety of the operation.Our research includes three parts as following:Chapter.1 The establishment of the surgical landmarks of the endoscopic endonasal approach to the ventral cranio-cervical junctionObjective:Through the anatomy study of the endoscopic endonasal approach to the ventral cranio-cervical junction,provide anatomical basis,find the structures which are relatively constant as well as easy to observe,and establish these structures as surgical marking points.At the same time,we describe its surrounding neighbors with a view to expediting this surgical method in clinical application.Methods:Twenty 10%formalin-fixed intact adult head-neck specimens were used to dissect and observe the anatomic features of this access in order to establish the surgical landmarks of the approach.Following the surgery approach,incised along the median sagittal line of head and neck specimens,in order to observe the the anatomic features of the approach,look for some of the relatively constant and easy to observe the anatomical structures,establish these structures as signs of the operation points,and make the use of instrument to make the anatomical observation of the anatomical landmark points on the around adjacent to the relationship.Results:Using twenty 10%formalin-fixed intact adult head-neck specimens observe the anatomic features of this access in order to establish the surgical landmarks of the approach,includ:middle turbinate,choana narium,Eustachian tube ostium,nasopharynx mucosa,longus capitis and longus colli,basion,tuberculum anterius atlantis.Make the anatomical observation of the anatomical landmark points on the around adjacent to the relationship.When operating the structures within the surgical field can be confirmed by identifying the anatomical location of structures around these points in order to complete resection of lesions,and also to reduce complications.Conclusion:In this study,we select constant and easy to confirm anatomical structures as surgical landmarks for the endoscopic endonasal approach to the ventral region of cranio-cervical junction,the structures around these points were described in detail.Intraoperative identification of these points as the basis,and to master the structures of these marking points around the adjacent relations can be targeted,that is,to achieve the purpose of effective disarmament lesions,and also essential to avoid injury in the structure,reduce or even avoid complications.Chapter.2 Applied anatomy of the endoscopic endonasal approach to the ventral cranio-cervical junctionObjective:Through the anatomical study,measurement the distance from the dependency structures to the median line,with a view to the entry road to provide anatomical basis and to establish the safe range of the operation.Methods:Twenty 10%formalin-fixed intact adult head-neck specimens were used,Along the midline sagittal incise corpse head to choanal and from the side of the mid-point of zygomatic arch inward cut coronal and sagittal tangent to the intersection,and separate the of two parts.An accuracy of 0.01 using the vernier caliper and compasses,measurement of the surgical approach important anatomical structure:①the distance from pterygoid canal,pharyngeal opening of eustachian tube,foramen lacerum,External entrance of the hypoglossal canal,Internal carotid artery at atlas or axis,Canalis caroticus,Vertebral artery at inferior border of axis, Vertebral artery at superior border of axis,Vertebral artery at inferior border of atlas, Vertebral artery at superior border of atlas,Anterior border of occipital condyle to the medial midline;②anterior nasal spine away from the pharyngeal tubercle, leading-edge midpoint of foramen magnum,anterior tubercle of atlas distance.And apply software spss 13.0 statistical data for measuring statistical treatment,the results of using(?)±s express,and the right and left data matching t test,P＜0.05 for significant difference.Results:The endoscopic endonasal approach to the ventral cranio-cervical junction completely exposed craniocervical junction,the shortest distance(89.75±2.80) mm;The anterior-inferior wall of sphenoid grinding to both sides of the pterygoid canal in addition to the scope of control for the sector,for the distance is midline(9.37±0.59) mm;under grinding in addition to the scope of the clivus to both sides of foramen lacerum for the sector,as from the midline(10.75±0.63) mm;At the atlantoaxial segment,vertebral artery at inferior border of axis of the shortest distance from the midline for(15.70±2.12) mm;left atlantoaxial department carotid artery is greater than the right distance from the midline (t=3.453,P=0.003),other targets of the right and left sides was no significant difference(P＞0.05).To reduce the postoperative cerebrospinal fluid leakage,set up the "U"-type flap,the lateral boundary should be for both sides of the pharyngeal opening of eustachian tube,away from the midline of about(10.82±0.92) mm.Conclusion:The endoscopic endonasal approach to the ventral cranio-cervical junction is the same as any other kind operation,there are certain indications for surgery.This approach will get better therapeutic effect when the lesions which located at the ventral craniocervical junction area near the midline,and while there is a larger deviation when the lesions from the midline of the risks.The approach can exposure the ventral craniocervical junction region safely and adequately by making full use of the surgical landmarks,and operating within important structures related to the lateral sector.Chapter.3 Endoscopic anatomy of the endoscopic endonasal approach to the ventral cranio-cervical junctionObjective:On the base of established the surgical landmarks and relatively safe operating range,simulate the approach following the actual operation of the clinical process on the specimens,validate the usefulness of surgical marking points have been established,and further prove the feasibility of the operation and safety.Methods:Using a diameter of 4mm,length 18cm of 0°nasal endoscope and the parts of endoscopic devices,five fresh and intact head-neck specimens injected with colored latex were used and completely simulant operation via endoscopic endonasal approach to the cranio-vertebral junction following the clinical course of the actual operation was performed in all cases.Results:Following the clinical course of the actual operation and making use of pre-established surgical landmarks,we can expose the ventral craniocervical junction area safely and fully.Craniofacial incision,bone and muscle injuries,brain tissue,as well as pull off the important nerves and blood vessels can be avoided.At the same time,compared with transoral approach:1.Get craniocervical junction more directly;2.without cutting the soft palate and hard palate can get all the clivus and craniocervical junction,even a broader regional areas;3.Endoscopic operation of a broader vision,a clearer surgical field;4.from the superior anterior arch of atlas and odontoid tip fate level slightly above the pre-show,there is probably only the tip part of odontoid resection,and the retention of some of the former arch of atlas or even atlantal not removed anterior arch,reducing the craniocervical junction stability.5.To avoid damage teeth,temporomandibular joints and tongue injury.Conclusion:In this study,on the base of established the surgical landmarks and relatively safe operating range,through simulate operation on the fresh corpse, confirmed the practicality of landmarks which were established prophase,and confirmed making use of established surgical marking point intraoperatively,and strictly controlling operation at the medial important surgery-related structures,can be fully exposed and deal with the ventral craniocervical junction epidural compressive lesion,and also can be revealed subdural structures,at the same time can avoid injury in the surrounding vital structures,thus avoiding serious complications.Further confirmed applying the endoscopic endonasal approach to the ventral cranio-cervical junction is feasible and safe.

Observation of Curative Effect of Recombinant Human Granulocyte-Colony Stimulating Factor for Children Infection Associated Neutropenia

Mon, 30 May 2011 13:08:29 +0000

Objective:Adopted the principle of clinical test with sequential design, summarized the clinical data of Infection Associated Neutropenia in children who met the inclusion criteria; Evaluated curative effect of rhG-CSF for Children Infection Associated Neutropenia.Methods:Infection Associated Neutropenia children who met the inclusion criteria visited our hospital during Feb, 2007 ~ Jan, 2009 were statistically analyzed in clinical data; All the patients were treated with rhG-CSF once, subcutaneous injection of 5μg/kg after admission, Observed improvement in clinical symptoms, reexamined the blood routine, evaluated curative effect of rhG-CSF and monitored the adverse reactions at the following day or the next day after admission.Results:(1) Accordance with the inclusive and exclusive criteria, we collected 102 children in our hospital during Feb, 2007~Jan, 2009, patients with virus positive occupied 55(53.9%), mycoplasma positive occupied 28 (27.5%), mycoplasma complicated by virus infection occupied 8(7.8%); 58 cases did throat swab culture, 6 cases(10.3%) positive; 85 cases did the c-reactive protein examination, 18 cases (21.2%) increased at different degree; 52 cases took the chest X-ray examination, 21 cases (40.4%) appeared abnormal.(2) Cases of this group, neutrophil deficiency occupied 20 (19.6%), 26 of them (15.7%) were accompanied by anemia, 6 cases (5.9%) with thrombocytopenia.(3) Cases of this group, primary diseases were infection, there were 41 cases with upper respiratory tract infection (40.2%), 22 cases with bronchitis (21.6%), 21 cases with pneumonia (20.6%), 7 cases with diarrhea (6.9%), 4 cases with infectious mononucleosis (3.9%), 3 cases with exanthema subitum (2.9%).(4) After the therapy of rhG-CSF, significant improvement in clinical symptoms (fever returned, ease fatigue, mental improvement, etc.). And absolute value of leukocyte increased to (7.5±2.58)×109/L from (3.53±1.31)×109/L; absolute value of neutrophil increased to (3.28±1.82)×109/L from (1.05±0.48)×109/L; absolute value of leukocyte and neutrophil were significantly different before and after treatment.(5) After the therapy of rhG-CSF, curative effect was obvious, total effective rate was 89.2%.(6) Adverse reactions were rare, 2cases with rash (2%), 1 cases with gastrointestinal reaction (1%).Conclusion:(1) The result in lab showed that virus infection was the common factor leading to neutropenia(53.9%), followed by mycoplasma(27.5%), primary diseases often occurred in respiratory and digestive tract infections.(2) After the treatment of rhG-CSF, absolute value of leukocyte and neutrophil increased, curative effect was obvious.

Effects of GABA on E_2 and P Secretions and Apoptosis by Human Ovarian Granulosa Cells in Vitro

Mon, 30 May 2011 13:08:29 +0000

The distribution of L-glutamate decarboxylase(GAD),a major biosynthetic enzyme for gamma-aminobutyric acid(GABA),is well-recognized function as a cerebral.Recently,people have discovered more than thirty kinds of peripheral tissues have GABA,and fallopian tube and ovary have higher GABA concentration than the CNS which indicated a physiological sibnificance of this amino acid in the female reproductive systems.Further study found that in rat ovary have GABA neurons,ovarian tissue has the specificity of GABA binding sites.Animal experiments prompted exogenous GABA on luteinized granulosa cell secretion of estradiol and progesterone have inhibitory effects,and this inhibition may be related to GABA receptor and adenylate cyclase system.The data suggested that GABA can promote cultured rat luteal spontaneous apoptosis of granulosa cells in vitro. Many scholars at home and abroad reported that apoptosis of granulosa cells increasing affected the IVF-ET cycles pregnancy outcomes.We used electrochemiluminescence technique to explore the possible mechanism of exogenous GABA effect on human luteinized granulosa cells secretion of estradiol and progesterone,and used TUNNEL technique to detect how exogenous GABA affect human luteinized granulosa cell apoptosis,and used ELISA technique to detect GABA concentration,to explore the impact of GABA on pregnancy outcomes.Part one Effects of GABA on E_2 and P Secretion by Human Ovarian Granulosa Cells in VitroRecent work suggests that exogenous GABA on rat luteinized granulosa cells secretion of estradiol and progesterone have inhibitory effects,and this inhibition may be related to GABA receptor and adenylate cyclase system.But exogenous GABA whether have the same inhibitory effects on human luteinized granulosa cells secretion of estradiol and progesterone,no one has reported.ObjectiveWe used electrochemiluminescence technique to explore the possible mechanism of exogenous GABA effect on human luteinized granulosa cells secretion of estradiol and progesterone.Methods1.Thirty-two barren patients were treated by IVF/ICSI-ET in southern hospital\' s reproductive center.They were less than thirt-five years old.Barren cause was fallopian tube factor or male factor.They were all at their first IVF/ICSI-ET cycle using down regulation long controlled hyperstimulation protocol. Centrifuging and separating luteinized granulosa cells,the isolated luteinized granulosa cells were divided into four groups to carry out in vitro for twenty-four hours,Discarded the culturing medium,replaced with serum-free medium, Interventing factors were added to the culture medium and the final volume was 1 ml,and GABA concentration was 4×10~(-6)mol/L,Bicuculine concentration was 1×10~(-6)mol/L,Forskolin concentration was 1×10~(-3) mol/L,HCG concentration was 0.11U/ml.After twenty-four hours cultured,the supernatants were collected.2.Divide into groups:(1) Adding GABA(4×10~(-6)mol/L)to the culturing medium,and observating the effect of GABA on E_2 and P secretion by human luteinsing granulosa cells in vitro(n=8),divide into two groups:control group;GABA group(2) Adding Bicuculine(1×10~(-6)mol/L)to the culturing medium,and observating the effect of GABA on E_2 and P secretion under the influence of Bicuculine by human luteinsing granulosa cells in vitro(n=8),divide into four groups:control group; GABA group;Bicuculine group;GABA+ Bicuculine group.(3) Adding Forskolin(1×10~(-3) mol/L) to the culturing medium,and observating the effect of GABA on E_2 and P secretion under the influence of Forskolin by human luteinsing granulosa cells in vitro(n=8),divide into four groups:control group; GABA group;Forskolin group;GABA+Forskolin group.(4) Adding hCG(0.1IU/ml) to the culturing medium,and observating the effect of GABA on E_2 and P secretion under the influence of hCG by human luteinsing granulosa cells in vitro(n=8),divide into four groups:control group;GABA group; hCG group;GABA+hCG group.3.Statistical methods to detect the concentration of estradiol and progesterone in supernatants by electrochemiluminescence technique.Comparing their variety of control group and treatment groups.Data were evaluated using SPSS13.0 software. Independment-sample T test was performed to analyse their variety.P＜0.05 was accepted as indicative of stastical significance.Results1.The GABA concentration of 4×10~(-6)mol/L could inhibit E_2 and P secretion by human luteinsing granulosa cells in vitro.2.Bicuculine concentration of 1×10~(-6) mol/L can promote E_2 and P secretion by human luteinsing granulosa cells in vitro,and GABA can inhibit E_2 and P secretion under the influence of Bicuculine.3.Forskolin concentration of 1×10~(-3) mol/L can promote E_2 and P secretion by human luteinsing granulosa cells in vitro,and GABA can inhibit E_2 and P secretion under the influence of Forskolin.4.HCG concentration of 0.1IU/ml can promote E_2 and P secretion by human luteinsing granulosa cells in vitro,and GABA can inhibit E_2 and P secretion under the influence of hCG.Summary1.GABA inhibit E_2 and P secretion by human luteinsing granulosa cells in vitro.2.the possible mechnism of GABA affect E_2 and P secretion by human luteinsing granulosa cells in vitro are mediated Through the GABA receptor on the membrane particles luteinsing granulosa cells in vitro.3.the possible mechnism of GABA affect E_2 and P secretion by human luteinsing granulosa cells in vitro are associated with the adenylate cyclase system;4.the possible mechnism of GABA affect E_2 and P secretion by human luteinsing granulosa cells in vitro are GABA and HCG with the corresponding receptor interaction may affect each other.PART TWO Effect of GABA on apoptosis by human ovarian granulosa cells in vitroMany studies reported that ovarian somatic cells and germ cells apoptosis were related to follicular atresia,and study on ovarian somatic cells apoptosis also focused on granulosa cells.ObjectiveTo invistigate GABA concentration in follicular fluid and how exogenous GABA affect human luteinized granulosa cells apoptosis in IVF cycles.Methods1 Forty barren patients were treated by IVF/ICSI-ET in southern hospital\'s reproductive center.They were less than thirty-five years old.Barren cause was fallopian tube factor or male factor.They were all at their first IVF/ICSI-ET cycle using down regulation long controlled hyperstimulation protocol.①Granulosa cells collected:Centrifuging and separating luteinized granulosa cells,the isolated luteinized granulosa cells were divided into two groups to carry out in vitro for twenty-four hours,Discarded the culturing medium,replaced with serum-free medium,GABA were added to treat group,the final volume was 1 ml,and GABA concentration was 4×10~(-6)mol/L.Continue to cultivate twenty-four hours,discarded supernatants,and collected adherent cells to smear.②follicular fluids collected:collecting matural follicular fluid(≥15mm) on taking ovum day,and follicular fluids were centrifuged in centrifugal machine,then to collect the supernatant.Supernatant were put the refrigerator.2 Divide into groups:(1) Divided into two groups:Pregnancy group(n=17) and non-pregnancy group(n=23),GABA concentration were measured in follicular fluid of the two groups,and comparison the GABA concentration difference of two groups.(2) Divided into control group and GABA group,comparison apoptosis rates.3 To detect the concentration ofγ-gamma-aminobutyric acid in follicle fluids by humanγ-gamma-aminobutyric acid ELISA kit.Comparing their variety of two group.Data were evaluated using SPSS13.0 software.Independment-sample T test was performed to analyse their variety.P＜0.05 was accepted as indicative of stastical significance.Results1.The concentration ofγ-gamma-aminobutyric acid in follicle fluid in pregnancy group was lower than one of non-pregnancy group (680.398±11.279 vs 713.832±9.517) ng/ml,and the difference was stastical significance(P＜0.05).2.Apoptosis rates of human luteinized granulosa cells in GABA group was higher than one of in control group(20.08±2.44 vs 29.27±2.02)%,and the difference was stastical significance(P＜0.05). Summary1.We found that GABA concentration of human follicular fluid in non-pregrant group was higher than one of in pregrant group,and the difference was stastical significance2.We added GABA to human luteinized granulosa cells in vitro,and found that GABA could promote apoptosis rate of human luteinized granulosa cells in vitro,and the difference was stastical significance CONCLUSIONS1.The experiment using electrochemical luminescence detection kit E2 and P concentration, observing the effect of GABA on E_2 and P Secretions by Human Ovarian Granulosa Cells in Vitro,explore the possible mechanism of exogenous GABA effect on human luteinized granulosa cells secretion of estradiol and progesterone.The results suggested:①GABA inhibit E_2 and P secretion by human luteinsing granulosa cells in vitro.②the possible mechnism of GABA affect E_2 and P secretion by human luteinsing granulosa cells in vitro are mediated Through the GABA receptor on the membrane particles luteinsing granulosa cells in vitro;are associated with the adenylate cyclase system;are GABA and HCG with the corresponding receptor interaction may affect each other.2.We used ELISA technique to detect GABA concentration of pregnancy group and non -pregnancy group in IVF cycles.The result was that the concentration of GABA in follicle fluid of non-pregnancy group was significantly higher than one of pregnancy group.And we used TUNNEL technique to detect the effect of exogenous GABA on human luteinized granulosa cell apoptosis,the result was that apoptosis rates of human luteinized granulosa cells in GABA group was higher than one of in control group.The experiment suggested that high concentrations of GABA may increase the rate of apoptosis of granulosa cells,thus affect the quality of oocytes,and reduce the pregnancy rate.

Functional Magnetic Resonance Imaging Research of Smokers Responses to Smoking Olfactory Cue

Mon, 30 May 2011 13:08:28 +0000

Objective:Make a paired test of the smoker’s response to the smoking olafctory cue under different physiological status (satiated vs. abstinent) by using the functional magnetic resonance imaging (fMRI),and make a comparison between groups of the smoker and nonsomker’s response to the smoking olafctory cue, we try to seek how the smoking olafctory cue impact the smoker’s physiological and psychological factors, localizing the smoking olafctory cue related brain region and analyse the related response,and complete the established theoretical system of the related brain region’s response to the smoking visual cue.Material and methods: Seventeen han smokers and five han nonsmokers underwent this study,they were right-handed, healthy and non-psychiatric disorder.All of them used cigarette as stimulus,paper for control,and the author put them on the subjects’nose.We adopted block design in this experiment. MRI examination was performed on GE signa 1.5T echospeed MRI using echo planner imaging(EPI)sequence, adopting dorsal position. A 8-channel head coil was used for transmission and detection of signal.Scaning parameters: TR/TE=3000/45ms , flip angle 90°, slice thickness 3mm,gap 0.5mm,NEX 1,FOV[field of view] 240×240mm,matrix 64×64,the whole scaning time is 9 minutes and 42 seconds,.The functional images were acquired in the same FOV and orientation as T1-weighted anatomic images.Collect and record fMRI datas to CD-ROM,and then input them into special data-processing computers.The raw fMRI datas were dealed with Matlab 6.5.1, SPM2 and Xjview4 tool software,P value was 0.001. Results: When smokers were exposed to the smoking olfactory cue in the abstinent state, we could find the activation on the left superior frontal gyrus(BA8、9、10、11)、left middle frontal gyrus(BA8、10、11、46)、left inferior frontal gyrus(BA9)、left medial frontal gyrus(BA6、8、10、11)、right superior frontal gyrus(BA10)、right middle frontal gyrus(BA8、9)、right inferior frontal gyru(sBA45、47)、right medial frontal gyru(sBA9、11)、left callosal gyrus( BA21、31)、right callosal gyrus(BA24、37)、right middle temporal gyrus( BA21) ,both sides of the thalamus and cerebellum.When smokers were exposed to the smoking olfactory cue in the satiated state,we could find the activation on both sides of the prefrontal lobe and cerebellar hemisphere,while there were no activation on the other brain regions. There was no activation on the whole brain of the nonsmokers,only a little small volume active points appeared in individual subjects,which we thought caused by the picture noise.After the paired test of the smoker’s response to the smoking olafctory cue under different physiological status (satiated vs. abstinent),we could find the the more notable activation on the prefrontal lobe、left callosal gyrus、right middle temporal gyrus、right thalamus in the smoking forbidden state(P4/12,for details see the following text table 2).There were no statistical differences between the satiated and abstinent status (P>0.1,for details see the following text table 2) in both sides of the cerebellum.After the comparison between groups of the smoker and nonsomker’s response to the smoking olafctory cue,we could find the activation on the the prefrontal lobe、left callosal gyrus、right middle temporal gyrus、right thalamus and both sides of the cerebellum(P4/12,for details see the following text table 4),it appeared only in the smokers,while there was no notable activation on the whole brain of the nonsmokers.Besides, in the abstinent state, there were no statistical differences of the craving value before and after the experiment(P=0.695).Conclusions:1.The Limbic system(cingulate cortex、thalamus)and the prefrontal loab and the paly an important role in the cue-induced smoking craving,and thiis experiment approved this theory from the new perspective of the olfactory.2.Compared with the previous experiments about the smoking visual cue,we found the brain region related to the memory and familiarity—right middle temporal gyrus active.3.Both sides of the cerebellar of the smokers were active in satiated and abstinent status ,while neither of the cerebellar of the nonsmokers were active, experiments showed that the activity of the cerebellar was not associated with activities including coordinating movement and maintaining posture and equilibrium,but associated with the cognitive fuction.4.It was a availible experiment method to research smoking addicts’brain activation and the role of neural mechanisms using olfactory stimulation in the clinical 1.5T MRI equipment.

Analysis of the Chemical Constituents Viola Tianshanica Maxim. and Study of the Antioxidative and Antibacterial Effects

Mon, 30 May 2011 13:08:28 +0000

Viola tianshanica Maxim.as a Uighur folk medicinal herb,is known and used by Uighur people since antiquity.It is of extensive pharmacological effects and is used for treating these diseases as cat fever, clearing away heat and toxic material,etc. There is a large amount of distribution on most of mountain areas of Xinjiang. This text has carried on comparatively detailed research to the effective components in Xinjiang. Antioxidative effects and antimicrobial of extracts of Viola tianshanica Maxim. were studied that can supply experimental data in food and other area.In chapter one, kind,shape,distributed,value and studying the situation of Viola tianshanica Maxim.,active constituent in natural medicine.Antioxidative effects and antimicrobial were made survey and introduction.In chapter two, volatile oil was extracted from Viola tianshanica Maxim.by steam distillation. The content of the volatile oil was 0.067%.Compositions of volatile oil of Viola tianshanica Maxim. were analyzed and examined by gas chromatography—mass spectrometry (GC/MS).60 constituents were separated and 47 compounds were identified. The relative content of the components were determined with peak area normalization method。the major constituents of the volatile oil are aliphatic hydrocarbons, alcohols, ketone, aldehyde, and esters.In chapter three ,the total flavonoids and alkaloids in Viola tianshanica Maxim. are determined by ultraviolet spectrophotometry with standard .Through studying two technological process: ultrasonic wave extraction and circumfluenc,a better technological condition to extracting total flavonids and alkaloids from Viola tianshanica Maxim. is obtained.In chapter four, determination of metal elements in Viola tianshanica Maxim. is studied by ICP-AES after HNO3-H2O2 digestion. The average recovery of the method for trace elements such asB,Cu,K,Mg,Mn,Se,Zn,Cr,Fe,V in Viola tianshanica Maxim. is found to be 95.4% -104.9% for the metal elements and the RSD\' s are less than 2.7%.The method is simple, fast, sensitive and accurate. The results obtained are satisfactory. It is of important significance to assess the value of the Chinese medicinal herb by the determination of trace elements in Chinese medicinal herb with ICP-AES.In chapter five, antioxidative effects of extracts of Viola tianshanica Maxim. were measured by Schall and Na2S2O3-I2 titrimetric method. Extracts obtained from the Viola tianshanica Maxim. by using different solvents were added to lard to study the effects of oxidation resistance. The extracts got by 85% ethanol had distinct antioxidative effects. The usage of extracts is in directly proportion to the antioxidative effect. When the addition amount of the extract was reached to 0.5%g/ml, the effect of oxidation resistance was better than 0.02%g/ml Vc,but the effects are not distinct. but VE(0.02%g/ml) have no antioxidant activity.In chapter six, the alkaloids and flavonoids of extracts from Viola tianshanica Maxim. with 85% ethanol were used as antimicrobial agents. Four species of bacteria and five species of fungu were taken as test-bacteria and had the best antibacterial effect on the tested bacteria. The minimal inhibition concentration(MIC) of antibacterial of extractions are respectively as follows: Klebsiella pneumonia 0.175g/ml ,Klebsiella ozaenae 0.15 g/ml and 0.175 g/ml, Pseudomona pseudoalcaligenes and Proteus mirabilis 0.20 g/ml .But effect was not obvious on fungus.

Expression of NDRG2 Gene in Breast Cancer Tissue and Its Effect on Proliferation of Breast Cancer Cell

Mon, 30 May 2011 13:08:27 +0000

The NDRG2 gene(N-myc downstream regulated gene 2) was first discovered and cloned from normal human whole brain cDNA library in 1999. This gene locates on human chromosome 14q11.2,including 16 exons and 15 introns, and its full mRNA is 2,024bp in length, encoding the 357 amino acid、40KD NDRG2 protein. NDRG1 was the first member found of NDRG gene family, which was isolated in N-myc mutant mouse embryos and found be repressed by N-myc and c-myc. Human NDRG gene family consists of 4 members: NDRG1, NDRG2, NDRG3, NDRG4, which has high similarity in nuclear acid or amino acid sequence. However, their expression levels in spatio-temporal distribution and periods of cells and tissues development are distinct.The knowledge about the function of the NDRG gene family came mainly from the studies on NDRG1. Now, many intracellular and extracellular factors have been found which could regulate the expression of NDRG1. Homocystein, p53, retinoic acid, oxidative and stress condition such as hypoxia, nickel and other reductive agents, et al, can upregulate NDRG1 expression. Inferring from all the references obtained, the functions of NDRG gene family may be involved in cellular differentiation events, maintaining the balance of cell redox potential, and counteracting cell malignant transformation and so on.Our preliminary studies showed that the mRNA expression patterns of NDRG2 gene was dominant in the white matter, gray matter and neuron of brain, salivary gland, skeletal muscle, and lower in bone marrow, testis, peripherial blood and placenta. However NDRG2 gene did not express in leukocyte, colorectal cancer and some tumor cell line such as Hela S3, leukemia cell, Burkitt’s lymphoma, adenocarcinoma SW-480, all above suggested that NDRG2 gene had the relationship with tumor, might be a new suppressor gene candidate. So, to study its function and regulation has important theoretical meaning and foreseeing clinical value, but up to now, there have been no reports about the precise cellular and molecular function of NDRG2 gene. Our research is aimed to observe the effect of NDRG2 gene on tumorigenesis of breast cancer, and explore its preliminary function. The main finding of this research include:1. Detecting mRNA and protein expression of NDRG2 gene in 80 fresh breast cancer tissue collections and corresponding tissue around breast cancer (including infitrating ductal carcinoma, n = 54; lobular carcinoma, n = 18; lobular carcinoma in situ, n =8) by immunohistochemistry, real time PCR, western blot and so on. Results indicate that mRNA expression of NDRG2 is lower in 63 breast cancer tissue collections compared with corresponding tissue around breast cancer, and is positive correlation to tumor grade; higher in 5 breast cancer tissue collections compared with corresponding tissue around breast cancer. In addition, there is not significant different in mRNA express of NDRG2 between 12 breast cancer tissue collections and corresponding tissue around breast cancer. So the results suggest that the express of NDRG2 gene is negative correlation to reproductive activity of cancer cells.2. To explore the effect of gene NDRG2 transfection on the proliferation in breast cancer cell. By the induction of liposome , pcDNA3.1- NDRG2 and eukaryotic expression vectors pcDNA3.1 ( +) were transfected into breast cancer cell line SK-BR-3. Then the subclone cell lines SK-BR-3- NDRG2 which expressed NDRG2 stably and higher were obtained by persistent G418 selection.. Which establish a solid ground for the subsequent in vivo and in vitro experiments. In order to observe the effect of NDRG2 gene expression on Bcap37 cell line,which expresses low level NDRG2 gene is transfected with recombinating adenovirus expressing high level NDRG2 gene to improve NDRG2 gene express level.3. The proliferation activity of the cell lines was detected by plate colony formation,MTT and flow cytometry. The results indicate that high expression level of NDRG2 gene inhibits cells growth and arrest cells in G1 phase ,which suggest that NDRG2 may inhibit cell proliferation by cell cycle arrest.To sum up, the express of NDRG2 gene in breast cancer tissue increases with breast cancer grade. Overexpression of NDRG2 in SK-BR-3 cells and Bcap37cells effectively inhibited cell proliferation and induced cell cycle arrest. These findings about the different expression and function of NDRG2 gene in breast cancer tissue and breast cancer cell lines is helpful to confirm the function of NDRG2 gene, which will provide theoretical base for mechanisms of breast cancer and gene therapy for breast cancer.

The Paracrine Effects of Insulin on Adiposederived Stem Cells and the Biological Effects of Its Paracrine Factors to Repair Cells

Mon, 30 May 2011 13:08:26 +0000

【Background】Recently, in view of many advantages of adipose-derived stem cells in harvesting and application, how to effectively apply ADSCs for the repair of various damaged tissues has become a research hot spot in surgery, especially in burns and plastic surgery. Nevertheless, so far, exact mechanisms about applications of ADSCs for the repair of various damaged tissues are unknown. Nowadays, most of studies are focus on cell differentiation mechanism.But lots of experiments indicate that there is one more important potential function, which is paracrine function. However, it is still unclear about paracrine mechanism of ADSCs for the repair various damaged tissues. In addition, insulin, which is one of important anabolic hormone of human body, can regulate nutrition metabolism of glucose, fat and protein. But it hasn”t been reported yet about regulation of inslin on paracrine secretion, Which could effectively promote ADSCs for the damaged tissues repair recently. 【Objective】1. To investigate the effects of paracrine function of adipose-derived stem cells on HaCaT cell.2. To investigate the effects of paracrine function of adipose-derived stem cells stimulated by insulin.3. Preliminary interpret the possible mechanisms of paracrine function of adipose-derived stem cells act in tissue repair by investigating the biological effects of the conditioned medium of ADSCs (ADSC-CM) unstimulated /stimulated by insulin on HaCaT cells.【Contentes】1. The effects of paracrine function of adipose-derived stem cells on HaCaT cell.2. The effects of paracrine function of adipose-derived stem cells stimulated by insulin.3. The biological effects of the conditioned medium of ADSCs (ADSC-CM) unstimulated /stimulated by insulin on HaCaT cells.【Results】1. The paracrine cytokines of adipose-derived stem cells could promote HaCaT cells on proliferation, migration, and anti-apoptosis.2. The concentration of 10~(-8)～10~(-7) mol/L insulin could significantly promote the paracrine function of adipose-derived stem.3. The ADSC-CM unstimulated /stimulated by insulin can promote HaCaT cells on proliferation, migration, and anti-apoptosis, and the concentration of 1×10~(-7) mol/L insulin was also the best one.【Conclusions】1. The paracrine function of adipose-derived stem cells plays an important role for the repair and regeneration of various damaged tissues.2. Insulin intervention could effectively enhance paracrine function of adipose-derived stem cells.3. Insulin could promote wound healing by intervention of paracrine function of adipose-derived stem cells, which can significantly promote HaCaT cells on proliferation, migration, and suppress apoptosis.4. The paracrine manner to promote wound healing may be one of important potential mechanisms that stem cells contributing to tissue repair.5. Reasonable and effective application of paracrine function of adipose-derived stem cells will open up prospect of research and therapy of wound healing.

Preparation and Study on Nano-dHA/PLA Drug Carrier for Sustained Release

Mon, 30 May 2011 13:08:26 +0000

Along with scientific and technological advancement,polymer materials are beginning to be broadly applied in modern medical science, called biomedical polymers. There are currents that bio-degradable materials will be the mainstream in the development of biomedical polymers. Poly lactic acid (PLA) is a kind of excellent biomedical poly-ester, which has good biocompatible feature and can completely degradable into H2O and CO2. It started to be highly anticipated as a kind of promising degradable biomedical polymer when its degradability and the high safety of its degradable products had been confirmed. Currently, PLA are mainly applied in the fields of inner-fixing of fracture, tissue-repairing materials, surgical seam and drug delivery system, etc.PLA has been prepared by such methods as direct polycondensation, ring-opening polymerization and melt/solid polycondensation and so on. Ring-opening polymerization is commonly used to get high-molecular-weight PLA. In this paper, Lactide, which is synthesized through lactic acid condensation, was polymerized to form PLA by ring-opened of lactide.With this method, the synthesized polymericular weight was about 7.6x104.Hydroxyapatite(HA), the main mineral composition of bone,can form strong bone-bonding with natural bone and has been used extensively for biomedical applications and bone regeneration based on its high biocompatible, bioactive and osteoconductive properties.It was reported by researcher that the HA in the bone was mainly nano-Calcium-deficient carbonated HA (nano-dCHA). In this paper, we fabricated an anti-infection nano-Calcium-deficient carbonated HA (nano-dCHA) and PLA compounds, providing a new local drug delivery system (DDS). TBMC/ nano-dHA/PLA were prepared by an in-water drying method using a W/O/W emulsion. The comparisons between the two different compounds are also made. It showed that the average sizes of TBMC/nano-dHA/PLA were 57.805μm. The encapsulation efficiencies was 42.85% and the drug loading was 54.72%. It was also proved that TBMC/nano-dHA/PLA contained the characteristic peaks of TBMC,HA and PLA by IR. The thermal behavior by differential thermal analysis indicated that TBMC,HA and PLA mingled each other. The determination of its release rate in vitro revealed its release process of the drug according with the two-phase release pattern and it was in sustained release except an abrupt release in the initial times, this because the drugs were released rapidly from the surface. Then the release speed became slower and even stopped in certain stage. Its cumulative release percentage in vitro was over 65%. Influence of some factors such as the content of drug, the compound size and the pH value of release media on the drug-releasing performance of the compounds were studied. The result showed that the greater the content of the drug and the bigger the difference of drug concentration between in and out of the compound, and the more quickly the drug released. The smaller the compound size, the more quickly the drug released. In the alkali medium, the PLA degraded more quickly and the drug became more stable, resulting in faster release of drug.

The Central Reactivity to Electroacupuncture Preconditioning in Aged Mice and Role of Wnt Pathway

Mon, 30 May 2011 13:08:25 +0000

Cerebrovascular disorders has exceeded cancer to become major reasons for death in China,especially in the aged population who are more susceptible to ischemic accidents. A various of methods including hypothermia,cryotherapy and NMDA receptor blocker have been used in the clinic to treat cerebrovascular disorders,but all have limitations and side effects. Therefore,looking for more safe and effective methods keeps being a hot spot of researches.Acupuncture treatment has been used for treatment of cerebral ischemic diseases for many years. It is widely accepted because of its effectiveness and safety. Electroapuncture(EA) is the combination of acupuncture and electric technology. Our previous researches revealed that EA at Baihui 30min a day for 5 days could alleviate the subsequent ischemic damage induced by MCAO in rats. But whether EA could protect the brain against forebrain ischemia hasn’t been reported yet. Moreover,the researches on EA preconditioning were done in adult animals,whether aged animals whom are more sensitive to ischemic damage could be protected is still unclear.In this study we tend to use BCCAO model of mice to investigate the protective effect of electroacupuncture and part of its machenism.Furthermore,though a number of evidences have been established about preconditioning,few of them have been translated to clinic. EA is safe and easy to take. Sufentanil is a widely used opioid. These two have great potential to be clinically used preconditioning methods. Therefore we evaluated the protective effect of sufentanil predosage as well as the clinical effect of EA preconditioning.Experiment 1. Electroapuncture preconditioning induces neuroprotection against forebrain ischemia/reperfusion injury in aged miceObjective:To investigate whether preconditioning with electroapuncture could alleviate forebrain I/R injury in aged mice. Methods:Twenty male C57BL6 mice aged 18m were randomly assigned to two groups (n=10 each). Animals in the control group underwent BCCAO for 20 min without preconditioning. Animals in EA group received electroacupuncture at the Baihui acupoint 30min a day for 5days. At 24h after the last preconditioning,the BCCAO model was induced for 20min in all animals.The neurological deficit scores(NDS)were evaluated at 24h after reperfusion and then mice were sacrificed under deep anesthesia. Number of necrosis neurons in the CA1 area was calculated after HE staining. Results:The neurologic outcomes in the EA group were better than those in the control group (P<0.05). The number of necrosis neurons in the EA group was significantly less than that in the control group(P<0.05).Conclusion:Electroapuncture preconditioning induces neuroprotection against forbrain ischemia/reperfusion injury in aged mice.Experiment 2. Change of Wnt pathway after electroacupuncture preconditioning in aged miceObjective: To investigate whether the benefical effect induced by eletroapuncture preconditioning against BCCAO is dependant on wnt pathway in aged mice. Methods: Twenty male C57BL6 mice aged 18m were randomly assigned to two groups (n=10 each). Animals in the control group underwent BCCAO for 20 min without preconditioning. Animals in EA group received electroacupuncture at the Baihui acupoint 30min a day for 5days. At 24h after the last preconditioning,the BCCAO model was induced for 20min in all animals. At 24h after reperfusion,mice were sacrificed and the hippocampus and cortex were dissected for Wnt signaling specified gene chip array. Results: Compared with the control group,the level of Wnt1,Wnt3,Wnt5a,Fzd4 and Fzd8 elevated while the level of Wnt7a decreased in the hippocampus in the EA group. However,in the cortex only the level of Wnt7a decreased. Conclusion: Wnt1,Wnt3,Wnt5a,Fzd4,Fzd8 and Wnt7a might be involved in the process of EA preconditioning in aged mice.Experiment3 The protective effect of electroacupuncture on patients undergoing craniotomy for brain tumorObjective: To investigate the effect of electroacupuncture preconditioning on serum S100βand NSE in patients undergoing craniotomy for tumor resection. Methods:32 patients with brain tumor scheduled for craniotomy under general anesthesia were divided into 2 groups(n=15 each).Patients in the EA group received electroacupuncture on Fengfu and Fengchi for 30min 2 hours before the operation.The stimulus is 1-4mA with a frequency of 2/15Hz. No pretreatment for the Control group. Anesthesia was maintained with remifentanil,vecuronium and propofol. Levels of the serum S100βand NSE were measured at 5 time points(before induction, before incision, tumor removal, end of operation,24h after operation) by ELISA. Results: The level of serum S100βand NSE did not change before incision. NSE increased significantly after taking away the tumor. And both of S100βand NSE increased at the end of the operation and 24h postoperatively.Compared with the control group, level of the two markers were significantly higher( P<0.05). Conclusion: Electroacupuncture at Fengchi and Fengfu for 30min before craniotomy could decrease the production of serum S100βand NSE.Experiment 4 Sufentanil pre-dosage induces protection against acute focal cerebral ischemia-reperfusion injury in ratsObjective:To investigate whether sufentanil pre-dosage could produce protection against middle cerebral artery occlusion( MCAO) induced ischemia-reperfusion injury in rats. Methods:Forty male SD rats were randomly divided into 4 groups(10 rats each):three sufentanil groups(group A,group B and group C) and a control group(group D). The four groups were intraperitoneally injected with sufentanil by the dose of 3μg/kg、6μg/kg、9μg/kg or normal saline respectively. 30 min after injection, all rats were subjected to the right MCAO for 120 min followed by 24h reperfusion. The neurological deficit scores ( NDS ) were evaluated at 24 h after reperfusion. Infarct volume, as a percentage of volume at normal cerebral hemisphere, was determined by 2,3,5-triphenyltetrazolium (TTC) staining. Results:The neurological deficit scores( NDS )in the group A(3μg/kg) at 24 h after reperfusion were higher than those in the group B (6μg/kg)、C(9μg/kg) and D (control) (P﹤0.05). The infarct volume in groupB、C and D was significantly larger than that in group A (P﹤0.05) at 24 h after reperfusion. There was no significant difference among NDS of group B、C and D . Conclusion:Sufentanil pre-dosage can protect rat brain against ischemia-reperfusion injury.Conclusion1.Electroapuncture preconditioning could alleviate forebrain ischemia reperfusion injury in aged mice.2. Wnt signaling pathway might be involved in the protective effect of electroapuncture preconditioning against forebrain cerebral ischemia injury.3. Electroacupuncture preconditioning before craniotomy could decrease the production of serum S100βand NSE,two markers of brain injury.4. Sufentanil pre-dosage can protect rat brain against ischemia-reperfusion injury induced by MCAO.

Comparison of Survival in Patients on Hemodialysis and Peritoneal Dialysis

Mon, 30 May 2011 13:08:24 +0000

Objective:To know basic characteristics of hemodialysis and peritoneal dialysis population in general hospital in china.Evaluating and comparing survival in patients on hemodialysis and peritoneal dialysis.And to explore the main risk factors affecting survival of dialysis.Methods:The demographic and clinical data of incident patients stating hemodialysis and peritoneal dialysis in Zhong Shan Hospital,Fudan University between 1 January 2005 and 31 December 2008 was retrospectively analysised.All patients were followed up through 31 March 2009.survival analysis was performed using Kaplan-Meier method, log-rank test and Cox regression model.The method of intention to treat analysis and as treated analysis were both performed in this study.Results:1.A total of 460 patients were enrolled.247 patients accepted the therapy of hemodialysis initially and others accepted the therapy of peritoneal dialysis.There was no significant difference of age,BMI,the history of renal insufficiency,eGFR, albumin,MAP,urine volume,cerebrovascular accident,cardiovascular accident, CHF and CCI between patients on hemodialysis and peritoneal dialysis at the beginning of dialysis(P＞0.05).2.Results of intention to treat analysis:Kaplan-Meier survival curve revealed that overall mortality rates were significantly lower among those being treated with HD initially(P＜0.05,log-rank test).But this risk was not constant over time. Mortality was similar in the first 1 year(P=0.14,log-rank test).In the period beyond 1 year,there was a significantly higher mortality among those treated with PD initially.When adjusted by age and DM,survival was better on HD(P＜0.05, log-rank test)in the group of nondiabetic patients old than 65 years.Although survival was slightly better on PD in the group of nondiabetic patients younger than 65 years,there was no significant difference of survival between HD and PD patients(P＞0.05,log-rank test).The multivariate Cox regression model indicated that there was no significant difference of survival between HD and PD patients when adjusted by case-mix differences(HR(HD:PD)=0.778,95%CI (0.483-1.254),P＜0.303) and age(HR=1.051,95%CI 1.030-1.073,P＜0.01)、cerebrovascular accident(HR=2.032,95%CI 1.125-3.670,P＜0.05)、CCI≥5 (HR=2.592,95%CI 1.230-5.465,P＜0.05)、PAB(HR=0.022,95%CI 0.001-0.768,P＜0.05) were the main risk factors to survival of dialysis.3.Results of as treated analysis was constant with those of intention to treat analysis except for PAB of which was no longer the main risk factors to survival of dialysis in as treated analysis.Conclusion:1.Survival was similar between HD and PD patients in the first 1 year of dialysis. In the period beyond 1 year,there maybe a significantly higher mortality among those treated with PD.Survival was better on HD in nondiabetic patients older than 65 years.2.Age,cerebrovascular accident and CCI≥5 were the main risk factors to survival of dialysis.

Clinical Studies on Interventional Therapy in Treating Early Stage Legg-Calve-Perthes Disease

Wed, 04 May 2011 01:24:06 +0000

Objective: To evaluate the clinical efficacy of the interventional radiology in treating early stage Legg-Calve-Perthes Disease,confirmed that interventional therapy in the treatment of early stage Legg-Calve-Perthes Disease was a scientific process.Methods: we collected the patients under 14 who were diagnosed to be avascular necrosis of the femoral head and treated in the traditional Chinese medicine hospital of Zhangzhou Fujian provence from 2005 to 2008. We divided them into two groups randomly with the random number table:the interventional therapy group cooperated with skin traction,the conservative treatment group cooperated with skin traction according to the following up order and the treated method. we compared the curative effect of every groups on pain,function,activity of joint and X-ray stages by more point of view and more levels to conclude the difference among various methods.Results: By 1-2 years\'following up,the success rate of therapy group,the control group was 92.35%, 80.95% respectively. By T test and Ridit test, we can see that there were notable difference between therapy group and control group in curative effect(p<0.05). the X-ray representation of the therapy group and the control group is improved(p<0.01); The prognosi under 7 years old is better than that above 7 years old; the prognosi was better when the course was no more than 3 months,while it was worse when the course was more than 3months; Sex has no effect on clinical curative effect.Conclusion: 1. Interventional therapy has provided an improving effect for the blood supply of the femoral head, for the avascular necrosis of vascular thrombosis provide good blood supply environment.2. There are close relatio between the prognosi of Legg-Calve-Perthes Disease and aging,the course. While sex has no effect on clinical curative effect.3. Interventional therapy can enhence the curative effect ,improve the pain,function, activity of joint and X-ray representation evidently, It\'s an effective way to cure avascular necrosis of the femoral head of children.

Clinical Study on Treatment of Pelvic Organ Prolapse (POP) with Prolift and Buzhongyiqi Tang

Wed, 04 May 2011 01:24:02 +0000

Objective:Through the use of polypropylene mesh combined with reconstructive pelvic surgery Buzhongyiqitang Chinese and Western Medicine methods pelvic organ prolapse, pelvic floor reconstruction to reduce postoperative recurrence rate and clinical complications and to achieve better treatment of pelvic organ the purpose of prolapse.Methods:1.the degree of pelvic organ prolapse and aboveⅡdiagnostic criteria for parallel mesh pelvic floor reconstruction surgery,the patients who would be in line with the standard figure of the randomized patients were randomly divided into the treatment group(polypropylene mesh pelvic floor reconstruction with Buzhongyiqitang) including 30 cases and the control group (polypropylene mesh reconstructive pelvic surgery) of 29 cases.2.The patients are respectively follow up in the out patient after operation of the sixth month to observe patients with clinical recurrence and complications.3.After 6-month operation,the patients were assessed by curative effect,quality of life score, different the indicatrix and traditional Chinese medicine changes in symptoms.Results:1.Compared the overall therapeutic effect of twe groupes of patients,The total effective rate of the treatment group was 96.7%,Thc total effective rate of control groups was 89.6%,There was signifcant effect between two groups(P＜0.05).2.Compared the relapse rate of twe groupes of patients,The relapse rate of the treatment group was 3.3%,The elapse rate of control groups was 83.7%,There was signifcant effect between two groups(P＜0.05).3.According to three chamber,there were noticeable difference in the middle pelvic(the impact of the vagina or uterus) and after the pelvic(the impact of bowel or rectum),and explained that treatment group are better the control group(P＜0.05).4.Whether an increase in vaginal discharged,vaginal itching or pain,whether or not sex life does not restored a sense of comparison,there was a significant difference between the treatment group and the control group(P＜0.05),Chinese medicine symptom score with treatment group than the control group,there is a significant advantage(P＜0.05).therefore the treatment group was significantly better than the control group.Conclusion: 1.It was definite clinical effect that prolift treat pelvic organ prolapse.2.The opration plus traditional Chinese drug was better than opration only.3.According to the three chamber,the degree of pelvic organ prolapse and clinical symptom quantified diffient chamber,the results showed that the treatment group was better than the the control group by reduced the Chinese medicine symptom score of the middle pelvic(the impact of the vagina or uterus) and after the pelvic(the impact of bowel or rectum).4.The oveall curative effect of pelvic organ prolapse,the Chinese medicine symptom score and parte of index in treatment group had significant advantage on treatment on pelvic organ prolapse,and can be applied in clinical.

Effects of Surgical Therapy and Conversative Treatment on Moyamoya Disease

Wed, 04 May 2011 01:24:01 +0000

Objective:Through using different therapeutic method(expectant treatment and operative treatment) to moyamoya disease patients were hospitalized in our hospital from November 2002 to August 2008,and long-term retrospective calling on their clinical data,we can control study the diference between therapeutic effect of expectant treatment and operative treatment.Methods:Total 35 moyamoya disease patients were hospitalized in our hospital from November 2002 to August 2008.They were divided into two groups:in groupⅠthere were 15 patients,who received surgery therapy; in group 11 there were 20 patients,who only received conservative treatment.Their clinical and image data were retrived and retrospective calls on were carried out for all patients transversely from September to December 2008.Results:In groupⅠ,the rate of re-haemorrhagia is 3.15%,the rate of Ischemia symptom is 4.93%;in groupⅡ,the rate of rehaemorrhagia is 6.15%,the rate of ischemia symptom is 6.13%.There are no significant diference between surgery and conservative treatment in present convalescent condition,rate of re-hemorrhage and rate of ischemic symptom occurrence.

Partition Map of Contusion of the Femoral Condyle under Magnetic Resonance and Its Clinical Significance

Wed, 04 May 2011 01:24:00 +0000

Objective: To investigate the MRI change of contusion in different regions of femoral condyle and its clinical significanceMethods: The knee joint MRI materials of 29 cases having femoral condyle contusion were analyzed, including the location and clinical characteristics of contusion. Of all the 29 cases, 46 areas of femoral condyle contusion were found, which included 42 bone injured cases in epiphysis. 31 areas of femoral condyle contusion were found in lateral condyle,and 11 areas of femoral condyle contusion were found in medial condyle. Of all the cases, 24 areas of femoral condyle contusion were found in weight loading areas, Which accounted for 52 percent of the total number of contusion. We recommended that patients with acute injury should restrict weight-bearing walking, and do functional exercise and physical therapy for three weeks. Cleanning-up, drilling decompression under knee arthroscopy were done to cure the four patients with symptoms over eight weeks after injury.Result:Of all the 25 cases of acute injury patients, 14 patients\' symptoms disappeared completely,6 patients\' symptoms improved more or less after three weeks\' conservative treatment. The 5 patients\' therapeutic effect was not obvious. While patients with obsolete injury began to do weight-bearing exersise in four weeks after surgery, Joint pain disappeared and had good range of motion. In seven months after surgery, 1 case underwent follow-up MRI scanning showed the contusion absorbed apparently, Integrity of cartilage filled completely under knee arthroscopy.Conclusion: The contusion of femoral condyle in the weight-bearing area occurred frequently. The time of full absorbtion was longer than that of expectation. schemic events might occur happen in long term. Contusion of femoral condyle, especially in the weight-bearing area, should be paid more attention to give early proper treatment.

The Clinical Analysis of Hypocalcemia after Thyroid Surgery

Wed, 04 May 2011 01:23:59 +0000

Objective: To investigate the prevention and treatment of hypo- calcemia after thyroid surgery .Methods: The change of serum calcium level of pre- and postoperative was analyzed in 49 patients with hypocalcemia after thyroid surgery . Hypocalcemia was defined as serum calcemia<2.20mmol/L . And serum PTH level was tested in 16 patients . The serum calcium level of postoperative changed with PTH level by statistical methods .Results: Hypocalcemia occurred in 49 cases (22.69%) after thyroid surgery . Of them , 5 cases were in total thyroidectomy , 36 cases in unilateral lobectomy with contralateral partial thyroid lobectomy , 7 cases in bilateral partial thyroid lobectomy , and 1 case in unilateral partial thyroid lobectomy . Transient hypocalcemia was identified in all patients , including 27 asymptomatic and 22 symptomatic which were serum calcemia <1.83mmol/L. There was a statistical significant different between pre- and postoperative serum calcemia in patients with total thyroidectomy, unilateral lobectomy with contralateral partial thyroid lobectomy and bilateral partial thyroid lobectomy(p< 0.05). The serum calcemia was the lowest point on second postoperative day. Serum PTH level was declined after operation but in normal range in 16 patients . Serum calcemia of postoperative reduced have relation with PTH declining.Conclusion:①Hypocalcemia often occurs in patients after thyroid surgery.②The function of parathyroid may be changed with thyroid surgery.③The lower PTH after thyroid surgery may be one cause of hypocalcemia after thyroid surgery.④Most patients were transient hypocalcemia and some of them occurred clinical symptoms.

The Treatment and Prognostic Factors of Primary Central Nervous System Lymphoma

Wed, 04 May 2011 01:23:58 +0000

Background and Objectives:Primary central nervous system lymphoma (PCNSL) is an uncommon form of non-Hodgkin\'s lymphoma(NHL) that has been increasing in incidence over the past three decades.PCNSL accountes for only 1%of lymphoma in all parts of our body,and accountes for only 0.3%～1.5%of all brain tumors.The long-term curative effect for most of patients is still poor because of the complexed clinical manifestation in the disease,and the difficulties in diagnosis and treatment.The patient\'s condition and survival time have been improved due to improvement of the therapy and the use of new therapy methods.There are many ways for the treatment of PCNSL,such as hormone therapy、surgery therapy、radiotherapy、chemotherapy、combined therapy and new methold,according to the symptom and condition of the patients.But there are fit and unfit for every kind of therapies in clinical treatment.One received、unified therapeutic regimen should be determined.The choice of treatment remains controversial.Analysis of the prognosis is important to judgement of patient\'s prognosis、choice of therapy method、plan for the treatment、distribution of the medical treatment resources reasonably.Approach of prognosis judgement remains to be analyzed.This study is to investigate the short-term and long-term survival of 56 cases patients with PCNSL in our hospital, and to evaluate the prognostic factor systematically,so as to make some help to clinical treatment. Materials and methods:56 cases patients with PCNSL between January 1997 and March 2009 are analyzed.All patients are diagnosed by pathology biopsy after surgery or stereotactic biopsy.There are 41 cases of B-cell lymphoma,and 15 cases ofT-cell lymphoma.38 cases have single-focus,18 cases have multi-focus.Tumors in 16 cases were completely resected.Tumors in 31 cases were partially resected,9 cases were received stereotactic biopsy.17 cases only received radiotherapy after surgery.8 cases only received chemotherapy after surgery.31 cases received combined thearpy with radiotherapy and chemotherapy,in which 18 cases received chemotherapy after radiotherapy,10 cases received radiotherapy after chemotherapy, 3 cases received radiotherapy and chemotherapy synchronously.Whole brain was radiated by 30～40Gy mainly in the radiotherapy,10～20Gy was delivered to GTV in some patients.Chemotherapy with CHOP schedule was used in 22 cases patients. 14 cases received high dose of MTX or combination of high dose of arabinosylcytosin(2—3g/m~2).7 cases received chemotherapy with HD-MTX mainly combined with whole brain radiotherapy.4 cases were performed(MTX 15mg, arabinosylcytosin 50mg,dexamethasone 5mg) by intrathecal injection when chemotherapy.Results:The medium follow-up time was 46.4 months(range,1.0～93.8 months). Of all the patients,CR was 21 cases(37.5%),PR was 14 cases(25.0%),NC was 10 cases(17.9%),PD was 11 cases(19.6%),respectively,and local control rate was 80.4%.LCR of group beyond 60 years old(53.3%)was lower than that of bellow 60 years old(90.2%),P＜0.05;LCR of multi-focus group(61.1%)was lower than that of single-focus group(89.5%),P＜0.05.LCR of group received combined therapy with radiotherapy and chemotherapy(93.5%)was higher than that of group without received combined therapy with radiotherapy and chemotherapy(64.0%),P＜0.05. The medium survival time(MST) was 22.6 months,and the 1-,2-,3-,4-,5-year OS rates was 66.1%、42.8%、34.0%、23.2%、respectively for all patients.5-year OS rate of the group beyond 60 years old was(0.0%)lower than that of bellow 60 years old(31.7%),P＜0.05;5-year OS rate of B-cell lymphoma group(0.0%)was lower than that ofT-cell lymphoma group(34.2%),P＜0.05;5-year OS rate of multi-focus group(0.0%)was lower than that of single-focus group(34.2%),P＜0.05.5-year OS rate of group received(HD-MTX+WBRT)(71.4%) was higher than that of group without received(HD-MTX+WBRT)(16.3%),P＜0.05.The MST of patients without receiving radiotherapy or chemotherapy was only 2 months.The MST of patients with radiotherapy alone after surgery、chemotherapy alone after surgery、combined therapy with radiotherapy and chemotherapy was 20.5、25.3、26.4 months respectively.The MST of patients receiving chemotherapy with HD-MTX and(HD-MTX+WBRT) was 32.5 months and 36.4 months respectively. We didn\'t think that the sequence of radiotherapy and chemotherapy would affect on the MST of patients.The MST of patients with radiotherapy after chemotherapy was 27.3 months,the MST of patients with chemotherapy after radiotherapy was 25.9months,there were not significant difference(P＞0.05).Conclusions:1.PCNSL is sensitive to radiotherapy and chemotherapy,but it is easy to recur and gives a poor prognosis to the patients.2.The curative effect of combined therapy with radiotherapy and chemotherapy is better than that of radiotherapy alone or chemotherapy alone.3.Chemotherapy with high dose of MTX is effective to the treatment of PCNSL.Chemotherapy with high dose of MTX combined with whole brain radiotherapy is the best choice for the treatment of PCNSL recently.4.The prognostic factors for PCNSL are age、pathologic category、single-focus or multi-focus and chemotherapy HD-MTX plus WBRT therapy or not.

Sensing Method Research Based on Porphyrins for Characteristic Breath of Lung Cancer

Wed, 04 May 2011 01:23:57 +0000

According to the situation about low survival rate of lung cancer patient after final diagnosis, this work proposed porphyrin-sensing method to detect lung cancer based on Breath Diagnosis. Through a research on trace volatile organic compounds of the breath gas from lung cancer patient, early noninvasive diagnosis and monitoring pathogenetic condition could be realized for patients.Sensing methods with VOCs about domestic and overseas investigation were introduced. Analyzing their advantage and disadvantage, new detecting method based on optical spectrum with porphyrin was proposed. First, several kinds of porphyrins and derivates were prepared. UV-Vis, Infrared absorption spectroscopy, and HNMR were used to characterize their structure and property. The synthetical compounds were confirmed for our target compounds.Secondly, sensitivity test on optical spectrum between porphyrins and VOCs of breath gas from lung cancer patients was studied. UV-Vis was used to scan successively with their mixed solution before and after reaction. Then, UV-Vis spectra change of single porphyrin exposed to single analytes, detection limit and featured interactional respond were determined, and some porphyrins were discovered that their limit of detection with target analytes was low to 1×10-8. The relationship between the structure of porphyrin and sensitivity, and their mechanism of action was elucidated simply according to the spectra change of P1 with nine kinds of target analytes.Cross-selecting of multitude sensitive materials can avoid the disadvantage of single material performance, and improve the specificity and selectivity for target analytes. Handiwork spotting was applied to prepared 6×6 porphyrin arrays with porphyrins and indicators. Owing to the colour change of porphyrin arrays, when porphyrins reacted with gas molecule, porphyrin array images before and after exposure to different concentrations of analytes were collected with porphyrin sensing micro-system designed by our lab, and gained the response characteristic spectrum of every analyte either. Principle Component and Hierarchical Cluster Analysis for featured responding RGB data of porphyrin array was conducted. The results showed that the detection had good repeatability and the nine analytes can be distinguished completely with good clustering in condition of low resemblance dimension.As porphyrins have such advantages of optical performance, the mechanism of action for porphyrins with gas molecule was identified for further study. When the stability and sensitivity keep well, porphyrin sensor with excellent performance could be developed for early diagnosis and ambulatory monitoring of lung cancer.

Design and Biomechanical Evaluation of Expansion Cannulated Screws

Wed, 04 May 2011 01:23:56 +0000

Femoral neck fracture is a common trauma, which is also particularly prevalent in the elderly. With the development of society, the incidence of femoral neck fracture in children and young adults is gradually increased. Many scholars design a variety of internal fixation materials for femoral neck fracture; however, given to special femoral neck fracture in terms of biomechanical properties, each has its own advantages and disadvantages. Traditional fixed appliances with the apparent shortcomings are as follows: stress at both ends of fracture is significant different, and the upper of the femoral neck has a higher stress, causing more impact on treatment and recovery. At the same time, the scope of application is limited than its predecessor owing to the lack of ability such as anti-compression, anti-bending and anti-reverse, and the screws are prone to unpinning process, rotation of the femoral head , cut-out of the screw caused by unstable fixed. Since the biomechanics complexity of the femoral head and the specificity of local blood supply, the incidence of avascular necrosis of femoral head (ANFH) was still as high as 20% ~ 40%. Femoral neck fracture in the elderly, due to the presence of osteoporosis, pull-out strength/holding power of fixation to the bone is weak, leading to screw loosening, fracture displacement easily, and also increased the rate of nonunion and ANFH. Therefore, without prejudice to the local blood supply, an increase of pull-out strength between bone and screw on the scope of femoral neck bone anatomy can improve the reliability and stability of the fixation. The purpose of this study is to explore the methods to provide superior biomechanical properties, and more applicable to cannulated bone, especially the femoral neck fracture fixation in the elderly.This subject contains the following three studies: 1) to design expansion cannulated bone screws (ECS). By way of screwing in the inter stick into the bolt, without increasing the diameter of screw in the femoral neck, the expansion cannulated screws could increase the diameter of screw tip that implanted in the femoral head, which can enhance the holding power; 2) on the femoral specimens of the adult, maximum axial pull-out tests, anti-rotation experiments and the maximum resist compressive strength tests were performed to compare with the hollow screw (cannulated compression screws, CCS), and the fixation reliability of the expansion cannulated bone screw was tested by biomechanical testing and evaluation; 3) a fatigue simulation was carried out, and 3-point bend tests were also performed to evaluated the static strength of expansive cannulated screws.Measure the largest diameter of expansive cannulated screws implanted in the femoral neck and head. The diameters of expanding screw tips measured by radiographs were ranged from 1 to 1.8mm, and an average increase is 1.3mm. After screws pull-out, the diameter can increase a range of 1.3 ~ 1.8mm and the mean increase is 1.5mm. All expansion of screws achieves the anticipated result.Biomechanical testing indicate that expansion cannulated screws are superior to the conventional after making use of the same number of screws for fixation in maximum compressive strength test and anti-rotation angle. The effect of 2 pieces of ECS and 3 pieces of CCS had no significant difference. The maximum axial pull-out strength was also significantly higher than the hollow compression screw.The mechanical properties tests indicate that all expansion cannulated screws could tolerate the periodic load without fatigue. A fatigue simulation using perpendicular loading up to 500,000 cycles of 0~2.1KN was carried out. The hollow structure of expansion cannulated screw did not have a significant effect on fatigue performance. In 3-point bending test, despite the expansion cannulated screw has a more feeble failure strength than that of the control, the anti-fracture strength is 0.98KN, which could withstand the maximum load.To analyze the experimental results, we can draw the following conclusions: 1) the expansion cannulated bone screw had a more reliable performance than hollow screws during the initial fixation; 2) in some specific circumstances, the use of two expansion cannulated bone screws were basically same to the traditional three hollow screws on effect; 3) there was no predisposition to failure of the expansion cannulated screws in fatigue-test; 4) Despite the static mechanical strength has been reduced in the expansion cannulated screws, the normal femoral neck coule withstand the maximum load.

A System of higher Speed Data Acquisition and Detection for Velocity of Blood Based on Linear CCD

Wed, 04 May 2011 01:23:55 +0000

Clinical research reveals that hemorheology parameter has strong inter-relationship with cardiovascular, artery, and hematic disease, therefore, it is a very important reference in medical research and clinical diagnose.The blood viscosity, regarded as the one of the most important hemorheology parameters, reflects the transporting and supplying condition of the blood and influences the flowing of the blood in the body. It can be seen that the velocity of blood is slower when the blood viscosity is low; whereas, the velocity of blood is faster when the blood viscosity is high. Consequently, it has the important meaning to reflect blood viscosity by measuring the velocity of the blood in the case of constant pressure.Charge Coupled Devices(CCD) is a photoelectric device with high performance, which is widely applied in many areas, including astronomical imagine, entertrainment and manufacturing, etc. The geometrical measure with CCD has an advantage on achieving high accuracy, dynamic detection and non-contact measurement. Therefore, the paper introduces a method of real-time measuring the velocity of blood in the use of TCD1702C image sensor.The focus of this dissertation research is to develop the hardware of linear-CCD data acquisition system based on PIC processor, and the related software to support the system. The principle of linear CCD and the scheme of whole system are introduced firstly in the dissertation. The system comprises three modules: the first is the CCD drive module, which is controlled with CPLD. Programming the CPLD to produce CCD drive pluses and synchronized communication signals. The second is data acquisition module. After preprocessing, when detecting the velocity of a drop of blood in the capillary glass,the output video signals of CCD are transmitted into high resolution ADC module, in which they are converted into digital signals and the useful digital signals is memorized by FIFO, and when detecting the velocity of continunous drops of blood in the capillary glass, the thresholding signal is produced by the comparator circuit and it leads that the useful pixel signal is captured by capture module of PIC singal-chip processor. The third is data processing and displaying. The data is processed in PIC singal-chip processor, and the velocity of blood is caculated, and then the data is transported into PC by serial communication. The interface graph is designed in the use of Visual Basic 6.0 software and the curve of the velocity of blood is showed. Finally, the debug process of the unit circuit is introduced in proper sequence. Compiling and silulating the system operation software. The experiment results are presented and analyzed.

Study of Software Phase Detection Platform and Data Acquisition in MIT System

Wed, 04 May 2011 01:23:50 +0000

Magnetic Induction Tomography (MIT) uses the theory of electrical magnetic field to measure the distribution of biological tissue’s electrical conductivity. As a non-invasive, contactless, miniature and continuous method, it is very important in monitoring and diagnosing the encephalic pathological changes.MIT mainly based on the theory of the eddy current. As the phase deviation has a linear relationship with the conductivity of biological tissue, the accurate detection of phase deviation will affect the imaging quality. The establishment of high-precision phase deviation detection unit plays an important role in improving the performance of MIT system.Now, most of MIT research groups mainly focus on designing hardware circuit or using down-frequency techniques to improve the precision of phase detecting unit. The precision is difficult to be improved for drift, noise and interference signals existing in the hardware circuit. The methods of software phase detecting, which mainly focus on algorithms and overcome the flaws in hardware techniques, make the detecting results more accurate.The thesis based on former researches of our group, aiming at improving the precision of phase detecting unit and reducing the interference signals, do a further research and design of phase detecting unit in MIT system with software methods. The software phase detecting platform was built using frequency spectrum analysis method, correlation method and phase sensitive detecting method. It can carry on real-time detecting of phase deviation.The main contents of this thesis are as follows:1. We analyzed the existing phase detecting algorithms and found frequency spectrum analysis algorithm, correlation algorithm and phase sensitive detecting algorithm were suitable for our system. We studied the theory of the algorithms and the influencing factors.2. We achieved the phase detecting algorithms and built software phase detecting platform with LabVIEW.3. We compared the software phase detecting methods with hardware phase detecting method by simulating experiments. We used evaluation indicators error and linearity to evaluate the software and hardware phase detecting methods.4. We detected the phase deviation of NaCl solution model, agar-NaCl model and cell model using the built software phase detecting platform and evaluated the results we detected.5. We built 16-channel data acquisition system.Finally, we established a whole software phase detecting platform with signal acquisition and processing. The phase accuracy of the software platform could be achieved at 0.01°, which was higher than the high-precision lock-in amplifier SR844 with phase accuracy of 0.02°. The paper laid a good foundation for further detecting of the encephalic pathological changes. These might be of great help for further cerebral edema-detecting work.

The Relationship between Apoptosis Proteins Bax XIAP and Apoptosis in Placenta and Their Significance in Pathogenesis of Preeclampsia

Thu, 31 Mar 2011 13:23:36 +0000

Objective:To observe apoptosis in placenta tissue of patients with preeclampsia and the relationship between BAX&XIAP and and their significance in pathogenesis of preeclampsia.Methods:30 Placenta samples were obtained from patients with preecl- ampsia(10 mild preeclampsia,20 severe preeclampsia),and control group were30 normal third-trimester pregnancies.Immunohistochemistry(S-P) me- thod was used to study the expression of BAX and XIAP in human placenta. Termianl deoxynucleotidyl transferase-mediated deoxyuridine triphosphate marker nick end labeling(TUNEL) was used to quantify the incidence of apoptosis.Results:(1)The apoptosis rate of preeclampsia group was higher than Control group.It was had significant difference between these two groups (P<0.001).With the aggravation of this illness the apoptosis rate was higher. It was had significant difference (P<0.001) (2)The expression of BAX in preeclampsia group was higher than control group (P<0.05). Preeclampsia group expression of Bax and the apoptosis rate were significant positive correlation (P<0.001)(3)The expression of XIAP of preeclampsia group was lower than control group Preeclampsia group expression of XIAP and the apoptosis rate were significant negative correlation (P<0.001)(6) Preeclam- sia group expression of BAX and XIAP were significant negative correlati- on (p<0.001)Conclusions:Apoptosis of the placental trophoblasts of preeclampsia human were heightened evidently .The expression of BAX increased and the expression of XIAP was reduced. The disequilibrium between BAX and XIAP may induce the occurrence of preeclampsia.

Relationship between Genetic Polymorphism of GSTP1 CYP19 and Alzheimer Disease

Thu, 31 Mar 2011 13:23:35 +0000

Introduction:Alzheimer\'s disease(AD),the major cause of presenile dementia and senile dementia,is a common progressive neurodegenerative disease in central nervous system with clinical manifestations as progressive memory decline,cognitive dysfunction,personality changes and language barriers.It was first discovered between 1906 and 1907 by Alois Alzheimer,a German neurologist.At present,20 or 30 million people are diagnosed with AD all over the world.Statisticas from Europe and the U.S.show that the prevalence rate of dementia in elderly population over 65 is from 3.0%to 5.0%and more than half of them with AD.In the United States,AD is considered as the fourth leading cause of death after cardiovascular diseases,cancer and stroke.In china,more than 100 thousand people are diagnosed with AD and the prevalence rate of dementia reported in elderly population over 60 is from 0.75%to 4.69%.With the world\'s aging population,SD is becoming one of the most toughest challenges ever in gerontology.Duing to its unclear etiology and pathogenesis,AD is generally considered as heterogeneity in complex disease and it may be caused by various factors(genetic and enviromental factors,neurotransmitters and immunocompetence,etc.).The transformation of exogenous chemical substances in the body is the result of joint action and active balance between multiple enzymes,and is dependent on toxicant-metabolizing enzymes. Such as cytochrome P450 enzyme,phaseⅠenzyme,,can alter the functional genes of substances or decompose them via oxidation,reduction and hydrolysis.The phaseⅡenzyme(like glutathione S-transferase and N-acetyltransferase) serves as catalyst activating the binding reaction between exogenous chemical substances and certain strong polar groups(like methyl and acetyl).Then the polarity and water solubility of those substances will be enhenced as a result,which will help them eject with urine and bile.The transformation of exogenous chemical substances in the body is the result of joint action and active balance between multiple enzymes.As a phaseⅡenzyme,Glutathione S-transferase(GST) is an important detoxification enzyme in vivo and serves as catalyst activating the binding reaction between electrophiles and reduced glutathione to disable electrophiles and make them eject with urine and bile.Thus it play an significant role in metabolizing environmental carcinogens and chemotherapy drugs.GSTs is a super-gene family with detoxification including membrane-type,cytosol-type,mitochondrion-type and LTC4 synthase.It is common in almost all cells and tissues but are found in highest concentrations in liver,colon and gonad.Population polymorphisms is revealed in GSTM1 and GSTP1.The relationshaip between the genetic polymorphisms of GSTs and environment afects our susceptibilities to certain diseases.Therefore,it is of great significance to correctly understsand their relationship to find out the role of GSTs in the incidence of deiseases and improve disease prediction.There is polymorphism in GSTP1 gene and it\'s exon 5(Ile105→Val).GSTP1 enzyme,the encoding product of GSTP1 gene,consists of 210 kinds of amino acid.It is reported that section 105 in amino acid sequence is vital for the biological functions of proteins as it determines the size and hydrophobicity of amino acid then finally impact enzyme thermal stability.GSTP1-Ile105 is reportedly about 2-3 times better than GSTP1-Val105 in thermal stability.Along with the gradual development of study on GSTP1 structure and function,it is realized that enzymes stem from GSTP1 allelic encoding are totally different.Therefore,the polymorphism of GSTP1 gene encoding is probably related with the genetic susceptibility of AD.Studies concluded that the ndogenous estrogen level in postmenopausal patients with AD is obviously lower than that of patients of the same age without AD.In AD patients,the local formation of estrogen has significant neurotrophic and neuroprotective effects and thus play a role in the promotion of neurotrophic factor expression Upregulation of neurotrophic factor receptor.On contrary,neurotrophic factors help cells conbined with estrogen to amplify signals and strenthen neurotrophic effects.The mechanism of estrogen in AD treatment(particularly in AD prevetion) is becoming a study hotspot both at home and abroud.Aromatase belongs to cytochrome P450 superfamily and is the last rate-limiting enzyme in estrogen synthesis.It\'s encoding gene CYP19 is located on chromosome 15----band 1 in long arm 21 region,with 123Kb of length.Studies showed that estrogen steming from aromatase is closely related with brain development and regeneration.Normally speaking,aromatase can be found in various tissues of human and other mammals, for example,in central nervous system,placental syncytiotrophoblast cells,ovarian granulosa cells,adipose tissue and its stromal cells and adrenal.In specific brain regions,aromatase helps androgen transforming into estrogen and it\'s activity is related with the estrogen levels.Studies in recent years revealed that CYP19 comprises many tissue-specific promoters mediating CYP19 expression and determining the tissue-specific pattern of CYP19 expression.There are few studies concerning the relationship between AD and aromatase.Population polymorphisms is revealed in CYP19 and thus the polymorphism of GSTP1 gene encoding is probably related with the genetic susceptibility of AD.Objective:The purpose of this experiment was to investigate the relationship between the genetic susceptibility of AD and CYP19 and it\'s polymorphism for the purpose of providing molecular biological indicators for AD diagnose.Methods:The blood samples were from 102 AD patients in Beijing Hospital and Military Nursing House and the blood samples in the control group were from 121 old healthy people over 65 with MMSE score higher or equal to 28 in community.As in accordance with ICD-10 diagnostic criteria,uspicious AD patients were taken as study object and MMSE and scale of cognitive function in the elderly were applied in this experiment.Imaging data were used for diagonose and over one year follow-up was conducted in all cases. Two millilitre whole blood was drawn from each testee and anticoagulated with sodium citrate.Peripheral blood leukocyte DNA was extracted with Phenol-chloroform.Polymerase chain reaction -restriction fragment length polymorphism technique was applied and 5\'-TGC TAT CGT GGT TAA AAT CCA AGG-3\'in upstream and 5\'-TTA CCT GAG AGG CCA AGA AAA ACA-3\'downstream were used for the amplificationof alleles located on Mfe I on cytochrome CYP19.Mfe I restriction endonuclease was applied to detect A→T polymorphism.5\'-ACC CCA GGG CTC TAT GGG AA-3\'in upstream and 5\'-TGA GGG CAC AAG AAG CCC CT-3\' in downsteam were used for the amplification of GSTP1 gene.BsmAⅠrestriction endonuclease was applied to detect the A→G polymorphism of GSTP1 gene.SPSS13.0 statistical package was employed in the comparison of genotype frequency gene frequency.Chi-square test was used to calculate the allele frequency and mutation frequency in AD patients and healthy population.Results:Mfe I restriction endonuclease was applied to detect A→T polymorphism.The result showed that 135 out of 204 AD patients were with allele m1(66.2%)and 69 were with allele m2(33.8%).In the control group,however,196 out of 242 healthy people were with allele ml(81.0%) and 46 were with allele m2(19.0%).There was significant difference between the two groups(χ~2=12.696, P＜0.05,95%CI:1.413～3.357).In the case group,the genotype frequency distributions of m1/m1,m1/m2 and m2/m2 in Mfe I locus of CYP19 were 44.1%, 44.1%and 11.8%respectively,while in the control group were 65.3%.31.4%and 3.3%.There was significant difference between the two groups(χ~2=12.384 P＜0.05). In the case group,the genotype frequency distributions of M1 and M2 were 73.4% and 26.6%respectively,while in the control group were 83.1%and 16.9%.There was statistically significant difference between the two groups(χ~2=6.134 P＜0.05,95%CI: 1.124～2.809).In the case group,the genotype frequency distributions of m1/m1, m1/m2 and m2/m2 in BsmAⅠlocus of CYP19 were 54.9%,36.3%and 8.8% respectively,while in the control group were 70.2%,25.6%and 4.1%.There was significant difference between the two groups(χ~2=6.062 P＜0.05).Conclusion:the genotype frequencies of Mfe I locus and BsmAⅠin CYP19 in the control group was almost consistent with the average frequency of healthy population.Significant differences of genotype frequency and gene frequency in Mfe I locus of GSTP1 gene were found in the case group and the control group,which indicates that carrying mutant gene T can increase that incidence of sporadic AD.Significant differences of genotype frequency and gene frequency in BsmAⅠlocus of GSTP1 gene were found in the case group and the control group,which indicates that carrying mutant gene G can increase that incidence of sporadic AD.

Effects of Medical Ozone on Cellular Immunity in Chronic Viral Hepatitis B Patients

Thu, 31 Mar 2011 13:23:33 +0000

BackgroundHepatitis B virus is one of the most serious and prevalent health problems, affecting more than 2 billion people worldwide.Although highly effective vaccines against hepatitis B virus have been available since 1982,there are still more than 350 million chronic carriers,75%of whom reside in the Asia Pacific region.People with hepatitis B are at increased risk of developing hepatic decompensation,cirrhosis,and hepatocellular carcinoma.The estimated worldwide mortality is 0.5 to 1.2 million deaths a year.In china,there are 120 million people with HBV infection,out of which 20 to 30 million people show chronic hepatitis B virus infection.Chronic infection remains a challenging global health problem.The outcome of HBV infection is closely related with host\'s immune status.The immune tolerance and defect of specific cytotoxic T cell(CTL) response are the important reasons of chronic HBV infection.There are about five immunological characteristics for chronic HBV infection.Firstly,in chronic HBV infection,it exists immune tolerance to some extent,and there is a large number of virus antigens in peripheral blood.The long-term stimulation of high concentration of viral antigens probably induced more Treg cells,and expression of immunosuppressive molecules such as PD-1 and CTLA-4 were increased,all of which may cause immune tolerance and chronic HBV infection.Secondly,The less quantity of antigen-presenting cells,especially dendritic cells(DC) and functional defects may affect the interaction of DC-T cells,so that the expression of HLA-Ⅱby HBsAg-induced T cells and T cells proliferation weakened,which should not activate Th cells to produce cytokines,or cytotoxicity of cytotoxic T cell(CTL).Thirdly,Patterns of HBV-specific TCR CDR3 are narrow and its peak shape is bias,and proliferation of T cells were monoclonal or oligoclonal,so that the spectral of identification of HBV antigen and response was narrow.Fourthly,Th1/Th2 response is being imbalance.The cytokines type secreted by T cells at the site of virus replication may affect the final outcome of HBV infection.When specific Th1 cell is dominant,they promote the cellular immunity which is in favor of the removal of virus;and when Th2 cell is dominant,although it\'s in favor of production of antibodies to HBV,easily lead to chronic infection.For chronic HBV infection,Th1-type cytokines(IFN-γ, TNF-α,IL-2,etc.) is relatively short,and the Th2-type cytokines(IL-4,IL-10,etc.) are being relatively high levels,thereby maintaining the chronic infection status.Fifthly,anti-virus-related cytokines(TNF-αand IFN-γ,etc.) may exist gene mutation, genetic polymorphism or signal transduction abnormalities.An improved understanding of hepatitis B virology,immunology,and the natural course of chronic infection,has identified hepatitis B virus replication as the key driver of immune-mediated liver injury and disease progression.The approval of potent oral antiviral agents has revolutionized hepatitis B treatment since 1998. Conventional and pegylated interferon alfa and nucleotide analogues(lamivudine, adefovir,telbivudine and entecavir,etc.) are widely authorized treatments,and monotherapy with these drugs greatly suppresses virus replication,reduces hepatitis activity,and halts disease progression.However,hepatitis B virus is rarely eliminated, and drug resistance is a major drawback during long term therapy.The development of new drugs and strategies is needed to improve treatment outcomes. In recent years,medical ozone for the treatment of hepatitis B has become a new kind of exploration.Medical ozone is a gas mixed ozone with medical oxygen,which can be used to treat a variety of diseases such as rheumatism,ulcerative colitis and protrusion of irtervertebral disc.The reaction of ozone with human whole blood could generate reactive oxygen species(ROS,mainly H_2O_2) and lipid peroxidation products (LOPs),the role of these two products is to separately act on the various components of the blood(monocytes,platelets,red blood cell,WBC) and endothelial cells and other organs,resulting in a variety of effects.In recent years,hydrogen peroxide (H_2O_2) is considered an important intracellular messenger in animal and plant cells, so ozone plays an indirect role through its reaction products.At the end of 2004 Chinese mainland start of ozone treatment for viral hepatitis B and has shown initial signs of efficacy,medical ozone treatment of chronic hepatitis B is expected to become a new strategy.However,the mechanism of treatment is not clear.Previous studies have inferred immunoregulatory role of medical ozone,but many things still need to be further confirmed,and therefore the study of immune mechanism of medical ozone has important clinical significance.Objective1.To explore the mechanism of cell-mediated immunity of medical ozone for the CHB treatment.2.To provide evidence of basic research for the treatment of CHB with medical ozone and promote the wide range of clinical applications of medical ozone.3.To look for new therapeutic strategies for the treatment of CHB and improve therapeutic effects.Methods1.IFN-γand IL-4 levels of PBMC culture supernatant after acting by medical ozone with Enzyme-linked immunosorbent assay(ELISA) in CHB patients:There are 11 cases of CHB patients in this study,eight cases of healthy donors as control.PBMCs are separated with density gradient centrifugation,and then add medical ozone of five concentrations(4, 10,20,40,80μg / mL) in the supernatants of PBMCs with the method of effervescency,then cultivated in 37℃with 5%CO_2 cell culture incubator for 72 hours.We set up medical oxygen(O2,acting with effervescency),positive (to PHA stimulation),negative and double-complex-hole self-control group. The IFN-γand IL-4 concentrations in PBMC culture supernatants are detected wirh enzyme-linked immunosorbent assay(ELISA).The experimental operation carried out in accordance with the kit instructions.2.Detection of proportions of IFN-γand IL-4 positive cells in CD4~+ and CD8~+ T cells with flow cytometry(FCM) in CHB patients:There are 8 cases of CHB patients in this study,six cases of healthy donors as control,PBMCs are separated with density gradient centrifugation,and then add medical ozone of five concentrations(4,10,20,40,80μg / mL) in the supernatants of PBMCs,after the medical ozone reacting completely,add protein transporter blockers(Brefeldin A),at 37℃with 5%CO2 incubator cultivate the cells 6 hours,set up medical oxygen(O2),positive(to PHA stimulation),negative and double-complex-hole self-control group.The cell surface molecules CD4 and CD8 and intracellular cytokines IFN-γand IL-4 are stained by different fluorescent dyes labeled antibody.The percentage of IFN-γand IL-4 positive cells of CD4 + and CD8 + T lymphocytes is detected with flow cytometry (FCM).The results are analyzed using CellQuest software.3.In this study,2×8 factorial design is used,which compared the overall differences of two groups(CHB patients and healthy volunteers) or eight kinds of different treatment(5 experimental groups and 3 control groups). Differences in the overall comparison between the two groups or eight different treatment groups are compared by factorial design analysis of variance data;the overall differences of treatment group within the two groups is in comparison by one-way ANOVA;between two groups comparison of each experimental group with control group use two independent samples t test;comparisons in each experimental group with control group use paired t test.All data are analyzed using SPSS 13.0 software for statistical analysis,and the testing standard less than 0.05(P＜0.05) is considered significant difference.Results1.The concentrations and percentages of IFN-γand IL-4 of CD8 and CD4 positive T cells were significantly higher in positive control group than that of other treatment groups(P＜0.05).2.The IFN-γconcentration of CHB patients was significantly higher than that of healthy volunteers(F = 12.097,P = 0.001),while IL-4 concentration was slightly lower than that of healthy volunteers in the overall level,though there was no significant difference(F = 1.822,P = 0.179).The overall level of IFN-γand IL-4 concentrations between each treatment groups was significantly different(P＜0.05) for both CHB patients and health volunteers. Low concentration(4μg/mL,10μg/mL) Ozone can slightly stimulate IFN-γand IL-4 secretion,while high concentration(40μg/mL,80μg/mL) has inhibitory effect.3.For CHB patients,the proportion of IFN-γ-positive cells within CD8 positive T cells was significantly higher than that of healthy volunteers(F=5.101,P= 0.026),and the proportion of IFN-γ-positive CD4 T cells was higher than healthy control group(except the positive control group),though there was no significant difference(F=0.034,P = 0.854).The percentage of IFN-γ-positive cells within CD8 positive T cells was significant both in CHB patients and healthy donors(P = 0.001 and P = 0.043),and the percentage of IFN-γ-positive cells within CD4 positive T cells was not significant(P＞0.05). The percentage of IL-4 -positive cells within CD8 and CD4 positive T cells were not significant(P＞0.05) in the overall between CHB patients and healthy donors,and there is no significant trend.The percentage of IL-4 -positive cells within CD8 and CD4 positive T cells were not significant(P＞0.05) for CHB patients,while that was significant for healthy donors(P = 0.019 and P = 0.009).When medical ozone concentration increased,the proportion of IFN-γand IL-4 positive cells gradually decreased,which shows that low concentrations of medical ozone had a slight stimulating effect on cytokine secretion of PBMC,while high concentrations inhibited it.Conclusions1.Low concentration of medical ozone in patients with CHB can enhance the function of cytokines secretion in PBMC,strengthens the body\'s cellular immune responses and will help to clear the virus;while high concentration of medical ozone shows inhibitory effect for the secretion of cytokines in PBMC.This discovery provides a theoretical basis for ozone application in clinical treatment.The effect of medical ozone on HBV-specific cellular immunity remains to be further researched.2.Effects of medical ozone on cytokines secretion of PBMC are closely related to ozone dose and the ozone response medium,and that is worth concerning.3.Even though this study found a very interesting phenomenon,it still needs many basic research,as well as multi-center randomized controlled clinical trial to further confirm the role of ozone.Medical ozone treatment of viral hepatitis is a new attempt and ozone treatment is an endless exploration.4.Though some comparisons could not form a significant difference,and some conclusions are not yet certain enough.That was possibly owing to our small sample.So we still need to carry out larger sample studies to get more meaningful results.

Clinical Observation of Applying Fibrin Glue in the Pterygium Excisions with Sutureless Amniotic Membrane Transplantation

Thu, 31 Mar 2011 13:23:32 +0000

Objective: To observe and investigate the clinical efficacy of fibrin glue for attaching human amniotic membrane after primary pterygium excision.Methods: 21 patients (26 eyes) with primary pterygium underwent pterygium excision with amniotic membrane transplantation. Patients were randomized to two groups: fibrin glue group and suture group. In fibrin glue group, 11 patients (13 eyes) underwent pterygium excision, and the bare sclera was covered with amniotic membrane using fibrin glue for graft adherence; while in suture group, 10 patients (13 eyes), after pterygium excision, the amniotic membrane grafts were attached to the sclera using 10-0 nylon sutures. All surgeries were performed by the same doctor and surgical time was recorded. Patients were followed up in six months and ocular signs and symptoms were recorded. The results were analyzed by statistic software of SPSS.Results: Average surgical time (13.6±3.5 min) in fibrin glue group was significantly less than suture group (22.3±5.2 min) (P <0.01). In 2 weeks after surgery (before sutures out), less regional symptoms, such as pain, foreign-body sensation, tearing, were observed in the suture group (P <0.01). There were no transplant losses or dislocations in both groups by the end of the follow-up. It was observed that human amniotic membrane was tightly attached to the scleral bed in fibrin glue group. Complete epithelialization occurred in fibrin glue group within one week, while that in suture group was delayed to 8-10 days. Effusion was found under amniotic membranes in 5 eyes of suture group. Grafs’healing in fibrin glue group was better and no regional scar observed, while scars were seen in suture group at different levels. No recurrence was observed in both groups at the end of 6-month follow-up.Conclusion: Our results showed that fibrin glue can replace of sutures for attaching human amniotic membrane graft in primary pterygium surgery. It not only significantly reduces the surgical time, minimizes postoperative discomfort, but also benefits in graft’s healing and decrease regional scar, thus leads to cosmetic aim.

The Study of the Immortalization of Human Bone Marrow Mesenchymal Stem Cells by Transducting Human Telomerase Reverse Transcriptase Gene

Thu, 31 Mar 2011 13:23:31 +0000

Objective: Human bone marrow mesenchymal stem cells (hBMSCs) have the multipotentials of differentiation into various cells,so they are considered as an important source of seed cells in tissue engineering. In vitro, hBMSCs have a limited life, and gradually lose their differentiation potential after several passages,which restrict their scientific and clinical application.The rapid development of transgenic technology makes the establishment of immortalized cells possible.By using the retroviral vector,we were trying to transduct exogenous human telomerase reverse transcriptase(hTERT) gene into hBMSCs in order to establish the immortalized hBMSCs.Methods:The retroviral vector containing hTERT gene (pLEGFP-C1- hTERT) was transfected into packaging cell PT67 through liposomes.Then cell clones positive for the virus were selected by G418.Three cell clones were chose and amplified culture respectively.NIH 3T3 cell lines were used to detect retrovirus titers.hBMSCs were extracted and amplificated. hBMSCs in passage 3(P3) were infected with the highest titer virus supernatant,then selected by G418.Three monoclones were selected and amplified respectively.The hTERT expressions of the transducted hBMSCs were tested by RT-PCR and Western Blot.We investigated whether the transduced hBMSCs(P3) can sustain the normal cellular properties in morphology,cell phenotype and chondrogenic induction.The cells growth curve and the telomerase activity were detected by MTT and TRAP-ELISA.The tumorigenesis of transducted hBMSCs was detected by the plate clone formation assay and subcutaneous tumor formation in nude mice.Results:The expression of hTERT gene can be tested in the level of mRNA and protein in transducted hBMSCs,while weak mRNA transcription of hTERT gene can be tested in untransducted hBMSCs.In the morphology, the transducted hBMSCs were spindle-shaped or fibrocyte-like shape, which as the same as the untransducted hBMSCs.The green fluorescent protein expression can be found in the transducted hBMSCs under fluoroscope. The phenotype of transducted hBMSCs were positive in CD29、CD71、CD106 and negative in CD34、CD45.The transducted hBMSCs maintain their differentiational capability,which was proved by induced into chondrocyte.In vitro the latter can express the type II collagen protein that was cartilage specific marker.The telomerase activity of transducted hBMSCs (hTERT-hBMSCs) was positive.The growth speed of hTERT-hBMSCs was faster than that of hBMSCs by MTT.Up to now,the passage of hTERT-hBMSCs is more than 50.The rates of hTERT-hBMSCs and hBMSCs cells colony formation were 0.83% and 0.67% respectively which were lower than the rate of normal cells.In the control group,the rate of malignant melanoma cells was 57.4%,which was much higher than that of tumor cell colony formation.In nude mice experiments,malignant melanoma cells can form local massess under skin after three weeks.It was proved that the mass was composed of malignant melanoma cells.There was no tumor formation in experimental groups.Conclusions:There is no change in cell morphology,surface marker and differentiation function in transducted hBMSCs.The hTERT gene activates the telomerase and extends the life span of hBMSCs.There is no malignant transformation in hTERT-hBMSCs.These results suggest that hBMSCs transduced with hTERT gene can be considered as a source of seed cells in tissue engineering because their prolonged life span and normal cellular properties in a long-term culture.

Association between P-selectin Gene Polymorphisms and Soluble P-selectin Levels and Their Relation to Thromboembolic Events in Patients with Nonvalvular Atrial Fibrillation

Thu, 31 Mar 2011 13:23:27 +0000

BackgroundAssociation between P-selectin gene polymorphisms and soluble P-selectin levels and their relation to thromboembolic events in patients with nonvalvular atrial fibrillation.P-selectin[also known as granule membrane protein-140(GMP-140),platelet activation-dependent granule-external membrane protein(PADGEM) and CD62]is a member of the selectin family of cell adhesion molecules expressed at the surface of activated endothelial cells and platelets,that mediates the interaction of activated endothelial cells or platelets with leukocytes.The protein is localized in the alpha granules of platelets and the Weibel-Palade bodies of endothelial cells,with a soluble form present in the plasma.Levels of plasma P-selectin(soluble P-selectin, sP-selectin) are mainly platelet derived and can be regarded a marker for platelet activatio.In humans,plasma P-selectin levels have therefore been used to substantiate active participation of platelets in thrombotic disease。The P-selectin gene is situated on chromosome 1q21-q24,spans＞50kb and contains 17 exons.In the ECTIM study,thirteen polymorphisms were identified:five in in the distal P-selectin promoter and eight in the exonic sequences.Some polymorphisms of the P-selectin gene have been reported that could affect the peptide sequence of the protein and its regulatory sequences.It is report that the P-selectin gene was identified as being significantly associated with levels of plasma soluble P-selectin(sP-selectin) and thromboembolic events.Several polymorphisms of P-selectin gene has previously been reported including three missense polymorphisms:2123 C/G and 1817 T/C in the distal P-selectin promoter within putative binding sites for a number of transcription factors including AP-1 and c-Fos13,Thr715Pro in exon 13 and plasma levels of P-selectin are especially determined by the P-selectin Thr715Pro polymorphism and high concentrations of soluble P-selectin are associated with risk of venous thromboembolism and the P-selectin Thr715Pro variant.These findings suggest the polymorphisms of P-selectin gene to be involved in the development of thromboembolic events by mediated the plasma soluble P-selectin levels.It is well known that atrial fibrillation is associated with high incidence of thromboembolic events,propably due to a prothrombotic or hypercoagulable state.Some genetic alterations of haemostatic markers in haemostatic risk factors may contribute to the increased risk of thromboembolism in nonvalvular atrial fibrillation patients.Recent studies indicate,the a-fibrinogen Thr312Ala polymorphism contributes to the pathogenesis of thromboembolic complications associated with the presence of atrial fibrillation.The contribution of genetic alterations in factors of the coagulation and fibrinolytic pathways(that is hemostatic risk factors) to the development of hypercoagulation state in atrial fibrillation requires clarification.At present,these is no report that SNPs in P-selectin is associated with thromboembolic events in patients with nonvalvular atrial fibrillation. ObjectivesTo investigate the association between the polymorphism of P-selectin gene in the position of 2123C/G,1817T/C,Thr715Pro and sP-selectin levels and their relation to thromboembolic events in patients with nonvalvular atrial fibrillation(AF) in Han Nationality of Southern China,we used polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) to determine the polymorphism of P-selectin gene in the position of 2123C/G,1817T/C,Thr715Pro.Methods:1.Clinic index of body mass index,blood pressure,plasma lipids,fibrinogen, C-reactive protein were measured in 95 nonvalvular AF patients with thromboembolic events and 106 nonvalvular AF controls without thromboembolic events,who were documented ischemic stroke by MRI and/or CT or venous thromboembolism(VTE) by colour doppler ultrasound.The presence of nonvalvular atrial fibrillation was established on clinical examination at the time of ischemic stroke or venous thromboembolism(VTE) and confirmed on a diagnostic 12-lead ECG and echocardiography.2.Samples were collected when the patients were admitted to hospital.The concentrations of the soluble P-selectin(sP-selectin) was quantified by ELISA3.Genomic DNA in white blood cell from venous blood was abstracted by reagent box.By polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),we analyzed P-selectin gene polymorphism in patients and controls and compared the distribution of genotype and allele in two group.Based on this,we compared the other result and clinic index including prevalence rate of hypertention and diabetes mellitus,BMI,level of serum cholesterol,triglyceride,high density lipoprotein cholesterol,lowdensity lipoprotein cholesterol,fibrinogenn and C-reactive protein. Results:1.Total cholesterol and C-reactive protein in patient group were significantly higher than those of controls with P value of 0.000 and 0.009 respectively while other clinic features such as age,the component of female,history of cigarette,age,BMI, the morbidity of hypertention and diabetes mellitus,triglyceride,high density lipoprotein cholesterol,low density lipoprotein cholesterol and fibrinogen show no difference between nonvalvular AF patients with thromboembolic events and nonvalvular AF controls without thromboembolic events(P＞0.05).2.The presence of G allele in 2123C/G locus was found to be a greater risk factor in nonvalvular AF subjects with thromboembolic events than in nonvalvular AF controls without thromboembolic events.The odds ratio(OR) of GG were 3.500 (1.401～8.744),when compared with CC genotype,(P＜0.05).however 1817T/C allele show no significant difference(P＞0.05).3.There were statistical differences in levels of sP-selectin between patients and controls(P=0.000).The concentration of the soluble P-selectin is significantly different between patients and controls.4.Soluble P-selectin levels have no correlation with serum cholesterol and C-reactive protein with P value of 0.095and 0.069 respectively.5.The 2123C/G polymorphisms were associated with plasma sP-selectin level and patients with GG genotype tended to present higher level of plasma sP-selectin level than those carrying GC or CC genotype(P＜0.05).Genotype and allele distributions significantly differed in 2123C/G locus with P value of 0.020 and 0.015 respectively between patients and controls.6.Soluble P-selectin levels were not related to 1817T/C allele(P＞0.05) and no significant difference was found in 1817T/C locus genotype and allele distribution frequency with P value of 0.443 and 0.207 respectively between patients and controls. 7.The Thr715Pro polymorphism was not found in Han Nationality of Southern China in this study.Conclusions1.The 2123C/G polymorphism of P-selectin gene was associated with thromboembolic events in patients with nonvalvular atrial fibrillation,the genotype of GG may be a genetic risk factor for the thromboembolic process in patients with nonvalvular AF.2.The 2123C/G polymorphism of P-selectin gene was associated with soluble P-selectin levels or thromboembolic events in patients with nonvalvular atrial fibrillation,however 1817T/C locus polymorphism was not associated.3.Thr715Pro locus SNP does not exist in Han Nationality of Southern China.4.This study revealed a relationship between P-selectin gene polymorphisms and serum sP-selectin levels or thromboembolic events.The P-selectin gene affect the development of the thromboembolic process in patients with nonvalvular AF by mediating the concentration and function of soluble P-selectin.

An Experimental Study of the Safe Controllability of 9L-rIL12-TK Rat Glioma Immune Gene Vaccine

Thu, 31 Mar 2011 13:23:24 +0000

Background and ObjectiveGlioblastoma(GBM) is one of the most frequent primary tumors in nervous system(NS),covering 40～60%of the intracranial tumors.Though the technology of the neurosurgery,diagnostic iamgingand,radiotherapy and chems have made great progress,the severity of the malignancy and the matastasis rate are very high,the tolerance to the radioactive ray of the brain and the resorption to the medicine of the blood-brain barrier are very bad.They all result in the unsatisfied clinic therapy.The cure rate of the GBM is very low and easy to recur,the average survival time is only 12～14 months.So it\'s necessary to study a safe and effective adjunctive therapy. With the further study of the immuno-character of the GBM,immunotherapy is considered to be the most promising.Many glioma immunothery approaches have been proposed,including administering nonspecific immune adjuv-ants(cytokines), antitumor antibodies(serotherapy),antitumor lymphocytes(adoptive immunotherapy),and antitumor vaccines(active immunotherapy).Among them,the therapeutic tumor vaccine is a hot field of research for glioma immunotherapy.We transduced the interleukin-12(rIL-12) plus herpes simplex viral thymidine kinase(HSV-TK) gene into glioma cell(9L),and got a 9L-rIL12-TK cell controllable vaccine.The live vaccine cell has no carcinogenicity but able to effectively stimulus cytokine in vivo.In the study,we choose and purify the cell with rIL-12 and HSV-TK gene including 9L-rIL12-TK cell、9L-TK cell.In vitro,we observe the stabilize of the cell proliferate,meanwhile,we will check the effect of the HSV-TK/GCV system.In vivo,the vaccine cell will be inoculated in subcutaneous of the nude mice which has no cellular immunity.We will observe the tumor of the vaccine cell and then give intervention of GCV,compare the change of tumor volume.In vitro and in vivo studies,we proved that HSV-TK/GCV system as a "death swtich" had the complete ability to contral the proliferation of vaccinal tumor cells.A promising hope is worthy to expectation.Materials and Methods1.Experiment materials9L-rIL12-TK cell、9L-TK cell and 9Lcell,DMEM medium,FCS, 0.2%(m/V)EDTA+pancreatin,GCV,MTT,DMSO,BSD,G418,25、75cm~2culture flask,24、96shadow mask,BABL/C nude mice(SPF),et al.2.Experimental methods2.1 Experiment of cells in vitro2.1.1 To detect the cells 9L-rIL12-TK、9L-TK and 9L proliferation in 24 plate.2.1.2 Lethal effect of HSV-TK/GCV system of the live tumor cell vaccine by MTT assay in vitro;2.2 To observe the safe controllability of the 9L-rIL12-TK cellular vaccine in vivo.BALB/C nude mice were divided randomly into 9L+saline group and 9L+GCV group,9L-TK+saline group and 9L-rIL12-TK+GCV group,9L-TK+saline group and 9L-rIL12-TK+GCV group,the cells were inoculated subcutaneously into the right axil of nude mice,For GCV groups,GCV was administered a day intraperitoneally for 7 day after s.c.tumor injection,control groups were given with saline.Tumor growth were measured once every 3 day(twice a week),and record the life time of the mice,tumors and organs were taken to analyze pathology.Results1.1 To detect the cell proliferation and lethal effect of HSV-TK/GCV system of the live tumor cell vaccine by MTT assay in vitro;9L-rIL12-Tk cells and 9L-TK cells were found to be stable,between cells and times there were significant diference (F=4.220,P=0.003),among the cells there were significant diference (F=8.761,P=0.017),among times there were significant diference (F=2316.347,P=0.000),4th day among cells there were significant diference (P=0.005),other times among cells there weren\'t significant different (P＞0.05).Meanwhile,in different density of GCV DMED the cells were culture,detect the living cells by MTT assay,9L-rIL12-Tk cells and 9L-TK cells were found to be more sensitive to the cytotoxic effect of GCV than 9L,between different density of GCV and the cells there were significant diference(F=407.699,P=0.000),among the different density of GCV the three cells there were significant diference(F=2830.488,P=0.000),among the three cells there were significant diference(F=6807.262,P=0.000),in 20～160ug/ml GCV among the three cells there were significant diference(P=0.000),but between 9L-rIL12-TK cell and 9L-TK cell there weren\'t significant diference(P＞0.05),in 10ug/ml GCV 9L-rIL12-TK cell and 9L-TK cell were half killed,when in 20～160ug/ml CGV 9L-rIL12-TK cell and 9L-TK cell were almost killed.We mainly observe the cell proliferation and the sensitivity Of GCV,to make sure of the cell growth cycle and FD50 of GCV for the next animal experiment.1.2 To observe the safe controllability of the 9L-rIL12-TK cellular vaccine in vivo.BALB/C nude mice were divided randomly into 9L+saline group and 9L+GCV group,9L-TK+saline group and 9L-rIL12-TK+GCV group,9L-TK+saline group and 9L-rIL12-TK+GCV group,the cells were inoculated subcutaneously into the right axil of nude mice,For GCV groups,GCV was administered a day intraperitoneally for 7 day after s.c.tumor injection,control groups were given with saline.Tumor growth were measured once every 3 day(twice a week),and record the life time of the mice,tumors and organs were taken to analyze pathology.We choosed four time-volumes of the tumor,including the before and after of intervention and the other two times,between different times and the tumor-volumes of different groups there were significant diference(F=47.004,P=0.000),among the different times the tumor-volumes of the six group cells there were significant diference (F=316.600,P=0.000),the tumor-volumes of different groups were significant diference(F=60.442,P=0.000),the 9th day the tumor-volumes of the different groups weren\'t significant diference(P=0.476),in different times 9L-rIL12-TK and 9L-TK cell-tumors weren\'t significant diference(P＞0.05),between the experiment groups and the control groups,both of the 9L-rIL12-TK and 9L/TK cell-tumors were significant diference(P=0.000),the tumor-volumes were obviously inhibited,relapse rates were 0 and 62.5%.9L-rIL12-TK and 9L-TK cell-tumors were obviously inhibted by GCV(p=0.000),and their long-term survivals were prolonged(LogRankX~2=15.782, LogRankX~2=15.665p=0.000),between 9L experiment and control groups their long-term survivals weren\'t prolonged(LogRankX~2=0.392,P=0.531).To observe the tumor,the heteromorphism of cells were manifest,in the tumors without rIL-12 there were more vessels,opposite,the spleens of nude mice were larger,there were not to transfer.Conclusions(1) The proliferation of the 9L-rIL12-TK cells modified by the genes was stable in vitro.The cells were sensitive to the lethal effect of GCV,which indicated the stable expression of the TK gene in vaccine cells. (2) 9L-rIL12-TK cells caused tumor in the nude mice,while could be inhibited and eliminated by GCV.(3) 9L-rIL12-TK cells and 9L-TK cells both caused tumor in the nude mice,while the tumor inhibition effects of GCV was obviously better in 9L-rIL 12-TK group than 9L-TK group.It may be a sign that rIL-12 played a role in the immunoregulation and elevated the effects of the tumor inhibition.(4) The spleens of the nude mice were notably bigger,meanwhile the lymph cells were increased,which indicated rIL-12 played a role in the humoral immunity.(5) 9L-rIL12-TK cells just caused tumor in some parts of the nude mice,and no signs indicated the tumor metastasis.All the conclusions above prove that 9L-rIL12-TK cello-vaccine is better proliferated and controlled.The effects of its immunotherapy need to be confirmed by the further researches.

The Study on Real-time Osborn Wave Detection Method for Holter Monitoring System

Thu, 31 Mar 2011 13:23:21 +0000

The Holter monitoring system can do a long-time continuous and real-time detection to the electrocardiosignal, which reflects the working states of the heart, and then make a routine analysis. But the system has not processed the emergency warning function to the acute and high-risk heart disease yet. Osborn wave is the abnormal formation of the J-point in the electrocardiosignal, which indicates the dangerous situation of some patients with heart diseases, such as sudden death. Now the Osborn wave diagnosis mainly depends on the doctors’subjective judgements. So it needs a further study on its real-time detection. According to the development of medical equipment and clinical medicine, this thesis studies the real-time detection methods of Osborn wave in the Holter monitoring system based on the principles of Holter monitoring system, the signature analysis of electrocardiosignal and the basic characteristics of acute and high-risk electrocardiosignal. Meanwhile, it also elaborates the results and the validity of the algorithms by parsing the contents of this subject study.In terms of the hardware, firstly, the signal acquisition, amplification and voltage conversion circuits are designed to achieve the functions of signal acquisition and processing by analyzing the characteristics of the electrocardiosignal like the amplitude and frequency range. Then, the interference suppression circuits are designed by analyzing the causes and characteristics of the undesired signals. Finally, the digital circuits using MSP430F1611 to be the central processor unit are designed to make the system operating under control.In terms of the software and algorithms, the system design is accomplished by using the development platform based on IAR Embedded Workbench for MSP430, programming the microprocessor in C language to achieve the functions of A/D sampling, digital filtering, R wave and S wave detecting, Osborn wave recognizing, the detection results indicating and the information transporting.The effects of the algorithms are tested by using Matlab, which simulates the algorithms to process the electrocardiosignal data in MIT-BIH database and the sample data of Osborn wave from related records. The tests show that it is effective to identify the characteristics of the R wave, S wave by adopting these algorithms and the check-out results are significant, which also can recognize Osborn wave effectively. The subject study is consisted of researches on the signal acquisition and conditioning circuits, microprocessor and its accessory circuits, the microprocessor procedure, the algorithm procedure for processing electrocardiosignal and the detection algorithms for R wave, S wave and Osborn wave. It also proves the effectiveness of the algorithms by analyzing the data in standard ECG database. Through clinical investigations, system development and experimental analysis, the system prototype can accomplish real-time electrocardiosignal acquisition and analysis in clinical trials. The real-time detection of Osborn wave can warn the doctor and nurses to do first aid or strengthen nursing in case of the serious consequences. This subject has higher scientific value and clinical significance, and is worth doing a further study in the future.

Study on Differences of Neutral Oligosaccharides in Breast Milk between Chinese and American Mothers and Gene Polymorphism of FUT2

Thu, 31 Mar 2011 13:23:19 +0000

PartⅠStudy on differences of neutral oligosaccharides in breast milk between Chinese and American mothersObjectiveTo investigate the concentration of oligosaccharides in Chinese mothers\' breast milk and its pattern of change at different time points;analyze the proportion of different oligosaccharides to total oligosaccharides concentration;to find out the differences of oligosaccharides at three time points through comparison with milk of American mothers\'.Methods51 pairs of Chinese infant and mother were recruited in this study.Breast milk was collected at 2,4,13 week postpartum and saliva sample at 2 week.American mothers were recruited according to the same standards.Neutral oligosaccharides concentration of breast milk was analyzed by High-performance liquid chromatography. Results1、There were not significant changes in the total concentration of neutral oligosaccharides in Chinese maternal breast milk postnatal 13 weeks,but the concentration of different types of neutral oligosaccharide varies at each visiting interval.The concentration of 3-FL and 2\'-FL increased from week 2 to week 4 (84.6-485.8μg/ml;579.6-787.8μg/ml;P＜0.05),but LNF-1 decreased from 596.9mg/ml to 304.5mg/ml2、According to the classification of the neutral oligosaccharides,29 mothers are Le(a-,b+),and 10 of them are Le(a-,b-) and 12 of them are Le(a+,b-)3、There were significant changes in the total concentration of neutral oligosaccharides in American maternal breast milk postnatal 13 weeks.LNF-1 decreased in the first 13 weeks(P＜0.05).The concentration of 3-FL increased(P＜0.05). The concentration of 2\'-FL,LDFH-Ⅰ,LNF-2,LNF-4 and LDFH-Ⅱwas stable.4、There were significant differences between Chinese and American maternal breast milk in total oligosaccharides concentration at 2,4,13 weeks.5、There was significant difference in concentration ofα1,2 linked oligosaccharides between Chinese and American maternal breast milk at 2 weeks.But there were no significant differences at 4 and 13 weeks.6、The concentration ofα1,3 linked oligosaccharides in American maternal breast milk was significantly higher than that in Chinese maternal breast milk at 2,4 and 13 weeks.Conclusion:1、The variation of neutral oligosaccharide in Chinese mothers\' breast milk is unique.The expression ofα1,2 linked oligosaccharides decreased;the expression of α1,2 linked oligosaccharides increased gradually in the first 13 weeks postpartum.2、The distribution of Lewis/Sec genotype matches with the distribution of general people group in China3、The concentration of neutral oligosaccharides in American maternal breast milk was significantly higher than that in Chinese maternal breast milk;the concentration ofα1,3 or 1,4 linked oligosaccharides was much higher in American maternal breast milk than that in Chinese maternal breast milk;there were no significant difference in the concentration ofα1,2 linked oligosaccharidesPartⅡGene polymorphism of FUT2 and its relationship with oligosaccharides concentration in American and Chinese mothersObjectiveIn order to explore the relationship between oligosaccharides differences and gene polymorphism,SNP spot on gene FUT2 was assayed and oligosaccharides concentration was analyzed based on SNP genotype.MethodSaliva were collected using Oragene KIT.DNA was abstracted from saliva and used for SNP assay.Result1、It can be A or G on SNP(rs601338).There are three genotypes as AA,AG, and GG.12 of American mothers are AA;24 of them are AG;15 of them are GG. 2、There are moreα1,2 linked oligosaccharides in genotype GG maternal breast milk than in genotype AA;there are moreα1,3 linked oligosaccharides in genotype AA maternal breast milk;the concentration ofα1,2 linked oligosaccharides andα1,3 linked oligosaccharides in genotype AG maternal breast milk lay between AA and GG.3、All of the 50 Chinese mothers are GG type.There were no obvious relationship between SNP genotype and neutral oligosaccharides concentration in this group.Conclusion1、There exists gene polymorphism of FUT2 SNP2、There was a clear relationship between SNP genotype and oligosaccharide concentration in American group.3、There was no definite relationship between SNP genotype and oligosaccharide concentration in Chinese group.

Research on Eukaryotic Expression and Reverse Genetics of Paramyxovirus Tianjin Strain

Thu, 31 Mar 2011 13:23:17 +0000

Paramyxovirus Tianjin strain is a new genotype of Sendai virus.Sendai virus causes outbreaks of high lethal pneumonia in mouse colonies.In 1999,the acute respiratory infectious disease outbroke among the common cotton-eared marmosets colonies in animal experiment center of Tianjin Medical University and most of marmosets died in 5 to 10 days even if promptly treated with kinds of antibiotics.We could see lung tissue\'s focal necrosis in autopsy analysis.However,the experimental mice in the same animal laboratory had not been suffering from respiratory disease during the epidemic period of respiratory infectious disease.Paramyxovirus Tianjin strain had been detected the in the bronchoalveolar lavage fluid(BALF) samples from the newborns and young children with severe lower repiratory tract infections by Sandwich ELISA assay.The positive rate was about 1.92%(2/104).All of these suggest that paramyxovirus Tianjin strain possess the unique characteristics in pathogenicity and host tropism,and it maybe the important pathogen of young children.More and more people pay attention to emerging pathogens,particularly Viruses coming from animals which cause emerging infectious diseases of human beings.It is important to make clear the mechanism of high pathogenicity and interspecies transmission of virus for prevention.Under natural condition,the interspecies transmission of paramyxovirus is limited by its host specificity.Although there are many factors influencing host specificity,the HN protein is important for recognizing and absorbing cell receptor.Therefore,we focus on paramyxovirus Tianjin strain by means of eukaryotic expression system to provide the basis for disclosing the mechanism of high pathogenicity and interspecies transmission of paramyxovirus Tianjin strain.We also do preliminary study on paramyxovirus Tianjin strain in reverse genetics to understand the interaction between Tianjin strain and host cells.In the experiment,we expressed nucleoprotein(NP) and hemagglutinin-neuraminidase (HN) of paramyxovirus Tianjin strain in eukaryotic cells,analyze the antigenicity and hemagglutination activity of HN and three truncated fragments HN1, HN2,HN3. First,HN gene and its segments HN1,HN2 and HN3 of paramyxovirus Tianjin strain was constructed into the eukaryotic expression vector pEGFP-C2-HN,pEGFP-C2 -HN1,pEGFP-C2-HN2,pEGFP-C2-HN3 and transfected into HEK293 cells, which is human embryonic kidney cells.The expressed proteins are identified by Western blot assay,in situ ELISA and hemadsorption assay.The results present the cells which were transfected by pEGFP-C2-HN,pEGFP-C2-HN1,pEGFP-C2-HN2 and pEGFP-C2-HN3 showed the green fluorescence.The expression of HN,HN1, HN2 and HN3 can be detected by Western blot assay.HN,HN1,HN2 and HN3 showed the diverse antigenicity in transfected HEK293 cells.The antigenicity ranks as HN＞HN2＞HN1＞HN3(precedence order).We also observed that HN and HN3 proteins could effectively adsorb the fresh chicken red blood cells,whereas HN1 and HN2 proteins were negative in the hemadsorption test.Second,we do further research of paramyxovirus Tianjin strain by reverse genetics. Paramyxovirus Tianjin strain was propagated in chick embryo allantoic liquid.We extracted RNA of paramyxovirus Tianjin strain and amplified NP gene and P gene by RT-PCR,then constructed the eukaryotic expression vectors which were named pcDNA3.1(+)-NP、pcDNA3.1(+)-P.We also amplified L gene by fusion PCR method. The transient and stable expression of NP gene in HEK293 cells which were transfected with the constructed vector could be detected by immunofluoresence assay and western blotting.These tests lay the base for further studies on NP protein function and high pathogenicity,as well as unique host tropism of paramyxovirus Tianjin strain by reverse genetics.

Study on Imaging Performance and Clinical Application of Femoroacetabular Impingement Syndrome

Thu, 31 Mar 2011 12:31:31 +0000

【Purpose】Femoroacetabular impingement has been recognized as an underlying cause of hip pain and secondary osteoarthritis.Several surgical options are available for treatment, and are guided by the severity and location of the disease,as well as the expertise and technical preferences of the surgeon.Accurate diagnosis of the impingement disease pattern and precise surgical technique are the basis of successful surgical care.On basis studying in abroad same kind of study,the purpose of our study is to probe the standard of early diagnosis of FAI,and the imageology appearance of radiography,spiral CT scanning and three-dimensional reconstruction,and MRI of progression of FAI patients.To study manifold imaging factors,imaging signs and clinical characteristic of femoroacetabular impingement,for improving the clinical cognition to this syndrome,and making a diagnose early and clear,and making a exact therapy.【Materials and mathods】1.Clinical informationFrom January 2007 to January 2009,75 hips being elected out according to FAI imaging definition,44 males,31 females among them,11 patients morbidity for pair of hips,randomly selected either side hip joint to study.age range 18～69 years,mean 37.5 years.All patients with complaints of the hip,groin or buttock pain,their duration was 6 months～30 years.To choose 50 volunteers who there are no the hip symptom carrying out contrast studies for observing the various imaging factors,age range 20～45 years,mean 31.9 years.26 males,24 females among them2.Examination methodsThe objects makes the examination,75 in X-ray,39 in CT and 17 in MRI.X-ray examine use DR imaging system,the routine pair of hips anteroposterior radiograph with the same method and condition.CT exmaine use GE Light speed 16 CT scanner with receive-only surface coil in axial scanning,coronal and oblique-axial reconstruction.MR examine use GE Signa VH/i 3.0T superconducting MRI scanner,MRI imaging at the position of coronal and axial plane apply FSE-T1WI、STIR-T2WI and FSE-T2WI-FS sequence.3.Imaging factors and data analyseThe clinical analyse of FAI hips included gender and age.The factors of X-ray,CT and MRI are measured and compared between FAI group and control group,the abnormal imaging findings of X-ray,CT and MRI are classified cohort studied.4.Imaging analyses methodsAll images were saved and sended to PACS.Three radiologists measured all the data of X-ray and CT image on PACS,and computed the average.2 radiologists examined the abnormal imaging findings,the final resul were acquired by the 2 radiologists.5.Statistics analysisStatistics analysis was performed with SPSS13.0 software.Contrast study of FAI group and control group,and three FAI types.Chi-square test in 2×2 table,Chi-square test in R×C table,One-way ANOVE,Binary logistic regression and Independent sanples t-test are used in study.Probability values＜0.05 were considered statietically signficant.to make sure of value of factors by contrasting abnormal hip morphology and feature.【Results】All FAI patients have carried out the short-term follow-up,exclude other disease possibility,such as abnormal imagings of nontraumatic hip demonstrate avascular necrosis,ankylosing spondylitis,inflammation,tuberculosis and traumatic osteoarthritis FAI is the category belonging to traumatic osteoarthritis,it is different from traumatic caused by acetabular dysplasia and injury,its theory basis and type,imaging findings shows obvious difference.1.X-ray examination factors:75 hips in FAI group and 50 hipsin control group are contrasted in Sharp angle, center-edge angle CCD angle,acetabular-head index,acetebular deph-to-width index, HTE angle and femoral head-neck offset.One-way ANOVE was used in the contrast study of these 7 factors of the FAI group and the control group.It shows four factors significant difference in FAI and control groups(P＜0.05).They are Sharp angle,HTE angle,acetebular deph-to-width index and femoral head-neck offset.Binary logistic regression and Independent Sample T-test was used in contrast study of three FAI types and the control group.75 hips are united into three groups according to abnormal femoral head or head-neck junction feature,named cam group,pincer group and mixed group.It shows five factors significant difference in FAI and control groups(P＜0.05).They are CE angle,Sharp angle,HTE angle,acetebular deph-to-width index and femoral head-neck offset.X-ray imaging features are ananlysed,and show 21 femoral heads with osteophyte or abnormal bony apophysis(the positive rate is 28.00%),27 reduced femoral head-neck offset(the positive rate is 36.00%),22 acetabular hypsokinesis(the positive rate is 29.33%),and 24 acetabular proliferations and sclerosis(the positive rate is 29.33%).Chi-square test in R×C table was uesd in the contrast study of these X-ray imaging features between three FAI types.It shows three festures significant difference.They are osteophyte or bony process,reduced femoral head-neck offse,acetabular hypsokinesis.2.CT examination factors:39 hips in FAI group and 20 hips in control group are contrasted in a angle.Independent Samples Test shows significant difference forαangle in FAI and control groups(P＜0.05).One-way ANOVE was used in the contrast study of three FAI types and the control group.There was statistical signficance inαangle between the C group and the P group.CT imaging features are ananlysed,and show 16 femoral heads with osteophyte or bony process(the positive rate is 41.02%),14 nonsmooth articular surfaces(the positive rate is 35.90%),15 acetabular proliferations and sclerosis(the positive rate is 38.46%)and 4 herniation pits(the positive rate is 10.26%). Chi-square test in R×C table was uesd in the contrast study of these CT imaging features between three FAI types.It shows osteophyte or bony process significant difference only.3.MR examination factors:17 hips in FAI group are analysedand summarized,it shows 15 hips with joint effusion(the positive rate is 88.24%),10 abnormal acetabular rims or labrums(the positive rate is 58.82%),7 femoral heads with abnormalsignal(the positive rate is 41.18%),8 acetabulums with abnormal signal(the positive rate is 47.06%),and 8 hips withabnormal articular cartilage(the positive rate is 47.06%).MRI detailed revelation to bone and cartilage can provide the definite evidence fordeveloped characteristic appearance of FAI lesion,some FAI hips have features only in MRI.4.Clinical examination factors:In 75 patients of FAI group,44 were men,31 were women.In 50 patients of control,26 were men,24 were women.Chi-square test in 2×2 table was uesd in the contrast study of sex between the FAI group and the control group.It shows that there wasn\'t statistcal significance(P＞0.05) in the differerca betwwen the FAI group and the control group.Chi-square test in R×C table was uesd in the contrast study of sex between the three FAI types.It shows that there wasn\'t statistcal significance (P＞0.05) in the differerca betwwen the three FAI types.The freauencies histogram and normal curve of the FAI group patients\' age shows the age was centered largely in the 25～45,73.33%of the total.[Conclusion]To date,imaging features of this condition have not been widely discussed in the radiolo gic literature.The radiographic findings of FAI are not widely discussed and recognized by physicians.This study have the summary and details in imaging part of FAI,and use part data to have verified the imaging haracteristics,on basis studying in abroad same kind study.there is obvious meaning for FAI diagnoses that osteop hyte or abnormal bony apophysis,reduced femoral head-neck offset,acetabular hypsokinesis,and acetabular proliferations and sclerosis in the X-ray imaging.C T scan examination and reconstruction of multi-directional display can be bette r to show the FAI characters.Theαangle in oblique-axial reconstruction is m ostly characteristic.Magnetic resonance(MR) imaging and can be associated w ith progressive damage to the adjacent cartilage,and has a higher detection sens itivity and specificity of the acetabular labrum and cartilage injury.MRI featur es of acetabular rims or labrums.The diagnosis of femoroacetabular impingeme nt(FAI) is based on the patient\'s clinical history and physical examination and is further supported by findings at radiography,computed tomography,and MR imaging.

A Study of 56 Cases of Torsion of Umbilical Cord Abstract

Thu, 31 Mar 2011 12:31:28 +0000

Objective:To investigate the prenatal diagnosis, abnormal clinical manifestation, relative management, and the relationship between the method of delivery、the fetus status and torsion of cord.Methods:56 pregnant women with torsion of cord cases reported from June 2007 to February 2009 were analyzed retrospectively, compared with 60 cases pregnant women without torsion of cord at the same time. To analyzed the diagnostic value of prenatal examinations in detecting torsion of cord, include the abnormal fetal heart, the abnormal fetal movement, the Fetal heart rate( FHR) monitoring, the color Doppler ultrasonographic assessment. The method of delivery and the effect to the fetus prognosis was evaluated. And two group were no different in symptoms of complications.Results:1.The incidence of torsion of cord in this research was 7.865%.2.About 96.4% torsion of cord cases can be projected by com- bining prenatal color Doppler ultrasonographic assessment with the FHR monitoring. 3.None of the umbilical blood S/D in the two groups was reduced. 7 cases were ascensused in the study group which occured in approximately 12.5% of all cases in the study group. The umbilical blood S/D in study group was significantly higher than those in normal group.4. The incidence of fetal distress, neonatal asphyxia,feta growth restriction in study group were significantly higher than those in normal group .5.The torsion of cord also led to the decrease of natural labor and the increase of cesarean sreduced.6.The abnormal clinical manifestation cases in the group with torsion of cord were different in the length of the two consecutive umbilical coiling. The length was 2.170±0.108 cm.Conclusion: The application of prenatal color Doppler ultrasonographic assessment, the fetal heart monitoring, the umbilical blood S/D, et al, we could find fetus in anoxia status earlier, and give evidence to the time and method of delivery.

Theoretic Study on Deficiency of Qi and Blood Stasis

Thu, 31 Mar 2011 12:31:27 +0000

From the "Neijing" creating the theory of Qi and blood, Yang Shiying\'s the "Qi is the commander of blood, blood is the mother of Qi " theory, Wang Qingren\'s "Yilingaicuo" about Qi and blood theory, Tang Zonghai "Xuezhenglun" of Qi disease and blood said, Zhang xichun\'s "deficiency of old leading blood stasis" theory, until the modern clinical research on "deficiency of Qi and blood stasis". Many doctors, scholars circumfuse "Qi and blood" theory to start development and research of reason, law, prescription, drug.In this study, through the collation and analysis of the literature, taking a combination of research methods, combing through ancient literature about the formation of Qi deficiency and blood stasis theory in history, the Chinese medicine theory of Qi deficiency and blood stasis began in the Pre-germination, the clinical application started in the Han and Tang Dynasties period, developed in the Song and Yuan Dynasties, populared in the Ming and Qing Dynasties. the modern study of Qi deficiency and blood stasis theory focused on evidence of clinical disease research, experimental research, as well as the biological basis of research studies.By studying the history books on the theory of Qi deficiency and blood stasis, retrieving the medical history of Qi deficiency and blood stasis history arguments, sentences, analysis, summarizing and organizing, clarifing the meaning of the theory of Qi deficiency and blood stasis. Qi deficiency and blood stasis, this is the disease, disease symptoms and a general permit. Qi is the commander of blood, blood is the mother of Qi. Qi-deficiency can be caused by blood stasis, blood stasis forms for a long time, It can influenced Qi\'s function, This clarities the Qi-deficiency and blood stasis syndrome logical relations. Original Qi, pectoral Qi, nutritive Qi and defensive Qi, visceral and meridian Qi, these is the Qi of human life. If virous Qi is vigorous, heart Qi\'s opening and closing, all vessels converging in the lung, spleen Qi controling blood circulation, liver Qi regulating the free flow of Qi, kidney Qi nourishing and warming. Qi\'s function contacts to blood stasis: from insecurity of defensive Qi and losing astringent therapy. from blood deficiency bringing blood stasis, from Yang-Qi losing warming function and cold accumulationg and blood stasis.Deficiency of Qi and blood stasis syndrome includes many manifests that Qi deficienc-syndromes hold aomasthenia, hypodynamia, asthenia qi, undertone, feeble breathing, blood stasis syndrome symptoms and objective signs, holding ache, hematocyanosis of bump、lip、claw、lingua. Deficiency of Qi and blood stasis\'s therapeutical principle finally needs replenishing Qi to invigorate blood, treating symptom and root cause simultaneously.Rearching on stroke, coronary heart disease, hypertension, tumor disease, diabetes, clinical research, as well as the experimental animal models, the biological basis of the progress of the study summarized, Qi deficiency and blood stasis in patients often accomparies high blood viscosity, high pour point, high-fiber protein, thromboxane high, high vascular reactivity and thromboembolic pathological features, confirmed the pathological nature of Qi deficiency and blood stasis, affirmed the existence of Qi deficiency and blood stasis and proved it in a wide range of clinical diseases.Experiencing in previous studies on the basis of modern health research experience and viewing of the origin of Qi deficiency and blood stasis, Mechanism, treatment, this study tries to provide an useful theoretical and clinical guidance to Qi deficiency and blood stasis syndrome treatment.

Alteration of Expression and Activity of Glutathione S-transferases π Associates with Drug-resistance in Human Lung Cancer Cell Lines

Thu, 31 Mar 2011 12:31:25 +0000

Objective: To investigate the relationship between the expression and activity of glutathione S-transferasesπ(GST-π) treated with ethacrynic acid (EA ) and resistance to anti-cancer drug cisplatin, adriamycin and VP-16 in vitro in lung adenocarcinoma SPCA-1, lung large cell carcinoma NCI-H- 460 and small cell lung caner NCI-H-446 cell lines.Methods: Immunofluorescence was used to detect the expression of GST-πof three cell lines treated by EA(concentrations were 60、80、100、160μmol/L respectively, for 24h)and the control cells. GST-πenzymatic activity was detected by the ultraviolet spectrophotometry. Cell viability was detected by the colorimetric MTT assay after being exposed to chemotherapeutic agents. Data was analyzed for statistical significance using the SPSS 11.5 software.Results: 1. The expression of GST-πat protein level decreased obvio- usly in SPCA-1 and NCI-H-446 cell lines inhibited by EA with a manner of dose-dependent on EA to a great degree assayed by immunofluorescence. The fluorescence intensity for the cells treated by EA was weaker than that for the control cell obviously. Whereas there was no difference between the EA-treated group and the control group in NCI-H-460 cell line. 2. Inhibiton ratios of GST-πactivity by treated EA in SPCA1, NCI-H-460 and NCIH446 cell lines were 23.96%, 30.80% and 24.12% respectively, showing no significant difference among the three types of cell lines. 3. The cell viability detected by MTT for the cells being treated by cisplatin or adriamycin showed that the viability of EA-treated cells was lower than that significant (P < 0.05), while there was no significant difference of the viability for the cells of the other two types of SPCA-1 and NCI-H-460.Conclusions: EA is effective in the inhibition of GST-πenzymatic activity in the three human lung cancer cell lines, which can decrease the expression of GST-πin SPCA-1 and NCI-H-446 cell lines, and subsequent increase the sensitivity of cisplatin and adriamycin to the NCI-H-446 cell line. Based on the trend of the change of cell viability and the level of GST-πin different disposed cells, we conclude that the reduction of expression and enzymatic activity of GST-πby EA could increase the sensitivity to cisplatin or adriamycin in the NCI-H-446 cell line. All these suggest that to decrease the expression and activity of GST-πby chemicals such as EA may be a new therapeutic approach to lung cancer.

Clinical and Pathological Characteristics of IgA Nephropathy and Analysis of Risk Factors Associated with the Renal Pathologic Lesions

Thu, 31 Mar 2011 12:31:23 +0000

Objective : IgA nephropathy(IgAN) is the most common form of primary glomerulonephritis worldwide,the clinical and pathological manifestations are diverse,but also the developing disease.The purpose of this study is to study the clinical and pathological characteristics of IgA nephropathy patients and analysis the risk factors associated with the renal pathologic lesionsMethod : 51patients with IgA nephropathy were collected from the inpatients who were diagnosed as IgA nephropathy by renal biopsy.Includeing 21 cases of male,female 30 cases,the average age of 33.03±10.1 years old.Except for purpuric nephritis,lupus nephritis,glomerular sclerosis,and HBV asociated nephropathy and other secondary kidney disease.And the clinical index of different severity of renal pathologic lesions was compared;the relationship between renal pathologic lesions and clinical features were studied.According to the severity of renal pathologic lesions are divided into A group(including Lee\'s grade I,Ⅱlevel,n=29),B group(including Lee\'s gradeⅢ,Ⅳ,V-level,n=22).Lee\'s classification standards by the literature [ 1 ] .All patients were carried out immunofluorescent IgG,IgA,IgM,C3,C4,C1q staining.Analysis of its clinical and pathological characteristics,according to the severity of renal pathologic lesions,the predicted factors were analyzed by step—wise multiple variance regrcssion an analysis.Results:1.Comparison of the clinical features:A group of 29 cases,7 cases of male,female 22 cases,the average age of 33.0±10.15,an average duration was 5 months;B group 22 cases,14 cases of male,female 8 cases,the average age of 32.0±9.17,The average duration was also 5 months.Between the two groups, there is no significant difference(P=0.703,0.573)in age and duration.In A,B groups the incidence of male repectively accounted for 24.13%、64.64%; hypertension accounted for 31.0%、40.9%;renal failure accounted for 3.4%、36.4%;nephritis syndrome accounted for 27.6%、27.3%;the incidence of ne- phrotic syndrome in A,B two groups repectively were 13.8%、9.1%;the incidence of microscopic hematuria repectively were 17.2%、0%;the incidence of asymptomatic proteinuria accounted for 31.0%、27.3%;the incidence of male,renal failure in group A was significantly lower than B group(x2 values were 8.058,9.326,P value0.05).2.In A group the level of serum creatinine,serum uricacid,24h Urinary protein excretion,was significantly lower than B group(P0.05).3.The degree of renal pathologic lesions in patients with IgA nephropathy, A group of 29 cases(56.87%),showed mild renal pathologic lesions,including Lee\'s16 cases of grade I-grade(31.4%),Ⅱgrade 13 cases (25.5%);B group 22 cases(43.13%),showed moderate to severe renal pathologic lesions,including Lee\'s8 cases of gradeⅢ(15.7%),12 cases (23.5%)of gradeⅣ,2 cases (6.0%) of grade V.4.Multi-factor risk analysis showed that serum creatinine,24h urinary protein excretion were independent predictors of the severity of renal path- ologic lesion in lgA nephropathy patients(0R values repectively were 1.033, 1.001;P value of 0.038,0.021).With the increasing of serum creatinine, 24h urinary protein excretion the renal pathologic lesion are more serious.Conclusion:Based on terms of age and duration in no significant difference in IgA nephro-pathy,the incidence of male is higher More than half of pathological lesions in patients with IgA nephropathy are mild, similar to minimal change nephropathy or mild mesangial proliferative glomerulonephritis, while the other parts of renal pathologic lesions are severe. Hematuria in patients with IgA nephropathy may be a better prognosis,but microscopic hematuria may be associated with renal pathologic lesions little The high level in Urinary protein excretion,serum creatinine are the risk factors for renal pathologic lesions in IgA nephropathy High blood pressure to renal pathologic lesions in IgA nephropathy may have little.

The Activity of Telomerase of Endothelial Progenitor Cells in Patients with Coronary Heart Diseases and the Effects of HGF on the Functions of Endothelial Progenitor Cells

Thu, 31 Mar 2011 12:31:22 +0000

Objective: 1. To investigate the relation between the proliferation, migration and adhering of endothelial progenitor cells(EPCs) in peripheral blood with coronary heart diseases cultured in vitro and the activity of telomerase. 2. To observe the effects of HGF on the proliferation, migration and adhering of endothelial progenitor cells cultured in vitro,and to investigate the relation between the concentration of HGF , the numbers of CD133~+ cells in peripheral blood and the biological function of endothelial progenitor cells.Methods: l.Mononuclearcells(MNCs) were harvested from peripheral blood by density gradient centrifugation,then inoculated in culture plates coated with gelatin. After cultured for 7d, the immunohistochemical method was performed to identify EPCs. The proliferation、migration and adhering of EPCs was measured by MTT assay、transwell insert and the measured experiment of adhering ability. 2. The activity of telomerase was detected by TRAP-PCR argentation. 3. 15 patients with coronary heart disease(CHD)and 15 matched control subjects were enrolled. To observe the change of the number and function of EPCs from patients with CHD. MNCs harvested by density gradient centrifugation were inoculate in culture plate coated with Gelatin. After culture for 4d, wash-out non-adherent cells and attached cells divide into HGF group and non-HGF group, then continue to culture for 3d, to observe the effects of 1ng/μl HGF on the proliferation, migration, adhering and apoptosis of endothelial progenitor cells. 4. MNCs in periheral blood were harvested by density gradient centrifugation, and marked with FITC-Conjugated Mouse Anti-human CD133 then the number of CD133~+ cells were detected by flow cytometry; The concentration of HGF in blood plasma was determined by the method of ELISA. Results: 1. EPCs were well isolated from human peripheral blood.The proliferation (P0.05) . EPCs apoptosis was not significantly different between CHD and non-CHD (P>0.05) . HGF had not a protecting effect on EPCs apoptosis (P>0.05 ) .2. Compared with patients with non-CHD,the number of CD133~+ cells in periheral blood of patient with CHD was lower (P<0.01) and the concentration of HGF in blood plasma was higher (P<0.01) .Conclusions: 1. The proliferation,adhering and migration activities of EPCs are decreased in patients with CHD. The weakening of EPCs biological function might be correlated with the attenuation of telomerase activity. 2. HGF could improve all biological functions of EPCs in patients with CHD and improve the ability of proliferating with non-CHD.but had not this effect of adhering and migrating in patients with non-CHD . 3. EPCs apoptosis was not significantly different between CHD and non-CHD. HGF had not a protecting effect on EPCs apoptosis. 4. It was promising auxiliary diagnosis and the way of examine for CHD to detect the number of CD133~+ cells and the concentration of HGF in periheral blood.

A Study on the Traditional Chinese Medicine Syndrome of 120 Type 2 Diabetes Combined with Hypertension

Thu, 31 Mar 2011 12:31:21 +0000

Objective:1.To study the distributive features of Traditional Chinese Medicine(TCM) syndrome of Type2 Diabetes combined with Hypertension(DH) patients.2.To analyze the correlations,such as FPG、CHO、TG、LDL-C、HDL-C,between the main syndrome elements of DH patients and physical and biochemical indexes.Methods:In this study,the distributive features of Traditional Chinese Medicine syndrome of type2 diabetes combined with hypertension(DH) patients are concluded by the collection of the general and the four TCM diagnostic materials of the public coincident with adoptive standards in the outpatients at The First Hospital of Xiamen or the patients engaged in the 3~(rd) National Diabetes General Survey of Xiamen district,the following TCM identification and theory of syndrome elements and combination of syndrome elements:the correlation between these indexes and patterns are studied,based on the examination of the various physical and biochemical indexes and comparing analysis of the indexes of main syndromes group between them.Results:1.The main patterns of DH were two pattern combinations with 47 cases(accounted for 39.2%) and three pattern combinations with 33 cases(27.5%).2.The main syndrome elements in the patterns of DH were syndrome blood stasis(79 cases, accounted for 65.8%).dryness-heat(63 cases,52.5%),qi deficiency(47 cases,39.2%)and phlegm-dampness(32 cases.26.7%).3.In 120 DH cases,there were 48 types of patterns,including excess pattern 47 cases accounted for 39.2%;deficient 11 cases accounted for 9.2%and deficiency-excess complex pattern 62 cases accounted for 51.7%. 4.Among the patterns in 120 DH cases,the most syndromes are dryness-heat(15 cases, accounted for 12.5%),the others are phlegm-dampness combined with blood stasis and dryness-heat(9 cases,7.5%),blood stasis(8 cases,6.7%),qi deficiency combined with blood stasis and qi deficiency with dryness-heat and blood stasis(respectively 6 cases,5%),qi deficiency with phlegm-dampness and dryness-heat,dryness-heat and heart deficiency combined with blood stasis(respectively 5 cases,4.2%).The patterns above account for 49.2%Of all cases.5.The BMI of dryness-heat groups was higher than that of the none dryness-heat groups(P＜0.05),the FPG of dryness-heat groups was significantly higher than that of the none dryness-heat groups(P＜0.01);BMI and FPG of phlegm-dampness groups were both higher than those of the none phlegm-dampness groups(P＜0.05);non-significant difference was observed by comparing the blood stasis groups with the none blood stasis groups,and qi deficiency groups with none qi deficiency groups(P＞0.05).Conclusions:1.The most common syndrome elements of DH are syndrome qi deficiency,blood stasis, phlegm-dampness and dryness-heat.2.The mechanism of DH featured by excess pattern,deficiency-excess complex and multiple pattern combinations is complex.3.The common patterns of DH are dryness-heat combined with blood stasis, phlegm-dampness combined with blood stasis and dryness-heat,blood stasis,qi deficiency combined with blood stasis,qi deficiency with dryness-heat and blood stasis,qi deficiency with phlegm-dampness and dryness-heat,dryness-heat and heart deficiency combined with blood stasis:and the uppermost pattern combination of MS is dryness-heat with blood stasis combined with other elements.4.BMI and FPG of Type2 DH patients with dryness-heat or phlegm-dampness pattern were both higher than the corresponding contrast,which showed that a high BMI or FPG could be the reference index of dryness-heat and phlegm-dampness,the two Chinese medical patterns of DH.

Establishing Methods and Evaluations of Experimental Stress Fracture Model in Rabbits

Thu, 31 Mar 2011 12:31:19 +0000

With the increasing intensity and subjects of military training, military training injuries are becoming more and more prevalent, which can depress the combat effectiveness severely. Especially, stress fractures are usually treated as the representative index of MTI for their high prevalence rate, excruciating pain and difficulty to be diagnosed by routine methods.During military training, stress fractures occurred in tibias take up a great proportion, which are high than 50%. Routine X-ray examination can’t discover the early sign of stress fractures, while skeletal scintigraphy is much sensitive to early stress fractures, and is regarded as gold standard of diagnosis.Stress fractures result from multiple factors, whose pathogenesis has not been identified. A reliable animal model is necessary for further studies on stress fractures.By developing an exercise system and carrying out related animal experiments, we established a stress fracture animal model, and made a preliminary progress on relationship between exercise intensity and stress fracture degrees.Major contents can be summarized as follows:1. Designed rabbit exercise experiment system based on MSP430 MCU, which can generate high voltage pulses to force rabbits to run and jump.2. Established an Exercise-induced stress fracture model in rabbits by using the exercise experiment system.3. Evaluated stress fractures degrees in rabbit tibias through skeletal scintigraphy.4 Through statistic methods, we analyzed effects exercise intensity contributes to stress fracture degrees.Conclusions can be categorized as follows:1. The rabbit exercise experiment system can adjust exercise intensity and quantity quantitatively with high reliability and expediently.2. The rabbit exercise experiment system and skeletal scintigraphy can be reliable to establish animal mode successfully in short time. On the exercise intensity degree of 10 times per minute and 400 times per day, exercise-induced stress fracture model in rabbits can be established in 1 to 2 weeks.3. Exercise frequency and quantity are positively correlated with prevalence rate of stress fractures in rabbit tibias, among which, exercise frequency plays a much more important role. So we can reach the conclusion that lower training frequency during military training courses may result in lower prevalence rate of military training injuries.

Experimental Study on Peripheral Blood Mononuclear Cells Transplantation and Bone Marrow Stem Cells Mobilization in Treatment of Heart Failure after Myocardial Infarction

Thu, 31 Mar 2011 12:31:17 +0000

Objectives:To compare effects of peripheral blood mononuclear cells (PBMCs) transplantation and recombinant human granulocyte colony-stimulating factor (rhG-CSF) to mobilize bone marrow stem cells on heart function and vascular proliferation after acute myocardial infarction (AMI) in rabbits.Methods:Forty-five healthy rabbits aged five months were randomly divided into three groups (n=15). GroupⅠwas control group, groupⅡwas PBMCs transplantation group, and groupⅢwas rhG-CSF treated group. Myocardial infarction was induced by ligating left anterior descending coronary artery in all rabbits. In transplantation group (groupⅡ), PBMCs were isolated by density gradient centrifugation and nuclei were stained by Brdu. PBMCs transplantation was performed 30 minutes after AMI. In mobilization group (groupⅢ), rhG-CSF (30μg·kg-1·d-1) was injected subcutaneously after AMI and every 24 hours for 5 days. Control animals (groupⅠ) didn\'t receive any treatment after AMI. Left ventricular end-diastolic diameter (LVEDd) and left ventricular ejection fraction (LVEF) were measured by echocardiography before operation and at the end of the first and sixth week after operation. All rabbits were sacrificed after 6 weeks to get the heart to make pathological section. The size of infarcted myocardium was measured by HE stain. Monoclonal antibodies against Brdu were used to locate the implanted cells, and the counts of microvessel on the centre and margin of the infarcted myocardium were evaluated by immunohistochemistry.Results:1. In groupⅡ, PBMCs were successfully isolated from peripheral blood by density gradient centrifugation.2. LVEDd of groupⅠ, groupⅡ, groupⅢwere 12.98±1.38 mm, 12.72±1.04 mm, 12.94±1.37 mm and LVEF were 79.84±3.55%, 79.80±3.59%, 80.31±3.64% respectively before operation. There was no significant difference among them (P>0.05). At the end of the first week after operation, LVEDd of groupⅠ, groupⅡ, groupⅢwere 17.77±1.59 mm, 16.87±1.28 mm, 17.02±1.64 mm and LVEF were 50.98±6.78%, 52.14±5.61%, 53.72±4.91%. LVEF decreased significantly at the end of the first week after operation among these three groups (P0.05).3. HE stain showed that the sizes of infarcted myocardium of groupⅠ, groupⅡand groupⅢwere 37.15±2.94%, 26.87±2.33%, 27.47±2.13% respectively. Compared with groupⅠ, the sizes of infarcted myocardium were significantly decreased in other two groups (P<0.01).4. Six weeks after transplantation, Brdu-stained cells were observed at the transplantation area in groupⅡ. The implanted cells mainly located in the local injection site and their shape different from the normal myocardial cells.5. The capillary density of groupⅠ, groupⅡ, and groupⅢassessed by immunohistochemical staining with monoclonal antibodies against factorⅧ-related antigen were 2.54±1.27, 8.40±1.96, 6.20±1.26 in the central area of infarction and 13.69±2.93, 25.33±3.75, 22.87±2.70 in the marginal area of infarction respectively. Compared with groupⅠ, the capillary density of infarcted myocardium were significantly increased in other two groups (P<0.01), especially in groupⅡ.Conclusions:1. The AMI model of rabbits can be successfully duplicated by ligating left anterior descending coronary artery.2. PBMCs transplantation and rhG-CSF mobilize bone marrow stem cells can promote vascular proliferation, decrease myocardial infarction size, inhibit ventricular remodeling after AMI and improve left ventricular function.

Initial Study on Domesticational Culture and Quality of Huperzia Serrata (Thunb.ex Murray) Trev

Thu, 31 Mar 2011 12:31:15 +0000

In this paper, the cultural adaptability, Huperzine A (Hup A) content and proteomics analysis were carried out, using Huperzia serrata (Thunb.ex Murray)Trev. from different areas as materials. The results were as follows:1、Through the allocation of resources, Huperzia serrata(Thunb.ex Murray)Trev. was more suitable for growing under the environmental conditions of higher degree shading and relative high humidity. The plants grown healthy,which was up to more than 20cm high, fast growing and developing,and much more spores on the place with higher humidity. While the plants grown weakly,which was about 10-15cm height, slow growing and developing, less spores . The result of soil treatment showed that the thehumic thicker layer sand soil was conducive to the growth of H. serrata.2、Through the introduction and acclimatization, the survival rate of H. serrata from different places was statistically significantly different (P< 0.05). The survival rate (65.5%) of the plant from Datian was higher than that from the other, but the plant had the lowest survival rate (14.3%) from Nanjing and weak growing potential. This discrepancy might be explained by the fact that H. serrata needs enough crown closure degree during the cultivation. The results also showed that the plant growth in loess had higher survival rate (75.0%) and strong growth potential, the plant growing in paddy field soil had lower survival rate (18.8%) and weak growth potential. As above description, the better environmental condition was loess and shade in the cultication, H. serrata from Datian was more suitable for the cultivation environment.3、The content of Hup A was determined by HPLC. The condition of HPLC was Waters C18 culumn, 1ml/min flow rate, mobile phase (MeOH:NH4OAc=30:70), detection at 310 nm, 30°C column temperature, 10μl sample size. The result showed that the content of Hup A in stem was three times higher than that in leave. Both contents of Hup A in stem (197.796μg/g) and leave (87.537μg/g) from Datian were the higher than that from Sanming and Nanjing. The content of Hup A in H. serrata from different places was obviously statistically significantly different (P< 0.01).4、The results of proteomics analyse showed that the electrophoretogram of protein in the plants from different places had 15 defferent mass points by Imagemaser 2D Elite 5.0 (image analysis software.). There were high abundances in 2, 4, 7, 11 mass point from Datian, high abundances in 3, 5, 8,14, 15 mass point from Sanming and the abundances in other six mass points from Nanjing.

An Experimental Study of Femoral Neck Fracture Fixed with Different Entry Angle of DHS

Thu, 31 Mar 2011 12:14:37 +0000

Objective:To study the feasibility of low entry angle fixation and the change of femoral neck fracture site fixed with different entry angle of DHS to provide an experimental basis for clinically applying DHS smartly.Methods:Twenty-four fresh,freezing cadaver femurs were obtained and assigned randomly to four fixation groups with different entry angles,group A was 135°fixation group,group B was 130°fixation group,C was 125°group,D was 120°group,six femurs in each group,the entry angle was determined by the angle between the guide pin and femoral lateral cortex.To manufacture a Pauwels\'Ⅲfemoral neck fracture model of 70 degree with a saw.After reduction,femoral fracture sites were fixed by different entry angle with a 135°DHS.After fixation,displacement and the angle between distal and proximal fracture ends were measured on cadaveric specimen.On anterioposterior and lateral X-ray plates,neck-shaft angle, tip-apex distance(TAD),Garden\'s align index,Parker\'s ratio were measured.In biomechanical test,displacement of fracture site and femoral head inferior displacement were measured.All data were statistically analyzed.Results:After DHS fixation,the neck-shaft angles in group A were no changed.In group B, C and D,the neck-shaft angle increased respectively 5.13°,9.97°and 14.67°,comparing with the neck-shaft angles before fixation,and there was a significant difference comparing with group A(P＜0.05).Garden\'s index in the four groups ranged averagely 160°～172°/179°～180°, there was a significant difference about Garden\'s index of anterioposterior X-ray plate among the 4 groups(P＜0.05).There was no statistic difference in TAD.Parker\'s ratio decreased when the entry angle decreased,lag screw\'s location trended to the inferior of the femoral head.The angle between the distal and proximal fracture surface in group A,B,C,D was respectively 0°,2.5°,4.5°and 8.5°,and there was had a significant difference among the four groups(P＜0.05).The entry angle decreased by 5°,so the entry point moved forward proximally by 5 mm.All specimen were finished the biomechanical test.The displacement of proximal fracture end in group B,C,D was gradually decreased during loading,however,the proximal fracture end in group A had a inferior displacement from no displacement.In static load test,femoral head inferior displacement among B,C,D had no statistical difference. There was a statistical difference between the three groups and the group D(P＜0.05). Comparing with B,C and D,axial stiffness of the group D was decreased respectively 31%, 29%,20%.In cycle test,there was no statistical difference among the group B,C and D in femoral head inferior displacement,which there was a statistical difference comparing with the group A. Conclusion:To the femoral neck fracture with neck-shaft angle of less than 135 degree,the fixation with the entry angle of less than 135°can make the neck-shaft angle rising, meanwhile,lag screw\'s location is easier to trend to the inferior of femoral head,displacement and angle between fracture surfaces satisfy the demand of functional(excessus) reduction or extroversion reduction,and a satisfied stability is performed in biomechanical test.So from the experimental outcomes,we suggest that a limited decrease of entry angle can be applied to attain a limited extroversion fixation for the poor stability of PauwelsⅢfractures,especially the smaller neck-shaft angle.

Violent Online Games Addictive Disorder and Adolescents Aggressive Related Research

Thu, 31 Mar 2011 12:14:36 +0000

Objectives To investigate aggressive of teen-age suffered from violent online games addictive disorder.Methods:5968 had been randomly drawn out from 8 junior middle schools from 4 districts in Dalian during August to December 2008. By Chen Internet Addiction Scale (CLAS) and Self-constructed games inve- stigation questionnaire, and met Internet addictive disorder diagnostic criterion for positive on the screening out Online Game Addiction 149(2.5%), According to gender and age, Screening out well-matched Violent Online Games Addiction and Entertainment Online Games Addiction each 66 students, of which 11 women, 55 men, average age of 14.36±0.81 years old. we also mach 66 students who are no-play online games students as third group. Methods All students from three groups have investigated with Eysenck personality questionnaire (EPQ), Minnesota Multiphase Persona- lity Inventory (MMPI-AF), Perceived Social Support Scale (PSSS), Egma minnen av bardndosnaupp forstran (EMBU), Aggression Questionnaire (AQ), (FES-CV). Statistics Database is stated with SPSS 12.0 package, database is analyzed with t test, one-way ANOVA, multiple linear regret- ssion analysis.Results:①Online games IAD is 149(2.5%)during the 5968 students, in which 71(1.18%) students had violent online games addictive disorder (VIAD,11 female, 60 male), 78(1.30%) students had entertainment online games addictive disorder (EIAD,23 female, 55 male).②Compare three groups, physio-aggression, language aggression, anger emotion, hostility cognition and aggression factors scores have significant difference(P<0.001). In the VIAD group, all factors scores higher than EIAD group and non-play online games group.③Compare three groups, ex-display anxiety, dependency and society of responsibility factors scores have significant difference(P<0.05). In the VIAD group, ex-display anxiety and dependency factors scores higher than EIAD group and non-play online games group. In the VIAD group, society of responsibility factor scores was lower than EIAD group and non-play online games group.④Compare three groups, affection express, contrary and control force factors scores have significant difference(P<0.05)during the three groups. In the VIAD group, affection express and control force factors scores were lower than EIAD group and non-play online games group. In the VIAD group, contrary factor scores were higher than EIAD group and non-play online games group.⑤Compare three groups, spiritual and neuroticism factors scores have significant difference(P<0.05). In the VIAD group, spiritual and neuro- ticism factors scores were higher than EIAD group and non-play online games group.⑥Compare three groups, family support factor score has significant difference(P<0.05). In the VIAD group, family support score was lower than EIAD group and non-play online games group.⑦Compare three groups, mother warm factor score has significant difference(P<0.05). In the VIAD group, mother warm factor score was lower than EIAD group and non-play online games group.⑧Multiple Linear Regression (Forward) of VIAD group: The dependent variable was aggression factor score, and the independent variables were Internet addictive scores and all other factor’s scores. The contrary score, religion morality score entered into Multiple Linear Regression equations.Conclusions①The incidence of secondary school students for online games Addiction Disorder 2.5%, VIAD rate was 1.18%, EIAD rate was 1.30%, Lower than expected, Game Addiction to multiple boys.②VIAD increased aggressive student, they often have a spiritual basis of the quality and neurotic personality, the lack of morality and social responsibility.③Over-reliance on them, there is the issue of emotional expression, and so it can easily lead to inner conflicts and anxiety, coupled with poor control.

Study on Warming Therapy in Treaties on Febrile Diseases

Thu, 31 Mar 2011 12:14:35 +0000

"Thermotherapy" is a treatment for cold syndrome with medicines of warm and hot nature.It is one of the "eight therapeutic methods".Cold-pathogen is one of the "six exopathic factors".In the olden days of Eastern Han, people think that cold-pathogen was the main cause of many diseases,therefore,cold syndrome had become the main research by a lot of medical scientists,and thermotherapy was commonly used.But after the Ming and Qing Dynasties,the existance of the science of epidemic febrile disease of traditional Chinese medicine was so popular that the cold syndrome and thermotherapy is neglected.People nowadays prefer to use cool and cold method and also cold-natured herbs.They are lacking in the cold-deficiency syndrome thinking and the usage of the thermotherapy.Shang Han Lun(Treatise on Febrile Diseases) is a classic book with brilliant achievements and important value in development history of Traditional Chinese Medicine, that must be read by every Chinese physician and the elements in the Shang Han Lun are mostly about thermotherapy.Till today,people still regard this book as a monograph.The thesis uses the following to discuss thermotherapy of Shang Han Lun:1.discuss the importance "yang qi".2.defination of cold syndrome.2.1.Etiologic and pathogenic factors of cold syndrome.2.2.main clinical characteristic of cold syndrome.3.importance of thermotherapy in Shang Han Lun.4.discuss on the basis of thermotherapy in Shang Han Lun.5.Segregate Shang Han Lun decoction into 10 types,namely rescuing.from collapse by restoring Yang method,warming and replenishing heart yang method,warming Middle-Jiao to dispel cold method,warming lung to reduce watery phlegm method,warming liver to dispel cold method,warming yang to induce diuresis method,warming yang and balance weiqi method,warming yang and relieving exterior syndrome method,warming yang and arresting discharge method,expelling pathogentic cold from channel and expelling wind and removing damness method.All these mentioned-above compare,systemize,analyze,compatible,decoct and consume.6.Summation of thermotherapy in Shang Han Lun in terms of usage and constraints.7.Explain the following:(1)《Synopsis of Prescription of the Golden Chamber》characteristics of thermotherapy(2) explain the usage and development of thermotherapy by various dynasty physicians(3) usage of thermotherapy in the disease caused by Pathogenic Warmth(4) modern mechanism of Action in thermotherapy8.discuss thermotherapy in the modern age of predominance and future expectation.

The Research of the Therapeutic System with Functional Electrical Stimulation

Thu, 31 Mar 2011 12:14:33 +0000

Functional Electrical Stimulation plays an important role in the treatment of neurological disorders and rehabilitation of nerve injury. It makes use of the nerve cells\' response to these stimulations to transfer our artificial controlling signals. Through the stimulation of adscititious electrical current, nerve cells are supposed to generate a nerve impulse that is the same as the one induced by natural inspiration. Then the nerve impulse contracts the corresponding muscle fibers which finally generates expected movement. With the development of micro-electronics technology, micro-nano technology and biological technology, FES is widely used in rehabilitation medicine. Now FES using not only has achieved great progress in control in the treatment of epilepsy, chronic pain, urinary incontinence, visual and listening impairment, but also has an attractive prospect in the upper and lower extremities\' move, the treatment of scoliosis, the assistant with breathing.The topic is about the application of the FES used in the upper and lower extremities\' move, proposing the method that using the sEMG signal picked-up from the health part of the body to stimulate the paralyzed part instead of using the electrical pulse signals. SEMG is biological signals accompanied by neuromuscular activities, it has a relation with muscles\' activity and functional status, so it reflect the activities of neuromuscular to some extent. The sEMG signal is complex and changeable; there are great differences between sEMG and the continuous wave, the bunch of shape wave and the dilatational wave used in traditional electricity stimulation treatment.In this paper, sEMG signal is picked-up from the health part of the body, then processed by matlab and stored as the stimulation source, considering the meridian phenomenon in traditional Chinese medicine, choose the appropriate points to stimulate. After introduced the EMG signals, the methods for EMG processing and the researches of FES, we detailedly introduced the hardware and software design of the system. The whole system is made up of three main parts, the Single Chip Micyoco, the D/A circuit and the signal enlarged circuit. Transform the EMG signals from digital signals to analog signals, and then enlarge them to the irritability intensity of the human body. Finally stimulates the human body with the analog signals.

Stady on CBS of Mices Brain after Carbon Monoxide Poisoning

Thu, 31 Mar 2011 12:14:32 +0000

Background: The carbon monoxide (carbon monoxide, co) is one kind of achromatic color, tasteless, not smelly, non-irritant virulent gas. It is in the life and the production causes the toxic death , the most common asphyxiating gas.It may cause organism multi-system harm, exhausts oneself by the cerebrum seriously; especially on central nervous system and cardiovascular system.Besides may create the acute toxic brain damage, have a part of patients to alleviate, consciousness after the toxicant symptom restores normally, probably experience 2-60 days performance normal or the basic normal dormant period, can appear a series of symptoms, including dementia、amentia and extrapyramidal system\'s symptom, is called the Delayed encephalopathy after carbon monoxide poisoning (DEACMP). The disease rate is 10-30% in civil,0.8-43% in abroad. The clinical manifestation is a group by the acute stupid primarily energetic nerve function disorder, is CO poisoned the most serious type. The related CO poison and the hangfire brain illness pathogenesis research, proposed until now some hypotheses but its pathogenesis still not extremely clear.20th Century lately, hydrogen sulfide(H_2S) was founded as the third gas of endogenous neuromodulator after NO and CO.It has important physiological action in brain. H_2S specifically potentiate the activity of NMDA receptor and alter the induction of long-term potentiation(LTP) in the hippocampus. H_2S can regulate the release of corticotropin-releasing hormone from the hypothalamus. H_2S acts as a vasorelaxant, to relax smooth muscle cells. H_2S has anti-oxidation function. Cystathionine- beta- synthase(CBS)and cystathionine-gamma-lyase(CSE) are key enzymes of the trans-sulfuration pathway but can also use L-cysteine as an alternative substrate to form H_2S. Both enzymes are pyridoxal-5-phosphate dependent enzymes .CBS seems to be the main H_2S-forming enzyme in the CNS. H_2S formation by brain CBS can be controlled. Calmodulin(CaM) is the fast regulated pathway to activate CBS and enhance H_2S formation. H_2S、NOandCO have similar property in many Pathophysiologiy processes and form complex regulating networks of gas signal in body.At present, the animal experimentation confirmed that H_2S was protective in cerebral ischemia model . But after the CO poison the H_2S of brain situation still not was explicit reported. We observed the expression of CBS and CaM in the mice brain tissue after carbon monoxide poisoning to study the mechanism of delayed encephalopathy after carbon monoxide poisoning.Objective: This experiment draws up in establishes the CO poison mouse model in the foundation, dynamic observes CBS expression change rule from the protein horizontal level,. to study the mechanism of delayed encephalopathy after carbon monoxide poisoning.Methods: 162 healthy male Kunming mice, body weight (18-22)g,were divided into the air control group stochastically (54), the contamination group (54), the butylphthalide group, each group divided into 6h, 1d, 2d, 3d, 4d, 7d, 14d, 21d, and 28d,9 subnit again.The contamination group according to the 170ml/kg dosage, establishes the CO poison mouse model through the single abdominal cavity injection 99.9% co gas law, the butylphthalide group given abdominal cavity butylphthalide injection(1ml/Kg,bid) after the CO poison mouse model, the air control group by the same level dosage air injection. After makes the CO poison mouse model the corresponding time spot, the execution mouse, beheads takes the brain, conventional. Use the immunity organization reduction to examine each group of CBS and CaM expression.Result1、The air control group mouse hippocampus area CBS expression is no difference. The contamination group mouse hippocampus area CBS IHS value starts in the contamination latter 6 hours to reduce, 2d to the lowest point, lower than the air control group. Then raising, the highest point at 14d. Then lower again. The butylphthalide group reduced gradually, 2d to the lowest point but higher than the air control group and the contamination group. Then raising gradually, 7d to the highest point value Then lower again. The air control group and CO group 6h、7d、14d、21d and 28d IHS value comparison has statistics difference (P<0.05). CO group and the butylphthalide group 1d、2d and 14d IHS value comparison has statistics difference (P<0.05).2、The air control group mouse hippocampus area CaM is no difference. The CO group mouse hippocampus area CaM presents two stages the changes, first reduce,2d to the lowest point, then raising , 14d to the highest point, then reduce again,gradually.. The butylphthalide group reduced gradually, then raised, 3d to the first peak, 4d reduced. 7d to the highest point, then reduced again. The air control group and CO group1d、2d、4d and 28d IHS value comparison has statistics difference (P<0.05). CO group and the butylphthalide group 6h、2d、4d、14d and 21d IHS value comparison has statistics difference (P<0.05).Conclusion1、CBS and CaM are expressed in normal mouse hippocampus area, after the CO poison High concentration H_2S is protective for the brain in the early period, but it is harmful in the later period if the concentration of H_2S is higher. Thereforce H_2S maybe participate the mechanisms of DEACMP.2、CaM is the fast regulated pathway to activate CBS, but they chang nonsynchronically.The reason maybe other factors participate the regulation of CBS. High concentration CBS is feedback inhibited the expression of CaM.3、Butylphthalide is protective for the brain, H_2S maybe participate the mechanisms of protection.

Expression and Significance of APC、β-catenin and Cyclin D1 in Breast Cancer

Thu, 31 Mar 2011 12:14:30 +0000

Objective: This study was to examine the expression of adenomatous polyposis coli (APC),β-catennin and Cyclin D1 in different breast tissues, and investigate their possible correlation with the occurrence, infiltration and metastases in breast cancer.Methods: The high sensitive SP immunohistochemical method was used to detect the protein expression of APC、β-catenin and Cyclin D1 in 52 breast cancer and 15 surrounding tissues. 13 specimens of ductal carcinoma in situ (DCIS) and 39 specimens of infiltrating duct carcinoma (IDC) were included.Results: Positive expression rate of APC in 52 breast cancer was 46.15%, and the positive rate in DCIS and IDC were respectively 76.92%, 35.90%. There was significant difference between them(P<0.05). Its positive rate in breast cancer tissues and surrounding tissues also had significant difference(P<0.05), and with the increase of TNM staging, APC\'s rate of positive expression reduced. Abnormal immunoreactiveityβ-catenin was observed in 29 cases (55.77%), abnormal expression was positively correlated with histological grades, TNM stages and axillary lymph node metastasis, but not with the age, tumor size, ER, PR and HER-2. The abnormal expression rates ofβ-catenin inⅢ～Ⅳgroup were signif- icantly higher than those inⅠ～Ⅱgroup,P<0.05.The over-expression of Cyclin D1 were found in 48.08% breast cancer cases and it was significantly correlated with ER statue, but was not correlated with age, histologic grade, degree of differentiation node statue and tumor size of patients. The positive expression of APC and over-expression of Cyclin D1 were respectively observed in 31.03%, 66.52% of these cases which showed abnormal staining ofβ-catenin, but only observed in 65.21%, 26.09% of these cases with normal membranous staining ofβ-catenin. There was a significantly positive correlation between the abnormal expression ofβ-catenin and the over-expression of Cyclin D1, but a negative correlation between the abnormal expression ofβ-catenin and positive expression of APC.Conclusions: The negative expression of APC, abnormal expression ofβ-catenin, over-expression of Cyclin D1 exists in breast cancer, which may play important roles in the carcinogenesis of the breast cancer.

Investigation on the Chemical Constituents of Saussurea Superba

Thu, 31 Mar 2011 12:14:28 +0000

Saussurea superba Anthony belongs to genus Saussurea DC of compositae family, wildly distributed in Northeastern and Northwestern mountainous regions of China. The genus Saussurea DC consists of about 400 species distributed throughout Asia and Europe, and more than 300 species were discovered in China. Plants of the genus Saussurea are commonly used as traditional Chinese medicine for anti-tumor, anti-inflammatory, anti-aging, and improving cardiovascular system functions.In this thesis, we summarize the plant classification and pharmacological activities of coumarins in plants and report the systematic phytochemical studies of the whole plant of saussurea superba, which was collected in Huzhu County of Qinghai Province, China. 26 pure compounds were isolated from the ethanolic extract by means of silica gel column chromatography (CC), preparative thin layer chromatography (prep-TLC), recrystallization, etc. The structures of 25 compounds were elucidated as triterpenoids, steroids, coumarins, fatty acids, etc., on the basis of spectral evidences, along with comparison to authentic samples or the data in the literature.In view of the four coumarins (umbelliferone, scopoletin, skimmin, scopolin) extracted from Saussurea superba were the dominant active ingredients in the plants, capillary zone electrophoresis (CZE) was explored for the measurement of these components. Our study is to set up a simple and rapid electrophoresis method for simultaneous determination of the four coumarins, and the current analytical methods have not yet been reported to isolate and identify this plant. The conclusions revealed that CZE was a useful, simple and reliable approach for the analysis of these components.The results of this thesis is of great significance to further research the chemical classification, pharmacological activities, the development and utilization of this genus.

Inhibitory Effects of Epirubicin and Liposome Doxorubicin on Human Breast Cancer Cell Lines and Dentritic Cells and Construction of Plasmid Interference Vector of CXCR4

Thu, 31 Mar 2011 12:14:26 +0000

Breast cancer is one of the most common malignancies in women.The incidence of breast cancer has increased steadily over the past few years,but breast cancer mortality appears to be declined,suggesting benefits from early detection and more effective treatment.The treatment of systemic disease with cytotoxic chemotherapy is one of the treatments of breast cancer.Anthracycline antineoplastic drugs,which include epirubicin and liposome doxorubicin,are the most important chemotherapy drug.Epirubicin is one of the first line chemotherapy drugs in the treatment of breast cancer.And liposome doxorubicin is a new antitumor drug,which through encapsulation of the drug in macromolecular,such as liposome,the volume of distribution is significantly reduced and the concentration of drug in the tumor is increased.The liposome protects the drug from metabolism and activation in the plasma and due to size limitations in the transport of large molecular or carriers across healthy endothelium,the drug accumulates to a reduced extent in healthy tissues. However,discontinuity in the endothelium of the tumor vasculature have been shown to result in increased extravasations of large carriers and,in combination with an impaired lymphatic,an increased accumulation of liposome drug at the tumor.Dentritic cells(DC)play important roles in tumor immunity.DCs phagocytose endogenous and exogenous antigens,and initiate the Ag-specific T cell proliferation with the capability of antigens presentation.It has been reported that in breast cancer,numbers of DCs are lower in sentinel lymph nodes(SLN).And the reason of this may be that tumor cells release a variety of cytokines and other biologically active materials.These various mediators must flow into the SLN,making it easy to imagine that they could influence the response to tumor cells within the SLN,for instance,via a reduction in DC number.CXCR4 is a G-protein coupled receptor,and it belongs to chemokines,a family of small pro-inflammatory cytokines.And chemokines and their eceptors regulate a variety of immune responses to infection,inflammation,and tissue repair.Recently,it has been established that cancer cells exploit signaling through chemokine receptors for several key steps in initiation and progression of primary and metastasis cancer.In particular,the chemokine CXCL12 and its receptor CXCR4 have prominent roles in primary and metastasis breast cancer, as well as a number of other important malignancies including lung,brain,and prostate.Signaling through CXCR4 activates a number of downstream effector molecules,including molecules that regulate key processes such as cell cycle control and apoptosis.Based on those above mentioned,our study carried out the research from the following two aspects: Experiment 1.Culture and identification of human dentritic cell.Successive adherence method was utilized to obtain mononuclear cells.The mature DC was induced in vitro by granulocyte-macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4)and tumor necrosis factor-a(TNF-α). Identification of DC included observing the utrastructure with electron microscope.Then,human breast cancer cell lines,Bcap37 and MDA-MB-231, together with human dentritic cells,were cultured with epirubicin and liposome adriamycin,respectively,under different concentrations(0、0.25、0.5、1.0、2.0、4.0、10.0μg/ml).The inhibitory effects were detected by MTT method after 24、48、72 h.The result was that epirubicin and liposome adriamycin could inhibited the proliferation of Bcap37 cells,MDA-MB-231 cells,and human dentritic cells.Liposome adriamycin exhibited a lighter inhibition on dentritic cells than on human breast cancer cell lines(Bcap37 and MDA-MB-231) (F=22.208,P＜0.01;F=20.534,P＜0.01).And compared with MDA-MB-231 cells,the proliferation of Bcap37 cells was inhibited more effectively by liposome adriamycin.In conclusion,breast cancer cell lines with lower malignant degree appears to be more sensitive to chemotherapy,as compared to those with higher degree.The toxicity of liposome adriamycin on dentritic cells is lower than epirubicin.Experiment 2.The construction of plasmid interference vector of CXCR4 -Psilencer3.1-CXCR4 was utilized the method of molecular cloning.It was detected to be correct with DNA sequencing.Then detecting mRNA and protein express of CXCR4 gene in human breast cancer cell line SK-BR-3.Results indicated that the express level of CXCR4 in breast cancer cell line SK-BR-3 interferenced by Psilencer3.1-CXCR4 was significantly lower than the control, SK-BR-3 without transfection.

Exposure of Rats to Genistein during Different Life Stages Alters Reproductive Development

Thu, 31 Mar 2011 12:14:25 +0000

Genistein is a phytoestrogen that is found in a wide variety of plant-derived foods especially in soybeans and soy-based foods. Genistein has potential as an alternative treatment for cancer, menopausal symptoms, osteoporosis and cardiovascular disease. However, it has been reported recently that genistein which presences in infant soy formula products has some adverse effects on the reproductive development of animals.Then, the scientists bigen to working on the toxic effect of genistein on animal and their infant, and whether there are benefits give it from early stage.ObjectiveThis study aimed to investigate the effects of dietary genistein on the reproductive development of rats during different periods.MethodsIn this study, we divided the rats into three groups: Control group, Middle Gen group and High Gen Group, and each group was fed a diet containing genistein at levels of 0, 20, or 100mg/kg per day.In the study of genistein exposure from early stage, we exposed genistein from pregnant day 10 to the age of 13 weeks. In the study of genistein exposure during adolescent and adult periods, we exposed genistein from 3 weeks old to 13 weeks old of rats. In the study of genistein exposure during adult period, we exposed genistein from 9 weeks old to 13 weeks old of rats. Body weight, food consumption and anogenital distance were recorded once during perinatal period and weekly thereafter. Vaginal opening and testicular descent were observed once a day after the pups’weaning. The plasma estradiol levels of females and testosterone levels of the males were measured with the Spectria Radioimmunoassay (RIA) kit.Histological slides of all of generative organ were coded and the pathological changes were observed under a microscope.The immunoblot was analyzed using a BIORAD gel documentation system and quantified using BIORAD Quantity One software.Results1. In the study of genistein exposuring during perinatal, adolescent and adult periods, the body weight and food consumption of High Gen group mother rats decreased postnatal day 21, and the plasma estradiol level increased compared with Control group. Also, the expression of estrogen receptor in uterus and ovary. Pup mortality increased in High Gen group, decreased anogenital distance(AGD) and ogran weight in female and male rats were observed. Histopathology showed that more vesicula graafianae appeared in ovary of 3 weeks old female pups in High Gen group. At 9 weeks old, the body weight and food consumption of female rats decreased, and the vaginal open time was earlier than control group. Body weight and food consumption of male rats increased at 9 weeks old. At the age of 13 weeks, the plasma estradiol level decreased. The cavity of uterus was larger than Control group. Treatment with genistein augmented the expression of estrogen receptor in uterus and ovary. And the plasma testosterone level of male rats increased.2. In the study of genistein exposuring during adolescent and adult periods, the plasma estradiol level of female rats increased and the plasma testosterone level of male rats decreased at the age of 9 weeks compared with Control group. At the age of 13 weeks, decreased body weight, food consumption, anogenital distance(AGD) and overy weight in female rats were observed. Also, the vaginal open time of female pups was earlier than control group. Histopathology showed that the cavity of uterus was larger than Control group. Otherwise, treatment with genistein caused reduction in the plasma estradiol level of female rats and the expression of estrogen receptor-αin uterus. There were few changes of male rats’body weight, testicular descent, testes weight and histopathology except that the increased food consumption from 10-week-old and plasma testosterone level.3. In the study of genistein exposuring during adult periods, compared with Control group, decreased body weight, food consumption, anogenital distance(AGD) and weight of ovary in female rats were observed. Also, treatment with genistein caused reduction in expression of estrogen receptor-αin uterus. There were few changes on male rats body weight, food consumption and the weight of testis.Conclusion1. Genistein exposure during early period could effect on rats development as estrogen. It could promote the development of pups’reproductive organ. However, this effect meight be harmful when they were growing up.2. Genistein exposure during adolescent period could effect on female rats’reproductive development more than male rats’. The effect of rats’bodyweight and food consumption was mainly after their growing up.3. Genistein exposure could effect on adult female rats development as antiestrogen. However, there were few effect on adult male rats development. It meight be have some effect on male rats if add the dosage.

The Protective Effect of Resveratrol on Cardiac Microvascular Endothelial Cells Injury Induced by Ischemia-reperfusion

Thu, 31 Mar 2011 12:14:23 +0000

Background and aimsCoronary heart disease (CHD) is one of the major diseases that threaten human health. Epidemiologic survey disclosed, French have a very low mortality rate due to CHD despite having a high-fat diet and smoking habbits. This so-called‘French paradox’has been attributed in part to wine consumption, particularly red wine. Resveratrol (trans-3,4’,5-trihydroxystilbene), a naturally occurring phenolic compound abundantly found in grape skins and in wines, has been thought to be responsible for the cardiovascular benefits associated with moderate wine consumption. Resveratrol has extensive physiological pharmacological effects, which include antioxygen, cleaning free radical, antiplatelet, regulating blood fat, preventing artherosclerosis, anti-inflammatory, anti-eumycete, anti-tumor, and hepatoprotective properties. The objective of this experiment is to study protective effects of resveratrol on cardiac microvascular endothelial cells (CMECs)injury induced by ischemia-reperfusion .Methods PartⅠ: The culture and identification of CMECs. 1. CMECs were isolated enzymatizally from left ventricle of SD rats, and cultured in DMEM supple- mented with 15% fetal calf serum. 2. Immunofluorescent co-stain with Dil-A c-LDL and DAPI to identify CMECs.PartⅡ: A cell model of ischemia-reperfusion injury was established. CMECs were randomly divided into three groups: Normal group (Con group), ischemia reperfusion group (I/R group) and resveratrol group (Res group). 1. Cell viability was detected by MTT colorimetry. 2. Flow cytometer based on Annexin V-FITC/PI double staining was used to detect apoptosis rates. 3. Colorimetry was used to detect caspase-3 activity. 4. The activity of superoxide dismutase and the Content of malondialdehyde were detected by ELISA. 5. The expression of TNF-a and IL-10 were detected by ELISA. 6. The positive rates of NF-кB were detected by immunofluorescence method.Statistical analysis was performed with SPSS version 11.0. One-way analysis of variation and post hoct (LSD-t) test were employed.Results1. The cells obtained showed double positive for Dil-A c-LDL and DAPI, according to the source of cells, they are CMECs .2. Compared with control group, Ischemia reperfusion injury could significantly decrease cell activity (P < 0.01); Compared with I/R group, resveratrol could significantly increase cell activity (P < 0.01).3. Compared with control group, Ischemia reperfusion injury could significantly increase cell apoptosis rates and caspase-3 activity (P < 0.01); Compared with I/R group, resveratrol could significantly decrease cell apoptosis rates and caspase-3 activity (P < 0.01).4. Compared with control group, Ischemia reperfusion injury could increase the activity of superoxide dismutase and the content of malondialdehyde. Compared with I/R group, resveratrol could significantly increase the activity of superoxide dismutase (P < 0.01), however decrease the content of malondialde- hyde (P < 0.01).5. Compared with control group, Ischemia reperfusion injury could increase the content of TNF-a and decrease the content of IL-10. Compared with I/R group, The TNF-a decreased, however the expression of IL-10 was increased.6. Compared with control group, Ischemia reperfusion injury could increase Masccline rates of NF-кB, but resveratrol could significantly decrease Masccline rates of NF-кB compared with I/R group.ConclusionResveratrol may exert protective effects by antioxygen, anti-inflammatory,decreasing CMECs apoptosis and decreasing positive rates of NF-кB.

Influence of Dexamethasone on Foxm1 Gene Expression in Pulmonary Cells Types Following Acute Lung Injry

Thu, 31 Mar 2011 05:04:01 +0000

Objective:To investigate the expression and the effect of protection of Foxm1 transcription factor and foxm1 gene in the lungs of mice with lipopolysaccharide(LPS), and the effect of dexamethasone on acute lung injury(ALI)induced by LPS.Methods: 72 mice were randomly devided into 3 groups: control group (A, n=24), model group(B, n=24), DEX treatment group(C, n=24 ). In the control group, mice were given 0.9% sline intraperitoneally(0.3ml). In the model group, mice were given i.p. injected with LPS at a dosage of 5mg/kg. In the DEX treatment group, the mice were given i.p. injected with LPS at a dosage of 5mg/kg, after 6 hours the mice were i.p. administered dexame- thasone at a dosage of 5mg/kg. Then the 3 groups respectively included 24h, 48h, 72h after LPS lung injury and DEX treatment. The level of foxm1 mRNA expression in lung was detected by reverse transcription polymerase chain reaction (RT-PCR). Foxm1 transcription factor in lung was detected by Immunocytochemica (IHC). Pathological changes in the lung were observed after the establishment of model.Results:When group comparison, at 24h, the expression levels of Foxm1 tanscription factor(8.881±0.817) in the mice lungs of the DEX treatment group were remarkably higher than model group(6.883±1.172) (P<0.05), and DEX treatment group also remarkably higher than control group(2.585±0.557)(P<0.05); the expression levels of foxm1 mRNA (0.608±0.094) in the mice lungs of the DEX treatment group were remar- kably higher than model group(0.268±0.015)(P<0.05), and model group also remarkably higher than control group(0.126±0.047)(P<0.05). At 48h, the expression levels of Foxm1 transcription factor(55.163±3.019) in the mice lungs of the DEX treatment group were remarkably higher than model group (15.804±1.555)(P<0.05), and model group also remarkably higher than control group (2.387±0.539)(P<0.05); the expression levels of foxm1 mRNA(1.190±0.087) in the mice lungs of the DEX treatment group were remarkably higher than model group (0.433±0.208)(P<0.05), and model group also remarkably higher than control group(0.133±0.119) (P<0.05). At 72h, the expression levels of Foxm1 transcription factor (36.773±1.610) in the mice lungs of the DEX treatment group were remarkably higher than model group(13.064±1.1641)(P<0.05), and model group also remarkably higher than control group(2.047±0.095)(P<0.05); the expression levels of foxm1 mRNA(0.894±0.010) in the mice lungs of the DEX treatment group were remarkably higher than model group (0.335±0.097)(P<0.05), and model group also remarkably higher than control group (0.097±0.095) (P<0.05). When comparison within group, for DEX treatment group, the expression levels of Foxm1 transcription factor (55.163±3.019) in the mice lungs at 48h were remarkably higher than 72h (36.773±1.610)(P<0.05), and 72h also remarkably higher than 24h (8.881±0.817)(P<0.05); the expression levels of foxm1 mRNA(1.190±0.087) in the mice lungs at 48h were remarkably higher than 72h(0.894±0.010) (P<0.05), and 72h also remark- ably higher than 24h(0.608±0.094)(P<0.05). For model group, the expres- sion levels of Foxm1 transcription factor (15.804±1.555) in the mice lungs at 48h were remarkably higher than 72h(13.064±1.164)(P<0.05), and 72h also remarkably higher than 24h (6.883±1.172)(P<0.05);the expression levels of foxm1 mRNA(0.433±0.208) in the mice lungs at 48h were remarkably higher than 72h(0.335±0.097) (P<0.05), and 72h also remark- ably higher than 24h(0.268±0.015)(P<0.05); For control group, the expression levels of Foxm1 transcription factor in the mice lungs at 48h (2.387±0.539) have no significantly variation compared with those at 72h(2.047±0.095) and 24h(2.585±0.557)(P<0.05), the expression levels of foxm1 mRNA in the mice lungs at 48h(0.133±0.119) have no significantly variation compared with those at 72h(0.097±0.095) and 24h(0.216±0.047) (P<0.05). The pathologic changes of the lung in mice of the DEX treatment group were wilder than those in mice of the model group.Conclusion: The expression levels of foxm1 mRNA and Foxm1 transcription factor increased significantly in lung in LPS-induced acute lung injury and LPS+DEX group. Administration of dexamethasone reduces injury of lung by protecting vascular endothelial cells (VECs) and alveolar epithelial cells (AECs) and increasing the expression of Foxm1 taranscri- ption factor.

Effect of the NF-κB(p65) Inhibitor on Endotoxin-Induced Uveitis

Thu, 31 Mar 2011 05:03:59 +0000

Objective:Immuning SD rats with the lipopolysaccharide(LPS),and establishing a uveitis model and this afford a worthy tool for experimental study of immunogenic and therapy of uveitis.Endotoxin-induced uveitis (EIU)is a model for acute panvueitis seen in human. We report a study of the effect of systemic administration of PDTC on the clinical presen- tation ,histopathologic change and the level of Toll- like receptor-4(TLR-4) on iris in the SD rats.Methods: The establishment of endotoxin-induced uveitis disease model.Female SD rats(180～200g) were selected between 6～8 weeks age. Twelve SD rats were randomly divided into the healthy control group (6 rats,group 1A) and the experimental group(6 rats, group 1B). Lipopoly- saccharides were dissolved in the sterile,pyrogen-free 0.9% saline at a con- centration of 2 mg/ml,and 50μl was injected into each hind footpad of Sdrats in the experimental group (200ug total does per rat).The rats in the healthy group were injected 0.9% saline 200μl per rat in the same way. Twenty-four SD rats were randomly divided into three groups,including the EIU group(6 rats, group 2A),PDTC group(6 rats, group 2B)and Dexa- meth- son group(6 rats, group 2C).200μg of Lipopolysaccharide were inject- ted into footpads of each SD rats in all groups. The rats in group 2A were subconjunctival injected with 0.9%salin at 1ml/kg twicely at the time of 0h and 6h after lipopolysaccharide injection.Both rats in group 2B with PDTC at 20μg/kg and group 2C with Dexa at 100μg per rat were injected in the same way.All samples were examined with light microscope in 6h,12h,18h, 24h after LPS injection.The numbers of leukocyt and concentration of pro- tein in aqueous humor were measured and TLR-4 in iris constitution were observed by immune-histochemistry after rats had been killed in 24h.Results:The data was denoted by x±s, and analyzed the data by SPSS 11.0 software.The EIU disease model was established successfully in SD rats in this study.The disease incidence was 100%.The clinical and histolo- gical features were stable.Obvious uveitic inflammatory signs such as flare,cells and protein exudates in anterior chamber,occlusion of pupil and vitreous opacity were obserbed.The quantity of leukocyt and protein obv- iously decreased in group 2B and 2C,p0.05.Conclusions:1.Uveitis with marked clinical signs is successfully indu- ced in the SD rats with injection of LPS.2. TLR-4 expression increased in the control group. TLR-4 probably participates in the formation and dev- eloppment of acute anterior uveitis.3.PDTC could relieve inflammatory reaction and TLR-4 expression decreased in the PDTC group. 4.Dexa- methason could obviously suppress the symptom of EIU relieving infla- mmatory reaction and could decrease the expression of TLR-4 on iris.

Carriage Frequency and in Vitro Virulence of Candida Spp. in the Oral Cavity of Chinese HIV-infected Patients

Thu, 31 Mar 2011 05:03:57 +0000

It was estimated that China would have more than 0.7 million people living with HIV/AIDS by 2007.Opportunistic mucosal infections caused by Candida species are common in individuals with human immunodeficiency virus(HIV) infection.Oral candidiasis remains the most commonly encountered oral complication in HIV/AIDS patients.Although episodes of mucosal candidiasis are rarely lethal,they contribute significantly to morbidity and reduced quality of life of patients with AIDS.However,only a limited number of studies have embarked on identifying the colonizing or infecting yeast species in China.The majority of candidal infections are caused by Candida albicans,however, reports of the emergence of other species of Candida have begun to appear.Despite the increased prevalence of Candida species and the increased frequency of candidiasis in HIV-infected patients,little is known about the changes.There are several factors that may contribute to the transition of Candida species from asymptomatic commensal to pathogen,including changes in the host and/or the virulence of the organism.Virulence factors that contribute to pathogenicity include dimorphism,increased adhesion,production of phospholipases, production of hemolysin,drug resistance,and antigenic modulation.Although there are many studies on the adherence of yeasts to mucosal surfaces, little is known of this critical step initiating fungal infection in HIV-infected individuals.While there have been a number of detailed studies on some of these hydrolytic enzymes,such as proteases,lipases,and phospholipases,little is known of the hemolytic activity exhibited by Candida species.Chapter One:Carriage frequency,species distribution of Candida spp.in the oral cavity of Chinese HIV-infected patients1.Objective:The present study was conducted to evaluate the frequency of colonization by Candida species and Candida species distribution in the oral cavity of HIV-infected patients.2.Methods:82 individuals(49 male and 33 female) diagnosed as HIV-infected patients by ELISA and Western-blot were included in the study.Oral rinses were collected and the yeast identification was performed by morphology in cornmeal agar,Gram staining reaction,CHROMagar Candida test and the API 20 C Aux,growth at 45℃.3.Result:Out of 82 HIV-infected patients studied,60(73.17%) had positive culture for Candida.The frequency of isolation of Candida in HIV-infected patients was:C. albicans,70%(42/60);C.glabrata,77.33%(44/60);tropicalis,1.67%(1/60);C. parapsilosis,3.33%(2/60);Candida pelliculosa,1.67%(1/60),others,3.33%(2/60). There were 51.67%(31/60) episodes of colonization by at least 2 different yeast species. Chapter Two:Carriage frequency,species distribution of Candida spp.in the oral cavity of Chinese healthy persons1.Objective:To evaluate the frequency of colonization by Candida species and Candida species distribution in the oral cavity of non HIV-infected persons.2.Methods:Oral rinses were collected in 82 individuals(49 male and 33 female) diagnosed as non HIV-infected persons.The yeast identification was performed by morphology in cornmeal agar,Gram staining reaction,CHROMagar Candida test and the API 20 C Aux,growth at 45℃.3.Result:Out of 82 HIV-free patients studied,17(20.73%) had positive culture for Candida.The frequency of isolation of Candida in HIV-infected patients was:C. albicans,64.71%(11/17);Candida parapsilosis,29.41%(5/17);others,5.88% (1/60).There were 1(5.88%)episodes of colonization by at least 2 different yeast species,Chapter Three:In vitro virulence of Candida spp.in the oral cavity of Chinese HIV-infected patients and healthy persons1.Objective:To examine the in vitro virulence of Candida species isolates from oral cavities of subjects with and without H IV-infection,including adhesion to healthy buccal epithelial cells,phospholipase activity and hemolytic activity.2.Methods:24 isolates were recovered from distilled water stored at -70℃.They were divided into 3 groups.A group:8 Candida albican isolates from oral cavities of subjects with HIV infection;B group:8 Candida glabrata isolates from oral cavities of subjects with HIV-infection;Cgroup:8 Candida albican isolates from oral cavities of subjects without HIV- infection.A loopful of the stock culture was streaked onto Sabouraud dextrose agar and incubated at 37℃for 18-24 h.(1) Adhesion to buccal epithelial cells.Human buccal epithelial cells(BECs) were used for the adhesion assay.The cells were collected by gently rubbing the cheek mucosa of healthy adult volunteers with sterile swabs and then resuspending the cells by rotating the swabs in 10 ml PBS.Equal volumes of BECs(1×10~5/ml) and yeast cell suspension(1×10~7/ml) were mixed and incubated at 37℃for 1 h on a shaking incubator at 1500 r.p.m.The cells were then filtered with a manifold filter apparatus through 12um pore polycarbonate filters.The filters were washed with 70 ml of PBS to get rid of unattached yeasts,removed carefully and then pressed gently onto glass slides cleaned with chromic acid.Afterwards the filters were carefully peeled off,thus leaving most of the BECs and the attached yeasts on the glass slide.The cells were air-dried and then stained by Gram\'s method.The number of yeasts attached to 100 epithelial cells was counted microscopically at a magnification of×400.Counting was undertaken randomly without prior knowledge of the source of the sample,and only uniform,unfolded epithelial cells were included.(2) Phospholipase detectionExtracellular phospholipase activity was detected using the egg yolk agar plate method.5 ul aliquots of the yeast suspension(approximately 10~8/ml) were inoculated onto the surface of the egg-yolk medium in quadruplicated samples,left to dry at room temperature and after incubation at 37℃for 48 h the diameter of the precipitation zone around the colony was determined.Each experiment was carried out on two separate occasions.The plates were read using a rule.Phospholipase activity(so called Pzvalue) was determined by the ratio of the diameter of the colony to the total diameter of the colony plus the precipitation zone.Thus,a Pz value of 1 indicates no activity,and less than one indicates the degree of phospholipase positivity.(3) Haemolysin production.Hemolysin production was evaluated using a modification of the plate assay described by Manns et al.The resultant cultures were harvested and washed with sterile saline,and a yeast suspension with an inoculum size of 10~8 cells/ml was prepared using hemocytometric counts.Ten microliters of this suspension was spot inoculated on a sugar-enriched sheep blood agar medium so as to yield a circular inoculation site of about 5 mm in diameter.The final pH of the medium so prepared was 5.6±0.2.The plates were incubated at 37℃in 5%CO2 for 48 h.The presence of a distinct translucent halo around the inoculum site,viewed with transmitted light, indicated positive hemolytic activity.The diameters of the zones of lysis and the colony were measured with a rule,and this ratio was used as a hemolytic index to represent the intensity of the hemolysin production by different Candida species.The assay was conducted in quadruplicate on two separate occasions for each yeast isolate tested.3.Results:H value of three groups were statistically significant(F=6.182,P=0.008).H value between A and C group were not statistically significant(P=0.682).H value between A and B group were statistically significant(P＝0.010).H value between B and C group were statistically significant P=0.004).Enzymatic activity was more pronounced in Candida albicans with 100%PL activity.In contrast,C.glabrata demonstrated only 25%PL.Pz value(The phospholipase activity) of three groups were statistically significant(F=25.562,P =0.000).Pz value between A and C group were not statistically significant(P= 0.660).Pz value between A and B group were statistically significant(P=0.000). Pz value between B and C group were statistically significant P=0.000).Hemolytic index of three groups were not statistically significant(F=0.284, P=0.756).Conclusion:1.Our results showed that the species isolated most frequently was Candida albicans and Candida glabrata in the oral cavity of HIV-infected patients.2.The species isolated most frequently was Candida albicans and Candida parapsilosis in the oral cavity of healthy persons.Asymptomatic oral Candida spp. carriage has been demonstrated in HIV-infected patients,and an increased incidence of asymptomatic oral Candida spp.carriage in HIV-infected patients compared to that in other at-risk groups has also been noted.Thus,a higher prevalence of oral Candida spp.colonization may be a predisposing factor for the subsequent development of clinical thrush.3.The pathogenetic isolates of Candida glabrata from HIV-infected patients were significantly lower than that of Candida albicans from HIV- infected patients and healthy persons in adhesion to buccal epithelial cells and phospholipase activities.There was no difference between Candida albicans isolates from oral cavities of HIV-infected patients and healthy persons in adhesion to buccal epithelial cells and phospholipase activities.And there was no significant difference in hemolytic index of Candida albicans isolates from oral cavities of subjects with and without HIV infection and Candida glabrata isolates from oral cavities of subjects with H IV infection.These results indicate that oral candidiasis was not correlated with some p redominant strains of Candida albicans with higher virulence, but was correlated with strains of Candida glabrata with lower virulence in H TV-infected patients.

Associations of Protein Tyrosine Phosphatase Nonreceptor 22(PTPN22) Gene Polymorphisms with RA in the Han Population of Guangdong in China

Thu, 31 Mar 2011 05:03:56 +0000

Research backgroundRheumatoid Arthritis(RA) is one of the most common systemic autoimmune disorders,showing the global distribution,afflicting up 0.32-0.36%of the population in our country,estimating five million people in china,causing enormous economy losses,and it is the main reason leads the workers lossing their labor ability and causes disability.The etiology of RA is complex and multifactorial,but genetic factors indicate a significant causal role.The etiology of RA is complex and multifactorial,genetic factors indicate a significant causal role,with heritability being estimated at 60%.Much of the research involving RA susceptibility factors has focused on the major histocompatibility complex(MHC) region on chromosome 6p21,which has been shown to account for approximately one-third of the genetic risk for RA.More recently,linkage data from genome-wide screens or mapping of multicase RA families identified a number of non-MHC chromosomal regions as possible RA susceptibility loci,and analyses of selected candidate genes within such regions revealed several gene variants as being associated with RA in some populations.Recently,a non-MHC gene has repeatedly and convincingly been found to be associated with RA and several other autoimmune diseases,namely the protein tyrosine phosphatase nonreceptor 22(PTPN22) gene,which be identifed as the key susceptibility gene of RA.The PTPN22(protein tyrosine phosphatase nonreceptor22) gene,located on chromosome 1p13,encodes a lymphoid-specific phosphatase(Lyp), including sixteen exons and fifteen introns,the PTPN22 variant(1858C→T) engenders a substitution in Arg620Trp that disrupts Lyp binding to Csk and would thereby predictably reduce or abrogate the inhibitory effect of the Lyp-Csk interaction on T cell activation.The PTPN22 1858T variant may be the etiologic relevance in multiple autoimmune diseases.Although the recent evidence showing that the 1858C→T SNP is susceptibility to multiple autoimmune diseases in European,the frequency of the 1858C→T SNP is almost absence in Asian and African people,showing a obviously genetic heterogeneity.By sequencing 21 exons, 16 introns,and the promoter region of PTPN22 on chromosome 1p13,Kawasaki etal found five novel SNPs,the snp of 1123G＞C(rs2488457) in the promoter region is associated with type 1 diabetes in Asian populations.The PTPN22-1123C variant may be another mutation associates with multiple autoimmune diseases and highly appears.Given the 1858C→T SNP relevants to the susceptibilities of multiple autoimmune diseases in European Caucasian populations,furthermore,both chinese people and japanese people have a similar genetic background,belong to Asia people of yellow race,we assume that the snp of 1123G＞C might also be etiologically relevant to the RA patients of china.Recently,there is no association data about the PTPN22 1123G＞C relevants to RA predisposition in yellow Asia people.To determine whether association of the PTPN22 1123G＞C variant with RA occurs in RA cohorts of the han population in the Guangdong province of china,the frequency of this variant was investigated by denaturing high performance liquid chromatography and DNA sequencing.The mechanism of the PTPN22 1123G＞C influences the transcription activity of PTPN22 is poorly understood.To investigate wheather this variant impacts the transcription activities of PTPN22,fluorescence quantitative PCR was used to assay wheather the gene expression of PTPN22 was difference between those carrying the-1123C risk variant and those not carrying the risk variant.The purpose of this study is to provide a theoretical basis to the deeply research of the exact mechanism how the PTPN22 1123G＞C SNP influences the transcription activities of PTPN22,and provides new ideas to the early diagnosis of RA.Finally,use the polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP) analysis and DNA sequencing technology to detecte the frequency of C1858T polymorphism in RA patients of the han population in the Guangdong provinc of china.Research Methods1 subjectsFor the RA patient cohorts,patients were recruited from South Medical University hospital from March 2007 to December 2008,all patients with RA met the American College of Rheumatology(formerly,the American Rheumatism Association) 1987 revised criteria for RA,includeing 27 males and 121 females,all the patients are from Guangdong province,except 10 people from Hunan Province;4 people from Jiangxi Province;and 3 people from Fujian Province.Control subjects for this study included 41 males and 111 females,none of the healthy anonymous volunteers with other inflammatory arthritis or severe heart,kidney and liver diseases.2 Research Methods2.1 Phenol-chloroform extracts the genomic DNA,designing specific primers,and PCR amplifies the fragment.2.2 Use the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP)analysis and DNA sequencing technology to detecte the C1858T polymorphism of PTPN22 gene. 2.3 Using the denaturing high performance liquid chromatography(DHPLC) method, combined with DNA sequencing technology to detect the G-1123C polymorphism of PTPN22 gene.2.4 Relative risk/odds ratios(ORs) excluded the impact of age and gender were calculated by using standard logisticregression methods and were processed with the SPSS 13.0 software.Subdivision of the RA patients into six groups,such as the group of X-ray positive,X-ray positive,RF factor positive,RF factor negative,age at onset more than 42 years old,or age at on set less than 41 years old,respectively compared to the healthy control group.Stratification nalysis was performed on the RA case cohort to investigate the effect of sex,disease severity(as assessed by the presence of erosive disease),RF status,and age at disease onset.2.5 There are only 52 patients have complete data of clinical characteristics among the 148 cases of clinical diagnosis RA patients.According to the genotype of PTPN22-1123 SNP,RA patients were divided into heterozygous mutations(GC type) RA group,homozygous mutant(CC-type) RA group,as well as the wild-type (GG Type) RA group,respectively assessing clinical similarities among the three different patient cohorts.2.6 We detected the PTPN22 gene expression levels in the patients with mutation and healthy control group to GAPDH housekeeping genes as reference genes by fluorescent dye SYBR Green quantitative PCR,calculating by△CT,using an independent samples t test for statistical analysis,finally,using 2~(-△△CT) to calculate the difference of gene expression between the two groups.2.7 Statistical MethodsThe frequencies of PTPN22 alleles were established by direct counting. Association was evaluated using the chi-square test.Relative risk/odds ratios(ORs) were calculated using standard logistic regression methods and were processed for the analyses using SPSS13.0 statistical software.A one-way ANOVA test was used to determine significance among multiple samples means,and a nonparametric Mann-Whitney test was used for assessing clinical similarities between the three patient cohorts.A P-value less than 0.05 was considered statistically significant. Results1,The PTPN22 gene G-1123C polymorphism is associated with RA in Chinese Guangdong Han population.RA patients were different from healthy control individuals in frequencies of PTPN22 G-1123C genotypes(P=0.016) and carded C allele(P=0.000;OR,1.837;95%CI,1.328～2.541).Excluding the impact of sex and age,logistic regression analysis suggested that there were still statistically significant difference in the frequencies of PTPN22 G-1123C genotype(P= 0.021).The presence of CT heterozygote was very strongly associated with RA((P= 0.006;OR,2.051;95%CI,1.234～3.410),but the CC homozygotes was not(P= 0.427).2,The frequency of PTPN22 gene C1858T polymorphism in Chinese Guangdong han population was 0,only one phenotype CC existed in the148 RA patients and 151 healthy control group members.Therefore,this SNP site was not associated with RA in Chinese Guangdong Han population.3,The PTPN22-1123 allele was association with the presence of RF.The allele frequency of PTPN22G-1123C was association with rheumatoid factor-positive (RF+) RA(P=0.041).4,None of the clinical characteristics was found significant difference among the three RA patient cohorts.All the P-value were more than 0.05.5,The result of real-time fluorescence quantitative PCR showed that the PTPN22 mRNA expression of mutation patient group(including the heterozygous and homozygous mutant-type RA patients) was 3.742 times to the control group (wild-type health control).Conclusion1,The polymorphism of PTPN22 gene C1858T SNP was not associated with RA in Chinese Guangdong Hart population,while the other SNP G-1123C may be related to RA susceptibility,suggesting that the polymorphism of PTPN22 G-1123C was possible another risk allele for RA,the OR of heterozygous mutation is 2.051 times to the control group,but CC homozygotes was no correlation,suggesting molecular heterosis.2,The allele frequency of PTPN22G-1123C was association with the presence of RF, and no association with the effect of sex,disease severity(as assessed by the presence of erosive disease),and age at disease onset.None of the clinical characteristics was found significant difference among the three RA patient cohorts. It is essential that the number of our inception cohort should be expanded in the future studies.3,The PTPN22 mRNA expression of mutant patients group was higher than the wild-type healthy control group,suggesting that it may be a compensatory mechanism to the function defect of mutant patients.Examining the DNA sequence around the -1123G＞C site,we could find it showed its perfect match with that of the binding site for transcription factor activatorprotein 4(AP-4),a member of the basic helix-loop-helix-zipper family of transcription factor on the antisense strand.The -1123G＞C is located at the critical region of the coremotif of the AP-4 binding consensus sequence(「1130-gcaaGCTGaa-1121;core motif is shown in uppercase, the position「-1123G is in bold) and the「-1123C allele did not correspond to binding elements of any known transcription factors.Thus,it is suggested that heterosis at the PTPN22 promoter may be related to the regulation of the expression of Lyp and confered the development of specific disease subphenotype. 4.Our research provides a new way of thinking for the further study of the patheogenesis of RA,and offers a theoretical basis to the further study of the exact mechanism of PTPN22 G-1123C SNP impact LYP expression.

Clinical Observations on the Immediate Effect of the Acupuncture Treatment of Arteria Vertebralis Type of the Cervical Spondylosis by Taichong Point

Thu, 31 Mar 2011 05:03:54 +0000

Objective:The research used the cervical vertigo symptoms and functional assessment scales for the assessment tools,to observe the immediate effect of the acupuncture treatment of arteria vertebralis type of the cervical spondylosis by Taichong Point.To study the clinical effect of Selection remote treatment of it.And to provide a few reference for clnical treatment by remote point selection.Method:Choosing 40 cases of the arteria vertebralis type of the cervical spondylosis based on strict diagnostic criteria and exclusion criteria.Then according to random number table 40 cases were randomly divided into 2 groups:observer group 20 cases and control group 20 cases,observer group acupuncture Taichong Point only,and control group acupuncture Jiaji Point.Score the degree of vertigo,neck pain and headache,which chosed from the cervical vertigo symptoms and functional assessment scales for the assessment tools, when before treatment and after that.Then analyse by statistics method.Result:(1) There was a notable statistical significance on vertebralis,on headache and on genenal condition between before and after treatment in observer group(p＜0.01),but no statistical singnificance difference on neck pain(p＞0.05).(2) There was a notable statistical signigicance on vertebralis,on headache,on neck pain and on genenal condition(p＜0.01)(3) There was a notable statistical signifinance on vertebralis,on headache and on neck pain between observer group and control group,but no statistical singnificance difference(p＞0.05).Conclusion:(1) Acupuncture Taichong Point can distinctly improve the condition of the arteria vertebralis type of the cervical spondylosis.(2) Acupuncture Taichong Point can distinctly relieve the vertigo and headache of the arteria vertebralis type of the cervical spondylosis. (3) Acupuncture Taichong Point can\'t distinctly relieve the neck pain of the arteria vertebralis type of the cervical spondylosis.(4) Acupuncture Taichong Point and acupuncture by Jingjiaji Point can get the effect for the arteria vertebralis type of the cervical spondylosis both,but acupuncture Taichong Point can get much more effect on vertigo and headache to acupuncture Jingjiaji Point,but short of the neck pain.On the whole,the effect is little difference between acupuncturing Taichong Point and acupuncturing Jiaji Point.

Activation of GABAergic Neurons in Mouse Brain during Anesthesia Induced by Propofol

Thu, 31 Mar 2011 05:03:53 +0000

General anesthesia has developed 160 years, but the mechanism underlying is still unknown, especially in central nervous system. The researches on mechanism of general anesthesia have always been the unode and focus.Poropfol has been used in induction and maintenance because of its pharmacological merits recent years. But the effect of poropfol on central nervous system is unknown. In vivo and ex vivo researches indicated that poropfol can enhance the binding of GABA and GABAA receptor. There are few research about the presynaptic mechanism. Whether poropfol has effect on presynaptic neurons is the focal point of this study.Objective: Emerging researches have focused on the participation ofγ-aminobutyric acid (GABA) neurons but there still lacks morphological evidence. To investigate the activation of GABAergic neurons during loss of consciousness induced by propofol in the GAD67-GFP knock-in mouse. Methods: 20 GAD67-GFP transgenic mice were used in the present study. The mice were grouped into two, control group and experimental group. Experimental group were grouped into three subsets, T1(5 min), T2(30 min), T3(1 h). Mice in experimental group were injected propofol i.p. 130 mg/kg, and saline was set as the negative control. Mice were graded by ethology while sacrificed at 5 min, 30 min and 1 h after injection, respectively. The whole brain was obtained and cut with a freezing microtome consecutively. Immunohistochemistry and double immuno-fluorescence were performed to observe the Fos expression and the coexistence of Fos and GABA in the whole brain.Results: 1 The scores of ethology were 13 at 5min, 14 at 30 min and 9 at 1h. It indicated that the mice were in the situation of moderate or deep anesthesia at 5 min and 30min, and various transition states at 1h. Expression of Fos protein increased remarkably in CA1, amygdaloid nucleus, thalamic paraventricular nucleus, hypothalamic paraventricular nucleus, hypothalamic ventromedialis nucleus, and periaqueductal gray compared with the control group at three time spots(p<0.05). 2 Neurons expressing Fos protein and GABA simultaneously could be observed in hypothalamic ventromedialis nucleus at 5 min, 30 min and 1 h, the proportion of which among Fos-positive neurons was 80.3%, 89.7% and 91.6%. The double staining neurons could also be seen in PaV, the proportion of which among Fos-positive neurons was T1:19.6%,T2:39.7%,T3:41.6%, C:8.6%。Conclusion: The activation of GABAergic neurons may be one of the mecha- nisms during loss of consciousness induced by propofol.

Establishment of Computerized Analysis System for the Arch Length Measurement

Thu, 31 Mar 2011 05:03:51 +0000

Dental cast is an important medium to record and preserve the information about patient’s teeth, dental arch, alveolar, basal bone, palate and occlusion. The result of dental cast measurement can help orthodontics diagnose and work out treatment plan. Dental crowding is one of important factors for the decision of extraction and non-extraction treatment. Dental crowding is defined as a discrepancy between tooth size and arch available length that results in a misalignment of the tooth row. Arch available length is an important part of dental crowding analysis. The accuracy of arch available length measurement directly affects analysis of dental crowding and treatment plan. Now the common measurement methods on clinic are wire and dividers. We find that the results measured by the two methods for a same cast are different, and the results measured by different people or at different time are different. Therefore, in this study, the differences of the repeatibility and reliability between results measured by the two ways were analyzed systematically. Then, we established digital models and computer program for arch length measurement to improve the reliability and repeatibility. And the established way was applied on the analysis of the Spee curve and the space requirement for levelling the Spee curve.The thesis consists of two parts:Part one is the comparative study of wire and dividers applied in orthodontic dental models of arch length measurement. This study shows that it’s necessary to establish the computer analysis system for arch length measurement. Methods: 80 pairs of dental models were selected, including 40 normal occlusion models and 40 crowded models. The arch length of every model was measured with the methods of wire and dividers respectively, and the casual error and paired t test was subsequently employed to compare the differences of the measurements obtained. Results: The variance coefficients of the dental length results measured by the dividers was larger than that measured by the wire no matter in the models with crowded or normal occlusion(P〈0.05). The dental arch length measured by the wire method was significantly larger than that measured by the dividers method(P〈0.05). The differences between the measurement results in the two methods were 4.4±3.0mm and 5.2±1.7mm in the dental models with crowded and normal occlusion, respectively. It is suggested that the repeatibility and reliability of the two ways are not enough for the clinical diagnosis needs. Therefore, we decided to establish a computerized analysis system for arch length measurement.Part two is the establishment of the computerized system for the arch length analysis. Methods: Dental cast was scanned by a 3D (three dimensional) laser scanner (Minolta Vivid 910) to form a digital model. The mathematical method was used to evaluate the dental arch length. Results: Digital dental model measurements could be analyzed in plane and in space. The results were more reliable and repeatable than clinical methods of wire and dividers. The calculation of the space for levelling the curve of Spee in the digital model was more accurate than clinical estimation.The study began with the problem of clinical methods of wire and dividers in arch length measurement. Then we established computerized system applied in arch length analysis of dental cast. Based on the system, the reliable information could be provided for the orthodontic treatment plan decision. The established system can also be used for preservation of dental model as 3D digital model, manifestation of 3D digital dental model, assistance of teaching, etc.

Calcium, Vitamin D and Risk of Colorectal Cancer in Canadian Populations

Thu, 31 Mar 2011 05:03:50 +0000

Objective Despite evidence from animal studies for a protective effect of higher calcium and possibly vitamin D and dairy product intake against colorectal cancer (CRC),epidemiologic studies have been inconclusive.We therefore examined the association between these factors and risk of colorectal cancer in a large case-control study from Newfoundland(NL) and Ontario(ON).Methods We analyzed data from a population-based case-control study of more than 4,000 participants from Newfoundland and Ontario,Canada.In NL incident cases diagnosed during 1999-2003 were identified through the Newfoundland Familial Colorectal Cancer Registry(NFCCR).In ON incident cases diagnosed during 1997-2000(PhaseⅠ) and 2003-2006(PhaseⅡ) were identified through the Ontario Familial Colorectal Cancer Registry(OFCCR).Both registries were used to identify newly diagnosed cases of colon or rectal cancer and aged 20-74 years.Controls were a sex and age-group matched random sample of the population of each province. Intakes of calcium and vitamin D from diet and supplements and consumption of dairy products were assessed with a baseline food frequency questionnaire(FFQ). Calcium,vitamin D and dairy products were categorized into quartiles based on the distribution among the study population with lowest quartitle as referent group. Multivariate logistic regression analysis was used to examine the association between these factors and risk of colorectal cancer after adjusting for age,gender,total energy intake,BMI,physical activity,and nonsteroidal anti-inflammatory drug(NSAID) et al.The trends were tested with the medians of the intake as a continuous variable in the logistic model.The P values for the trends were two-sided.Results Total calcium intake(from diet and supplements) was significantly associated with lower risk of colorectal cancer in NL and ON,particularly among women(ORs highest vs lowest quartile were 0.49,95%CI:0.26-0.92,P_(trend)=0.04 in NL;and OR=0.53,95%CI:0.37-0.76,P_(trend)=0.02 in ON).Total vitamin D intake(from diet and supplements) was inversely associated with risk of colorectal cancer in NL (OR=0.52,95%CI:0.36-0.74,P_(trend)=0.04).Dietary Calcium and vitamin D intakes were associated with reduced risk in ON women(OR=0.64,95%CI:0.44-0.93, P_(trend)=0.03 for dietary calcium;OR=0.68,95%CI:0.49-0.94,P_(trend)=0.04 for dietary vitamin D),while the use of calciumlcium and vitamin D supplements were associated with lower risk in NL men(OR=0.53 for calcium supplements;OR=0.60 for vitamin D supplements) and NL women(OR=0.46 for calcium supplements; OR=0.46 for vitamin D supplements) and ON men(OR=0.69 for calcium supplements;OR=0.76 for vitamin D supplements).Consumption of high fat dairy products was associated with increased risk of CRC in NL(OR=1.54,95%CI:1.09-2.17),while consumption of non-fat milk and yogurt was associated with reduced risk in ON(OR=0.53 for non-fat milk;and OR=0.60 for yogurt).The inverse association for total calcium intake was strongest in cancer of rectum(OR=0.56,95%CI:0.32-1.00 in NL;OR=0.74,95%CI:0.55-1.00 in ON), while the inverse association for total vitamin D intake was limited to cancer of colon (NL:OR=0.65,95%CI:0.42-0.98).The inverse association for calcium supplements use was strongest among men with lower intake of dietary calcium(OR=0.29 in NL, OR=0.67 in ON).The inverse association was strongest among women with higher intake of total calcium and total vitamin D together(NL:OR=0.49;ON:OR=0.38)Conclusions High calcium and vitamin D intakes,particularly from supplements,are associated with a reduced risk in NL population.High calcium and vitamin D intakes, particularly from food,are associated with a reduced risk of CRC in ON females, while the inverse association was strongest for calcium from supplements in ON males.Consumption of high fat milk may increase risk,while consumption of skim milk or yogurt may decrease risk.The inverse association for total calcium was strongest in rectum cancer,while the inverse association for total vitamin D was limited to colon cancer.Calcium supplements use appears to have further benefits among males with lower intake of dietary calcium.Women with higher intakes of calcium and vitamin D together had the lowest risk of colorectal cancer.In conclusion, calcium and vitamin D intakes were inversely associated with colorectal cancer risk.

Preparation, Storage and Experimental Conditions of Biofluid Samples for NMR Based Metabonomics

Thu, 31 Mar 2011 05:03:48 +0000

The NMR-based metabonomics approach evolved from the pioneering work of Nicholson and co-workers in 1999 has become a novel analytical technique.At present,main targets of NMR-based metabonomics experiments are biofluids,and the experiment results are sensitive to the experimental conditions,sample storage and preparation.Study of these external conditions is important to keep the stability and comparability of biofluid sample data.In this paper,we investigated the effect of experimental conditions of human urine and serum samples,the effect of storage and preparation of human saliva samples.Some parameters are fully researched and optimized.The main contents of this study are as follows:1.Investigations of the effect of experimental conditions to human urine sample NMR spectra.Resonances of acetyl,lycine and histidine in urine sample shift as the temperature changes;signals whose chemical shifts are closed to that of water would decrease their intensity with the saturation power rising.Those results demonstrate that trade-off between the water suppression and signal decrease must be taken into consideration;External method is suitable.2.Investigations of the effect of experimental conditions to human serum sample NMR spectra,intensity of resonance signal of LDL,VLDL,valine and choline would increase obviously in the serum spectra with the experimental temperature increasing.Those evidenccs implied that it is important to acquire the 1H NMR spectra of the samples in the same temperature;internal method is suitablc.3.Investigations of the effect of storage and preparation conditions to human saliva sample NMR spectra.The results indicated that human saliva should be stored at or below -25℃,as no changes in the ~1H NMR fingerprints have been observed during 6 weeks. Formation of phenylalanine and formic acid,probably because of microbe activity,was observed in samples stored at 4℃.Addition of NaF is effective to keep saliva sample at -25℃,whereas the increase of acetate in saliva sample with NaF at the 6th week reflects it is not suitable for long term storage.A study evaluating the effects of phosphate buffer concentration on signal variability led to the conclusion that a higher concentration is adequate for normal saliva.For the comparability of NMR spectral data,making a standard of sample storage and experiment progress is useful and this work needs us to keep on researching in the future.

The Mechanism of IL-6 Effects on Glucose Metabolism in Skeletal Muscle of SD Rat with T2DM

Thu, 31 Mar 2011 05:03:47 +0000

ObjectiveThe objective of this study was to explore the mechanisms of Interlukin-6(IL-6) influencing the glucose metabolism and the expression of the insulin-sensitive glucose transporter 4(GLUT4) in skeletal muscle of type-2 diabetic rat model.MethodsThirty-six healthy male Sprague Dawley(SD) rats were randomly divided into 4 groups:group N was control rats with routine diets(n=10);group A,B and C were experimental groups fed with high-glucose-fat diets and induced type-2 diabetic rat models by injection of low-dose of streptozotocin(STZ) on the seventh week.Then, group A was introceliac injected IL-6 diluents for 5 days(n=10);Group B was introceliac injected PBS for 4 days and IL-6 antibody diluent on the following day (n=8);group C was introceliac injected PBS for 5 days.The blood samples were collected to determine blood glucose,serum insulin and lipid at the end of eight weeks and then the rats were sacrificed.HE staining was used to evaluate the skeletal muscle pathological changes by light microscope.Muscle glycogen was tested with colorimentry.The expression of GLUT4 in the rat skeletal muscle was tested with immunohistochemistry analysis.All data were analyzed by using SPSS 13.0 PC program for windows.ResultsAt the end of seventh week,there was no significant difference in blood glucose in the sense of statistics(P＜0.05) between group A,B,C and N respectively;the serum insulin of group A,B and C was significantly higher than group N(P＜0.05).Before the rats were sacrificed,there was no significant difference in serum insulin between group A,B,C and group N(P＜0.05);the serum TG,TC,LDL,FFA of group N was significantly lower than group A,B and C(P＜0.05).The content of muscle glycogen in the rat skeletal muscle was higher in group B,N compared with group A and C (P＜0.05).The expression of GLUT4 protein in group N was also significantly higher than that in group A,B,C(P＜0.05).Strikingly,the expression of GLUT4 protein in group C was dramatically increased compared with group A and B after IL-6 antibody treatment(P＜0.05).Conclusions1,High-glucose-fat diets for 7 weeks may cause high insulin concentration and insulin resistance in rats,but not high blood glucose.2,GLUT4 protein expressed lower in the skeletal muscle of STZ induced DM rats than in controls.3,IL-6 blockage increased the expression of GLUT4 protein and content of muscle glycogen in skeletal muscle of STZ induced DM rats,but muscle glycogen was not increased in corelation with the expression of GLUT4 protein.That is to say,GLUT4 protein has internal vitality.

Risk Factors in Patients with Degenerative Lumbar Spondylolisthesis

Wed, 30 Mar 2011 17:01:19 +0000

【Background】Degenerative lumar spondylolisthesis(DLS) is the slippage of one vertebra relative to the adjacent vertebrae due to spinal degenerative changes.The main manifestations are mechanical low back pain,the radicular pain with numbness, paraesthesia,and sensory or motor deficit,and neurogenic claudication.Unlike isthmic spondylolisthesis,degenerative spondylolisthesis is not identifiable defect in the posterior neural arch.It has traditionally been considered as one of the major causes of low back and leg pain among the elderly.When symptomatic,the resultant neurogenic claudication often leads to a diminished quality of life.Spondylolisthesis in the absence of a pars defect was first described as early as in 1930 by Junghanns.The term "degenerative spondylolisthesis" was coined by Newman in 1955,who noted that slippage of the vertebrae with an intact neural arch was the result of degenerative arthritis of the lumbar facet joints.Previous studies showed that an overall incidence of degenerative spondylolisthesis was 8.7%.This condition typically presents in individuals older than age 50 years and is four times more common in females than in males.Predominantly affects the L4-L5 level and much less frequently L5-S1 or L3-L4 levels,and the degree of slip rarely exceeds 30%.Degenerative spondylolisthesis(DS) was first described by Junghanns in 1930, many studies have been undertaken in an attempt to explore its etiology and pathomechanisms,but so far this are still controversial.It has been thought that excessive mechanical stresses at the L4-5 level may cause degenerative changes in the disc and lumbar facet joints,and with disc and facet joints degeneration,which eventally lead to segmental instability in the sagittal plane.Degenerative changes causing wear and anterior deformation of the articular processes,and the facet joints orientation are gradually more sagittally and horizontally,eventually allow the 4th lumbar vertebra to slip forward.But the pars interarticularis of a vertebra is not defect, so this type of spondylolisthesis is also called "pseudo-spondylolisthesis".Pregnancy, generalized joint laxity,and oophorectomy are thought to be predisposing factors of this condition and its predominance in females.Sagittal orientation of the facet joints have been described as predisposing factors.However,it is unclear whether the facet joints orientation is a primary cause or a secondary effect,and this hypothesis does not explain the mechanism of anterior vertebral slip in patients with a coronal orientation of the facet joints.It may be additional predisposing factors involved in the process of vertebral slipping.Degenerative lumbar spondylolisthesis is not an isolated disease,which is an integral part of the spinal degenerative changes and often accompanied by lumbar hyperosteogeny,lumbar disc bulging or herniation,and facet joint\'s disease at a latter stage.Spondylolisthesis often lead to spinal stenosis,lateral recess stenosis,the ligamentum flavum hypertrophy,and a series of pathological changes,which increase the symptoms and affect the patients quality of life seriously. Therefore,early prevention,diagnosis and to take reasonable measures to this condition may alleviate patient\'s pain and improve the quality of life,and it\'s important clinical significance,many studies have been taken to explore the etiology and pathogenesis of Degenerative Lumbar Spondylolisthesis in the past years,but these studies often be carried out from One aspect of this field.Because the causes and pathogenesis of degenerative lumbar spondylolisthesis are extremely complex and are the result of the interaction of various factors,the study of single factors maybe exaggerate the role for some certain,and the credibility of the conclusion is not enough.In current study,a retrospective case-control study,using single-factor screening,multi-factor logistic regression analysis to explore the risk factors of degenerative lumbar spondylolisthesis and to provide a theoretical basis for prevention and treatment of degenerative lumbar spondylolisthesis,and to lay the foundation for further study.【Objective】1.To find out the risk factors which contribute to degenerative lumbar spondylolisthesis.2.To explore the correlation between age and orientation of the lumbar facet joints.3.To provide a theoretical basis for prevention and treatment of degenerative lumbar spondylolisthesis,and to lay the foundation for further research.【Methods】1.From March 2004 to March 2009,200 cases of degenerative lumbar spondylolisthesis(DLS) in our hospital were review.200 cases without spondylolisthesis were studied as control group.Age,gender,body height,weight, body mass indes(BMI),history of diabetes,imaging indicators,including the facet joints orientation,the severity of degeneration of facet joints,the pedicle-facet angle,the pelvic incidence,pelvic tilt,sacral slope,disc height ratio, lumbar intervertebral disc angle,lumbar lordosis,lumbosacral angle and intercrestal line using single-factor screening and multi-factor Logistic regression analysis.2.300 spinal disease-free people coming for periodic physical examination during May 2007 and May 2008 in our hospital accepted lumbar CT scanning and were randomly selected.All the subjects were divided into 6 groups based on age (GroupⅠ:less than 30 years of age,GroupⅡ:31～40 years,GroupⅢ:41～50 years,GroupⅣ:51～60 years,GroupⅤ:61～70 years,and GroupⅥ:greater than 70 years of age).CT scans of the lumbar spine were stored in digitized form for measurement and analysis.The orientation of the lumbar facet joints at the L4-5 was measured on axial CT scans,and the intersection angle of the midsagittal line of vertebra to the facet line represent the orientation of the facet joint. 3.Statistical analysis:Statistical analysis was performed using SPSS13.0 statistical software.Measurement data using(?)±SD.Comparison between two groups,if they were measurement data,unpaired t test would be used,if they were enumeration data or ranked data,we would apply Mann-Whitney.Comparison between the groups and multiple comparisons were conducted using variance analysis and LSD respectively.The correlation analysis was performed with Pearson method.All the independent variables were analyzed with the single factor analysis and multifactor logistic regression analysis.The criterion for statistical significance was P＜0.05.【Results】1.The general clinical data and imaging measurement indicators of DLS and control groups.Total cases-400,there were 200 cases in DLS group,200 cases in control group.Age,DLS group(61.11±10.25) years old(37～82ys),control group (59.82±10.02)years old(33～80ys).(P=0.206,t=1.268 ).Gender,DLS group,male 37 cases,female 163 cases,control group,male 41 cases,female 159 cases.(P=0.705,χ~2=0.255).History of diabetes,DLS group 51 cases,control group 15 cases.(P=0.000,χ~2=23.517).Body height,DLS group(159.24±6.29)cm,control group(160.31±5.88)cm. (P=0.080,t=1.758)Weight,DLS group(66.99±11.09)Kg,control group(52.76±7.03)Kg.(P=0.000, t=15.335).Body mass index(BMI),DLS group(26.25±2.75)kg/m2,control group (20.49±2.19)kg/m2.(P=0.000,t:23.155).Facet joints orientation(FJO),DLS group 33.53°±8.75°,control group 47.74°±5.52°.(P=0.000,t=19.417).Pedicle-facet angle(PFA),DLS group 115.52°±55.28°,control group 102.51°±4.54°.(P=0.000,t=26.428)Pelvic incidence(PI),DLS group 58.98°±7.39°,control group 51.32°±7.60°. (P=0.000,t=10.215).Pelvic tilt(PT),DLS group 22.06°±7.90°,control group 11.21°±2.75°. (P=0.000,t=18.347).Sacral slope(SS),DLS group 36.92°±6.03°,control group 40.12°±8.36°. (P=0.000,t=4.384).Lumbar lordosis(LL),DLS group 27.90°±6.40°,control group 40.58°±7.63°. (P=0.000,t=18.008).Lumbosacral angle(LSA),DLS group 39.63°±3.46°,control group 36.21°±2.38°. (P=0.000,t=11.529).Disc height ratio(DHR),DLS group 0.23°±0.05°,control group 0.27°±0.48°. (P=0.000,t=8.068).Lumbar intervertebral disc angle(LIDA),DLS group 4.13°±2.51°,control group 12.38°±4.08°.(P=0.000,t=24.350).The severity of degeneration of facet joints,DLS group,Mean rank 266.26, control group,Mean rank 134.75.(P=0.000,Z=11.898).Intercrestal line(ICL),DLS group,Mean rank 218.18,control group,Mean rank 182.82.(P=0.000,Z=3.612).The degree of spondylolisthesis of DLS Group was 8%～37%,with an average 18.81%±6.29%.2.Result of binary logistic regression.DLS--YPFA--X1 FJO--X2BMI--X3PI--X4Equation,Y=-606.689+5.504X1-2.397X2+9.098X3-11.300X4(R=0.75,P＜0.001).Analysis of this model suggests that PFA,FJO,BMI and PI exert significant influences to DLS.The greater of P-F angle,BMI and PI,and the smaller the angle between the sagittal plane of vertebral body and facet joint line are more susceptible to the DLS.【Conclusions】1.In recent study,the history of diabetes,weight,body mass index,the facet joints orientation,the pedicle-facet angle,pelvic incidence,pelvic tilt,sacral slope, lumbar lordosis,disc height ratio,lumbar intervertebral disc angle,the severity of degeneration of facet joints as well as the intercrest line comparison between DLS group and the control group were significantly different.2.The binary logistic regression model suggested that the increase of P-F angle, BMI and PI were risk factors of DLS.3.Our study found that with advancing age,the coronal orientation of the lumbar facet joints gradually decreased.The coronal orientation of the L4-5 facet joints was negatively correlated to age.

Studies on the Measuring Accuracy of Root ZX and Propex

Wed, 30 Mar 2011 17:01:18 +0000

Objective: With the teeth in vitro as research objects, some factors which may have influence on the measurement accuracy of Propex and Root ZX would be studied, including completely dry root canal, irrigate liquids and the size of the apical foramen, besides, the comparative study on the measurement accuracy of the two devices would also be brought into the research scope.Methods: Experiment 1: Thirty teeth in vitro were implanted into the model in vitro and then measured orderly by Root ZX (A) and Propex (B), both of which were operated according to their own instructions. The working length of root canal (LR or LP) was obtained by EAL (Electronic apical locator) when the root canals were filled with different irrigate fluids, including completely dry, 3% H2O2, 0.9% NaCl and 2.5% NaClO. For the teeth which have been measured, the micro-observation of root apex was done and then the actual working length of root canal (LW=L-D) was obtained by means of software calculation. Finally, the difference value between LW and LR/LP was calculated, and the statistical analysis for the difference value was done. Experiment 2: The 40 teeth in vitro were randomly divided into four groups (A, B, C, D) with 10 in each group. To get the ones with different diameter (0.4 mm, 0.6 mm, 0.8 mm, 1.0 mm), the root canal were pretreated orderly by different size K file ( type 15~40、type 15~60、type 15~80、type 15~100). Under microscope, for the four groups (A, B, C, D), different type K files (type 40, type 60, type 80, type 100) were respectively used to measure the actual length of the root canal (L). And then the teeth in vitro were implanted into the model in vitro and then measured orderly by Root ZX (A) and Propex (B), both of which were operated according to their own instructions. And the length of root canal (L_(R0) or L_(P0)) was obtained by EAL. Finally, the difference value between L and L_(R0)/L_(P0) was calculated, and the statistical analysis for the difference value was done. Results: The results showed that the measuring deviations of Root ZXand Propex had no statistical significance (p>0.05) when the root canals were filled with different irrigate fluids, including completely dry, 3% H2O2, 0.9% NaCl and 2.5% NaClO. But the measuring deviation between Root ZX and Propex had statistical significance (p0.05).Conclusions: 1. When the root canals were completely dry or filled with the same irrigate fluid, the measuring deviation of Propex was smaller than that of Root ZX, but the differences between them had no statistical significance. 2. The size of apical foramen and the size of file could affect the accuracy of Root ZX and Propex, respectively. The bigger the size of apical foramen and the smaller the size of file, the greater the deviation. So files which corresponding to root canals should be recommended to the clinical application, when measuring the working length of root canals.

Study of the Essence of the Spleen QI Deficiency Based on Metabolomics of Urine

Wed, 30 Mar 2011 17:01:17 +0000

Objective: The study of spleen qi deficiency essence is an important element in syndrome essence. Traditional Chinese Medicine believes that spleen qi deficiency could lead to dysfunction of transportion, and stagnation of body fluid which can produce dampness, phlegm, drink as pathological products, thus triggering a variety of diseases. As the theory goes, " internal injury of spleen and stomach can cause a lot of diseases" "spleen is the source of sputum crudum ". In modern medicine, the spleen is a unit of multi-systems, multi-functional organs, which is based on the digestive system, including energy metabolism and water metabolism, and is closely connected with blood, endocrine, immune and nervous system. Metabolic syndrome(MS) is a disease which is seen as central task of central obesity, it involves a variety of metabolic abnormalities, including blood pressure, blood sugar, blood lipids, and it is associated with cardiovascular disease closely. The main etiology of MS is obesity. Phlegm-dampness is an important pathological factor in obesity, which is also the center of MS, so the phlegm-dampness is the central pathology factor in the main syndromes of MS.This shows that the spleen deficiency which causes the dysfunction of transportion is important in the pathogenesis of MS. Therefore the morbidity of MS is inseparable with the spleen. Metabolomics is an emerging science and is applied widely in the medicine domain. It measures small molecules of the metabolite of body fluid and tissue qualitatively or quantitatively by modern instrumental analysis methods such as nuclear magnetic resonance (NMR), chromato- graph(GC), mass spectrum(MS), reveals the physiological and pathological state in disease process systematically, however this systematic character- ristic observation is similar to holistic concept in the Traditional Chinese Medicine. At present, metabolomics has already been seen as a new technology to apply in diagnosis of clinical disease. We investigated the small molecular metabolite in urine of Metabolism syndrome patients as the foundation of spleen qi deficiency using method of disease and syndrome with combination and liquid chromatography-mass spectrometry technology. We expect to discover the change pattern or characteristic peak in metabolome through comprehensive and integrated analysis, attempting to express qualitatively or quantitatively in diagnosing spleen qi deficiency of Traditiongal Chinese medicine or evaluation index. Then further reveal metabolome characteristic in spleen qi deficiency and also bring to light the modern research target of spleen essence in Chinese medicine.Methods:The cases come from physical examination center of second hospital affiliated with Dalian Medical University.The age is from 40 to 65. Collection of physical examination personnel\'s early morning urine or second morning urine , then lay them aside in -20℃. Diagnosis standard for metabolism syndrome was promulgated by international diabetes alliance (IDF) on April, 2005.Traditional Chinese medicine New Medicine Clinical research Guiding principle" promulgated in 2002 is taken to diagnose spleen qi deficiency. The cases were divided into three groups:including cont group, metabolism syndrome group, and metabolism syndrome combined spleen qi deficiency group. The urine were storaged at -20℃for one week,and then were storaged at -80℃.The ultra performance liquid chromatogram-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF- MS)is utilized to analysis ingredient in urine, and to express the different metabolite qualitatively. Partial least squares-discrmininant analysis (PLS-DA) is used to process the data.Results:After analysis of metabolomics, from the PLS-DA schema chart we discovered that metabolism outline had the satisfactory discri- mination in every two groups of urine, and we obtained the corresponding latent designated object: 130.048m/z unknown compound and 310.198m/z meat alkali are latent tracer in urine of Metabolism syndrome group,the density in urine decreased compared to cont group; the density of 229.137 m/z leucylproline was increased, but the density of 265.115m/z pheny- lacetylglutamine was decreased in urine of metabolism syndrome and spleen- qi-deficiency compared to normol and metabolism syndrome.Conclusions:1. Urine metabolism profiling has the satisfactory discrimination between the spleen qi deficiency connect with metabolism syndrome and the normal group.2. We had discovered some latent tracers of spleen qi deficiency, such as leucylproline and phenylacetylglutamine, this is helpful for clinical doctor to practice the standar of western medicine\'s microscopic diagnosis of spleen qi deficiency, and it played a standard role in quantification of the Chinese medicine spleen qi deficiency card.3. The systematic characteristic of metabolomics observed is similar to holistic concept in traditional Chinese medicine., this can unifies the syn- drome essence of traditional Chinese medicine and the modern research better.

Synergistic Effect of TRAIL and Adriamycin on Apoptosis of Osteosarcoma Cell Line MG-63 in Vitro

Wed, 30 Mar 2011 17:01:15 +0000

Objective:Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of tumor necrosis factor (TNF) family. TRAIL could specifically induce apoptosis in various tumor cells, and is nontoxic to normal tissues and cells. Because not all tumor cells are sensitive to TRAIL, more and more researches are focusing on the method of combination application of TRAIL and radiation or chemotherapeutic agents recently. One aim of this study is to investigate the effect of TRAIL inducing apoptosis on osteosarcoma cell line MG-63 in vitro. Another aim is to determine if there is a synergistic effect of combined TRAIL with adriamycin (ADM) and to find out the possible mechanism, in order to provide a new strategy in treating osteosarcoma.Method:The osteosarcoma cell line MG-63 were cultured in the regular condi- tion. Respectively inhibition rate and apoptosis rate were analyzed by MTT and Flow Cytometry (FCM). The morphologic changes and characters of apoptosis were observed by inverted phase contrast microscope. The mRNA expression of cell-surface receptors DR4, DR5, caspase-8, Bcl-2 were examined by RT-PCR. Statistic analysis was processed by SPSS13.0 software.Result:The result of MTT and FCM showed that both TRAIL and ADM can inhibit MG-63 cells. There was a synergistic effect of combined TRAIL with ADM. Under inverted phase contrast microscope,some cells became to be small and round with intact cellular membrane 24 hours after treated with TRAIL. Cellular fragments can be seen when treated with TRAIL combing ADM. The result of RT-PCR showed that, compared with TRAIL alone group,ADM could up-regulate the level of DR4 DR5 and caspase-8 mRNA expression in MG-63 cells in combination group(P0.05).Conclusion:1.TRAIL could inhibit MG-63 cells by inducing apoptosis in the dose dependent manner.2.TRAIL combining with ADM had a synergistic effect in inducing apoptosis in MG-63 cells.3 .The cause mechanism of synergistic effect may because of the up-regulating of the level of DR4 DR5 and caspase-8 mRNA expression.

Changes of Serum and Urine Monocyte Chemoattractant Protein-1 in Children with Henoch-Schonlein Purpura Nephritis

Wed, 30 Mar 2011 17:01:15 +0000

Objective1.Measuring the change of serum and urine of monocyte chemoatractant protein-1 in children with Henoch-Schonlein purpura,in order to probe into the clinical significance of monocyte chemoatractant protein-1 in children with Henoch-Schonlein purpura nephritis.2.At the same time comparison of the relevance of serum and urine of monocyte chemoattractant protein-1,to investigate the impact of both of them in children with Henoch-Schonlein purpura nephritis.3.Selecting 25 cases with Henoch-Schonlein purpuranephritis in recovery duration, three months post-discharge follow up,monitoring urine MCP-1 consistence dynamically, assessing prognosis.Methods1.Sera and urine of MCP-1 were measured by enzyme linked immunosorbent assayin 50 patients with Henoch-Schonlein purpura(HSP),(25 of them patients with which renal injury),and other 25 healthy children,in comparison of changes of serum and urine of MCP-1.2.At the same time serum urea nitrogen,creatinine,urinary albumin,urine NAG, urineβ_2-MG,24 hours urinary levels of protein were investigated with HSPN,analysing of the correlation between these indicators and urine MCP-1.3.Urine of MCP-1 in HSPN group were measured in recovery duration,and were compared with urine of MCP-1 in HSP group and HSPN group in acute duration.Results1.The expression of sera MCP-1 was higher in HSPN group in acute duration than in HSP group and healthy controls,but no siginicant difference in the three groups (P＞0.05).The expression of urine MCP-1 was significantly higher in HSPN group in acute duration than HSP group and healthy controls,and significant difference were found in three groups(P＜0.05).2.Urine MCP-1 level were associated with proteinuria in children with HSPN in acute duration,but serum MCP-1 levels has nothing to do with HSPN.3.There was a close correlation between urine MCP-1 expression and urinary albumin, urine NAG,urineB2-MG and 24 hours urinary levels of protein in acute duration,but the expression of urine MCP-1 level was not correlated to the serum urea nitrogen and creatinine.4.There were statistically significance of urine MCP-1 in acute and recovery duration with HSPN group(P＜0.05).ConclusionsMCP-1 was one of the inflammatory response factor in acute phase with HSPN,which may play a role in inflammatory injury;urine MCP-1 might be one of the feasible predictor to evaluate the degree of renal injury in children HSP.

Effect of Nutritional Intervention on the Treatment of Nasopharyngeal Carcinoma

Wed, 30 Mar 2011 17:01:13 +0000

[BACKGROUND]Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in southern China with annual incidence of 10-25 per 100,000 people, it accounts for more than 70% of head and neck malignant tumors in this region. And because the incidence is highest in Guangdong province, the carcinoma is also called "Cantonese tumor". Radiotherapy is currently the most effective method in treating this kind of carcinoma with the five year total survival rate of 40-70%. Furthermore, comprehensive chemotherapy is able to improve the therapeutic effect on the patients in advanced stage, increase local control rate and reduce distant metastasis rate. This therapy, however, increases the severity of side effects, thereby compromising the quality of life of partial patients as well as causing weight loss and even malnutrition.Most cancer patients develop clinical malnutrition. According to variations of sites and clinical stages of the malignant tumor, the incidence of malnutrition is between 31% and 87% among tumor patients. The nutrition requirement of tumor patients is higher than the recommendations for general population, including daily nutrition requirement and additional nutrition requirement by tumor growth, infection, anemia and treatment. But food intake of the patients is insufficient due to anorexia, food preference and dietary restriction, psychological factors, surgical operations, radiotherapy and chemotherapy. Unsal et al. used Subjective Global Assessment (SGA) to evaluate the nutritional status of the patients with head and neck tumors and found that the incidence of malnutrition was as high as 88%. Beaver et al. found in a study that 60% patients with head and neck tumors showed significant weight loss after radiotherapy. Study in nasopharyngeal carcinoma has reported that concurrent chemotherapy group, when compared to sole radiotherapy group, showed significantly more grade 3-4 adverse reactions; specifically, incidence of acute mucositis increased by 13%; incidence of gastrointestinal reactions increased by 12%; hematological toxicity increased by 14.8%; incidence of weight loss had exceeded 10% and increased by 23.6%.Nutrition is the fundamental substance for human body\'s growth and development, tissue repairing and maintenance of normal physiological function, and is essential for the recovery of patients. Malnutrition may cause dysfunction of the immune system, increase the probability of infection, and thus increase mortality rate and prolonging the rehabilitation period and hospitalization time, and influence treatment and prognosis of the disease. To investigate, analyze and regulate the nutritional status of tumor patients is gaining more and more attention from both doctors and patients. Whether or not ingested nutritional substances potentially promotes tumor growth, is a common concern for clinical doctors. As a matter of fact, the ability to obtain nutrition of tumor cells is far stronger than that of normal cells in human body. Even though we don\'t provide the patients with nutritional substrates, tumor cells could still compete with the host in ingesting limited nutritional substances and manage to obtain adequate nutrition they need for growth. Damage on the host body produced by limiting ingestion of nutritional substances for the tumor patients far outweighs its benefit of inhibiting tumor growth. Numerous studies have found that percentages of tumor cells at S stage and proliferative stage significantly increased while cell apoptotic rate decreased in patients who had received supportive nutritional therapy, which promoted tumor growth and proliferation;however, concurrent chemotherapy increased treatment response by allowing bringing into play the effect of cell-cycle-specific chemotherapeutic drugs; treatment response of nutrition+chemotherapy group was significantly better than that of sole chemotherapy group. Meanwhile, some studies have also suggested supportive nutritional therapy, especially those with additive special nutritional agents, such as glutamine, arginine, methionine,ω-3 fatty acids, could not only achieve the goal of inhibiting tumor growth, but also attain the purpose of improving nutritional status of the patients.Currently, tumor patients with malnutrition are not able to receive sufficient nutritional support during clinical treatment of tumor. And there are shortages or deficiencies with respect to the evaluation and monitoring of nutritional status, selection of the best time and duration of nutritional support treatment. In early stage when adverse reactions of radiotherapy and chemotherapy start to appear, most patients simply experience subtle decrease of ingestion and indistinct physical weakness. Because symptoms and signs of malnutrition in such patients are not obvious, they are often neglected by clinical doctors and the patients per se, which leads to the absence of early treatment. Repeated and multiple chemotherapy and radiotherapy are often given when the patients are starving and nutritionally deprived. In most cases, supportive nutritional treatment is started only when the patients experience extreme physical weakness or show decreased biochemical markers or decreased tolerance to radiotherapy and chemotherapy. However, the optimal opportunity for nutritional support treatment may be lost at the time and the malnutrition is difficult to be cured.For the time being, no studies have been reported as whether or not evaluation and monitoring on nutritional status and early systemic nutritional interventions at the beginning of and during the treatment for nasopharyngeal carcinoma, could alleviate treatment related adverse reactions, improve quality of life and promote healing for the patients while not affecting treatment response or even increasing treatment response.The present study aimed to reveal clinical evaluation and monitoring on nutritional status in tumor treatment and preliminarily investigate the effect of nutritional status on the treatment and prognosis of nasopharyngeal carcinoma by retrospective analysis. In addition, a prospective study was conducted to investigate the effect of nutritional intervention on the adverse effects caused by radiotherapy or chemotherapy, quality of life, therapeutic effect and the prognosis, and thus provide clinical evidence for the nutritional intervention during tumor therapy.[OBJECTIVE]We retrospectively analyzed the cases and understood the current conditions of nutritional evaluation and monitoring on NPC patients during chemotherapy and radiotherapy, and preliminarily discussed the effect of nutritional status on the treatment and prognosis of nasopharyngeal carcinoma in various stages. Then, a prospective study was conducted to investigate the effect of systemic nutritional intervention on the adverse effects caused by radiotherapy or chemotherapy, quality of life, therapeutic effect and the prognosis of NPC patients, and thus provide clinical evidence for the nutritional intervention during tumor therapy. [METHODS]1 A total of 298 NPC patients without distant metastasis admitted to our hospital for initial treatment from October 2003 and October 2005 were included in the study and clinical data such as weight changes, nutritional evaluation and monitoring and treatment conditions were recorded. The present study retrospectively analyzed 191 NPC patients who met the following conditions: age≤60 years; no significant cardiac, pulmonary, liver and renal dysfunctions, no severe endocrine or metabolic diseases and with complete information. These patients were divided into two groups according to the nutritional status during treatment (control group and malnutrition group). Single factor and multi-factor methods were used to analyze the relationship between nutritional status and the three-year survival rate, free from distant metastasis survival rate and free from local recurrence survival rate of NPC patients in I-II stage and III-IV stage, respectively.2 A prospective study was conducted in 60 NPC patients who admitted to our hospital from December 2005 to September 2006 and met the following conditions: (1) Treatment-na(?)ve patients with Undifferentiated non-keratinizing nasopharyngeal carcinoma (by WHO classification) as confirmed by pathological examination; (2) aged 20-65; (3) distant metastasis was ruled out by CT scan, chest X-ray, B-ultrasound examination and bone ECT scan; (4) with normal liver and kidney function and no significant heart or pulmonary dysfunction; without severe endocrine diseases or metabolic diseases; (5) KPS score≥70 points, ECOG≤grade 2.These patients were randomized into nutritional group (30 cases, radiotherapy and chemotherapy plus nutritional intervention) and control group (30 cases, radiotherapy and chemotherapy only) to investigate the effect of systemic nutritional intervention on the patients\' nutritional condition, quality of life, adverse effects caused by radiotherapy or chemotherapy, therapeutic effect and the prognosis. 3 Statistical analysis was conducted by SPSS 15.0 package. Differences of clinical factors and ranked data between the two groups were analyzed by X~2 test; quantitative data were expressed as mean number±standard deviation (x±s) and processed with t test; the survival rate was calculated by life table method and the survival status was compared by Kaplan-Meier method and tested with Log-rank. The multi-factor analysis of the prognosis was conducted with COX proportional hazard model. All statistical data were processed with SPSS 15.0 package, and data were recognized as significant when the Ps were less than 0.05 (bilateral).[RESULTS]1 Retrospective study1.1 Among 298 NPC patients, 93.3% cases lacked food and nutrition intake records; only 73.2% had complete records of weekly body weight or recent body weight changes; 43% patients received nutritional support treatment, including vitamins therapy, among these patients, 18.1% received fat emulsion and/or intravenously received amino acid for nutritional therapy, most of them were elderly patients with significantly less food intake after severe radiotherapy and chemotherapy, treatment duration≤seven days, and without record of orally taken enteral nutrients. The incidence of malnutrition of NPC patients during treatment was as high as 49.7% (95/191).1.2 The incidence of malnutrition of NPC patients during the treatment was 49.7%(95/191). The univariate analysis showed that the three-year survival rate, distant metastasis-free survival rate and local relapse-free survival rate in stage I and stage II NPC patients in control group were higher than those in malnutrition group, but there was no statistical significance between them. However, the three-year survival rate and distant metastasis-free survival rate in stageIII and stage IVa NPC patients in control group were higher than those in malnutrition group, possessing a statistical significance (P 0.05).[CONCLUSION]Incidence of malnutrition of NPC patients during treatment is rather high, which is one of the factors influencing treatment and prognosis of the disease. The nutritional support treatment performed during radiotherapy and chemotherapy is effective and safe. We proposed that the rational nutritional intervention should be an important part of the overall treatment scheme. The early systemic nutritional intervention is able to effectively improve nutritional status of tumor patients, promote the quality of life, strengthen immune ability and anti-tumor competent, enhance the tolerability to the radiotherapy and chemotherapy, decrease complications and adverse effects, promote recovery and healing and facilitate the treatment process.

Effects of Ginsinoside Rd on Neurogenesis in the Dentate Gyrus of Normal and Infrasound-exposed Adult Rats

Wed, 30 Mar 2011 17:01:12 +0000

Ginsenoside is one of the effective ingredients in ginseng.At present,more than 40 kinds of ingredients have been separated from ginsenoside.As one of the most important ingredients,ginsenoside Rd shares many pharmacological effects.Ginsenoside can facilitate the acquirement,consolidation and reappearance of learning and memory,and improve neural plasticity.It is said that the learning and memory ability of adult animals has close relation to neurogenesis in the dentate gyrus,so it is possible that ginsenoside Rd can affect the process of neurogenesis.Neurogenesis is defined as the process of proliferation,survival and differentiation of NSCs.Recently,it has been investigated that,neurogenesis exists in the brain of many kinds of adult animals,including mammals,and new neurons and glial cells are generated through this process.Dentate gyrus is one of the regions being studied most.It is said that the neurogenesis in adult hippocampus participates in the formation of learning.In our experiment,to study the effects of ginsenoside Rd on neurogenesis,ginsenoside Rd in different dosage was given intraperitoneally to normal rats,then the proliferation, differentiation and migration of NSCs in SGZ was observed by immunostaining at different time after Rd treatment.Many environmental factors affect the level of neurogenesis.We have investigated that infrasound of 8 Hz at 130 dB can inhibit the proliferation of NSCs in normal adult rat SGZ.Infrasound is usually defined as the sound with frequeny below 20 Hz,which comes from mechanical vibration.A large variety of natural and man-made phenomena can produce infrasound.Infrasound at high intensity does harm to our bodies and plays an important role in noise pollution.As a significant target organ,brain is very sensitive to infrasonic damage.Experimental studies have reported that many parts of brain changed after infrasound exposure in some aspects,such as structure,function and neurotransmitter metabolism.This process is complicated and has special mechanism of action.In our study,we observed the effects of ginsenoside Rd on neurogenesis in SGZ in rat brain after infrasound exposure to investigate the safety action of ginsenoside Rd to infrasound injury and to provide experimental evidence for the prevention and cure research of infrasound.Experiments were carried out to observe the effects of ginsenoside Rd on neurogenesis in the dentate gyrus of normal and infrasound-exposed adult rats. Experiment 1:Adult normal male Sprague-Dawley(SD)rats were treated intraperitoneally with ginsenoside Rd and killed at different time after that. Before the rats were sacrificed,three pulses of BrdU were given intraperitoneally to label the S-phase cells,which were detected by anti-BrdU immunostaining.Rats killed at 3 d,7 d and 14 d were used to observe the proliferation of NSCs,and those killed at 34 d were used to observe the differentiation and migration of NSCs.Experiment 2:Adult male SD rats were randomly divided into a normal control group,a sham control group,an infrasound group and a infrasound+ginsenoside Rd group.Rats of the infrasound group and the infrasound+ginsenoside Rd group were exposed to infrasound of 16 Hz at 130 dB for 2 h/d in 7 consecutive days.Observation was carried out after infrasound exposure and the method was identical to experiment 1.The results are showed as follows:1.Effects of ginsenoside Rd on the proliferation of NSCs:BrdU positive cells in each group appeared as irregular shaped single or clusters located exclusively in the SGZ.Experiment 1:3 d and 7 d after Rd treatment,BrdU positive cells increased markedly,but the number of dividing cells appeared to return to control levels at 14 d after Rd treatment.Experiment 2:After Rd treatment,BrdU-positive cells were significantly increased in the SGZ at 3 d after infrasound exposure,compared with those of the infrasound group,and the difference was more obvious at 7 d.However,14 d after infrasound exposure,a large number of BrdU-positive cells were reappeared in the infrasound group,and the difference disappears accordingly.2.Effects of ginsenoside Rd on the differentiation of NSCs:From the double labeling immunostaining,we found in each group the percentage of NeuN-expressing BrdU immunoreactive cells is about 70%.However,the BrdU labeled cells scarcely expressed astrocyte markers.So,there is no influence of ginsenoside Rd on the differentiation of NSC.3.Effects of ginsenoside Rd on the migration of NSCs:At 34 d treatment group,the location of BrdU positive cells changed.Some of the cells were still in the SGZ,but many were found throughout the GCL.There was also a marked change in the morphology of BrdU positive cells.By the 34 d,most nuclei in the SGZ and GCL appeared much bigger and round,just like those of surrounding granule neurons.After Rd and infrasound treatment,there was no ectopic granule cell migration.The following is the conclusion of our experiments:Ginsenoside Rd markedly increases the neurogenesis in the dentate gyrus of normal and infrasound-exposed adult rats,and the differentiation and migration of new NSCs are not affected after Rd treatment and infrasound exposure.

Clinical Effectiveness of Sequential Therapy with Methylprednisolone and Nebulised Pulmicor Respules in the Treatment of Acute Moderate/Severe Asthma

Wed, 30 Mar 2011 17:01:11 +0000

AIM: To compare the effect of methylprednisolone and nebulised pulmicor respules in sequential therapy of acute moderate/ severe asthma.METHODS: twenty-seven patients(16-70 years) with acute moderate/ severe asthma were randomly divided into three groups:group A.B and C.Group A were treated with methylprednisolone 80 mg,iv,bid for 3d,then oral methylprednisolone was substituted 8 mg bid for 4 d;Group B were treated with methylprednisolone 80 mg,iv,bid for 3d,then nebulised pulmicor respules was substituted 2 mg bid for 4 d;Group C were treated with nebulised pulmicor respules,4mg,bid for 3d,then nebulised pulmicor respules 2 mg bid for 4 d.RESULTS: The day- and night- time asthma symptom scores decreased significantly after one day in all group (P< 0.05). Compared with the first day, the peak expiratory flow(PEF) of daytime and nighttime measurements both increased significantly(P<0.05) in group A and B in the third day, but for C group, there only existed significant difference in daytime PEF. And the results were similar in the seventh day. FEV1 % levels were increased from (48.89±22.17).(55.11±16.09)and(54.22±17.56)to(77.56±17.02).(79.33±16.09)and(74.22±24.05) when completing the treatment, there only existed significant difference in group A and B. The ACQ scores decreased significantly after seven days’treatment in all group (P< 0.01). The fractional exhaled nitricoxid(FENO) decreased at the 8th day in all group(n=14). There was a significant suppression of Plasma cortisol at the 8th day in group A (P<0.05). In group C, the patients felt discomfortable in upper airway when inhale pulmicor 4mg and sputum was increased, one of which got worse again and required additional systemic steroids.CONCLUSIONS: For acute moderate/ severe asthma, the therapy with methylprednisolone iv is more effective than nebulised pulmicor respules, as it can improve the symptoms significantly after one day and the pulmonary function after two or three days. The effects of intravenous methylprednisolone with sequential nebulised pulmicort and oral methylprednisolone are similar. and it has no influence on the hypothalamic-pituitary-adrenal axis; however, it costs a lot. The fractional exhaled nitricoxid(FENO) decreases at the 8th day in all group(n=14), which suggests that FENO be used to monitor the airway inflammation of acute asthma.

Long-standing Effects and the Mechanisms of Ischemic Tolerance Induced by Hyperbaric Oxygen Preconditioning in Spinal Cord of Rabbits

Wed, 30 Mar 2011 17:01:10 +0000

Background Since 1986 the concept of ischemic preconditioning was proposed by Murry, several strategies (such as low temperature, electro-acupuncture, etc.) as well as drugs (such as excitatory amino acid antagonists, free radical scavengers, minocycline, etc.) have been proved to induce ischemic tolerance in the organs. However, most observation time of these studies’did not exceed 72 hours after reperfusion. It was suspected that preconditioning didn’t permanently prevent cell death after ischemic/reperfusion injury, but only postponed the evolution of the progress of cell death. Whether the protective effects will gradually weaken, disappear or not change in a long-term observation is a question disputable but very important that need us to answer. Our previous study showed that hyperbaric oxygen (hyperbaric oxygen, HBO) preconditioning can simulate "ischemic preconditioning" and induce ischemic tolerance in spinal cord ischemic/reperfusion injury. This phenomenon also exists in focal cerebral ischemia model of rats. Another study showed that apoptosis does exist in ischemia/reperfusion injury in the central nervous system, and the delayed cell injury of ischemia/reperfusion in spinal cord is mainly caused by apoptosis rather than necrosis. Hyperbaric oxygen preconditioning can reduce early apoptosis of neurons and may even reverse apoptosis. Therefore, this study take "the ischemic tolerance induced by hyperbaric oxygen preconditioning in rabbit spinal cord" as the breakthrough point, to study the neuroprotective effect of HBO preconditioning for long-term observation and the relationship between the long-term effects and neuron apoptosis.PartⅠLong-standing ischemic tolerance induced by hyperbaric oxygen preconditioning in spinal cord of rabbitsObjective To determine if the ischemic tolerance induced by hyperbaric oxygen preconditioning in spinal cord of rabbits is long-standing. Methods In experiment 1, sixteen male New Zealand rabbits were randomly assigned to 2 groups (8 in each group) : animals in control group received room air and normal ambient pressure (1 h/d, 5 d); animals in HBO group received HBO pretreatment (100% O2, 2.5 atmosphere absolute [ATA], 1 h/d, 5 d). At 24 hours after the last treatment, the spinal cord ischemia was induced for 20 min by infrarenal aortic cross clamping. At 1day, and 2, 7, 14, 21 days after reperfusion, the neurologic function was scored by the Tarlov criteria (in which 4 was normal and 0 was paraplegia). In experiment 2, thirty-two animals were also randomly assigned to HBO group and control group (16 in each group). Animals underwent the same treatment as in experiment 1. Then eight animals in each group were sacrificed at 2 days and 21 days after reperfusion, respectively. And their spinal cords (L5-7) were removed immediately for histopathologic examination. The neurologic functions were scored before they were sacrificed.Results The neurologic function scores in HBO group were significantly greater than in control group at all time points (P0.05). The numbers of normal neurons in the anterior spinal cord in HBO groups were also significantly greater than in control groups at 2 days and 21 days, which had no statistical differences between 2 days and 21 days in HBO groups. There was a strong correlation between the neurologic function scores and the numbers of normal neurons in the anterior horn of the spinal cords (r2d =0.903, P0.05). The numbers of TUNEL positive neurons in HBO groups were significantly less than that in control groups at all time points(P0.05). Conclusion The long-standing ischemic tolerance induced by hyperbaric oxygen preconditioning doesn’t significantly related to later stage apoptosis after reperfusion in spinal cord of rabbits.

Identification of a Novel ERM Family Molecule and Its Functional Role in Cell Morphology

Wed, 30 Mar 2011 17:01:09 +0000

AIM: To generate polyclonal antibody of N-terminal 160 amino acids of hErmin (hErmin160), and to identify expression of hErmin in human brain and make further research about its role in cell morphology. METHODS: The coding sequence of hErmin160 was amplified by PCR from pBluescriptR plasmid encoding with full length of hErmin cDNA sequence, and then cloned into prokaryotic expression vector pET41-b(+). The plasmid was transformed into E.coli BL21(DE3)pLysS to express hErmin160 fusion protein in response to IPTG induction. The products were analyzed by SDS-PAGE and Western-blot. Expressed protein was purified by Ni2+ metal-chelating chromatograph and then injected into rabbits to generate polyclonal antibody. Blood serum after injection was purified using affinity chromatography. The polyclonal antibody was identified by Western-blot. hErmin was identified by Western-bolt and immunohistochemistry experiments, in which anti-hErmin160 antibody was used as primary antibody. After transfection, cells overexpressing hErmin were observed by immunocytochemistry experiments. RESULTS: DNA sequencing confirmed that the coding sequence of hErmin160 was completely concordant with the original sequence (BC026345, hErmin’s GenBank accession number). SDS-PAGE showed that the relative molecular masses (Mr) of the expressed, purified products were about 51ku, which was in accordance with the predicted. Grayscale scanning showed that the expressed hErmin160 fusion protein accounted for 10% of the total bacteria protein in the supernatant, and the purity of purified product was 92%. Fusion protein was identified by Western-blot analysis. Blood serum after injection and purified polyclonal antibody can both bind hErmin160 specifically in Western-blot. The purified polyclonal antibody can specifically bind hErmin expressed by eukaryotic system and the expression of hErmin in human brain was confirmed. hErmin overexpression promoted cell arborization. CONCLUSION: We acquired high purification soluble hErmin160 fusion protein through the E.coli prokaryotic expression system and purified polyclonal antibody of hErmin160 protein. We have identified hErmin as a novel molecule of ERM family, which is expressed in adult human brain. hErmin overexpression promoted arborization of cultured cells.

Prelimilary Experimental Study of Three-demensional Engineered Skeletal Muscle Tissue in Vitro

Wed, 30 Mar 2011 16:57:08 +0000

The development of skeletal muscle tissue engineering will provide some treating advice for many skeletal muscle diseases such as muscular dystrophy,spinal muscular atrophy,etc.Besides,plastic surgery for the treatment of trauma,tumor resection and denervation-induced loss of muscle tissue will also benefit from skeletal muscle tissue engineering.Even though tissue engineering research has made significant progress in many artificial tissues such as skin,bone and cartilage have been used in clinical treatements,but skeletal muscle tissue engineering still faces immense challenges and it has difficulties in the high density at the cell proliferation and cell differentiation,three-dimensional polar culture,construction of the larger the size of the muscle tissue,etc.However,many scholars are still committed to skeletal muscle in vitro and alternative research,attempt to create a function of the muscle tissue. After many years of exploration and efforts,a number of authoritative research team made considerable achievements,and constantly push forward the study of skeletal muscle tissue engineering progress:Powell~([1,2]) used collagen and matrix to build a successful three-dimensional skeletal muscle and added mechanical stimulation to tissue,making sure the formation of muscle tissue,differentiation and the formation of contractile function in vitro;Dennis and Kosnik~([3,4]) explored three-dimensional skeletal self organized model and added electric stimulation to the muscle tissue Myooids in the horizonl it would produce a sustained contraction of the excitatory and function.In Germany Bach\'s team~([5]) had co-cultured myoblasts fibrin and scaffold in three-dimension and obtained myotubes that had the multi-polar differentiated functional.The key technologies of skeletal muscle construction in vitro include implanting largly amplified in vitro target cells such as adult skeletal muscle satellite cells or myoblast cell line into the three-dimensional matrix of different materials and making them grow and differentiate into skeletal muscle in vitro analogues that have shapes and contraction ability and could survive in a long-term.To realize the true meaning of skeletal muscle in vitro construction,foreign scholars have improved skeletal muscle tissue construction technology,at the same time,detect expression of key regulatory molecules(such as the myogenic transcription factors,contractile proteins, muscle creatine kinase,acetylcholine receptor,etc.) systematically in muscle tissues that differentiate in vitro and composite with different matrix.They have tried to explore whether the differentiation,the corresponding expression of specific transcription factors and control of the signal system in muscle tissues in vitro are coincided with growth and differentiation of the body\'s skeletal myoblast.The results of above skeletal muscle construction study provide clues and ideas for future skeletal muscle tissue engineering researchers.Through analyzing home and abroad skeletal muscle research data we found that though it is still difficult to construct in vitro large skeletal muscles that can mimic the features and function of structure of muscle in vivo,the existing and exploring in vitro construction muscle tissues are able to mimic the differentiation and development of in vivo tissues, express contractile proteins,produce contraction effect and avoid the interference from other cells or tissues,making them become ideal in vitro model of studying the development of skeletal muscle and skeletal muscle myopathy.However,there are no coverages of success three-dimensional construction of muscle tissue in vitro at home. Sylgard 184 belongs to silicone rubber elastic material and has two-component mixed-curing,solidification at room temperature,the surface of it is smooth and has a certain degree of flexibility.As a basal layer,benift to detect the cell\'movement and traction force generated by load~[6].Therefore,Sylgard 184 has been widely used in skeletal muscle tissue engerring,and more scholars pay attention to it~([7,8,9]).In view of the above research background,this study selected Sylgard 184 (silicone elastic material) as the growth of stents that co-cultured with myoblasts in vitro and explored the practicability of constructing in vitro three-dimensional model of skeletal muscle.As the effective contact of scaffold material and seed cells decides cell proliferation,differentiation and the function expression we first coate Sylgard184 surface with different matrix components(collagen,laminin,etc.),and then analyze growth and differentiation of myoblasts at different matrix materials Sylgard184 surface-modified,finally,select the best surface-modified ingredients;As the keys to construction of three-dimensional muscle tissue in vitro,are emergence and maintenance of polar parallel differentiated myotube,this study further modified to optimize the Sylgard184 indentation to the mold through the guide groove cast,as well as promote growth and scalability of the Matrigel matrix,obtain the polar parallel differentiated muscle tissue,and analyze and detect muscle characteristics and systolic function of polarity of muscle tissue,this method will set the foundation to successful construction of three-dimensional model of muscle tissue in vitro.Two parts of our research.Part one:Biocompatibility of Sylgard 184 coated with different matrix materials and C2C12 cellsObjective:An attempt was undertaken to acquire the ideal matrix material that can enhance the compatibility between silicone rubber elastomer(Sylgard 184) and murine C2C12 cells.Methods:Two-component of sylgard184 admixture to the ratio of 10:1 then mixed into 4 holes of 6 Orifice,solidification at room temperature,and the remaining 2 holes without coating were taken as blank control(group A).The surfaces of 4 holes were coated by collagen typeⅠ,laminin and poly-lisine,respectively as group B,C and D,and the hole of uncoating as group E,every group has six samples.Each group C2C12 Cells morphology was observed by the inverted microscope.Flow cytometry (FCM) was used to detect the cell generation cycle of C2C12 cells after enrichment culture 48h.RT-PCR was used to test MyoD,Myogenin mRNA expression of proliferation or differentiation cultured for 48 h C2C12 cells respectively that growed on coated or uncoated sylgard 184 surfaces.Results:Material Sylgard 184 has cytotoxicities as vaccination C2C12 cells in group E all float deadly in 24h and most cells in group D become dead,only a small number of adherent cells are survival.While the toxicity of Sylagard 184 in group B and C are reduced significantly after materials are coated and the compatibility of C2C12 cells\' surface are also enhanced.Besides,the percentage of cells in the synthesis phase,the quantity of MyoD in proliferating phase,and Myogenin differentiation gene mRNA expression in group C are significantly higher than that in group A and B(P＜0.05).Conclusions:The data suggests that Sylgard 184 modified by laminin is superior to that modified by other biomaterials,in terms of which can enhance the proliferation and differentiation of cultured C2C12 cells in vitro. Part two Construction and detection of highly organized 3dimensional muscle tissue in vitroObjective:To cultivate C2C12 cells on the modified mold-grooves of Sylgard 184 and induce C2C12 cells differentiation to highly organized 3-dimensional muscle tissue,then detect the growth and functional properties of differentiation myotube.Methods:Mix bicomponents Sylgard 184 by the ratio of 10:1 uniformly and poured the mixture into 6-well plates.After curing by standing at room temperature(The width of each mold groove is 0.8mm,depth is 0.15mm,length can adjust by requiring),carve grooves at the central basal Sylgard 184 and indentate in the groove at the bottom of the mold.After washing grooves with Hank\'s fluid,pave matrigel matrix and collagen (mixing ratio 1:6) mixture evenly at the bottom of grooves and add 0.3ml mixture into each groove,making sure that the cell matrix material of coverage is uniform. Finally,place them in biological safety cabinet and under UV disinfection.Inoculate C2C12 cell suspension whose density is 1.0 -1.2×10~5 /ml,remove the entire medium when there are 80%cells proliferations,then add 2%horse serum containing DMEM/F12 and differentiation cultivate 7d,the inverted microscope to observe X-ray film morphology and continue to divide after the 21d,the construction of skeletal muscle tissue was detected by immunofluorescence assay muscle-specific protein expression,RT-PCR was used to detect Myogenin,Desmin mRNAs expression,while the use of electron microscopy to watch myotubes\' morphologies and inter-connections.Results:C2C12 cells were differentiation cultivated in the mold-grooves of Sylgard 184 for 7 days,polarity-differentiation and myotube fusion were observed under the inverted microscope.After 21 days,it showed that myotubes stand closely and connect with each other deteced by the scanning electron microscopy.The thickness of the membrane-like structure with the character of three-dimensional was 0.15mm. Detect Myogenin,Desmin mRNAs of positive expression in constructed skeletal muscle tissue by RT-PCR,and there is differentiation-specific gene protein Myogenin and Desmin myoblast-specific protein expression by immunofiuorescence assay and DAPI nuclear staining.Results show that Myogenin expressed in differentiated myotubes and there is a large number of Myogenin in nucleis.While the positive expression of fluorescent protein showed that has the big differentiation potential. Desmin are skeletal muscle cytoskeleton protein and intermediate filament-specific protein,which mainly distributed in the cytoplasm of skeletal muscle cells,and are also one of the earliest indication of skeletal muscle.It expresses in the differentiated myotube in our results,which indicated that the constructed three-dimensional differentiated muscle tissue polarity analogues have the same physiological characteristics with mature skeletal muscles in vivo.Conclusions:The modified mold-grooves of Sylgard 184 is able to guide the differentiation of C2C12 cells and promotes them to form parallel multinucleated myotubes.Myotubes could overlap with each other and form three-dimensional skeletal muscle tissue.

Study of Time-Resolved Fluoresence Immunoassay System

Wed, 30 Mar 2011 16:57:07 +0000

Time-Resolved Fluorescence Immunoassay(TRFIA) processes the advantages of high—sensitivity、specificity、high—stability and nonradioactivity etc.,so preclinical medicine、clinic—detection and molecular biology paied much attentions to TRFIA.In the thesis,the theory and several immunoassay methods and measurement techniques of TRFIA widely used in medicinal researches are discussed in detail.Through experiments and theoretical analysis,the characteristics of excitation spectrum of lanthanide Eu and Eu~(+3) chelaes are studied.Based on the research mentioned,the TRFIA system are designed、developed and tested.Finally,three functions are realized,they are the measurements of fluorescence lifetime、time-resolved spectrum and concentration of the detected solutions. The specifications of the system are:sensitivity of 10~(-13)mol/L、lower limit of detection 10~(-14)mol/L、range of linearity 10~(-13)-10~(-11)mol/L.The system is nonradioactive、full automatic and ultramicro concentration detectable.

Effects of Pet Dogs on the Empty Nester Elder

Wed, 30 Mar 2011 16:57:06 +0000

Objects: To investigate the influence of pet dogs on the empty nester elders.Methods Object: A simple of 314 elders which randomization and stratification by age was drawn from four districts of Dalian, including 102 males and 212 females, 157 of them breed pet dogs as study group. Methods SF-36 (Short Form 36 Health Survey Questionnaire), MUNSH (Memorial University of Newfoundland Scale of Happiness), LES (Life Event Scale), PSSS (Perceived social support scale), NEO-FFI (NEO Personality Inventory Short Form) and LAPS (Lexington Attachment to Pets Scale) were employed as research tools. Statistics t-test, Chi-square test, Mann-Whitney-Wilcoxon rank sum test, Pearson correlation, and Multiple Linear Regression were used to analyze the data.Results:⑴There were significant differences between pet-owners and non-owners in perception of out-family support, perception of social support, agreeableness of NEO-FFI. There were no significant differences between pet-owners and non-owners in LES. There were differences between them in physiological operation, physiological function, spirit, social function, emotional function and reported health transition, pet-owners group were better than non-owner group in these aspects.⑵There were positive correlations between Attachment to Pets Scales and perception of social support, Attachment to Pets Scales and extraversion of NEO-FFI, Attachment to Pets Scales and openness of NEO-FFI.⑶Multiple Linear Regressions: There was a close relationship between pet-breed behavior and physiological operation, physiological function, social function, emotional function and reported health transition. So pet-breed behavior could be a good positive forecast factor for physiological health. Conclusion:1. The pet-breed empty nester elders are less in agreeableness of personality.2. The pet-breed empty nester elders are less in perception of out-family support.3. The pet-breed empty nester elder are better in quality of life than common nester elder; The pet-breed behavior can improve empty nester’s quality of life.

Distribution and Toxic Effect of Intracameral Injection of Bevacizumab in Rabbit Eyes

Wed, 30 Mar 2011 16:57:05 +0000

Objective: To observe distribution of bevacizumab (avastin) in eye tissue and toxic effects of the anterior segment in rabbits which received intracameral bevacizumab 1.25mg/0.05ml.Methods: Twenty New Zealand albino rabbits were divided into two groups randomly, the left eyes in experimental group received intracameral injection of bevacizumab 1.25mg/0.05ml; the left eyes in control group received intracameral injection of balanced salt solution 0.05ml. The anterior segment of eyes and ocular fundus were examined by slit-lamp microscope and direct ophthalmoscope. Intraocular pressure and corneal endothelial microscope were measured before and after injection. Five rabbits of the two groups were randomly killed in the first day, the fourth day, the seventh day, the fourteenth day, the thirtieth day after injection, and the enucleated eyes were observed by the light microscope. The eyes of the fourth day and the thirtieth day were observed by the transmission electron microscope, paraffin sections and then immunofluorescence staining were used to observe distribution of bevacizumab in the tissues of eyes.Results: All experimental animals were healthy survival during the observation. No any signs of infection, corneal opacity, cataract, vitreous opacities, retinal hemorrhage were observed in eyes after surgery by slit-lamp microscope and direct ophthalmoscope. No significant difference regard to intraocular pressure before injection and the two hour, the first day, the seventh day, the fourteenth day, the thirtieth day after injection compared with the control group(P=0.982). No significant difference regard to corneal endothelial cell density before injection and the first day, the seventh day, the thirtieth day after injection compared with the control group(P=0.889). No obvious structural change of cornea, lens, chamber angle, iris, ciliary body and retina in the first day, the fourth day, the seventh day, the fourteenth day, the thirtieth day after injection in experimental group and control group under light microscope. No obvious ultrastructural change of cornea, lens, iris, ciliary body in the fourth day, the thirtieth day after injection in experimental group and control group under transmission electron microscope. Bevacizumab were distributed in the anterior chamber angle, iris, ciliary body, choroid and retina in injected eyes and fellow eyes after intracameral injection. Staining in fellow eyes was weak than injected eyes, bevacizumab was mainly distributed in the vascular tissues, such as the vessel wall and lumen. Strongest staining in iris was seen from the first day to the fourth day in injected eyes, then gradually becoming weakened, weak staining still existed in the thirtieth day. Strongest staining in iris was seen in the seventh day in fellow eyes. Staining in ciliary body was stronger than in iris in the fellow eyes and injected eyes. Strongest staining was seen in the seventh day, then gradually becoming weakened, and the weak staining still remained in the thirtieth day. Strongest staining of chamber angle was seen from the fourth day to the seventh day in the injected eyes, then gradually becoming weakened. The staining was disappeared in the thirtieth day, weak staining of trabecular meshwork was seen in the seventh day and the fourteenth day in the fellow eyes. Strongest staining of retina was seen in the seventh day in the fellow eyes and injected eyes, then gradually becoming weakened, weak staining still existed in the thirtieth day. Staining in choroid was seen from the first day to thirtieth day in the fellow eyes and injected eyes; The control eyes did not show positive staining.Conclusions: There is no obvious toxic effects of intracameral inject- tion with bevacizumab 1.25mg in the normal anterior segment of rabbits,which provides a reliable basis for the clinical safe application of Beva- cizumab. Bevacizumab were quickly distributed in the anterior chamber angle, iris, ciliary body, choroid and retina in injected eyes after intra- cameral injection of bevacizumab 1.25mg, bevacizumab accumulates par- ticularly in the vascular tissue, our findings support and explain the clinical observation of that iris neovascularisation was rapid regressed after intracameral injection and treatment would not remain less than one month; Distribution of bevacizumab in the anterior chamber angle, iris, ciliary body, choroid and retina in the fellow eyes illustrate systemic circulation of bevacizumab. Anyway, intracameral injection of bevacizumab may be a new strategy or a joint strategy for iris neovascularisation, even may be the preferred strategy.

Sample Pool Development and Application of Solid-Phase Time-Resolved Fluorescence Immunoassay Analyzer

Wed, 30 Mar 2011 16:57:04 +0000

Solid-phase Time-resolved Fluorescence Immunoassay is a new immunoassay technology established,which is used to substitute radio immunity immunoassay,and have been widely applied in the basic medicine,clinic field and life science.To meet Solid-phase Time-resolved Fluorescence Immunoassay requirement.A key technique of direct solid-phase TRFIA is to design and prepare opaque solid-phase reflection immunologic reaction pool and increase S/V ratio of sample pool.It can increase absorption between polystyrene plate and detection protein,modify polystyrene plate and increase detection sensitivity.Nylon-6 is dissolved with 22mL acetic acid and 10mL m-methyl phenol is added polystyrene plate after it is soaked by pyridine:acetone(1:29) solution.Polystyrene plate smear layer is even when it is rotated at room temperature after 0.4mL pyridine is added.The labeled rate of plate and antibody is good after glutaraldehyde and hexamethylene diamond solution are connected arm by turns with the plate which is modified by alkylation. Sensitivity of it 3 times higher than routine physical adsorption method,CV＜10%and stability of it is that term of validity is three months longer.Operation of the method is simple and cost of it is cheap.The method has general practical value and is applied TRFIA, RIA and ELISA.It has an important significance to study development of protein link technique.It has practical value to study conjugate of polybenzene chelate,legend and protein.

Effect of Constant Magnetic Fields on Chemotherapeutic Phlebitis

Wed, 30 Mar 2011 16:57:03 +0000

Venous chemotherapy is one of effective treatments of Malignant Tumor. However, the injected veins are resulted in vein sclerosis or nearby tissues have such symptoms: rubefaction, swelling, aches, caused by many chemotherapeutics made by alkaloids, by repeating punctures which cause the mechanical stimulation, and by medicines of high density as the intensive stimuli to some issues. It is reported that 50%~80% of patients treated by venous chemotherapy suffer from phlebitis of varying degrees on clinical practice. Phlebitis is so severe that some tissues of patients are festering and non-viable, which results in patients’suffering and hindrance of the implementation of treatment regimen.Currently, there are some prevention and treatments to phlebitis caused by chemotherapy, for instance, external used medicine, infrared physiotherapy. Magnetic field as the physical agent is widely used to cure diseases in clinic field and biological effect are catching much attention in the field of biomedical engineering.Based on researches on the treatment home and abroad on phlebitis caused by chemotherapy and the biological effects of magnetic fields, the current research attempts to futher investigate the effect of uninterrupted magnetic filed on chemotherapeutic phlebitis , from the approach to the mechanism of occurrence of chemotherapeutic phlebitis, by the means of the athermal biological effects of magnetic fields and with rabbits as experimental subjects. The significance of present research is to offer an innovative treatment and prevention for chemotherapeutic phlebitis.Three research tasks are accomplished:1. Based on the researches at home and aboard in the field and in contrast to the treatment by using Magnesium Sulfate, the effectiveness of constant magnetic field on treating chemotherapeutic phlebitis is explored.2. The model of animal trauma was successfully constructed in the experiment similar with humanbeings’, which has the significanec in aspects of the occurrence, deterioration, prognosis mechanism of chemotherapeutic phlebitis and exploration of its treatment.3. Through the general observation and examination of pathological section, the present study investigated the symptom of chemotherapeutic phlebitis affected by the constant magnetic field with different intensity on chemotherapeutic phlebitis. Through preliminary screening and optimizing magnetic intensity, a new kind of physiotherapy in clinic filed is obtained on treatment and prevention of chemotherapeutic phlebitis.The result demonstates that magnetic therapy, a new kind of physical therapy, provides a promising therapeutic method for clinic prevention and treatment of chemotherapeutic phlebitis. However, the mechanism of constant magnet is required to investigate further.This experiment has several innovative points:1. Based on athermal biological effect of magnet, a new kind of physical therapy is achieved for the treatment and prevention of chemotherapeutic phlebitis in the clinic.2. The means of constructing the animal model of chemotherapeutic phlebitis is improved.

Change of Glucogen in Detrusor of Rats with Partial Bladder Outlet Obstruction and Their Implication

Wed, 30 Mar 2011 16:57:02 +0000

Objective:To explore the change of glucogen in detrusor of rats with partial bladder outlet obstruction(PBOO) and their implication.Materials and Methods:A PBOO rat model was created,a control group had a sham operation for comparison.Individual urodynamic data were obtained weekly,then were killed at different levels of bladder dysfunction.Bladder sections were stained with Periodic Acid-Schiff(PAS) to assess overall glycogen granules density and disposition location, scored from 0 to 3.Results were demonstrated with the 5%,25%,75%, 95%percentile.To analysis the results with the methods of Mann -Whitney U test.Results:Glycogen granules were seen in the detrusor,as their number increased their location expanded from only close to the serosa(glycogen score 1) through the detrusor(score 2) up to the urothelium(score 3); The week of obstruction and bladder humid weight were increased accompany with the glycogen score increased(P＜0.05);The max bladder pressure and volume of the score 2 and 3 group were higher than that in score 0 and 1 group(P＜0.05),there was no statistical significance between score 2 and 3 group(P＞0.05).For the bladder compliance of score 2 was decreased compared to score 1 group(P＜0.05),and was also no statistical significance between score 2 and 3 group(P＞0.05).Conclusions:The occurrence of glycogen deposits in the detrusor near the serosa is an indicator of a bladder that has more potential for functional recovery;This finding may exemplifies a important parameter for functional analysis of bladder detrusor.

Determination of Serum B-type Natriuretic Peptide in Diabetic Patients with Left Ventricular Dysfunction

Wed, 30 Mar 2011 16:57:02 +0000

Background:Diabetes is a syndrome with chronic high blood glucose as the main features. Cardiovascular diseases are main complication of diabetes and main cause of its death. The pathological changes of heart disease in diabetes included coronary heart disease and cardiomyopathy lesions. This could lead to cardiac failure. Cardia failue develops gradully, the pathology and physiological changes of which is begun with LVD. In the early stages of LVD, ejection fraction of heart is normal.In this stage, nowadays diagnosis methods are difficult. It would be benefit to control the progression of LVD, to improve cardia compensatory mechanism, if it be diagnosis and reasonable treatment on time. So early identificating the type 2 diabetes patients with cardiac diastolic dysfunction is clinically important.BNP is a peptide mainly secreted from the ventricles. When ventricular wall tension is overload or increasing, its secretion increases. BNP can reflect cardia insufficiency. Some researches show that the diagnosis of LVD is important. But there are just a few reports for the diagnosis of cardiac function in patients with diabetes. In this study, the concentration of BNP was detected in type 2 diabetes patients with LVD, to see if it is helpful in the diagnos of these patients.Objective: Serum B-type natriuretic peptide (BNP) level was detected in type 2 diabetes patients with LVD, to explore its value in diagnose this disease.Methods: Out-patient and hospital patients in Xijing Hospital from January 2008 to January 2009 were randomly choosed as research subjects. 80 cases of type 2 diabetes mellitus with left ventricular dysfunction grouped as T2DM + LVD patients, 40 type 2 diabetes mellitus with no heart changes grouped as T2DM patients, while 30 cases of normal persons and 32 cases of patients with non-diabetic left ventricular dysfunction grouped as control and LVD patients respectively. Left ventricular ejection fraction was measured with echocardiography, while serum BNP concentration was determinated by enzyme-linked immunosorbent assay.Results: BNP levels in T2DM + LVD patients (221.1±54.1 pg /ml) higher than in other 3 group patients (P<0.01), serum BNP levels in LVD (140.1±27.4 pg/ml) was higher than that in T2DM patients (82.4±29.4 pg/ml) and normal control (60.9±25.7pg/ml), with statistical significance (P< 0.05); BNP level in T2DM patients and normal control group had no difference. Receiver operating characteristic curve analysis showed that BNP cutoff point at 90pg/ml for diagnosis of LVD having sensitivity as 87%, specificity as 92% and accuracy rate as 88%. In The serum BNP levels positively correlated with age (r= 0.3326, P <0.05); and with fasting blood glucose (r = 0.2754, P <0.05). HbA1c and serum BNP level had no correlation (r = 0.2028, P = 0.0853).Conclusion: Left ventricular dysfunction patients had high serum BNP levels, while diabetic patients with left ventricular dysfunction had higher levels, suggesting that determination of serum BNP concentration can be an objective assessment of cardiac function, it is helpful to identify asymptomatic patients with left ventricular dysfunction in early stage. Detection of serum BNP is a simple, cheap and reliable screening indicator in diabetes patients with left ventricular dysfunction.

Construction, Expression and Purification of Human Anti-BP180 Single-chain Fv Antibody

Wed, 30 Mar 2011 16:57:01 +0000

Bullous pemphigoid(BP) is the most common and chronic autoimmune blistering skin disease characterized clinically by tense bullae that may develop on normal or erythematous skin, which is seen predominantly in the elderly. Up to recently, BP is usually treated with systemic glucocorticosteroids, frequently in combination with other immunosuppressants. Although the condition can be temporarily relieved, the severe side effects of glucocorticosteroids and the frequent recurrence of the disease remain to be the most serious problems[1, 2].It has been confirmed that the pathogenesis of BP is a complicated course initiated by the binding of autoantibodies, and mediated by complements and other inflammatory factors[3,.4]. The major histologic feature of BP is subepidermal blisters with variable degrees of dermal inflammation. The immunopathological tsets reveal linear deposition of C3, IgG, and/or IgE along the basement membrane zone. A majority of the patients with BP demonstrate circulating autoantibodies reactive with autoantigens located in the region of the basement membrane zone.BP180, also referred to as type XVII collagen or bullous pemphigoid antigen 2 (BPAg2), is a transmembrane protein that is associated with hemidesmosome and is thought to harbor the pathogenic autoantigen responsible for the initiation of BP. The extracellular domain especially the 16th non-collagenous A domain (NC16A) is the main pathogenic target epitope for BP autoantibodies[3, 5, 6].The pathogenic autoantibodies of BP bind to dermal-epidermal junction (DEJ) components and activate the complement system that mediates a series of inflammatory events including dermal mast cell degranulation and generation of neutrophils, eosinophils, monocyte rich infiltrates and proteinases and inflammatory mediators released, result in the loss of cell-matrix adhesion structure, and finally, subepidermal blister formation. Newly developed humanized BP animal model further elucidated the pathogenic evects of the autoantibody in vivo[7]. It was confirmed that the activation of the complement cascade via the classical pathway is required for the development of the disease. It is therefore rational to develop new therapeutic strategies by preventing complement activation via blocking autoantibody binding to the corresponding pathogenic autoantigen using targeted scFv antibody fragments.In our previous studies, phage antibody library was constructed using the the lymphocytes from BP patients and a Fab monoclonal antibody (clone P10), which specifically bind to BP180-NC16A, was isolated[8]. The purpose of the present study is to generate anti-BP180-NC16A scFv.Objective: To construct and express anti-BP180-NC16A scFv by genetically engineering technology, and to identify the binding activity and specificity of the expressed scFv.Methods: Variable region genes of VH and Vλwere cloned from Fab fragment against BP180-NC16A separately, linked with a linker sequence by SOE PCR method, and then ligated with the vetor pGEMT. After rapid screening and identification, the correct ligasion of VH-linker-Vλwas inserted into the prokaryotic expression vector p3MH, and transformed to E.coli by electroporation. The recombinant was identified by restriction double-enzyme digestion and sequence analysis. After that, soluble scFv express vector was obtained by removing the gene III fragment in the phagemid vector. Anti-BP180-NC16A scFv was expressed, and was purified using Ni affinity chromatography method. The reactivity and specificity of the expressed scFv with rhNC16A and other control antigens were tested by ELISA and Western blotting. In order to evaluate the binding of anti-BP180-NC16A scFv to the naive BP180 molecule, indirect immunofluorescence (IIF) was performed on the frozen section of normal human skin.Results: Anti-BP180-NC16A scFv was successfully constructed. DNA sequence analysis showed that there was no any mutation in Vλand VH genes of the constructed scFv. The soluble anti-BP180-NC16A scFv was successfully expressed and was purified by Ni affinity chromatography. SDS-PAGE analysis showed that the purity of scFv products was higher than 89%. By ELISA and Western blot characterization, the isolated anti-BP180-NC16A scFv was found to have a good antigenic specificity as well as excellent binding activity with the NC16A domain of human BP180. Indirect immunofluorescence (IIF) staining showed linear binding of the scFv along the dermal-epidermal junction (DEJ).Conclusion: Our success in generating the anti-BP180-NC16A scFv makes it possible to create a novel specific therapy for BP. In addition, it would be also useful in furthering the studies on the pathogenesis of the disease.

Skin Barrier Function and Its Related Influential Factors in 160 Healthy Chinese

Wed, 30 Mar 2011 16:56:59 +0000

【Objective】To greater understand the possible age,gender,regional and seasonal variation of skin barrier function in healthy Chinese.【Methods】The stratum corneum(SC) hydration,sebum content,skin surface pH and transpedermal water loss(TEWL) were measured seasonally for a year on the forehead,left forehead,nose,chin,zygomatic area,cheek,perioral area,dorsal hand(sun-exposed site) and volar forearm(sun-protected site) of the same 160 Chinese subjects with MoistureMeter SC,Skin analyzer SHP88 and VapoMeter.The SC and epidermal thickness were measured seasonally on the forehead,zygomatic area,dorsal hand and volar forearm of 49 Chinese volunteers with confocal microscope,Vivascope 1500.【Results】TEWL were significantly different(P＜0.05) in all age groups in the nose,the zygomatic area,the cheek,the dorsal hand and the volar forearm. Male TEWL was significantly(P＜0.05) higher than female except the volar forearm.TEWL exhibited regional difference(P＜0.05).The nose TEWL values were the highest,followed by the perioral area,chin,left forehead,forehead, zygomatic area,dorsal hand and volar forearm.Seasonal variations in TEWL were found(P＜0.05) and could be generally ranked as follows:winter＞spring＞autumn＞summer.SC hydration were not identical(P＜0.05) among different age groups in all tested regions.SC hydration of the age 50～60 group was significantly lower than other age groups in all studied sites.In the left forehead,and volar forearm,the male SC hydration were higher than the female,while in the chin and zygomatic area,SC hydration was higher in the female(P＜0.05).SC hydration were the highest in the zygomatic area,followed by chin,left forehead,forehead,perioral area,nose,cheek,volar forearm and dorsal hand. SC hydration showed significant(P＜0.05) seasonal variations:Summer＞spring＞autumn＞winter.The sebum content of almost all tested areas(except the cheek) were significantly influenced(P＜0.05) by age.The sebum content decreased with age.The sebum content in all investigated areas was significantly(P＜0.05) higher in the male group than the female group.Anatomical differences of sebum content were found(P＜0.05) and could be ranked as follows:nose,chin,left forehead,forehead,perioral area,zygomatic area,cheek,dorsal hand,and volar forearm.The sebum content differed seasonally(P＜0.05):summer＞spring＞autumn＞winter.The skin surface pH were affected by age(P＜0.05),with pH of the age 50～60 group the most alkaline in comparison to other groups.In the forehead, zygomatic area,cheek,perioral area,dorsal hand,and volar forearm,the female pH were higher than the male(P＜0.05).The skin pH indicated regional variations(P＜0.05) and could be put in the following order:cheek,perioral area,cheek,zygomatic area,dorsal hand,volar forearm,chin,forehead,nose, left forehead.The pH in the winter and autumn were higher than in the spring and summer(P＜0.05).The male forehead and dorsal hand SC thicknesses were higher than the female (P＜0.05).SC thickness differed locationally:dorsal hand＞volar forearm＞forehead= zygoma。The SC thicknesses were not influenced by age and seasons. The forehead epidermal thickness of the age 40～49 and 50～60 group were thicker than that of the age 20～29 group.The volar forearm epidermal thickness of the age 50～60 group were thicker than that of the age 40～49 group.The male dorsal hand epidermal thickness was thicker than the female(P＜0.05).The dorsal hand epidermal thickness was the thickiest,followed by the zygomatic area, forehead,and volar forearm.Seasonal variations were not found in the epidermal thickness.Parameters correlated with each other.TEWL correlated positively with the sebum content in the forehead,chin,left forehead,zygomatic area,cheek, perioral area,and dorsal hand.Negative correlations were detected between the sebum content and skin pH in the zygomatic area,dorsal hand,and volar forearm.TEWL correlated with SC hydration positively in the forehead,nose, left forehead,cheek,and volar forearm,but negatively in the chin and zygomatic area.Negative correlations were found between the SC hydration and skin pH in the forehead,nose,chin,left forehead,zygomatic area,cheek, perioral area and volar forearm.The forehead and dorsal hand SC hydration correlated negatively with the SC thickness.The SC hydration correlated negatively with the epidermal thickness in the dorsal hand and volar forearm. The zygomatic,dorsal hand and volar forearm SC hydration correlated positively with the corresponding epidermal thickness.【Conclusions】The SC hydration,sebum content,skin surface pH and TEWL showed age,gender,regional and seasonal variations in Chinese healthy skin. Parameters correlated with each other to a certain extent.Age,gender, anatomical sites and seasons as well as related biophysical parameters must be considered comprehensively in the selection of more appropriate skin care products and medications.

Correlation Study between the Severity of the Symptoms and the CT Signals of the Lumbar Disc Protrusion Sufferers

Wed, 30 Mar 2011 16:01:19 +0000

Objectives1. To approach the correlations between the severity of the symptoms and the CT signals of nucleus pulposus herniation( NPH)of the lumbar disc protrusion sufferers and to find the closed CT signals to the severity of the symptoms of the lumbarl disc protrusion sufferers.2. To approach the correlations between the severity of the symptoms of the lumbar disc protrusion sufferers and massage therapy .It is helpful to choose the appropriate patients for the massage doctors.Methods1. data and case 129 patients who are diagnosed with lumbar disc protrusion sufferers and checked by CT from Nanfang Hospital.2. inclusion and exclusion criteria Patients who met the criteria of lumbar disc protrusion sufferers are checked by clinical and CT examinations in Nanfang Hospital. If the patients have more than one lumbar intervertebral protruding discs, we will choose the most serious one which is the most fit to the symptoms. Excluded patients such as: pregnant womman, psychiatric patients, severe osteoporosis patients, spondylolisthesis patiens, bone fracture patients, cancer patients, tuberculosis patients, spinal stenosis patients, data incompleted or lost. 3. score criteria of the severity of the symptoms and signs To use the low back pain assessment scale from Japanese Orthopedic Association. It have ten items including low back pain, numb,tenderness, strength of flexion and extension of hallux muscle, straight leg raising and the strengthening of the experiment, radiating pain, bending and lifting weights, walking distance or time, in bed time, working ability. Every item have 4 levels, the highest level is 3 points, the lowest level is 0 point. Getting more points means more serious. The total points of the assessment scale is 30. The mild level points are less or equal 10 points. The moderate level points are less or equal 20. The severe level points are more than 20 points. After massage therapy, if the symptoms of the lumbar disc protrusion sufferers are gone, it means the patients has been cured. If the points are less 10 points than prior-treatment, it means the effect is excellent. If the points are less 0-10 points than prior-treatment, it means the treatment effect is valid. If the points have no changes than prior-treatment, it means the treatment effect is invalid.4. CT signal According to the literature and clinical experience, there are ten signals including 31 performances which relate to the lumbar disc protrusion sufferers. Such as schmorl nodule or not, the NPH through bone or not, the Patho-type(Protrusion, Prolapse, Liberation, Shrink back), the NPH density(≤80,≤100, > 100), the NPH shape(arc, the edge is smooth; the edge is not smoth; cauliflower-like, island-like), the size of NPH of sagittal plane(1 level, 2 level, 3 level) , the size of NPH of horizontal plane(a domain, b domain, c domain, d domain), the size of NPH of frontal plane(1 zone, 2 zone, 3 zone, 4 zone) , the oppression of NPH to nerve root(close to nerve root, compression to nerve root, adhesion to nerve root), the oppression of NPH to dural sac(fat gap smaller, fat gap disappears, deformation to dural sac). 5. Measuring equipment of CT signals To scan L3-L4, L4-L5, L5-S1 vertebrals with Lightspeed 16 layer spiral CT scanner of GE company. The thickness of scan is 5mm, the layer spacing of scan is 0.1mm. To complete the measurement of the image data in the computer with the help of the PASC system of the image center in Nanfang hospital.6. Measurement of the CT signal (1)Schmorl nodul: the nucleus pulposus into the vertebral body. It appeared low-density lesion around with vertebral bone sclerosis at CT. (2) The NPH through bone: Free bone at the edge of vertebral. The degenerative nucleus pulposus has penetrated between epiphyseal and vertebral, which makes epiphysis free.(3). The Patho-type of protrusion:CT showed a arc-shaped soft tissue shadow with intact border connected with the disc, whose density consistent with the disc, prominents partially to the marginal median or the side of lumbar posterior and its edge and the posterior of annulus intersect into an acute angle . Prolapse: nucleus pulposus breakes annulus fibrosus into vertebral canal, protrution edge makes into acute angle with annulus fibrosus. liberation fragment: liberation fragment makes angulated or polyps high-density in the vertebral canal, intervertebral discs posterior border seems normal or posterior process slightly. Shrink back: The NPH is dehydration or ossification.(4). The shape of nucleus pulposus herniation To be soft tissue shadow of epidural gap. The density of nucleus pulposus herniation is higher than that of spinal dura mater and fat of epidural gap. The essence of soft tissue shadow of epidural gap is nucleus pulposus herniation. (5). The density of nucleus pulposus herniation When nucleus pulposus herniation appears, the change of posterior border of intervertebral discs is from flat to partly prominent. Measure the CT value of the protrusion. The normal situation is 50-100Hu. If CT value is higher than 100, it means the tissue has been ossification. (6). Anteroposterior axes level :There are 3 level in the anteroposterior axes. The intervertebral discs level is 1 level. The upper tier of intervertebral discs is from the inferior thyroid notch of upper vertebral body to the upper bound of intervertebral disc which we called 2 level. The underlayer of intervertebral discs is from the lower bound of intervertebral disc to the inferior thyroid notch of lower vertebral body which we called 3 level. (7). Horizontal position region There are 4 regions behind the posterior border of the vertebral body. To divide 3 equation between two pedicles of vertebral arch, the middle equation is called 1 region,the left or right one called 2 region.3 region is from exterior to interior of pedicle of vertebral arch. 4 region is outside the lateral pedicle of vertebral arch. (8). Z axes region To divide radius vector which is from center line of posterior border of the vertebral body to anterior border of vertebral plate of spinous process into 4 equation . We called it a,b,c,d domain separately from the front to the back.. (9). In the normal situation, density of nerve root is as seem as that of tissue contrast by epidural fat. It is in the intervertebral foramina of vertebral canal. The left one is symmetry to the right one. if nerve root has no shift in the lumbar disc protrusion sufferers, we call it\'no change\', if nerve root moved, we call it \'root compression\', if nerve root are covered by NPH, we call it \'nianlian\'. (10) The density of Dura mater and its contents is lower than that of the disc,the fat is filled with the district from the membranous sac and edge of the discs. The interfacing is clear because of the density difference. We divide 3 levels (the fat gap become smaller, the fat gap become disapear, spinal dura mater shift because of oppression)acording to the position relation between nucleus pulposus herniation and spinal dura mater.7. The massage (1). The patients is in the prone position,the doctor massage on their back from up to down with rolling and rotating maneuvers, especial low back living acupuncture point, giving consideration to acupoints and the Bladder Meridian of Foot-Taiyang and its points. The doctor relax the patients with rolling and rotating maneuvers along sciatic nerve, from Chengfu to Weizhong to Chengshan to Kunlun. The time is 10-15mins.(2). Patients is in the lateral position, with located in the down natural extension of the lower limb and lower limb above the knees hip flexor. The doctor stands facing patients, with one hand pressing the patient\'s shoulder anterior part, with another elbow resisting the patient\'s buttocks, For the slowly push on the opposite direction, which makes lumbar vertebrae moving slowly. When there is resistance, pull the last time by a substantial, pulled on the control of sudden. The right and the left would be down one time. We massage every patient two times every week and the treatment will last for 2 months.8. Error Control At single-blind conditions, CT signals are measured for 3 times, then averaged the results. It is not allowed to view the clinical assessment scale in the measurement, and we will select the most serious disc to measure if two or more intervertebral discs are prominent in patients.9. The course of the experiment According to the clinical data and experience, we maked sure CT signals related to the the lumbar disc heriation signs. At first we taked physical examinations to the patients from chinese medical department, learned the gravety of the symptoms,according to the waist assessment scales recommended from Japanese Orthopedic Association, we maked groups acccording to 3 degrees:light, moderate and heavy, and then we make a correlated-test and a logistic-test. We take the massage medical treatment, two mounths later, we maked a score on the gravety of the symptoms according to the waist assessment scales recommended from Japanese Orthopedic Association. The score of prior-treatment and post-treatment was tested by correlate-test to watch if massage therapy was effective . According to the results we choose the appropriate patients for the massage doctors.Results 1. There is no statistics significance between the schmorl and the severity of the symptoms of the lumbar disc protrusion,so is the NPH through bone and the size of NPH of sagittal plane.(p>0.05). But the patho-type,the NPH density, the NPH shape, the size of NPH of horizontal plane, the size of NPH of frontal plane, the oppressio of NPH to nerve root, the oppression of NPH to meninx fibrosa have the statiscs significance with the severity of the symptoms and signs of the lumbar disc herniation.(p<0.01).2. The size of NPH of horizontal plane, the size of NPH of frontal plane, the oppressio of NPH to nerve root have the statiscs significance with the severity of the symptoms and signs of the lumbar disc herniation. Compared to d domain, a domain and b domain and c domain could made the lumbar disc protrusion sufferers less serious(OR is0.003, 0.002, 0.008 separataly). Compared to 4 zone, 3 zone could made the lumbar disc protrusion sufferers more serious(OR is28.560). Compared to the adhesion to nerve root, compression to nerve root could made the lumbar disc protrusion sufferers more serious(OR is 8.882).3. 129 patients are carried by massage therapy. Based on the assessment scale, the severity of symptoms and signs for the light has 44 cases of patients, of which 15 were cured, 16 markedly effective, 8 effective, 5 null and void,the efficiency is 88.64%. The severity of symptoms and signs for the middle level has 61 cases of patients,of which 9 were cured, 23 markedly effective, 17 effective, 12 null and void, the efficiency is 80.16%. The severity of symptoms and signs for the severial level has 24 cases of patients, of which 2 were cured, 8 markedly effective, 6 effective, 8 null and void, the efficiency is 66.67%. The effect of the 3 groups is obviously different(P为0.007).Conclution1. The patho-type, the NPH density, the NPH shape,the size of NPH of horizontal plane, the size of NPH of frontal plane , the oppressio of NPH to nerve root, the oppression of NPH to meninx fibrosa effect the symptoms and signs of the lumbar disc herniation.2. The smaller is the size of NPH of frontal plane, the smaller is OR. 3 zone , the oppressio of NPH to nerve root could made the lumbar disc protrusion sufferers more serious.3. Massage is effective to lumbar disc herniation. The medical treatment is closely related to the gravity of the lumbar disc heriation signs. The lighter are the severity of the symptoms and signs, the better are the medical treatment effect.

The Clinical Research on External Fixation Porsture of Supination-eversion Fracture of Ankle

Wed, 30 Mar 2011 16:01:17 +0000

Objective:To explore the best external fixation posture of ankle fracture of Lauge-HansonⅢ°andⅣsupintion-eversion after manipulative reduction with a view to heightening the clinical curative effect and reducing the incidence of traumatic arthritis.Method:according to our selection standards,42 patients,followed up with periodical visits, were selected for this test and were randomly divided into two groups.One was fixed to the posture with inversion,internal rotation and plantar flexion,the other was fixed to the posture with inversion,internal rotation and dorsiflexion.Each patient was followed up with visits for 6 to 18 months.The AOFAS Scoring was adopted to evaluate the clinical outcomes of the two groups. Moreover,through a statistical comparison of complications,the short-terms clinical effects of these two groups and their relevant factors come up for evaluation.Results:Both groups were followed up with visits for 6 to 18 months.According to the AOFAS Scoring,for the group of plantar,results were as follows:excellent in 16 cases,good in 3 cases,fair in 2 cases,poor in 0 case,with the excellent rate 90.48%;while for the group of dorsiflexion:excellent in 9 cases,good in 8 cases,fair in 4 cases,poor in 0 case,and the excellent rate being 80.95%.So there was obvious difference between two groups(P＜0.05).The same was in the case of their AOFAS Scores,as follows:the group of plantar was 91.62±6.53,and the group of dorsiflexion was 85.24±8.43.The complications in the near future:there were 2 cases with stiffness of joint in the plantar flexion group;and there were 3 cases ached and 1case with stiffness of joint in the dorsiflexion group.So there was obvious difference(P＜0.05).Conclusion:In terms of the near-future clinic curative effects,the posture with inversion, internal rotation and plantar flexion exceeds the one with inversion,internal rotation and dorsiflexion with plaster after manipulative reduction.

Aspirin Enhance the Expression of VEGF in Ischemia/Reperfusion in Rats

Wed, 30 Mar 2011 16:01:16 +0000

Objective: Vascular endothelial growth factor (VEGF), which is also called vascular permealility factor, is a powerful factor to promote an- giogenesis and a neurotrophic factor in vivo. VEGF can directly affect vascular endothelial cell to enhance its multiplication,and can directly or indirectly participate in Angiogenesis and enhance vasopermeability. It can also directly affect cellula nervosa, and bring neurotrophy and neuro- protection function into play by IP3K/ Akt/NF-κB and MAPK/ ERK signal mediate, including reducing apoptosis, advancing neural stem cell mu- ltiplication, migration and differentiation and improving nerves function. Aspirin(ASA) has been known as a classic non-steroidal anti-inflammatory drug and is widely used in clinics as an antipyretic, analgesic, anti- inflammatory agent, antirheumatic medication and antithrombotics. Apart from its preventive actions against stroke due to its antithrombotic properties, recent data in the literature suggest that ASA also exert direct neuroprotective effects by inhibiting excitatory amino acid release, NF- kappaB translocation to the nucleus, inducible nitric oxide synthase (iNOS) expression, decrease of brain ATP levels, production of free radical and inflammatory and facilitate neuron nitric oxide synthase(nNOS) expression and Bcl-2/Bax. suppressing the expression of Fas-Lt caspase-3 , and inducing the expression of NGF, bFGF and BDNFMethods: The rat model of MCA-CI/RP were established with suture occlusion technique according to modified Zea Longa method and the rats of 1-3 score(Bederson measuring scale) was selected into the 10 groups at 6 hours after CI/RP. Some cerebral slices were taken for TTC staining. The rats were divided into five groups according to the time of reperfusion in control groups: A(0h), B(6h), C(24h), D(48h), E(3D), F(5d), G(7d), so did the experimental groups (A\'~E\'). ASA were given to the experimental groups with the dose of 80mg/kg via an i.p. route, while the same dose of diaminocaproic acid were given to the control groups after the CI/RP and the next three days till killed at assigned time. The neurologic detect scale were used by Bederson scores. The VEGF expression in CI/RP were detected by immunohistochemistry technique.Results: Comparing with the control groups, the Bederson measuring scale of experiment groups were conspicuously reduced, but only at the 3 days after CI/RP had statistical significance(P<0.05), and there was more significance at the 7 days (P<0.01). After CI/RP, VEGF could be mainly seen around Ischemia area in infarct side, and also be seen in neurons, gliacyte, and kytoplasm of vascular endothelial cell. The positive cell immunohistochemisty staining was brown. In ASA and comparing groups, the trend of VEGF expression were the same. 6 hours after CI/RP, the expression increased. After 24 hours, it increased obviously (P<0.05). It reached the hightest at the 48 h, and then gradually decreased. The VEGF expression of ASA group was higher than the comparing group. There was significant difference between 24 hours to 3 days (P< 0.05). The di- fferences were more obvious at the 24h and 48h (P<0.01).Conclusion: 1. ASA could obviously improve the rat’s neurologic impairment after CI/RP. 2.After CI/RP, the expression of VEGF was increased and reached the highest at the 48 h. 3. ASA could increase VEGF expression after CI/RP, but it would not chang the peak time. 4. It was one of the ASA neuroprotection to add ischemia brain tissue VEGF expression.

The Clinical Study of the Chairside CAD/CAM Made Restorations

Wed, 30 Mar 2011 16:01:15 +0000

The chairside CAD/CAM system is a trend of stomatology ,because it is a dental restoration system that allow dental professionals to deliver high-quality esthetic and presise restorations to their patients at chairside without laboratory intervention. At present, in China, the application of it is not that extensive for kinds of reasons. The more appliance of CAD/CAM technology is the design and manufacture of aluminum-oxide and Y-TZP based FPD frameworks in laboratory which are not that proficient for making a restoration.The high cost of the machinable ceramic blocks is one of the main reasons that limit the application of the chairside CAD/CAM system. These machinable ceramic blocks now are all made abroad and very expensive. Furthermore, the restoration made by it has long been thought to be formed in a process meaning that once it has a fault, it has no remedy and has to be disposed. The cost then is increased much. Therefore, cutting down the cost of material is the key to this problem.This issue is composed of three experiments. First, make a 8-year follow-up study of 24 Cerec2 CAD/CAM all-ceramic crowns to evaluate their clinical performances, which can be the base for the further study. Second, investigate shear bond strength between Vitablocs markⅡand four kinds of commercial veneering ceramics so as to find a suitable veneering ceramics to make up the imperfect restorations. Thrid, make restorations of Vitablocs Mark II and make a short-term follow-up study so as to accumulate the operation experiences for further extensive application. At the same time, try to make restorations of self-made ceramic blocks to evaluate their clinical applicability.Experimental results1. After the 8-year follow-up, 3 of the 24 restorations failed as results of ceramic crack, loosing and extraction of the abutment teeth, root fracture of the abutment teeth. The failed occurred respectively at 2 months, 43 months and 53 months after restoration and all the failed cases’abutments are molor. While 86% of the other 21 restorations’marginal fitness, surface texture, shape and color which still function well achieve the standard of level A. The results of the satisfaction questionnaires show that all the targets’satisfaction rates achieve 100% except the expenses. According to Kalplan - meier analysis, the success rate of Cerec2 crowns was 87.5% after 8 years’function in oral.2. the shear bond strengths of four groups of E-max ceram、Vintage ZR、Cercon ceram kiss、Vita VM9 are 41.07±12.42、43.44±11.32、35.81±8.69、37.35±9.71(MPa) respectively .More than 67% of the specimens′fracture originated and propagated in the base ceramics .The result of one-way ANOVA (F=0.641,P>0.05 )shows that there is no statistic differences among the data of four groups. Comparing with ISO9693: 1999 standard using one group t-test, all of the data of the four groups were higher than the standard. The results of SEM showed that the combination interface between 4 kinds of veneering porcelain and base porcelain are smooth without obvious air bubbles and therefore can indicate a satisfied combination.3. 27 Cerec restorations were fabricated using the Vitablocs Mark II ceramic blocks, of which three restorations failed as results of ceramic crack and crown fracture of the abutment teeth. The failed occurred respectively at 1 month, 3 months and 4 months after restoration. The results of the satisfaction questionnaires show that all the targets’satisfaction rates achieve 100%. While more than 87% of the restorations’marginal fitness, surface texture, shape and color which still function well after one-year follow-up achieve the standard of level A. 4 Cerec restorations were fabricated using the self-made ceramic blocks. 2 restorations failed during the veneering and staining process, the cause may be the unsuitable sintering temperature or the dismatching between veneering porcelain and base porcelain. The other two restorations without veneering and staining still function well after more than two-month follow-up.Conclusion1. The Cerec2 chairside CAD/CAM system can allow dental professionals to deliver high-quality esthetic and presise restorations to their patients at chairside without laboratory intervention. The only shortcoming of the restoration is the occlusion appearance which need to be modified.2. The data of the shear bond strength between Vitablocs markⅡand four kinds of commercial veneering ceramics are higher enough to meet the requirement of clinic use. All of the four kinds of veneered porcelain can be used as the individual character veneering ceramics for Vitablocs markⅡ. 3. The crowns of Vitablocs markⅡmade by Cerec2 CAD/CAM system can be kept in oral and function well for many years.4. Comparing with the Vitablocs Mark II ceramic blocks, the self-made ceramic blocks have satisfied artherectomy ability, but unsatisfied color which need to be improved in the further study.

Experimental and Clinical Studies on the Bonding Performance to CAD/CAM Feldspathic All Ceramic Restoration

Wed, 30 Mar 2011 16:01:14 +0000

Tooth defect is common and frequently-occurring disease of human being,People need for restorations and treatments of inlay,crown and bridge. Traditional production processes are time-consuming, patients have to take many appointments to complete treatment.The technology of CAD/CAM which integrates with mathematics,optics,electronic technology,image recognition and processing,automatic control and manufacture has advantages with precise,rapid,efficient and single visit.The most representative CAD/CAM chairside dental restoration system is CEREC,which can fabricate all ceramic restorations in a single visit.Feldspathic machinable ceramic blocks are used in CEREC.The restorations are bonded to dental tissues with resin cement,the bond performance between feldspathic ceramic and resin cement is greatly meanful for the durability and fine function. Vita MarkⅡblock and four types of resin cements are selected in the present study(RelyX Veneer,Panavia F,RelyX Unicem,FL-BOND+Beautifil Flow).The purpose of this study was to evaluate the effects of different ceramic surface treatments and thermo-cycling on the bonding strength and bonding interfaces between feldspathic machinable ceramic and four types of resin cements.Eighty-six restorations are assessed using modified USPHS criteria.The results from this study could present to provide some references for clinic procedures.According to the results,the following conclusions could be achieved.1.The bonding strength of silicane group utilizing four types of resin cements were 12.18±2.86 MPa,12.47±3.54 MPa,11.71±3.14 MPa,7.27±3.13 MPa,which were significant lower than the acid group(15.26±3.02 MPa,16.72±3.10 MPa,15.53±3.47 MPa,10.68±3.22 MPa),it illustrated that simple chemical combination was weak,higher bonding strength could be achieved to mechanical combination.2.The highest bonding strength can be seen in alliance group,they were 20.61±2.89 MPa,22.02±4.03 MPa,20.50±3.18 MPa,18.96±2.73 MPa,it can be conclude that alliance of silicane and acid significantly enhance the bonding strength of feldspathic machinable ceramic.SEM observation revealed that bonding interfaces of alliance group were dense between ceramic and resin cement.3. The shear bond strengths of the groups of RelyX Veneer,RelyX Unicem and FL-BOND+Beautifil Flow dropped markedly from 19.86±3.39MPa、20.66±3.08MPa、21.18±3.30MPa to 16.42±1.90MPa,16.28±3.45MPa and 13.29±2.47MPa after thermo-cycling of 5,000 cycles.Panavia F showed the greatest bond strength after thermocycling among the four resin cements used. 4. According to modified USPHS criteria after one year,the restorations which reached Alpha were 85.7% in marginal adaptation,83.9% in anatomic form,88.9% in surface texture,85.7% in color match.There was no significant difference between four types of resin cements.

The Immunohistochemic Expression of DACH1 and Its Clinical Significance in Papillary Thyroid Carcinoma and Hashimotos Thyroiditis

Wed, 30 Mar 2011 16:01:13 +0000

Background and Objective: DACH1 is the analog of Drosophila dachshund gene in vertebrate, it is inhibited oncogene-mediated oncogenesis, blocking cancer epithelial cell DNA synthesis , colony formation, growth in Matrigel and tumor growth in mice . The role of DACH1 gene expression in papillary thyroid carcinoma and Hashimoto’s thyroiditis was unknown. Our experiment is in order to investigate the immunohistochemic expressions of DACH1 in the tissues of the papillary thyroid carcinoma, the Hashimoto’s thyroiditis and the nodular goiter, to explore the relationship between their expressions and the carcinogenesis and development of the papillary thyroid carcinoma.Methods: 30 cases of papillary thyroid carcinoma, 10 cases of Hashimoto’s thyroiditis and 10 cases of nodular goiter were involved. Immunohistochemic SABC method was used to detect the expression of DACH1. 30 cases of papillary thyroid carcinoma were divided into 5 subgroups according to clinical features such as the size of tumor, with metastatic lymph nodes or not, age and sex. The correlations of the expressions of DACH1 with the clinical pathological parameters of the papillary thyroid carcinoma were analyzed.Results: (1) The immunohistochemic expression of DACH1 in papillary thyroid carcinoma and nodular goiter is orientated as the cytomembrane and cytoplasm. The positive rate of DACH1 is 76.7% in papillary thyroid carcinoma, notably higher than that in nodular goiter, later is 30% (P<0.01). (2)The positive rate of DACH1 is 60% in Hashimoto’s thyroiditis, also higher than that in nodular goiter (P<0.01).(3)The positive rates of expression of DACH1 protein in papillary thyroid carcinoma with lymph nodes metastasis was 89.5%, while it was 54.5% in group of without lymph nodes metastasis. The differences are significant (P<0.05).Conclusions: The DACH1 protein is immunohistochemic overexpressed in papillary thyroid carcinoma and Hashimoto’s thyroiditis. There is no obvious relationship among age、sex、the size of tumor and TNM of the positive expression of DACH1,but there is obvious relationship between the degree of infiltration and metastasis of lymph node, indicating that DACH1 may play a role in the carcinogenesis and development of papillary thyroid carcinoma.

The Effect of Long Term Repeated Hyperbaric Oxygen Preconditioning on Spinal Cord Ischemia Reperfusion Injury in Rabbits

Wed, 30 Mar 2011 16:01:11 +0000

The concept of preconditioning was first proposed by Murry in 1986. It is an interventional strategy that is given before ischemia to prevent the following ischemic injury. The internal environment changes caused by preconditioning could accommodate the organism to the ischemic insult and alleviate the damage. This phenomenon is known as ischemic tolerance. After it has been successfully applied to protect the heart from ischemic injury, various studies have confirmed its protective effect in the central nervous system. And the way of preconditioning has been expanded from moderate ischemia to hypoxia, hyperbaric oxygen (HBO), inhaled anesthetics, electric acupunture, hypothermia and several drugs. And the mechanisms involved in this protective effect include inhibiting excitatory toxicity, oxidative stress and inflammation during ischemia, affecting signal transduction and neurovascular pathophysiology, and preventing cell death and gene damage. HBO has been applied clinically treating many neurological diseases. It has been proved that HBO treatment could reduce ischemia reperfusion injury in the central nervous system. And we have reported that 2.5-3.0 ATA, 100% O2, 1h·d-1, 3-5 d of HBO preconditioning could induce ischemic tolerance in rat’s brain or rabbit’s spinal cord. However, the effect of long term repeated HBO preconditioning on ischemia reperfusion injury in the central nervous system has never been evaluated before. Therefore, the following two experiments are designed to investigate whether long term repeated preconditioning could induce more potent ischemic tolerance in the rabbit spinal cord, and to investigate the relationship between the neuroprotective effect produced by HBO preconditioning and neuronal apoptosis/degeneration. This research may provide essential information for protocol design of HBO preconditioning when coming to clinical use.Experiment 1 The effect of long term repeated hyperbaric oxygen preconditioning on the spinal cord ischemia reperfusion injury in rabbitsObjective: To investigate the effect of long term repeated HBO preconditioning on spinal cord ischemia reperfusion injury in rabbits.Methods: 45 New Zealand rabbits were randomly divided into 5 groups. Sham group (SHAM, n=5): open the abdominal wall and dissect the abdominal aorta without cross clamping. Control group (CON, n=10): animals received no pretreatment. The abdominal aorta was cross clamped for 20 min to produce spinal cord ischemia reperfusion injury. Hyperbaric oxygen preconditioning groups (HBO5, HBO10 and HBO20 group, n=10 respectively): animals in each group received 2.5 ATA, 100% O2, 1 h·d-1 of HBO preconditioning for consecutive 5 d, 10 d or 20 d, respectively. And spinal cord ischemia reperfusion injury was induced at 24 h after the last session of HBO preconditioning. Neurologic function was evaluated at 4, 8, 12, 24 and 48 h after reperfusion according to modified Tarlov score criteria. The spinal cords were collected after the last neurologic function evaluation and stained with hematoxilin and eosin. The number of normal neurons in the anterior horn of the lumber spinal cord was calculated.Results: Animals in HBO5 and HBO10 groups scored higher than the animals in CON group (P<0.05 and P<0.05, respectively) in the neurologic function evaluation. But the modified Tarlov score of animals in HBO20 group is similar to that of animals in CON group. All groups but HBO5 group had fewer normal neurons in the anterior horn of the lumber spinal cord as compared to SHAM group (P<0.05). But more normal neurons were preserved in HBO5 group and HBO10 group (P<0.05, P<0.05) compare to CON group, whereas no significant difference in this evaluation was found between HBO20 group and CON group. And the number of normal neurons in the spinal cord of animals in HBO20 group was less than that in HBO5 group.Conclusion: 5 d or 10 d of HBO preconditioning induce ischemic tolerance whereas 20 d of HBO preconditioning fails to protect the spinal cord from ischemia reperfusion injury. 5 d of HBO preconditioning is the most effective protocol in this research.Experiment 2 The effect of long term repeated hyperbaric oxygen preconditioning on neuronal apoptosis and degeneration in the spinal cord of rabbits after ischemia reperfusion injuryObjective: To investigate the effect of long term repeated HBO preconditioning on neuronal apoptosis and degeneration in the spinal cord of rabbits after ischemia reperfusion injury.Methods: 25 New Zealand rabbits were randomly divided into 5 groups. Sham group (SHAM, n=5): open the abdominal wall and dissect the abdominal aorta without cross clamping. Control group (CON, n=5): animals received no pretreatment. The abdominal aorta was cross clamped for 20 min to produce spinal cord ischemia reperfusion injury. Hyperbaric oxygen preconditioning groups (HBO5, HBO10 and HBO20 group, n=5, respectively): animals in each group received 2.5 ATA, 100% O_2, 1 h·d~(-1) of HBO preconditioning for consecutive 5 d, 10 d or 20 d respectively. And the spinal cord ischemia reperfusion injury was induced at 24 h after the last session of HBO preconditioning. Spinal cords were collected at 48 h after reperfusion and stained with TUNEL or Fluoro-Jade B. The number of TUNEL positive neurons and Fluoro-Jade B positive neurons in the anterior horn of the lumber spinal cords were calculated and analyzed.Results: TUNEL positive neurons and Fluoro-Jade B positive neurons in the spinal cords of animals in HBO5 group and HBO10 group were fewer than that in CON group (P<0.05 and P<0.05, respectively). But no significant difference was found between HBO20 group and CON group in these two evaluations. And the number of Fluoro-Jade B positive neurons in HBO20 group was more than that in HBO5 group or HBO10 group (P<0.05 and P<0.05, respectively). Conclusion: 5 d or 10 d of HBO preconditioning prevented apoptosis and degeneration of neurons in the ischemic spinal cord. But 20 d of HBO preconditioning had no protective effect on these patterns of neuronal death, which might be a major cause for the inability of HBO preconditioning to induce neuroprotective effect when prolonged to consecutive 20 d.

The Influence of Resistance Training on Serum Leptin and Adiponectin

Wed, 30 Mar 2011 16:01:10 +0000

Objective: The purpose of the present study was to investigate the different feature of leptin、adiponectin in resting or after acute resistance training by Enzyme-Linked Immunosorbent Assay determined method. And analyzed the relationships between adipocytokines and other body composition and biochemical factors, the mechanism of resistance exercise in energy balance and obesity development,so as to provide theoretical evidence and guidance to make exercise prescription to lose weight and control weight.Methods: Twelve healthy volunteer male wrestling(training year3.25±1.18),as A group; fourteen bodybuilding fans as B group;twelve sports kinetics students of WuHan institute of physical education as C group.Overnight fasting,blood samples were collected in venous(5ml) before and immediately after exercise, and analysis with Automatic Biochemical Analyzer, statistical analysis were performed by using SPSS11.0 software.Results: 1.The leptin levels in resing:A group4.94±1.03ng/ml more than B group4.73±0.89ng/ml,compared with C group, Both A and B significantly more than control group(P<0.01)2.the adiponectin levels in resting:A group16.19±2.92μg/ml is highest,compared with control group,Both A and B significantly more than control group(P<0.01)3. Immediately after exercise, Serum leptin concentration had decreased significantly(P<0.01),However,serum adiponectin did not decreased significantly.4. leptin has positive linear relationship with Fat%(P<0.01)、Fat mass(P<0.05)、BMI(P <0.05), but adiponectin has negative linear relationship with Fat%(P<0.05)、TG(P<0.05), positive linear relationship with muscle(P<0.01)、fat-free body weight(P<0.01).Conclusion: 1.Chronic resistance training can increase serum leptin and adiponectin levels. However,considered fat mass, resistance training has more effect on adiponetin than leptin.2.Acute resistance exercise don’t significantly affect serum leptin and adiponetin.3.The present indicated that leptin and adiponectin have linear relationship with fat% significantly.4.In addition,it is limited to take BMI as a parameter to investigate the correlation of adiponectin and leptin.

The Experimental Study of Suxiao Jiuxin Pellet on Effect the Experimental of SDF-1/CXCR4 and Anti-Atherosclerosis

Wed, 30 Mar 2011 16:01:08 +0000

Objective:Through establish model of rat with atherosclerosis(AS),This experiment designed to observe the effect of Suxiao Jiuxin pellet(SX) on pathology of morphological changes in AS rat aortic wall and the expression of stromal cell-derived factor(SDF-1),its receptor CXCR4(CXC chemokiner re ceptor-4) on aortic wall,and to further explore it\'s possible mechanism of anti-AS.Method:Experimental atherosclerosis in rats was produced by feeding with atherogenic-diet and vitaminD3 administration(600,000 IU/Kg by intraperitoneal inject at one time) for 12 weeks.60 male mature Sprague-Dawlays rats were devided into 6 groups randomly and equally:①normal group(N):normal diet;②AS model group:atherogenic-diet;③low dosage SX group(SXL):aterogenic-diet and SX60 mg.kg~(-1).d~(-1);④mediume dosage SX group(SXM):aterogenic-diet and SX600 mg.kg~(-1).d~(-1);⑤high dosage SX group(SXM):aterogenic-diet and SX1800 mg\'kg~(-1).d~(-1);⑥Astorvastain group(ATO):aterogenic-diet and ATO4 mg\'kg~(-1).d~(-1).In the beginning, all groups were adaptive fed by normal diet for one week.Then SX or Astorvastain was given by gavaging from the beginning of establish AS model to the end of the experiment.At the same time,normal saline was given to the rats of normal group and model group with the same volume and the same way.After 12 weeks,all rats were killed by breaking away from the spinal cord.Aortas were taken and sectioned quickly.Hematoxylin and eosin(H&E),masson staining were used for morphological investigation by light microscope.Measured intima thickness(IMT)、media thickness (MT) of artery stained by HE and artery collagen area(CA)、vascular cross-sectional area(CVA) as well as the ratio(CA/CVA) of artery stained by masson respectively with Pro Plus 6.0 image analysis system.The expression of SDF-1 and CXCR4 were examined by immunohistochemistry and western blot.Results:1.Pathomorphological changes in aorta(1) Pathological staining:HE stain:In normal group,the structure of the aortic wall was well-constructed.The intima was thin and endothelia were intact.The smooth muscle cells didn\'t proliferate.In the model group,Some endothelial cells were lost and the intima thicked.The smooth muscle cells proliferated and lined up in disorder. The structures and arrangements of elastic fibers were in disorder and the fibrous tissues proliferated.Compared with the model group,the pathological changes lessened in the SXM,SXH and ATO groups.Masson stain:In the normal group,the structures and arrangements of elastic fibers were intact and in order.In the model group,the vascular smooth cells and collagen fibers proliferated obviously in the SXM、SXH and ATO groups.(2) Image Analysis Results:The numerical value of intima-media thickness(IMT), media thickness(MT),collagen area(CA),vascular cross-sectional area(CVA) as well as the ratio(CA / CVA) were higher than the normal group,the differences were statistically significant(IMT,MT,CA,CA / CVA:P 0.05),while the level of IMT、MT and collagen fiber content were declined in the SXM,SXH and ATO group,especially the SXH group and the ATO group(P 0.05).2.Immunohistochemistry Results:The expression of SDF-1 and CXCR4 in the model group were increased when compared with the normal group(P 0.05).3.Western Blot Results:The results are basically the same with immunohistochemistry.The expression of SDF-1 and CXCR4 in the model group were increased when compared with the normal group(P<0.05);Compared with M, the expression of SDF-1 and CXCR4 were not influenced in the SXL group(P <0.05), while,both were increased in the SXH and ATO groups(P <0.05).Conclusions:The rat model of atherosclerosis can be established successfully by atherogenic-diet and vitamin D3 administration(600,000 IU/kg by intraperitoneal injected at one time).The result suggest that Suxiao Jiuixn pellet can ameliorate AS pathological changes of rats\' artory by resisting and inhibiting the proliferation of AS rats\' endarterium、membrane,and collagen fibers,thus delay the process of being AS. The possible mechanism is Suxiao Jiuxin Pellet can inhibit chemokine SDF-1 / CXCR4 axis function.

Toll-like Receptor 4 Ligand Can Differentially Modulate the Release of Cytokines by Human Platelets

Wed, 30 Mar 2011 16:01:07 +0000

ObjectiveThis study examined the immune activation and the immunomodulatory factors released by platelets with the stimulation of Escherichia coli lipopolysaccharide(LPS) after Toll-Like Receptor 4(TLR4) engagement on their surface,which lead to the activation of platelet and the difference of the release of sytokines.This study shows that the plates are involved in the course of inflammation and Toll-like receptor 4 ligand can differentially modulate the release of cytokines by human plateletsMethodsPlatelet-rich plasma(PRP) were got from blood of 15 healthy person.Platelets were incubated with the anti-human FcγRⅡMonoclonal anti-body to saturate the free FcγRⅡand block FcγRⅡengagement.The PRP were grouped under many parts:1.Stimulating the PRP with LPS which have different concentration(0μg/ml、1μg/ml、3μg/ml and 5μg/ml).Platelet surface expression of TLR4 was determined by flow cytometric analysis.2.The platelet were stimulated with TLR4 ligand LPS,in the presence or absence of a blocking anti-human TLR4 MoAb.And the release of IL-8,β-TG,TNF-αand sCD40L were measured by specific enzyme-linked immunosorbent assay.Results1.IL-8 release was independent of platelet activation after TLR4 engagement(P=0.657);sCD40L、β-TG、TNF-αwere present in large concentration in the releaseate of platelet stimulated by TLR4 ligand(P＜0.01 ).2.Platelet surface expression of TLR4 were decreased after the engagement of LPS,and it was independent to the LPS with different concentration(P＞0.05).3.The effects of LPS from E.coli on the modulation of secretory factors were attenuated by preincubation of platelets an anti-TLR4 Monoclonal antibody.ConclusionWe propose that platelets adapt the subsequent responses with polarized cytokine secretion,after TLR4 involvement.TLR4 ligand bound platelet TLR4, which differentially modulates the release of cytokines by platelets.

The Contrast Analysis of Pathologic Results with Ultrasound and Radioisotope Scanning in Solitary Thyroid Nodules

Wed, 30 Mar 2011 15:31:33 +0000

BackgroundNodular thyroid disease is very common and more prevalent in women than in men.Thyroid nodules were found in 4%to 7%of the symptomatic population,in 17% to 27%of cases on ultrasonography,and in 50%to 70%of cases at autopsy.Thyroid nodules can be divided into malignant and benign according to the pathology.Benign nodules include thyroid adenoma,nodular goiter,phlogotic nodules et al.Malignant ones mainly thyroid carcinoma,fleshy tumors are rare.Refer to the clinicopathologic classification criteria raised by Hazard and Woolner,thyroid carcinoma is routinely classified into four types:papillary carcinoma,folliclar carcinoma,medullary carcinoma and anaplastic carcinoma(including squamous carcinoma).Although most thyroid nodules are benign,approximately 5%to 15%of such nodules are malignant,of which about 80%are well-differentiated ones.Some reported that solitary nodules were mostly malignant while multiple nodules were considerd to be a feature of benign lesions.But more and more research indicates that there is no difference between solitary and multiple nodules in the prevalence of carcinoma. Folliclar carcinoma is often found in multiple nodules,and 20%of papillary carcinoma is multipleceters.Surgery is applied to deal with most of thyroid solitary nodules,however is still hard to distinguish malignant nodules from benign ones.Thus to elevate preoperative diagnosis accuracy is essential for the selection of operandi modus and for prognosis.The purpose of imaging examination in thyroid is to detect lesions, discriminate malignancy in thyroid nodules and estimate the extent correctly so as to provide a guidance to choose therapeutic regimen in clinic.The diagnostic method of imaging in thyroid are mainly radionuclide image and ultrasonic imaging method.Radionuclide examination take a important place in imaging diagnosis of thyroid nodules.It was principal in diagnosing thyroid disease especially thyroid tumor. before the 20th century,70s.It assessment the character according to the biological activity and the function.of lesions.Meanwhile the thyroid morphous and texture can be displayed directly to the clinician,also make them to know the state of fuction of nodules.This method is of high sensitivity and low specificity.Ultrasonic inspection is widely used because of its non-harm,convenience,low expense and non-radioaction.With the development of high-frequency probe,color Doppler and increase of the experience of inspectors,the diagnosis of thyroid nodules has improved rapidly making ultrasonography(USG) the initial auxiliary examination of diagnosing thyroid nodules.In all imaging method of detecting throid nodules ultrasonography is the most sensitive.It can acquire lesions around 2～3mm in diameter and the detecting ratio of small lesions which are less than 2cm in diameter is higher than anyother methods.However the specificity is low respecting the complexity of ultrasonogram due to the multi-source of pathology of thyroid nodules.The current practice guidelines published by The American Thyroid Association (ATA) and The American Association of Clinical Endocrinologists(AACE) broadened the role of USG into more diagnostic fields,such as the initial screening procedure for the presence,location,and nature of the nodules,monitoring the change of size during observation,and setting the priority for aspiration among multiple thyroid nodules based on suspicious USG features.The sonographic criteria for detecting malignant thyroid nodules is still controversial at present.In this study, solitary thyroid nodules were studied with pathology as gold standard.Part one,the nodules were classified according to the separate criteria recommended by Hae Kyung and Stacul et.al of ultrasonography,and analysis the fuction of nodules assessed by radionuclide imaging to estimate the accuracy of combined ultrasound with radionuclide imaging in diagnosing solitary thyroid nodules.Part two,scored each nodule based on its ultrasonic features to evaluate the diagnosis value of ultrasonic semi-quantitive scores in thyroid nodules.Objectives1.To estimate the value of ultrasonography,radionuclide scanning,combined ultrasound with radionuclide imaging for detection of thyroid malignancy in solitary thyroid nodules.2.Evaluate the value of ultrasonic semi-quantitive scores in the differential diagnosis of solitary thyroid nodules.3.Analysis the correlation between ultrasonogram,radionuclide image and pathology.Materials and Methods1.Study objectPart one,98 patients with solitary thyroid nodules(male 27,female 71),mean age 41.8 years old.They were all underwent ultrasonography and ~(99m)TcO_4~- scan before operation,and confirmed by post-operation pathology.Part two,165 patients with solitary thyroid nodules(male 40,female 125),mean age 40.4 years old,all were underwent ultrasonography and finally diagnosed by pathology.2.Equipment and Method2.1 Ultrasonic inspection instrumentation is Philips IU22 and Philips HDI5000 color ultrasonoscope.The frequency of the linear array is 7～12MHz.The predetermined condition of thyroid examination was used in all patients.The patient takes supinely lie,shoulder department pad pillow,made multi-cross section scan in the thyroid.Sonograms were interpreted in term of nodular size, morphous,margin,shape,with or without a halo,echogenicity and calcification.2.2 Nuclein scanning inspection is made in GE company MG2000 SPECT with a high resolution parallel aperture collimator.An anterior static scanning was carried out 15 minutes after an intravenous injection of 185MBq(Smci) 99mTc-pertechnetate(~(99m)TcO_4~-).Depending on the pattern of uptake ~(99m)TcO_4~-,nodules are classified as nonfunctioning(cold), hypofunctioning(cool),normal functioning(warm),or hyperfunctioning(hot).2.3 Part one,the nodules were assessed based on their sonographic appearance and radionuclide image,and the results were comparatively analyzed with pathology. The three diagnostic method of ultrasonograpy,radionuclide scanning,and combined ultrasound with radionuclide imaging were compared after the sensitivity,specificity and accuracy of each method were calculated respectively.Part two,scored each nodule based on its sonographic features and compared with pathology.The receiver operator characteristic(ROC) curve was drawn according to the results.2.4 The data were analyzed by SPSS13.0,P value of less than 0.05 was considered to be statistically significant.McNemarχ~2 test was used to evaluate the three methods in the differentiation of solitary thyroid nodules.Wilcoxon rank sum test andχ~2 test was used to compare the ultrasonographic score of these benign and malignant nodules.ROC curve was drawn with sensitivity as Y-axis and 1-specificity as X-axis.Results 1.Part one,(1) The sensitivity,specificity and accuracy of ultrasonography, radionuclide scanning,and combined ultrasound with radionuclide imaging was 91.7 %、77.0%、80.6%,95.8%、37.8%、52.0%,87.5%、87.8%、87.8%,respectively; (2) There was a statistically significant difference between radionuclide scanning and combined ultrasound with radionuclide imaging(P=0.000);(3) The difference was suggested to be statistically significant between ultrasonography and combined ultrasound with radionuclide imaging(P=0.039).2.Part two,(1) Significant difference of the total scores existed between benign and malignant nodules(P=0.000).Malignant nodules got a higher score than benign ones;(2) Significant difference of the scores in each index existed between benign and malignant nodules(P＜0.01);(3) The incidence of calcification between benign and malignant nodules has a significant difference(P=0.000),but there was no significant difference between the two groups in the mode of calcification(P=0.065); (4) The area under ROC curve was 0.917.If the cut-off value of the total scores was equal or more than 4,the sensitivity,specificity and accuracy was 88.2%,94.6%, 93.3%,respectively.Conclusions1.Combined ultrasound and radionuclide was more valuable in the diagnosis of malignant nodules when compared with ultrasonography or radionuclide scanning was used alone.Both ultrasound and radionuclide scanning were effective and painless in the diagnosis of the thyroid nodule and each has its own superiority.If both methods were integrated,the accuracy will be improved.2.Ultrasonic semi-quantitive scores can improve the accuracy of diagnosis by comprehensively analyzing the sonographic features of solitary thyroid nodules.

The Effects of Zi-Bu-Pi-Yin Recipe on the Insulin Resistance in Hippocampus of Spleen-Yin Deficiency Diabetic Encephalopathy Rats and Possible Mechanisms

Wed, 30 Mar 2011 15:31:31 +0000

Objective: Diabetic encephalopathy (DE) is the central nervous system complication of diabetes, which is chronic and progressive. Its clinical symptoms are mild to moderate cognitive impairments, such as the decline of learning, memory and problem-solving abilities. When the diabetic encephalopathy occurs, there are corresponding changes on brain morphology, nerve electrophysiology and molecular biology. Although researchers have studied DE from various angles, the pathogenesis of DE is still unclear. Recent studies found that insulin was a neurotrophic factor of nerve cells. The biopotency changes of insulin in the brain play an important role in the central nervous systems of diabetes patients. The insulin signaling pathway begins with the combination of insulin and insulin receptor substrate. The insulin signaling pathway disturbance will lead to insulin resistance (IR) which means that the biopotency of normal dose insulin is lower than normal. Our previous studies indicated that endoplasmic reticulum stress (ERS) played an important role in Alzheimer\'s disease (AD) and its pathological products. The concentration of insulin is increased in the cerebrospinal fluid of AD patients, compared to the control ones. The insulin receptor density in the occipital regions is increased and the tyrosine kinase activity is reduced. Furthermore, evidences indicated that there exists a close relationship between endoplasmic reticulum stress and insulin resistance. Based on these, our experiment toke endoplasmic reticulum stress as a starting point to investigate the pathogenesis of diabetic encephalopathy which may be a very important work. Diabetes mellitus is named Xiaokebing in the Traditional Chinese Medicine. Its basic pathogenesis is yin-deficiency and inner heat. Although lung, spleen, and kidney are the organs related to the pathogenesis of Xiaokebing, the point that spleen-deficiency contributes to Xiaokebing draws more and more attention now. Zibu Piyin Recipe (ZBPYR) is created by Professor Libin Zhan. It could reinforce spleen and nourish spleen yin. It was established to treat spleen-yin deficiency syndrome.Previous experiments showed that ZBPYR had notable effects on neuroprotection and anti-aging of brain, and could cure the spleen-yin deficiency in Alzheimer’s disease animal models and primary cultured rat hippocampus neurons against Aβ1-40 toxicity. Therefore, this study observed the role of ZBPYR in changing insulin signaling pathway in hippocampus and its possible mechanisms in spleen-yin deficiency diabetic encephalopathy rats.Methods:1.Healthy adult male Sprague Dawley (SD) rats were divided randomly into control group, spleen-yin deficiency group, diabetic encephalopathy group, spleen-yin deficiency diabetic encephalopathy group and spleen-yin deficiency diabetic encephalopathy+ZBPYR group, four rats each group.2.After the acclimatization, the control group and spleen-yin deficiency group were feed with foundational diet. Other groups were feed with high fat diet. 4 weeks later, the rats which were feed with high fat diet intraperitoneal injected with 1% STZ (30mg/kg) once. The rats whose radom blood glucose was more than 16.7mmol/L were identified as diabetes mellitus models after 72h. We detected the level of fasting serum insulin (FSI), oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) after intraperitoneal injection with 1% STZ on the 4th、8th、12nd day. We induced spleen-yin deficiency encephalopathy model rats by defatigation, hungry-full diet and drenching the drug of damaged yin. ZBPYR group were induced by the intervention of ZBPYR in 16 days and decapitated for collecting hippocampus.3.Morris water maze test and step-down test were used to evaluate the behavioral changes.4.Using the Western Blotting method to detect the protein expression of JNK, phospho-JNK, IRE1αand IRS-1, phospho-IRS-1 in hippocampus. Results:1.After STZ injection, the diabetes mellitus rats blood glucose was significantly higher than nondiabetes rats (P0.05).2.The spleen-yin deficiency group rats showed increased water drinking (P<0.01), and rectal temperature (P<0.05). The results of Morris water maze test and step-down test showed that the learning and memory abilities of the diabetic encephalopathy group and the spleen-yin deficiency diabetic encephalopathy group were reduced significantly (P<0.05), but that of the ZBPYR treatment group were increased significantly (P<0.05).3.Western blotting results showed that compared with the control group and spleen-yin deficiency group, the hippocampus phospho-IRS-1 phospho-JNK and IRE1αprotein expression of diabetic encephalopathy group and spleen-yin deficiency diabetic encephalopathy group were increased (P<0.05), but these of ZBPYR treatment group were weakened.Conclusion:1.The spleen-yin deficiency diabetic encephalopathy rats model was successfully established through the way of combination of syndrome and disease. The results of Morris water maze test and step-down test showed that the learning and memory of the diabetic encephalopathy group and the spleen-yin deficiency diabetic encephalopathy group were reduced.2.ZBPYR can be significantly improved learning and memory abilities of spleen-yin deficiency diabetic encephalopathy rats .3.The function of ZBPYR improved learning and memory abilities of spleen-yin deficiency diabetic encephalopathy rats may be associated with insulin signaling pathway in hippocampus. ZBPYR can protect and maintenance conduction of insulin signaling pathway.4.ZBPYR can reduce the phospho-JNK and IRE1αprotein expression level and improve the endoplasmic reticulum stress to regulate the insulin signaling pathway.

Development of Chinese Version of Patients Learning Needs Scale and Preliminary Application in Patient with Coronary Heart Disease

Wed, 30 Mar 2011 15:31:29 +0000

Objective: To develop the Chinese version of Patient Learning Needs Scale(PLNS), and evaluate feasibility, validity and reliability of the developed scale. Supply an instrument to measure patients\' perception of learning needs at time of discharge for clinical nurses. To explore the learning needs of patients who have received treatment for Coronary Heart Disease (CHD) before their discharge home from hospital.Methods: (1) Development Chinese version of PLNS: The standard procedure of cross-culture adaptation was used to develop the Chinese version PLNS. A total of 385 adults hospitalized with a medical or surgical illness completed the Chinese version PLNS. The feasibility and psychometric properties of the scale such as internal consistency, split-half reliability, content validity and construct validity were evaluated.(2) Preliminary Application in Patient with Coronary Heart Disease: The Chinese Version of Patient Learning Needs Scale was completed by 135 CHD patients within 72 hours of their intended discharge. Quantitative descriptive and inferential analysis was conducted using Statistical Package for Social Sciences.Result: (1) The Chinese version of PLNS was developed which consist of 40items and 5 subscales:①Support and Care in the Community (10 items);②Medications (8 items);③Treatment and Activities of Living (8 items);④Complications and symptoms (8 items);⑤Illness-Related Concerns (6 items).These are evaluated using a five-point likert scale, responses ranging from 1 (not at all important) to 5 (extremely important).(2) The Chinese version of PLNS has satisfactory feasibility, validity and reliability. Questionnaire recovery rate of the investigation was 98.5%, the completion rate was 97.7%. Average time of completing was 13.75±4.36min. The overall Cronbach\'sαcoefficient was 0.95 and the value ofαin 5 subscales of PLNS ranged from 0.75 to 0.86. The overall split-half reliability coefficient was 0.95 and the value of split-half in 5 subscales of PLNS ranged from 0.802 to 0.876. the CVI for content validity was 0.86. The construct validity was confirmed by factor analysis with 55, 08 % variance was explained by 9 factors.(3) CHD patients indicated how important it was to know about each of 40 information items before discharge from hospital. The 5 item which scores are all higher than the others are all belong to the dimensions of symptom and medication. Subscales relating to medications, complications and symptoms, treatment and activities of living were rated highly.Univariate analysis revealed a significant association between patients\' leaning needs and patients\' age, education level, marriage status, ways of payment, times of hospitalization, ways of treatment, clinical type of CHD and medication.Stepwise canonical discriminant analysis found that three factors, times of hospitalization, clinical type, ways of payment, were the important influencing factors of total score. The important influencing factor of Support and Care in the Community is patients\' education level. The important influencing factors of Medications are ways of payment, types of medication, marriage status and times of hospitalization. The important influencing factor of Treatment and Activities of Living are times of hospitalization and ways of treatment. The important influencing factors of Complications and Symptoms are ways of payment and ways of treatment and. The important influencing factor of Illness-Related Concerns is clinical type.Conclusion: (1) An acceptable psychometric property of the Chinese version of PLNS was indicated. PLNS could be used as positive aspects measurement of general patients\' perceptions of learning needs to manage their care at home at time of discharge from hospital to home for further clinical study and nursing care.(2) The perception of learning needs of CHD patient is existed in every dimensions. The main influencing factors of patients\' learning needs are the following such as patients\' education level, ways of payment, types of medication, marriage status, times of hospitalization, hospitalization days, clinical type of CHD and ways of treatment. Further should be taken to meet patients\' needs for information and continuing support. Comprehensive and scientific health education based on individual differences in patients with CHD should be carried out to meet the needs of patients in support of health information.

The Study of the Disequilibrium and Modulation of T-cell Specific Transcription Factors T-bet/Gata-3 in Acute Lung Injury

Wed, 30 Mar 2011 15:31:27 +0000

Objective: To identify the expression of T-cell specific transcription factors T-bet/Gata-3 in acute lung injury,Further we interfere in acute lung injury with Dex and Lipo-PGE_1 respectively to probe what effect they have on transcription factors T-bet/Gata-3 and the blance of Th_1/Th_2.Method: Sixty male BALB-C mice were randomly dived into four groups:A:normal group (Sodium Chloride group);B:model group(Lps group);C: Dex group(Lps+Dex group);D:PGE_1 group(Lps+Lipo-PGE_1 group).ALI was induced by intravenous injection of LPS.In C or D group,PGE_1 or DEX was administered 1 hour after LPS deliver.After 6 hours,lung wet-to-dry ratio and lung morphology were studied.The expression of Th_1 and Th_2 pheno- type cytokines were detected by flow cytometry.Use SYBR Green I real -time fluorescent quantitative PCR technology to establish T-bet and Gata-3 standard product and their standard curve,and the mRNA expressions of T-bet and Gata-3 were detected by SYBR Green I real-time fluorescent quantitative PCR.Result:1、The animal model of acute lung injury was successfully established. Lung hemorrhage,edema,exudation and neutrophil infiltration were con- firmed in pulmonary artery,in addition,the lung wet-to-dry weight ratio of the LPS group were significantly elevated compared with those of the control group(5.74±0.31 vs 4.79±0.27, P﹤0.05).DEX and PGE_1 can lessen the change above mentioned compared with model group.2、Compared with the control group,Th_1、Th_2 cells decreased﹙16.65±1.7 vs 23.67±2.1, 9.4±1.25 vs 12.17±1.8, P﹤0.05﹚ and the ratio of Th_1/Th_2﹙1.733±0.03 vs 1.945±0.04, P﹤0.05﹚were markly decreased in LPS group.3、Compared with the LPS group,not only Th_1 and Th_2 cells﹙12.52±1.5 vs 16.65±1.7,7.5±0.8vs9.4±1.25, P﹤0.05﹚ decreased but also the ratio of Th_1/Th_2﹙1.667±0.05 vs 1.733±0.03, P﹤0.05﹚。by DEX treatment, while PGE_1 treatment increased them﹙20.31±2.2 vs 16.65±1.7, 10.5±0.8 vs 9.4±1.25,1.933±0.05 vs 1.733±0.03, P﹤0.05﹚.4、The imbalanced T cell–specific transcription factors T-bet/Gata-3 contributed to both low expression of Gata-3 and T-bet﹙1.1831±0.49452 vs 3.4393±0.55731, 0.6927±0.28513 vs 1.2027±0.23827, P﹤0.05﹚,and the ratios of gene expression of T-bet/Gata-3 were low﹙1.7132±0.13057 vs 2.8569±0.28988, P﹤0.05﹚ in LPS group.Dex treatment inhibited the mRNA expression of T-bet﹙0.7415±0.2489 vs 1.1831±0.49452, P﹤0.05﹚,and the ratios of T-bet/Gata-3 were decreasing﹙1.2605±0.15206 vs 1.7132±0.13057, P﹤0.05﹚.Compared with LPS group,Lipo-PGE_1 resul- ted in increasing T-bet﹙ 1.8265±0.70469 vs 1.1831±0.49452, P﹤0.05﹚,and the ratios of gene expression of T-bet/Gata-3 were markly increasing﹙2.5024±0.35236 vs 1.7132±0.13057, P﹤0.05﹚.Conclusion:1.In LPS group,the expression of T-bet and Gata-3 were low,also the ratios of T-bet/Gata-3 were low,and above mentioned confirmed that imblanced mRNA expression of T-bet/Gata-3 existed in acute lung injury.2、Compared with LPS group,Dex treatment inhibited the mRNA expression of T-bet,thus decreased the ratios of T-bet/Gata-3,that may hunt Dex regulated the ratio of Th_1/Th_2 by upriver mechanism in acute lung injury3、Compared with LPS group,Lipo-PGE_1 resulted in increasing T-bet, thus increased the ratios of gene expression of T-bet/Gata-3,Lipo-PGE_1 committed T helper cell to a Th_1 phenotype.by modulating key master switches T-bet. and Gata-3

The Expressions of Intercellular Adhesion Molecule-1 in Experimental Pulpitis of the Rat

Wed, 30 Mar 2011 15:31:26 +0000

BACKGROUNDIntercellular adhesion molecule-1(ICAM-1) is belonging the immunoglobulin gene superfamily.Through mediate adhesion between cells and cell,cells and extracellular matrix,ICAM-1 can participate in a series of physiological and pathological processes.ICAM-1,a single-chain transmembrane glycoprotein with a molecular weight of 80-110 KDa,consists of five Ig-like domains,a hydrophobic transmembrane domain and a short cytoplasmic C-terminal domain.And NF-κB, plays the leading role in the activation of ICAM-1.Trough the special interaction of protein-protein,NF-κB participate in the expression adjustment of ICAM-1 with other transcription factors and syn-activator activates together.Ligand of ICAM-1 includes lymphocyte function- associated antigen-1(LFA-1) and macrophage antigen-1(Mac-1).When LFA-1 is increased,resting affinity can be regulated.First,the affinity can be increase by an addition of the aggregation degree of integrin distribution on the membrane.And,integrins change configuration,so that its intrinsic affinity with ligand is increased.Under physiological conditions,ICAM-1 is expressed at a low level in endothelial cells and epithelial cells or constitutively on the surface of the cells,providing the underlying molecular basis for cell recognition, activation,proliferation,differentiation and motility,and thereby helping to stabilize the internal environment of the body.Moreover,ICAM-1 also plays a key role during pathological conditions,such as inflammatory reaction etc.For these reasons,a comprehensive and objective understanding of ICAM-1 is needed.Inflammatory cells leave the circulation at sites of local inflammation through a series of distinct sequential steps.The leukocyte-endothelinm interaction,the fundamental event of inflammation is orchestrated by the action of some cell adhesion molecules(CAMs) found both on leukocytes and endothelial cells.The initial adhesive event is the margination characterized by rolling of leukocytes along the luminal surface of the endothelium.Each member of the selectin family(E-, P-L-selectin) has been shown to support leukocyte rolling under flow conditions. Once the marginated leukocytes form a stationary adhesion with endothelial cells,it is stimulated by chemotactic factors to downregulate the selectin-based adhesion and upregulate adherence dependent on integrins.Subsequent stages of leukocyte immigration are mediated by the interactions between integrins and members of the immunoglobulin superfamily:intercellular cell adhesion molecules(ICAM-1, ICAM-2,and ICAM-3),vascular cell adhesion molecule(VCAM-1) and platelet cell adhesion molecule(PECAM-1).Thus,teukocytes rapidly immigrate into surrounding tissues.And according to literature review,ICAM-1 is believed to allow not only recruitment of naive lymphocytes and rapid accumulation of leukocytes at injured and infected sites,but also recirculation of lymphocytes stimulated by inflammatory agents.ICAM-1 is already playing the influential role in the cell adherence,it is also a player in the actual transmigration of leukocytes through the vessel wall (diapedesis).OBJECTIVESDental pulp is surrounded by dentin and isolated from other tissue,like an in vitro system,and it is almost always exposed to inflammatory agents entering through dentinal tubules in the case of dental caries and through the apical foramen in periodontitis.The pulp develops inflammation as a basic protective biologic defense mechanism in response to any type of injury.As a result,dental pulp could be a model for studying the roles of adhesion molecules on the vascular endothelium in transendothelial migration of leukocytes both with and without inflammation.Despite the extensive information on the tissue distribution of ICAM-1 in healthy and pathological conditions in other fields of medicine to date,there is limited scientific data concerning the expression of this molecule by the vascular endothelium of the oral tissues and particularly of the dental pulp.Therefore,the present study was undertaken to determine the expression of ICAM-1 on the vascular endothelium of healthy and inflamed dental pulp.Materials and methods1.Animals and reagentsWe used 60 female Sprague-Dawley rats(n=60,200-250 g body weight) for this study.The rats were divided equally into six groups included one control group and five experimental groups.The rats were purchased from the Center of Experimental Animals of Southern Medical University.ICAM-1 antibody was purchased from Wuhan Boster Biological Technology Co.,Ltd;SP-9001 Histostain~(TM)-Plus Kits and DAB complex kit was supplied by Zhongshan Golden Bridge Biotechnology Corporation,Beijing,China.2.Sample preparationIn 50 female rats of five experimental groups,following anesthesia with 10% chloral hydrate,the roof of the pulp chamber of the right first and second maxillary molar teeth was removed using a 005 round bur in a high-speed handpiece.The pulp chambers were left open to the oral environment.10 female rats were control group and were untreated in all animals.After experimental pulpal exposure,the rats were killed at various time points: 1h,1d,3d,5d and 7d;each n=10.The teeth and the part of maxillary bone were rapidly removed to determine quantitative changes in pulpitis induced ICAM-1 activity.3.Observation under light microscopeThe specimens were fixed in 4%paraformaldehyde for 48 hours,and further trimmed and transferred into 15%EDTA for decalcification for 15 days and then embedded by routine paraffin.Slices were continuously sectioned parallel to the long axis of tooth with a thickness of 5μm.One paraffin section of each sample was selected and stained with HE,and then the dental pulp histological and morphological changes were observed.4.Immunohistochemistry examinationImmunohistochemical SP methods were employed in this study.After being dewaxed and reacted with 3%H_2O_2 for 10 min,the slices were put in citrated saline buffer liquid(pH 6.0±0.1) and heated in the microwave stove adjusted to grade"3"at 92～98℃for 10min.And then the slides were pre-incubated with 10%goat serum for 15 min at room temperature.Then immune-stained were began with adding monoclonal antibodies to ICAM-1,for 12 hours at 4℃in refrigerator.The slides were sequentially incubated with biotinconjugated secondary antibodies followed by horse radish peroxidase-conjugated streptoavidin.The first step lasted 15 min and the second step lasted 10 min at 37℃in thermostat with 5min PBS-buffered saline washes three times each step.The reactions were revealed using DAB substrate, slides were then washed in running tap water.Contrast with hematoxylin and mounted in balsam,observed with light microscopy.Blank staining was performed similarly with elimination of the primary antibody.5.The method used to quantify labelingEssentially,computer-assisted image analysis software(Image-Pro Plus,Version 5.0;Media Cybernetics,Silver Spring,MD,USA) was used to create a digital image from the microscopic image.The OD value of ICAM-1-labelled tissue was then determined within each field of analysis.6.Statistical analysisAll the data were shown as mean±S.D.Statistical analysis of data was performed by applying one - way analysis of variance(ANOVA) with SPSS V.13.0. The q-test was conducted after the test for homogeneity of variance and analysis of variance,and the correct t test was employed in the two-two comparison.In all statistical comparisons,P＜0.05 or P＜0.01 was considered significantly difference statistically.RESULTS1.HE stain:In the normal control group,pulp tissue have a small amount of blood vessels and a great quantity of fibroblasts,and at the edging of the medullary cavity there are a layer of columnar odontoblast cells.The typical pulpitis was observed in experimental groups.The dental pulp inflammation may appear in 1 hour group,and aggravates gradually along with the time development.Leucocyte appears in early stage of pulpitis,but lymphocyte appears latterly from the third day.Blood vessel hyperplasia and dilatation and vascular engorgement are also main manifestation of pulpitis.2.Immunohistochemistry examination results: Very less positive expressions of ICAM-1 were found in microvascular endothelial cells of the dental pulp regions in thenormal control group,but in the five experimental groups,the expression was higher than those in the normal control group,especially at the first and the third and and fifth day.The seventh day was gradually reduction.3.Statistical analysis results:There were showed significant difference,indicating that the ICAM-1 positive cell expression in microvascular endothelial cells increased sharply in the dental pulp of the pulpitis.CONCLUSIONSThis study showed that the method to establish a rat pulpitis induced by exposure of dental pulp was feasible and ICAM-1 had a significant increase in the experiment pulpitis compared with normal dental pulp tissue.ICAM-1 may play a role in the process of dental pulpitis.ICAM-1 guides the stable adhesion between leukocytes and vascular endothelial cells,which enables leukocytes to be recruited and anchored on the inflamed tissue. But,for recruitment of lymphocytes,ICAM-1 played the very minor role.There must have different regulatory mechanism for recruitment of lymphocytes in the process of dental pulpitis.Through the animal experiment,we can observe the entire dynamic process of pulpitis from start at the molecular level,further clarify the mechanism of ICAM-1 involve in the development of pulpitis.

Lethal Effects and Mechanism of Nicotinamide for None-Focused Ultrasound on Different Pulmonary Adenocarcinoma Cell Lines in Suspension

Wed, 30 Mar 2011 15:31:23 +0000

BackgroundMalignant pleural effusion is a common complication of patients with cancer, especially lung cancer.In most cases,pleural effusion is the rusults of disease progression and recurrence and indicates unfavorable prognosis.Lung cancer (about 35%) is the most common reason,predominantly lung adenocarcinoma. Treatment aimed directly at hydrothorax can abate or eliminate symptoms and raise or improve survival quality of cancer patients.The commonly used method,local injection of chemotherapeutics after chemotherapy all over the body coupled with thoracentesis,is not effective to malignant hydrothorax caused by lung adenocarcinoma.This method may cause sever injuries to normal cells unselectively while killing cancer cells,which may jeopardize cancer patients. Repeated application of chemotherapeutics results in the resistance of tumor cells to drugs,which is an important factor for effectiveness of chemotherapy.Multidrug resistance is one of the main reasons of chemotherapy failure.As a result,novel treatment with good selectivity and minor side effects has become the hot spot of research.Ultrasound treatment is a novel developed technique for tumor treatment after physical treatments like tumor thermotherapy,freezing and microwave.Searches on tumor treatment by high intensity focused ultrasound(HIFU) have been widely reported.However,when tumor cells are within the hydrothorax,hydroperitonia, or diffused in tissues,it is not likely to kill these cells by HIFU.Previous researches conducted in our laboratory indicate that using NFU to kill tumor cells in suspension is an effective method.During the spread of ultrasound in tissues,its energy can decrease exponentially with the increase of the spreading distance, which causes decreased energy in target areas and decreased therapeutic effects.Nicotinamide(NA),an acid amide derivative of nicotinic acid,and moity of coenzymeⅠand coenzymeⅡ,plays a role in hydrogen transmission during biological oxidation,and promotes biological oxidation and metabolism.It has a fairly wide rage of safety.Only large dosage can lead to vitaminosis.In recent years,it has drawn special attention used as a kind of radiosensitizer.Can we enhance the lethal effects of NFU to tumor cells in suspension using NA?What is the killing mechanism of the unification? Are the lethal effects in conformity? Dose it cause injury to normal cells while killing cancer cells? What is the effectivenese compared with chemotherapeutics? In order to elucidate these questions,we design this study answer these questions.Objective and SignificanceExplore whether NA can enhance the lethal effects of NFU to tumor cells and the killing mechanism,and identify the specificity.Compare the effectiveness of NFU sensitized with NA and four chemotherapeutics for lung cancer.Provide experimental bases for the clinical treatment of malignant pleural effusion with NFU sensitized with NA.Methods1.Lethal effects of NFU sensitized with NA on human pulmonary carcinoma cell line A549 in suspension(1)Material and groupsHuman pulmonary adenocarcinoma cell line A549 Blank control group;NA group;NFU group;NA sensitization NFU group(2)Determination of experimental doses through measuring the inhibition ratio and sensitization ratioWe observe cytostasis ratio with MTT colorimetric method.The concentrations of NA were 0,1,2,3,4 and 5mg/ml respectively.The dosage of ultrasound wave irradiation was increased from 0W/cm~2×5s to 70W/cm~2×5s. SPSS13.0 was used to obtain the Inhibitory concentration 50(IC_(50)) and sensitization ratio(SER).(3)MethodsWe observed the morphologic changes of A549 both directly and after HE stain with inverted microscope.Instant and 24 hours\' survival rates of A549 cells treated by different methods were determined based on the trypan blue exclusive assay.Growth rates were detected by MTT after 24 hours.The percentage changes of apoptosis and the distributional percentage of cell cycle were analyzed with flow cytometry.The capability of cell colony formation was detected by clone formation test.2.Lethal Mechanisms of NFU sensitized with NA on human pulmonary carcinoma cell line A549 in suspension(1)Material and groupsHuman pulmonary adenocarcinoma cell line A549Blank control group;NA group;NFU group;NA sensitization NFU group(2)MethodsNucleolar organi-zer regions staining were used to observe the argyrophilic nucleolar organi-zer regions(AgNOR)grains.The apoptotic changes of four groups were analyzed with Hoechst33342 fluorescence staining,AO/EB double fluorescence staining and transmission electron microscope.Apoptosis DNA Ladder was studied by Agarose Gel Electrophoresis.Changes of cell:count was obtained by observing cell culture fluid color and counting chamber.Immunocytochemistry was used to detect expressions of p53,Bcl-2 and Ki-67.Protein changes in cell culture fluid and with in cells were detected by BCA method and protein gel electrophoresis. 3.Lethal Effect Comparisons of None Focused Ultrasound sensitized with nicotinamide on different human pulmonary adenocarcinoma cell Lines and immortalized bronchus epithelial cells(1)Cell LinesFive cell lines were chosen.Human pulmonary adenocarcinoma cell line A549, GLC-82,GLC-82/CDDP(resistant to cisplatin),mouse Lewis pulmonary adenocarcinoma cell line LLC,and human immortalized bronchus epithelium HBE.(2)GroupsBlank control group;NA group;NFU group;NA sensitization NFU group(3)MethodsShock and intermitant method of large dose were used to induce GLC-82/CDDP cells.MTT was used to measure the inhibition ratio of GLC-82 and GLC-82/CDDP in response to different concentration cisplatin was calculated IC_(50) value and Resistance Index(RI).Instant and 24 hours\'survival rates treated by different methods were determined based on the trypan blue exclusive assay. Growth rates were detected by MTT after 24 hours.The percentage changes of apoptosis were analyzed with flow cytometry.Survival rates,growth rates and apoptosis rates of five cells were compared after different procedures.4.Comparisons of lethal effect between None Focused Ultrasound sensitized with nicotinamide and four chemotherapeutics(1)Cell LinesHuman pulmonary adenocarcinoma cell line A549,GLC-82,GLC-82/CDDP and human immortalized bronchus epithelium HBE.(2)Chemotherapeutics and concentrationCDDP:concentration in vitro(plasma drug concentration)13.6μg/mlPTX:concentration in vitro(plasma drug concentration)119μg/mlGEM:concentration in vitro(plasma drug concentration)850μg/mlBLM:concentration in vitro(plasma drug concentration)3μg/ml(3)GroupsSix groups were divided.NA-NFU 10s group;NA-NFU 5s group;DDP group; PTX group;GEM group;BLM group(4)MethodsInhibition ratios were detected by MTT.Lethal effect were according to the inhibition ratios.Results1.Lethal effects of NFU sensitized with NA on human pulmonary carcinoma cell line A549 in suspension(1) We found microscopically that large amount of cell debris and charring-like materials were produced after NA-NFU treatment.Cells became shrinked,round,low refractive or floated.Obsered after HE staining, morphological changes of A549 were very obrious.The cells become slender with scarce cytoplasm.The number of pyknosis and karyolysis was increased.The pathological mitosis was decreased dramatically.(2) The lethal effect of NFU was improved significantly when NA concentration was≥3mg/ml.The sensitization and dosage were positively correlated in a certain range(2-5mg/ml).Under the circumstances of 30W/cm~2×5s NFU sensitized with 3mg/ml NA,IC_(50) was near 50%with an inhibition ratio of 51.48%,which was settled as experimental dose.(3) We found that there was no significance among the instant survival rate,24 hours\'survival rate,growth rate;clone formation capability,apoptotic rate,cell cycle changes and the negative control group when NA(3mg/ml) was used solely (P＞0.05).(4) The instant survival rate,24 hours\' survival rate,24 hour growth rate and clone formation capability were significantly different among four groups(P＜0.05). The instant survival rate,24 hours\' survival rate,24 hour growth rate and clone formation capability were decreased significantly in groups treated with NA combined with NFU compared with the other three groups.Twenty-four hours\' apoptotic rate was increased dramatically(P＜0.05).2.The lethal Mechanism of NFU sensitized with NA on human pulmonary carcinoma cell line A549 in suspension(1) Endonuclear AgNOR particles were changed markedly both in quantity and distribution after NA-NFU treatment.Quantity was reduced or absent. There were statistical significances compared with control group,NA group and NFU group(P＜0.05).Growth rate was descended(P＜0.05).Growth curves were flat.NA was interacted with NFU.NA could enhance the inhibition of NFU to A549 cells.(2) Typical apoptosis changes could be seen under fluorescence microscope with Hoechst33342 fluorescence staining,AO/EB double fluorescence staining and transmission electron microscope after 24h of NA-NFU treatment.Typical DNA Ladder strip was obtained from Agarose Gel electrophotogram.(3) There were more cells in NFU and NA-NFU before treatment than those after treatment(P＜0.05).There was no signiticance between the control group and NA group(P＞0.05).(4) Immunohistochemisty revealed that:p53 was expressed in small amount of cells in NFU group and NA-NFU group.The expressions in control and NA were negative.Bcl-2 was negative in four groups.Ki-67 was positive in four groups: NA-NFU＞NFU＞Control＞NA.This was different from MTT.We considered this as false positive results due to the presence of antigens after cell injuries.(5) Protein concentration was raised in cell culture fluid but descended intra-cellularly.This was more obvious in NA-NFU group.This showed that NFU could destroy cell membrane or increase permeability.3.Lethal effect comparisons of None Focused Ultrasound sensitized with uicotinamide on different human pulmonary adenocarcinoma cell line and immortalization bronchus epithelium(1) Differences of inhibition ratio between GLC-82 and GLC-82/CDDP were significant(P＜0.001).IC_(50) of GLC-82 was 0.190μg/ml CDDP.IC_(50) of GLC-82/CDDP was 1.444μg/ml CDDP.RI was 7.6.(2) Survival rates,growth rates and apoptosis rates of five cells were not significant after NFU only.After sensitized with nicotinamide survival rates and growth rates were significantly different(P＜0.05).Survival rates of human immortalization bronchus epithelium HBE was higher than that of human pulmonary adenocarcinoma cell line A549,GLC-82,and GLC-82/CDDP(P＜0.05).Survival rates of mouse Lewis pulmonary adenocarcinoma cell line LLC was lower than those four cells above(P＜0.05).4.Comparisons of lethal effect between None Focused Ultrasound sensitized with nicotinamide and four chemotherapeutics(1) The inhibition ratio in NA combined with NFU 10s group was higher than NA combined NFU 5s group.The inhibition ratios with CDDP and PTX were higher than NA combined with NFU.GEM and BLM were lower.(2) The inhibition ratio in HBE as a control was a little lower than pulmonary adenocarcinoma cell line A549,GLC-82,and GLC-82/CDDP after NFU treatment. The inhibition ratio in GLC-82/CDDP was lower than other three cells obviously. HBE was lower than A549 and GLC-82 only after PTX treatment,while the difference was not significance with other three chemotherapeutics.(3) The highest inhibition ratio was exhibited on the first day,then lower on the second and third day.The opposite situation was shown in chemotherapeutics.Conclusions1.NA-NFU has lethal effects on some pulmonary carcinoma cell lines,but it is not better than chemotherapeutics.NA-NFU shows advantages over resistant cells. It also causes injury to normal cells while killing cancer cells,which are slight, compered to chemotherapeutics.2.NA(3mg/ml) has little lethal effects on cells.NFU can reduce survival ratio, suppress cell multiplication,increase permeability of cell membrane and induce apoptosis.Some cells are made into fragmentation directly.This is related to mechanical effect,cavatition erosion and thermal effect of ultrasound.NA plays sensitizing effects mainly through inhibiting injury and repair abilities of cells.3.The lethal effect of NFU is improved significantly when NA concentration is≥3mg/ml.

A Preliminary Investigation on the Mechanism of Reverse Redistribution in Thallium-201 Myocardial Perfusion Scintigraphy in Patients with Coronary Artery Spasm

Wed, 30 Mar 2011 15:31:22 +0000

Background:Our previous studies found that reverse redistribution(RR) in ~(201)T1 myocardial perfusion scintigraphy was one of the characters in patients with coronary artery spasm(CAS).Being non-invasive criteria by combining of the rest chest pain,negative ECG exercise test and RR in ~(201)T1 myocardial perfusion scintigraphy,the sensitivity and the specificity for diagnosing CAS were 98%and 90%,respectively.However,the mechanism of RR is not clarified.Objective:This study was aimed to explore the possible mechanism of RR in thallium-201(~(201)T1) myocardial perfusion scintigraphy by comparing the changes of plasma endothelin- 1(ET- 1 ),nitric oxide(NO) levels,ET- 1/NO ratio,coronary artery blood flow and myocardial blood flush during stress tests in patients with CAS.Materials and methods1.Study population The patients who underwent ECG exercise tests and were proposed to receive ~(201)T1 myocardial perfusion scintigraphy were included.According to the final diagnosis,the patients were divided into following three groups.(1) control group(n=30):①A normal presentation on ~(201)T1 myocardial perfusion scintigraphy.②Coronary heart disease(CHD) was clinically excluded.(2) CAS group(n=30):①RR was present on ~(201)T1 myocardial perfusion scintigraphy.②With rest chest pain.③And a negative ECG exercise tests.④No angiographic stenosis(≥50%) was found by coronary artery angiography (CAG).(3) CHD group(n=30):①Ischemic perfusion defect on ~(201)T1 myocardial perfusion scintigraphy.②Consistent with Denomination and Diagnosis Standard for Ischemic Heart Disease by International academy of Cardiology and International Association of Cardiology as well as WHO Clinic Denomination Standardization Associated Experts Group in 1979.③CAG showed stenosis of 50%or above in one or more vessels.2.Methods(1) ~(201)T1 myocardial perfusion scintigraphyVasodilators like Calcium channel blockers and long-acting nitrates were discontinued for 24 h,short-acting nitrates for at least 12h before ~(201)T1 myocardial perfusion scintigraphy.Dipyridamole was injected intravenously at a rate of 0.14mg/kg·min for 4min.Followed by intravenous 111 MBq of ~(201)T1,the initial images were taken at 10 min,and the delay images at 3h,after the injection.Semi-quantitative intergration was used in the analysis,the left ventricle was divided into 9 portions:apical,anterior,basal anterior,anterolateral,posterolateral, ante-septal,post-septal,inferior,and posterior segments.Each segment was assessed using a five-point scoring system(0=normal;1=slightly reduced perfusion; 2=moderaterly reduced perfusion;3=severely reduced perfusion;4=no perfusion). The summed different score(SDS) was calculated as the difference between summed stress score and summed rest score.Myocardial ischemia was assigned if SDS≥2. Conversely,RR was defined if SDS≤-2.(2) Detection of ET-1 and NO:Venous blood was collected before stress test, immediately and 3h after stress test.NO was measured by nitrate reductase method, and ET-1 was measured by the homogeneous competitive radioimmuno assay method.(3) Coronary angiography and dipyridamole testEighteen patients with CAS were undergone coronary angiography and dipyridamole test.Coronary angiography was performed with conventional methods. In particularly,at least four projections for left coronary artery and two perpendicular projections for right coronary artery were taken during CAG.Nitroglycerin and other vascular dilators were prohibited during CAG After the initial CAG,dipyridamole was injected intravenously at a rate of 0.14mg/kg·min for 4min,and then repeated CAG.Anlysis of images:Coronary artery diameters,corrected TIMI frame count (CTFC) and TIMI myocardial perfusion grade(TMPG) were calculated automatically by the software and compared between RR related vessels and non-RR related vessels.Results1.comparison of blood ET-1,NO and the ratio of ET-1/NO during ~(201)T1 myocardial perfusion scintigraphy.(1) The dynamic changes of plasma ET-1 levels during heart stress test1) Before stress test,the plasma ET-1 level was higher in CAS groups(163.53 ±8.58 pg/ml) than in CHD group(137.73±8.32 pg/ml) and in control group(116.15±6.65 pg/ml,P＜0.01).2) In control group,there were no significant differences in plasma ET-1 levels between different time points(F=0.138,P=0.871 ).3) In CSA group,the plasma ET-1 level went down at immediately after stress test(151±9.15 pg/ml) compared with those before stress test(163.53±8.58 pg/ml,P＜0.01 ),and went up at 3h after stress test(164.76±7.14 pg/ml).There were no significant differences between before and at 3h after stress test(P=0.571).4) In CHD group,the plasma ET-1 level went up at immediately after stress test (147.00±9.68 pg/ml) compared with those before stress test(137.73±8.32 pg/ml, P＜0.01 ),and went down at 3h after stress test(138.25±8.78 pg/ml).There were no significant differences between before and at 3h after stress test(P=0.822).(2) The dynamic changes of serum NO levels during heart stress test1) Before stress test,the serum NO level was higher in control group(70.10±6.90μmol/L) than in CHD group(62.64±7.45μmol/L) and CAS group(60.32±8.20μmol/L,P＜0.01).2) In control group,the serum NO level increased at immediately after stress test (83.14±8.31μmol/L) compared with those before stress test(70.10±6.90μmol/L, P＜0.01 ),and decreased at 3h after stress test(68.06±6.21μmol/L,P＜0.01 ).There were no significant differences between before and at 3h after stress test(P=0.117).3) In CAS group,the serum NO level inceased at immediately after stress test (63.78.±8.30μmol/L) compared with those before stress test(60.32±8.20μmol/L,P＜0.01 ),and decreased at 3h after stress test(46.70±8.79μmol/L) which were lower than those before stress test(P＜0.01).4) In CHD group,the serum NO level increased at immediately after stress test (67.04±7.99μmol/L)compared with those before stress test(62.64±7.45μmol/L, P＜0.01 ),and decreased at 3h after stress test(60.33±7.92μmol/L,P＜0.01 ).There were no significant differences between before and at 3h after stress test(P=0.473).(3) The dynamic changes of ET-1/NO ratio during heart stress test1) Before stress test,the ratio of ET-1/NO was higher in CAS group(2.75±0.39) than CHD group(2.22±0.31) and control group(1.67±0.18,P＜0.01).2) In control group,the ratio of ET-1/NO went down at immediately after stress test(1.40±0.12) compared with those before stress test(1.67±0.18,P＜0.01 ), and went up at 3h after stress test(1.71±0.18,P＜0.01 ).There were no significant differences between before and at 3h after stress test(P=0.562).3) In CAS group,the ratio of ET-1/NO went down at immediately after stress test(2.41±0.36) compared with those before stress test(2.75±0.39,P＜0.01), and went up at 3h after stress test(3.36±0.78,P＜0.01 ) which were higher than those before stress test(P＜0.01).4) In CHD group,there were no significant differences in the ratioes of ET-1/NO between different time points(F=1.168,P= 0.316).2.Comparison on coronary artery diameters,CTFC and TMPG before and after dipyridamole test during CAG.(1) Blood flow was slower in RR related vessels while compared with non-RR related vessels before dipyridamole test.After injection of dipyridamle,with heart rate increasing and blood flow going faster,the coronary artery diameters increased by 16%±8%in RR related vessels and 14%±12%in non-related vessels,respectively.(2) Before dipyridamole test,CTFC was higher in RR related vessels(36±6) than those in non-RR related vessels(29±7,t=2.982,P=0.009).After intravenous dipyridamole injection,CTFC decreased remarkable in RR related vessels(26±7,t =4.466,P=0.001) as well as in non-RR related vessels(25±5,t=2.308,P= 0.038). (3) Before dipyridamole test,TMPG was lower in RR related vessels(2.02±0.39 ) than those in non-RR related vessels(2.56±0.31,t=2.832,P=0.018).After intravenous dipyridamole injection,TMPG increased remarkable in RR related vessels(2.92±0.12,t=2.832,P=0.018) as well as in non-RR related vessels (2.92±0.12,t=4.261,P=0.001).Conclusions1.The abnormal ratio of ET-1/NO indicates endothelial dysfunction which results in a slight coronary artery constriction under rest state in patients with CAS.2.Under the rest state,endothelial dysfunction induces a slight coronary artery constriction in patients with CAS which results in slower blood flow and presents as a perfusion defect in ~(201)T1 myocardial perfusion scintigraphy.However,exercise or dipyridamole induces coronary artery relaxation by changing the ratio of ET-1/NO and increases coronary blood flow which presents as normal myocardial perfusion.It may be one of the possible mechanisms of RR in ~(201)T1 myocardial perfusion scintigraphy in patients with CAS.

The Expression of MIF in the Serum、Decidua、Chorion of UERSA Patients

Wed, 30 Mar 2011 15:31:19 +0000

Objective:Understand the level of macrophage migration inhibitory factor (MIF) of unexplained early recurrent spontaneous abortion patients and normal early pregnancy women in serum,decidua,chorion.Discuss the relationship between the expression levels of human serum,decidua,chorionic MIF and the unexplained early recurrent spontaneous abortion patients,while test the clinical significance of the expression level.Methods:The clinical research was a control clinical study.The number of the two group were 30.The research group was patients with unexplained early recurrent spontaneous abortion(UERSA) while the control group was patients with normal early pregnancy women(normal pregnancy):1.ELISA(enzyme-linked immunosorbent assay) assayed concentrations in peripheral blood of research group and control group and statistic comparison was performed.2.Real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR) detected the level of MIF in both research group and control group,and then compared the differences.Results:In serum,decidua,chorion,the level of MIF in research group and control group:1.The peripheral blood concentrations of MIF in research group and control group:The peripheral blood concentrations of MIF in research group was 10.98±3.02ng/ml,while the peripheral blood concentrations of MIF in control group was 17.53±5.34ng/ml,the level of serum MIF in research group was less than which of control group,P<0.05.2.The level of decidua MIF in research group and control group:In research group,the decidua△Ct、△△CT、2~(-△△CT) were 8.84±0.93、2.41±0.93、0.97±0.58. While,in control group,that were 6.44±1.09、0.00±1.09、1.20±0.85 respectively.That was to say,the level of decidua MIF in research group was less than which of control group,P<0.05.3.The level of chorionic MIF in research group and n control group:In research group,the chorionic△Ct、△△CT、2~(-△△CT) were 14.66±1.79、0.22±1.79、0.022±0.001, respectively;while,in control group that were 8.43±1.24、0.00±1.24、1.29±1.11.That was to say,the level of chorionic MIF in research group was less than which of control group P<0.05.4.The correlation analysis of serum and decidua,chorionic MIF:Between serum MIF and decidua or chorionic MIF,had positive correlation,r was 0.78,0.69, respectively(P<0.01).Conclusion:1.The level of decidua、chorionic MIF in research group was less than which of control group.2.The peripheral blood concentrations of MIF in research group was less than which of control group.The most important was,the level of serum MIF was positively related to MIF-mRNA expression of villi,decidual tissue,it may be a effective monitoring incidence of UERSA.

Influence of Xingnaojing on the Edema and the Expression of Matrix Metalloproteinases 9 and Complement 3 Around Hematoma after Intracerebral Hemorrhage in Rats

Wed, 30 Mar 2011 15:31:17 +0000

Objective1.To study the relationship between metalloproteinases-9,complement 3 and the edema after intracerebral hemorrhage(ICH);2.To explore the effect and the mechanism of Xingnaojing on edema induced by ICH and to investigate the underlying clinical mechanisms.Methods1.Seventy-two healthy adult male Sprague-Dawley(SD) rats were randomly divided into three groups,ICH group,ICH with Xingnaojing treatment group(XNJ group) and sham-operation group.2.Saline(1ml/kg)(for sham-operation group and ICH group) and Xingnaojing injection(1ml/kg)(for XNJ group) were injected respectively through vena caudalis right after operation;3.The rats were anesthetized with intraperitoneal injection and fixed on the stereotactic frame.A microsyringe was inserted stereotaxically into the right caudate nucleus(coordinates:0.2mm anterior,5 mm ventral,and 3mm lateral to the bregma)of the rat,and 50ul auto-non-anticoagulated blood were given(XNJ group and ICH group) or nothing was given(sham-operation group);4.The neurofuction of rats was evaluated immediately and at 24,48,72 hours after operation;5.The water contents in the area of peri-hematoma were examined respectively at 72 hours after operation in every group.6.EB solution,which cycles in vivo for 2 hours,was given through vena caudalis separately at 70 hours post-operation,and then the rats were killed and the brain were removed for the detection of the contents of EB in brain tissue.7.Four percentage paraformaldehyde was injected into the heart and cycled in vivo body,then the rats were decapitated.The expression of MMP-9 and C3 around the hematoma were measured with immunohistochemistry technology.Results1.Neurobehavioral scores were decreased in the rats with intracerebral hemorrhage. As compared to the sham-operation group,neurobehavioral scores were lower in the ICH group at every time points(P＜0.05).However,neurobehavioral scores increased gradually and peaked at 72h after the treatment of xingnaojing injection,.At 48h and 72h post-operation,scores of XNJ group was higher than that of ICH group (P＜0.05).2.At 72h after the operation,water content of ICH group was higher than that of sham-operation group(P＜0.05).The water content in brain tissue of XNJ group was lower as compared with ICH group(P＜0.05).3.The permeability of blood-brain barrier increased in ICH group at 72h post-operation. The content of EB in the area of perihematoma in ICH group was higher than that of the sham-operation group(P＜0.05),as well as XNJ group(P＜0.05).4.The number of positive cell labeled with MMP-9 and C3 in the ICH group was higher than that of the sham-operation group(P＜0.05),and the expression of MMP-9 and C3 around hematoma in XNJ group lower than in ICH group(P＜0.05).5.The gedtalt form of the brain:the bilateral hemispheres were appozimately symmetric in the sham-operation group.The enlarged hemisphere,the extended gyri and the shallow sulci were observed in the right cerebral hemisphere of ICH group. While the appearance of the bilateral hemispheres of XNJ group changed slightly.6.HE staining:No obvious pathological changes were observed under Light microscope in the sham-operation group.In ICH group,the nervous tissues around hematoma were loose and inflammatory cells transmigrated into the area of peri-hematoma.,The neural ballooning degeneration and necrosis were observed in the brain tissue of ICH group.While the pathological changes in the brain tissues of XNJ group were not as serious as that of ICH group. Conclusion1.The preparation of this ICH rat model,established by stereotaxic injection of auto-non-anticoagulated blood,was relative easy,reliable and practical.2.It is proved that the breakdown of Blood-Brain barrier induces cerebral edema after incerebral hemorrhage.3.The expression of MMP-9 and C3 in the area around hematoma was increased with the ICH.4.Xingnaojing Injection may protect blood-brain barrier and alleviate cerebral edema by decreasing the expression of MMP-9 and C3,and finally improve the neural function.

A Clinical Trial about Treatment with Nr-CWS for Cervical High-risk HPV Infection

Wed, 30 Mar 2011 15:31:16 +0000

Objective:To seek the clearance effect of Nocardia Rubra Cell Wall Skeleton (Nr-CWS) for treating the high-risk HPV infection of the lower female genital tract.Methods:Select 144 cases of those who do not have clinical symptoms,and the test results of HPV subtypes testing by HybriMax are positive(including single high-risk HPV positive,mixed high-risk HPV positive,and both high-risk & low-risk HPV positive),all cases are excluded high grade cervical lesions by colposcopy and cervical biopsy.All cases were random divided into 4 groups:the first group was treated with Nocardia Rubra Cell Wall Skeleton(Nr-CWS)(n=39) every other day, 120μg a time,10 times a procedure;the second group was treated with 50% trichloroacetic acid(n=38),one time per week,about 8～10 times a procedure, according to the disappearance of acetic white epithelium;the third group was treated with microwave therapy apparatus(n=46) to contact treat on the surface of cervix for 2～3 minutes in orde to make the cervical surface tissue solidified and turned into yellow or white after menses clean and without acute inflammation of intemal or external genitals;and the patients without any trerapy as control group(n=21).All patients get HPV subtypes testing by Hybribio flow-through hybridization(short for HybriMax) before and 6 months after treatment.Compare the negative conversion ratio of high-risk HPV of each group and roughly analyse the clearance effect for HPV subtypes.Result:The negative conversion ratio of Groupl,Group2,Group3 and control group are 71.8%,63.2%,60.9%and 28.6%.The negative conversion ratio between groupl and control group is significant difference(χ~2=10.385,P=0.001),the conversion ratio of groupl is higher than that of control group;the negative conversion ratio between group2 and control group is significant difference(χ~2=6.474,P=0.011),the conversion ratio of group2 is higher than that of control group;the negative conversion ratio between group3 and control group is significant difference(χ~2=6.017,P=0.014),the conversion ratio of group3 is higher than that of control group.No statistical significance of negative conversion ratio in 3 treatment groups(χ~2=1.197, P=0.551),and we do not think there is difference among the negative conversion ratio of the 3 groups.There is no statistical significance of negative conversion ratio between single HPV subtype positive and mixed HPV subtypes positive(P=0.731).There is also no statistical significance of negative conversion ratio among single HPV-16 subtype,HPV-16 mixed other HPV subtype and non-HPV-16 subtype(P=0.682).Conclusion:This clinical research proves that there is clearance effect of Nocardia Rubra Cell Wall Skeleton(Nr-CWS) in clearing the high-risk HPV infection of the lower female genital tract.Nr-CWS has pharmaclolgical action on HPV without selectivity to its subtype.Compare with the clearance effect of 50%trichloroacetic acid and microwave therapy,there is clearance effect of all these 3 therapy methods to clear the high-risk HPV infection.But Nocardia Rubra Cell Wall Skeleton(Nr-CWS) is more simple,convenient,safe and effective without any side effects,and is easy to accepted by the patients.It has very important clinical applied cost.

Effect of Plasma IL-6, C-reactive Protein on Post-thoracotomy Patients Combined Intercostal Nerves Cryoanalgesia with Xuebijing and Clinical Observation

Wed, 30 Mar 2011 15:01:37 +0000

Complications after thoracotomy are mainly atelectasis, hypoxemia, pulmonary infection, arrhythmia, etc. These postoperative complications are closely related to systemic inflammatory response and the extent of incision pain. Interleukin-6 (IL-6) and C-reactive protein (CRP) are two major inflammatory response in the media, both in the inflammatory response in patients with the occurrence and development of the process has played a very important role, the recent study showed that perioperative IL-6 and CRP changes is closely related to the occurrence of postoperative complications. Because incision pain after thoracotomy, the Patient thoracic exercise is limited, decreased respiratory function, leading to pulmonary atelectasis, pneumonia, lung abscess, such as the occurrence of pulmonary complications, which cause hypoxemia , arrhythmias, so that preoperative occult ischemic heart disease sudden attack. Either directly or indirectly, these complications increase levels of inflammatory mediators, thereby exacerbating the inflammatory response in patients, cause multiple organ dysfunction (MODS). Therefore, lower plasma levels of inflammatory mediators and effective analgesia on the prognosis of patients has a very active role. Intercostal nerves cryoanalgesia after thoracotomy surgery as a way of its simple, safe and effective, has been widely used clinical. Xuebijing as an effective pro-inflammatory factor antagonist, is widely used in sepsis and multiple organ dysfunction syndrome treatment, as well as in lung disease, renal transplantation has been applied.Objective Through the combined with intercostal nerves cryoanalgesia and Xuebijing on Post-thoracotomy patients with plasma IL-6, C-reactive protein and clinical observation, to explore the effect of this combination of Traditional Chinese Medicine with Western Medicine method for thoracotomy in patients with inflammatory mediators and their clinical significance.Methods Selected close to the surgical site, surgical cases in the same surgical methods as to minimize the differences in observed indicators because of Surgical factors caused by, Therefore select line left thoracotomy in patients with esophageal carcinoma 60 cases, they were randomly divided into 3 groups, control group, cryoanalgesia group (frozen group), combined with cryoanalgesia and Xuebijing group (United group). Severity of pain was graded by Visual Analogue Scale (VAS), recorded after 48h in patients with VAS score situation. Plasma IL-6 concentration levels by peripheral venous blood in vitro were measured before 1d and at 2h, 6h, 24h, 3d and 6d after operation. Plasma CRP concentration levels by peripheral venous blood in vitro were measured before 1d and at 2h, 24h, 3d and 6d after operation, on the measured results were compared to observe the different methods effect of patient in perioperative plasma IL-6 and CRP concentration levels. Record three groups postoperative complications and compared. Observation of combined with Intercostal Nerves Cryoanalgesia and Xuebijing on the impact of complications after thoracotomy and clinical significance.Results The day before surgery, each group preoperative plasma IL-6, CRP concentration was no significant difference. After thoracotomy in patients with plasma IL-6, CRP concentrations were significantly higher than preoperative. IL-6 level after operation in all patients was significantly higher than that before surgery, reach its peak at 2h after operation. At 6h after operation plasma IL-6 started to decline, after 3-6d plasma IL-6 dropped to close to preoperative levels. CRP level was no significant change at 2h after operation. CRP level at 1d after operation in patients of all groups was signifieantly higher than preoperative, 3-6 days after operation it\'s significant downturn. IL-6, CRP level were similar in each groups, but the increased rate were different. Patients with frozen group at each phase of the IL-6 levels below the control group, and at 2h, 6h, 24h after operation measured level of IL-6 has significantly differences with the control group (P<0.05). In united group patients the level of IL-6 at each phase is lower than the control group and the frozen group, and at 2h, 6d after operation measured level of IL-6 has signifieantly differences with the control group (P<0.05) .And at 6h, 24h, 3d after were very signifieantly different with control group (P<0.01) . At 2h after patients in each group detected no signifieantly difference in CRP, patients with frozen Group at after operation 24h, 3d measured level of CRP was lower than the control group (P<0.05) . In united group, postoperative 24h, 3d measured concentration of CRP was significantly lower than control group (P<0.01) . At 6d after operation measured concentrations of CRP was lower than the control group (P<0.05) . At 48h after operation, the VSA score of Control group was 7.25±2.33, the total effective rates were 35%. Frozen group was 2.1±2.22, and the total effective rates were 95%. There was significant difference between the frozen group and the control group (P<0.01) . United group was 2.25±2.24, the total effective rates were 90%. There was significant difference between the united group and the control group (P<0.01), with no difference between frozen group. Control group has three cases of postoperative complications in patients, the incidence of complications was 15%; frozen group has two cases, the incidence of complications was 10%, which is lower than the control group; United group without postoperative complications is lower than the frozen and control groups, but each group had no statistical significance.Conclusion Freezing intercostalnerves during thoracotomy can effectively prevent postoperative chest pain, also can reduce post-operative plasma IL-6, CRP levels, reduce the occurrence of postoperative complications. Combined with intercostal nerves cryoanalgesia and Xuebijing can further reduce the peripheral blood of patients with IL-6 and CRP levels .Tips combined with Intercostal Nerves Cryoanalgesia and Xuebijing this Traditional Chinese medicine with Western Medicine method on prevention of complications after thoracotomy has been effective.

The Clinical Study on Curative Effects of Halitosis with H.pylori Infection by Traditional Chinese Medicine Combined with West Medicine

Wed, 30 Mar 2011 15:01:34 +0000

Purpose:The purpose of the this study is to investigate the clinical curative effect of halitosis with Hp infection by liquor of xianzhuli with trigeminy therapeutic resisit Hp.Method:Choose 60 patients in the Secondary Hospital of Fuzhou City diagnosed with halitosis with Hp infection,belonging to damp-heat in spleen and stomach,during the Nov.2007 and Nov.2008.All the cases are divided into 2 groups at random-treatment group and comparison group,30 for which respectively.The Treatment group is given "by liquor of xianzhuli with trigeminy therapeutic resist Hp (Rabeprazole+Clarithromycin+arilin)",the other group is given "trigeminy therapeutic resisit Hp(Rabeprazole+ Clarithromycin+ arilin)".Both group Take 7 days as a period.Reexamine the halitosis numerical value after 1week to observe the short-term curative effect;Reexamine the halitosis numerical value after 4 week to observe the long-term curative effect..Result:1.Comparison of general effects on halitosis between the treatment group and comparison group:first,therapeutic effect in the near future,23 are completely cured and 6 are partly cured,1 is successfully affected and 2 is wasted in the former group,of which the general effective power is 96.66%.In comparison group,the corresponding data is 14,11,5 and the effective power is 83.33.%. secondly,after one month observe the long—term therapeutic effect.There is obvious difference between the efficacy of the two kinds of methods(P0.05);3. Comparison of general effects on chief symptoms between the treatment group and comparison group the symptoms in treatment group:There is obvious difference between the efficacy of the two kinds of methods on halitosis、bitter taste of mouth and belch(P＜0.05),but no obvious difference on stomachache、gastric upset、anorexia and stool irregularity(P＞0.05).Conclusion:1.The research is to improve the relation between Hp and halitosis;2.The effect of the traditional Chinese drug of zhuli with trigeminy therapeutic resist Hp is more great than trigeminy therapeutic resist Hp only;3.According to the differentiation of symptoms and signs in TCM halitosis due to weishihejiang,zhuoqishangni,with the syndrome of bad breath,sticky,Dry mouth and thirsty,the traditional Chinese drug of zhuli has cold nature,can clear heat、eliminate sputum and dissipating dampness;On the basis of west medicine pharmacology,zhuli can eliminate inflammation,antibiosis,furthermore accelerate enterokinesia to promote the effluence of turbid pathogen.Therefore the halitosis and the curative of Hp both will make a great improvement..In addition,xianzhuli can be safety taken at low cost without any side effects,which is worthy of being wide applied.

The Analysis of the Syndrome Type of Chronic Hepatitis B Patients with Anticiral Therapy in Fuzhou

Wed, 30 Mar 2011 15:01:32 +0000

Objective:By the people at the second hospital in Fujian Province,Fuzhou Hospital for Infectious Diseases to conduct anti-hepatitis B virus(HBV) in patients with chronic hepatitis B treatment of the symptoms,signs and supporting a comprehensive assessment examination, analysis of chronic hepatitis B in Fuzhou region etiology,pathogenesis,as well as the distribution of TCM,Fuzhou region for the combination of the actual situation of Chinese and Western medicine to Chinese medicine or combination of prevention and treatment of chronic hepatitis B study to provide reference.Methods:According to TCM syndrome of chronic hepatitis B study reported in the literature, select 1 January,2005-2008 in the period between December 31 in Fujian Province People\'s Hospital,the second anti-HBV treatment in patients with chronic hepatitis B,as well as in 2008 March 1 -2008 in the period December 31 visit hospital for infectious diseases in Fuzhou,anti-HBV treatment of 232 cases of patients with chronic hepatitis B,combination of symptoms,signs to Syndrome Differentiation,analysis of Fujian,the cause of chronic hepatitis B patients,pathogenesis,as well as the distribution of TCM.Results:1、Fuzhou region of chronic hepatitis B patients treated with anti-virus-type distribution of TCM in order to humid type of resistance was the most common followed by the remaining type of kidney-yin deficiency of liver,spleen liver depression type,blood type resistance network.See the survey identified six cases of illegal immigrant can be no spleen yang-deficiency type.2、patients with chronic hepatitis B as a common tongue for the Red tongue,yellow and greasy tongue dark-red moss.3、ALT,AST and other auxiliary inspection target,and its points are the level of heat resistance of the highest type,and compared to other card-type,the difference was significant(P＜0.05).Conclusion:1、hepatitis B virus is the cause of chronic hepatitis B,as one of Exogenous evil, evil drug humid nature.Fuzhou area and timing,and are hot and humid air,the combined effect of the human body,healthy energy dispute settlement,as throughout the entire process of the basic contradictions in the pathogenesis.2、hepatitis B virus as a heat evil drug,acting on the human body heat can be more different from the vicissitudes of human factors such as righteousness and show different clinical manifestations and evidence-based.3、Chinese medicine,traditional Chinese and western medicine on patients with chronic hepatitis B in Fuzhou to carry out anti-HBV therapy,to Qingre Huashi a result of the treatment,according to Syndrome Differentiation different rationale for combining liver spleen,Qi Yin,and dredging collaterals blood differentiation,such as treatment,could achieve good curative effect,pending further study.

Influence of Huajiyiganjian on Expressions of Fas、FasL and C-erbB-2 in Asciteshepatomacell Line H22

Wed, 30 Mar 2011 15:01:30 +0000

Objective:To study the resistant function of HuajiYiganjian(HJYGJ)to ascites he- patic carcinoma, influence to the expression of Fas、Fasl and C-erbB-2, and discuss the possible mechanism of tumor resistanc.Methods:After injecting ascites hepatoma cell line into their right oxter ,48 mices were divided into four groups:normal saline(NS)、Huajiyiganjian(Chinese traditional medicine)、5-Fu(medicine)、Huajiyiganjian and 5-Fu (combine) groups. After 14 days obtaining tumor and accounting the inhibiting rate. Fas、Fasl and C-erbB-2 were examined by using immunohistochemistry straining method in the tumor tissue.Results:1. The experiment of QLYGJ resistance to growth of primary hepatic carcinoma of mice shows:Tumorweight of control group (n=8) is 2.7638±0.7311g, while the tumor weight of QLYGJ group (n=8) is 1.8763±0.7028 g,the resistant rate is 32.11%, itis obviously lower than control group (P0.05)。Conclusion: 1.HJYGJ、combine groups have apparently inhibiting effects on ascites hepatoma cell line H22,and combine group is higher than HJYGJ grou- p.2.HJYGJ group makes The expression of Fas increased and promoted tumor cell apoptosis; it makes the expression of FasL decreased and promoted immunological cytoactive,thus it hold-backs the development of tumor3. HJYGJ makes the expression of C-erbB-2 is decreased;it inhibits the generation of cancer cell .4. HJYGJ and 5-FU group degrade drug tolerance of hepatoma, relieve adverse effect, thereby it can implement motive of attenuation and synergism;

Study on Left Atrial Systolic Function in Patients with Uremia Using Strain Rate Imaging and Real-time Three Dimensional Echocardiography

Wed, 30 Mar 2011 15:01:25 +0000

Objective: To evaluate left atrial(LA) systolic function in patients with uremia by strain rate imaging(SRI)and real-time three dimensional echocardio graphy(3-DE).Methods: The uremic patients as research subjects, 30 uremic patients as the experimental group A,which the internal diameter of LA (LAD) were normal with tow-dimensional ultrasound measurement; 44 uremic patients as experimental group B,which mass index of left ventricle(LVMI)were normal. 30 healthy volunteers as the control group.The parameter of the conventional echcardiographic、SRI and 3-DE were repectively measured to the experimental group A,B and the control group,obtained parameters include: parameters of doppler echcardiographic:the peak E velocity(E), and the peak A velocity(A), E/A ratio, isovolumic relaxation time(IRT),velocity time integral of A wave(A-VTI) ; parameters of SRI:the left atrial systolic strain rate at left atrial wall top segment of Atrial septal, lateral, inferior, anterior, posterior(SRa);parameters of 3-DE:the maximun left atrial volume (LAVmax),mininum left atrial volume(LAVmin), left atrial volume before systole(LAVp) were determined by 3-DE. calculate left atrial ejection fraction(LAEF) and left atrial systolic volume(LASV). the internal diameter of LA(LADd) were measured to the experimental group A and the control group by tow-dimensional; posterior wall of interventricular septal thickness(IVST),the internal diameter of left ventricle(LVDd)and left ventricle (LVPWT) were measured to the experimental group B and the control group by tow-dimensional. calculate mass index of left ventricle (LVMI). Results: 1. LAD、E/A、IRT、A-VTI were not significantly different between the experimental group A and the control group,but the left atrial strain rate(SRa),left atrial systolic volume(LASV) and left atrial ejection fraction(LAEF) are significant differrent (P < 0.05). 2. E/A、IRT、A-VTI were not significantly different,mass index of left ventricle(LVMI) were at the normal value range and had no significant differences between the experimental group B and the control group,but the left atrial strain rate(SRa) , left atrial systolic volume(LASV) and left atrial ejection fraction(LAEF) are significant differrent (P < 0.05). 3. parameters of SRa, LAEF and LASV in the experimental groups are higher than those of the control groups.Conclusions: The results show the LADd were normal and LVMI no change in patients with uremia,and left ventricular diastolic function did not significantly change by the conventional ultrasonic:but left atrial systolic function can happen obvious changes by SRI and 3-DE in patients with uremia.Left ventricular diastolic dysfunction were predicted in directly.Ear- ly diagnosis may guide clinical treatment taken timely and effectively.It was an important reference that early clinical diagnosis and timely treatme- nt and prognosis .

The Expression and Significance of B7-H4 in Ovarian Epithelial Tumors

Wed, 30 Mar 2011 15:01:23 +0000

Objective: Ovarian tumors is one of the common major tumors of female reproductive system, the epithelial tumors are the most common ovary tumors, account for 50%～70% of primary ovarian neoplasms and 85%～90% of all ovarian malignancies. Ovarian cancer is predominantly seen in middle and aged women, the symptoms are insidious in onset, the early diagnosis is difficult and the mortality is high. Recent years,the transvaginal ultrasound (TVUS), measurement of serum CA125 levels and radical surgical cytoreduction followed by platinum and taxane-based polychemotherapy in various combinations have become the early diagnostic and effective therapeutic methods for ovarian cancer. However, complete surgical excision of advanced ovarian cancer is very difficult and conventional chemotherapy generally associated with a large number of toxic side effects, the 5-year survival rate of advanced ovarian cancer is still very low. Clearly there is a need to discover the tumor genesis in order to develop novel therapies for the treatment of ovarian cancer, tumor development and progression is related with host immune system. T cell is at the central position in immune response of body versus tumor cells, which would be a most effective and most safe approach to eliminate tumor if it can arouse the immune system. The activation of T lymphocytes needs double signals: one is antigenic peptide-MHC complex, which is recognized by T cell receptor; one is co-stimulatory signal. Thus, co-stimulatory molecule is very important to the initiation of immune system.B7-H4 is an new member of the co-stimulatory family, recent study confirmed it can inhibit influence the activation and function of T response. At present, there are few experiments about the expression of B7-H4 in ovarian epithelial tumors. This study was designed to evaluate the expression of B7-H4 protein in ovarian epithelial tumors and its relation with ovarian cancer clinicopathological variables, and discussed its role in occurrence process of ovarian tumors, in order to find out a new method for diagnosis and therapy of ovarian cancer.Method: The expression of B7-H4 in 23 cases of serous cystadeno- carcinoma,8 cases of borderline serous cystadenoma,18 cases of serous cystadenoma,10 cases of mucinous cystadenocarcinoma,6 cases of border- line mucinous cystadenoma, 11 cases of mucinous cystadenoma, 6 cases of endometrioid carcinoma and 8 cases of clear cell carcinoma tissue were assessed by immunohistochemical staining. The data was analyzed by SPSS 17.0, and examined the correlation between B7-H4 expression and histological type, tumor grade, operation-pathological stage and age.Results: 1 Immunohistochemical staining: The positive expression of B7-H4 was buffy grains and mainly stains in tumor cell cytoplasm and circum- ferential membrane.2 B7-H4 positive expression was detected in primary serous (95.7%,22/23), endometrioid (100%,6/6), and clear cell carcinomas (87.5%, 7/8).By contrast,B7-H4 positive expression was detected in only 1/10 mucinous carcinomas, the positive rate was 10%,the differences were statistically significant﹙P<0.001﹚. The total positive rate of B7-H4 in ovarian was 76.6% (36/47).3 The proportion of positive expression was greater in ovarian carcinomas than in borderline cystadenoma(28.6%,4/14) and benign tumors(24.1%,7/29),the differences were statistically significant﹙P<0.001﹚.4 No correlation was observed between the proportion of B7-H4 positive expression in ovarian carcinomas with tumor grade, operation- pathological stage and age.Conclusion: The expression rate of B7-H4 was higher in ovarian cancer than in borderline cystadenoma and benign tumors, this suggests that B7-H4 should be further investigated as a new diagnostic marker for ovarian cancer, and the blockade of B7-H4 in ovarian cancer might be one strategy to pursue for future immunotherapy in ovarian cancer.

Analysis of Hazard Factor Effects of the Early Mortality about Penetrating Cardiac Trauma after Operation

Wed, 30 Mar 2011 15:01:22 +0000

Penetrating Cardiac Trauma(PCT) was considered to be "unabled medical" lethal damage because of its heavy state,fast development,high mortality and difficulty treatment.Until 19 century\'s ends(in 1896),Germany surgeon Ludwing Rhen successed to suture patching on a patient whose right ventricle was punctured by knife.In the development about 100 years after that,the PCT\'s prognosis had gradually improved.However,the PCT still is the critical illness in the range of chest trauma,according to the late-1990s overseas related report,the PCT total mortality rate was 25%-65%.Along with the science of medicine technology,the communication and the trasportation development,the more state seriously injured patients may deliver the hospital to obtain the rescue.Although the heart penetration wound\'s rescue success ratio improves,but the mortality rate was quite still high.Surgery is the main treatment for PCT,the outcomes of the PCT operations are greatly diferent due to the injury mechanism,operation spot,and the other clinic conditions.Some preoperative or operative variables related to the diseases or the operations may influence the early mortality after operations significantly.But the effects of these factors on the early mortality after PCT operations aren\'t consensus. At present there is no systematic review of the quantitative analysis for such clinic studies aimed at such clinic problem in China and abroad.On the basis of the retrospective review of the clinic data of patients who underwent PCT operations in our hospital and analysis of the risk factors influencing early outcomes after PCT operations,we applied a new pattern,reliable method of synthesis and appraisal of the literatures(meta-analysis) to analyses the risk factors that may affect the early mortality after PCT operations,identify the effects of these factors on early postoperative mortality,and expect to guide the treatment for PCT in the future. Objective:Records of the 27 patients suffering penetrating cardiac trauma over a twenty-year period were retrospective reviewed.To identity the risk factors for early mortality after PCT operations.Then,the experimnt applies the Meta-analysis to quantitative study the variables which affects the early mortality after PCT operation, by merger and synthesis of the results in many small sample studies.It will be proposed more reliable conclusion to prevent the early death after PCT,and established more powerful evidence about evidence-based medicine,to further direct the clinic work.Methods:In the first part of the study,the clinic data of the PCT patients who underwent opewration in our hospital during the 20 years was collected,concomitant procedure were selected to identify the risk factors for early mortality after PCT operation by univariate analysis.In the second part of the study,all literatures related to analysis of the risk factors for early mortality after PCT operations in MEDLINE database,EMBASE database and CBMdisc were searched for and appraised critically to undertake meta-analysis of risk factors for early mortality mortality after PCT operations and identify the effects of these variables on early after PCT operations.The variables which were used to undertake meta-analysis included injury mechanism,operation spot,number of chambers to be injuryed,pericardial tamponade,combined injury,Revise Trauma Score,blood pressure on admisson, physiology index.Results:1.Results of the first part:Overall early postoperative mortality was 41%(11 deaths).Injury severity score (ISS)＞25(p=0.03),The Revised trauma score(RTS) at presentation≤2(p=0.002),the combination with aortic injury(p=0.004),the quantity of metachysis at the first 24 hours≥12 unit(p=0.0203),exploration at the emergency room(p=0.0002) were significant univariate risk factors for early mortality after PCT operation.2.Results of the second part:2.1 Results of the effect of injury mechanismThe results of meta-analysis of 11 retrospective studies and 1 prospective study indicated that the gun shoot group and the stab group comparsion of early mortality in PCT patients after operation with the pooled OR was 4.56(P＜0.00001,95%CI, 3.54-5.88).There was no statistical heterogeneity among studies included in the meta-analysis(P=0.50).2.2 Results of the effect of operation spotThe results of meta-analysis of 3 retrospective studies and 1 prospective study indicated that the emergency room group and the operation room group comparsion of early mortality in PCT patients after operation with the pooled OR was 22.26 (P＜0.00001,95%CI,13.87-35.73).There was no statistical heterogeneity among studies included in the meta-analysis(P=0.17).2.3 Results of the effect of the number of chambers to be injuryedThe results of meta-analysis of 3 retrospective studies and 1 prospective study indicated that the one chamber to be injuryed group and the more than two chambers to be injuryed group comparsion of early mortality in PCT patients after operation with the pooled OR was 0.19(P＜0.00001,95%CI,0.11-0.32).There was no statistical heterogeneity among studies included in the meta-analysis(P=0.32).2.4 Results of the effect of pericardial tamponadeThe results of meta-analysis of 4 retrospective studies indicated that the pericardial tamponade group and the no pericardial tamponade group comparsion of early mortality in PCT patients after operation with the pooled OR was 0.68(P=0.93, 95%CI,0.47-0.97).The test of heterogeneity showed that there was statistical heterogeneity among studies included in the meta-analysis(P=0.005).The synthesis was completed by random effect model.2.5 Results of the effect of the combined injuryThe results of meta-analysis of 4 retrospective studies indicated that the with the combined injury group and the no combined injury group comparsion of early mortality in PCT patients after operation with the pooled OR was 3.31(P=0.02,95% CI,1.17-9.35).The test of heterogeneity showed that there was statistical heterogeneity among studies included in the meta-analysis(P=0.02).The synthesis was completed by random effect model.2.6 Results of the effect of Revise Trauma Score(RTS)The results of meta-analysis of 2 retrospective studies and 1 prospective study indicated that the RTS=0 group and the RTS=1+ group comparsion of early mortality in PCT patients after operation with the pooled OR was 5.26(P=0.0008,95%CI, 2.00-13.83).There was no statistical heterogeneity among studies included in the meta-analysis(P=0.51).2.7 Results of the effect of the blood pressure on admissonThe results of meta-analysis of 4 retrospective studies and 1 prospective study indicated that the blood pressure on admisson＜90 mmHg group and the blood pressure on admisson＜90 mmHg group comparsion of early mortality in PCT patients after operation with the pooled OR was 6.52(P＜0.00001,95%CI, 3.39-12.52).There was no statistical heterogeneity among studies included in the meta-analysis(P=0.27).2.8 Results of the effect of the physiology index(PI)The results of meta-analysis of 4 retrospective studies indicated that the(PI≤10) group and the(PI＞10) group comparison of early mortality in PCT patients after operation with the pooled OR was 0.10(P＜0.00001,95%CI,0.06-0.17).There was no statistical heterogeneity among studies included in the meta-analysis(P=0.07).Conclusions:1.From the experience of surgical treatment for PCT in our hospital,Injury severity score(ISS)＞25(p=0.03),The Revised trauma score(RTS) at presentation≤2 (p=0.002),the combination with aortic injury(p=0.004),the quantity of metachysis at the first 24 hours≥12 unit(p=0.0203),exploration at the emergency room (p=0.0002)may affect early postoperative mortality significantly.2.The results of the meta-analyses indicated that variables related to the gun shoot,emergency room operation,with combined injury,RTS=0 and the blood pressure on admission＜90 mmHg are the risk factors for the early death after PCT operation,while the number of chambers to be injuryed(n=1),pericardial tamponade and the physiology index(PI≤10) are the protective factors for the early death after PCT operation.3.By merger and synthesis of the results of the same kind studies, meta-analysis can increase the sample size,enhance the statistic efficiency.In particular,when the results of diferent studies aren\'t consensus or there aren\'t statistic significance in results,meta-analysis could obtain statistic analytic result that approachs the real situation.4.The results of this study would advantage the systematic evaluation of the operative risk of the patient with Penetrating Cardiac Trauma in the future clinic work,take reasonable measures to relieve risk factors to improve surgical outcome. They would also benefit the prediction of the prognosis of the patient undergoing operation.Therefore,this study is helpful for the clinic work in the future.

Expression of NTs in Human Dental Pulp Stem Cells and Its Effect on Proliferation and Differentiation of the Cells: An Experimental Study in Vitro

Wed, 30 Mar 2011 15:01:19 +0000

It was thought that human dental pulp stem cells (hDPSCs)were progenitors existing in dental pulp.They could be activated by dental damage and terminally differentiated into functional odontoblasts. Neurotrophins (NTs) were important growth factors and of multiple functions, and thay are proved to be expressed during tooth development. NTs is an important part of network regulation of tooth development and plays an important role in the development of tooth. Studies showed that NTs promoted proliferation, differentiation and calcification of dental pulp cells in vitro,and they may play a role in the differentiation of progenitors of odontoblasts. However, whether human dental pulp stem cells express NTs, and whether NTs could enhance the proliferation and differentiation of human dental pulp stem cells in vitro had been poorly investigated. In this study, we preliminarily cultured human dental pulp stem cells, investigated the expression of NTs, and explored the effects of NTs on the proliferation and differentiation of human dental pulp stem cells. It provided experimental evidence for further research of role of NTs in the tooth development and regeneration ,as well as proliferation and differentiation of human dental pulp stem cells.This study includes three parts:1. Cell culture and identification: Dental pulp tissue of healthy third molars was digested with mixture of collagenase and dispase and the stem cells were isolated by limited dilution of cultured cell for single cell clone. Their morphology and growth condition were observed under inverted light microscope. Immunohistochemistry procedure was employed to detect surface marker of the cloning cells. Results: hDPSCs exhibited self-renewal and clonogenic cell population.The incidence of colony forming cells was 4-28 colony/104 cell plate. Immunohistochemical analysis showed that cultured cells expressed vimentin and STRO-1 while keratin were absent in hDPSCs.2. Expression of NTs in hDPSCs: RT-PCR, immunohistochemistry and western blotting were employed to detect mRNA and protein expression of NTs in hDPSCs, respectively. Results: RT-PCR, immunohistochemistry and western blotting showed that hDPSCs expressed BDNF in the level of both mRNA and protein. Western blotting showed NT-4/5was also expressed in hDPSCs, while other NTs were absent.3. Effects of NTs on proliferation and differentiation of hDPSCs: Cells between the third and fifth passages were used in this study. The optical density was obtained by MTT assay in cells stimulated by NTs in different concentrations. Effect of NTs on differentiation of hDPSCs was indirectly measured by the activity of alkaline phosphatase of the cells.Results: When compared with control group, NGF of 25ng/mL and BDNF of 25ng/mL stimulated the cell proliferation significantly (P<0.05) .However, NT-3 and NT-4/5 showed no effect on cell proliferation. When compared with control group, the ALP activities of hDPSCs were obviously increased with NGF of 100ng/mL(P<0.05), BDNF of 100 ng/mL(P<0.05). NT-3 and NT-4/5 of 25ng/mL(P<0.05). It is concluded that different concentrations of certain NT and different kind of NTs have different effects on proliferation and differentiation of hDPSCs.

Screening and Study on the Mechanism of Elicitors of Tripterygium Wilfordii

Wed, 30 Mar 2011 15:01:17 +0000

Tripterygium wilfordii is the important poisonous natural resources in our country , Its main active material are Total alkaloid and triptolide . but the content is too low for every per tree.for this , on the basis of the researches at home and abroad , through the different drugs spraying on the tree of Tripterygium wilfordii,this paper analysed the impacts of different storage methods on triptolide and characteristics of physiology and biochemistry,studied physiological ecological mechanisms on triptolide through adopting correlative analysis and regression analysis and trend forcast and analyzing the impacts of the main indexex of triptolide and physiology and biochemistry on triptolide and the characteristics of physiology and biochemistry.the results were as follows:1.There were significant differences on the effects of different treatments to triptolide.the treatments which adopted the 12th day of A1 treatment method,the 12th day of A3 treatment method , the 20th day of A6 treatment method , the 30th day of A11 treatment method , the 15th day of A12 treatment method , were able to maintain higher volume of triptolide of Tripterygium wilfordii leaf than others . the content are : 6.215、8.813、5.48、5.715、4.75(μg/g). but the content of triptolide of Tripterygium wilfordii leaf of The control group is only about 1.502μg/g.2. There were significant differences on the effects of different treatments to the main indexex on physiology and biochemistry of Tripterygium wilfordii .between the osmolytes, the Comparison between the non-induction and induction for the proline and MDA content,the result is the induction content is much more than the non-induction content.but the SPC content is opposite,the induction content is much less than the non-induction content; For Chlorophyll, the Comparison between the non-induction and induction for Chlorophyll a and Chlorophyll b content, the result is the induction content is much more than the non-induction content. but the Carotenoids content is opposite,the induction content is much less than the non-induction content,although the result is not Significantly;For the activity of Enzyme, the Comparison between the non-induction and induction for the SOD content,the result is the induction content is less than the non-induction content. the CAT content is opposite,and the Change of POD content is not Significantly. the induction content is not much less than the non-induction content;3.The content of triptolide in the fresh leaves、roots and branches of Tripterygium wilfordii were measured by this Experiment , but it did not change significantly in the roots and branches, except the content of triptolide of method A6 branch has improved obviously , it has also improve slightly in Method A9、A10、A11、A12 root . the content are: 2.863、2.588、2.25、2.33(μg/g).but the content of triptolide of Tripterygium wilfordii roots and branches of The control group are only about 0.5725 and 1.5567μg/g. 4. The main indexes of physiology and biochemistry had important influences on triptolide of Tripterygium wilfordii . In these main indexes , SOD and POD had the most influences than other indexes on triptolide of Tripterygium wilfordii leaf . showed negative correlation with triptolide of Tripterygium wilfordii leaf , the Correlation coefficient are -0.3487and-0.3709 . Chlorophyll a and Carotenoids had more influences than other indexes on triptolide of Tripterygium wilfordii leaf, showed positive correlation with triptolide of Tripterygium wilfordii leaf ; the Correlation coefficient are 0.3303 and 0.3057 ; In addition , soluble protein showed negative correlation with triptolide of Tripterygium wilfordii leaf. the Correlation coefficient is -0.3298 , but the proline、MDA、CAT and Chlorophyll b are all showed positive correlation with triptolide of Tripterygium wilfordii leaf. the Correlation coefficient are 0.2051、0.2166、0.1659、0.2531(μg/g).

Typing of X-Chromosome SNP Markers with SNPlex System

Wed, 30 Mar 2011 15:01:14 +0000

Objective:To type 48 SNP loci in X chromosome by SNPlex platform and evaluate their application values for kinship testing.Methods:A 48 X-SNP pool selected from 167 X-SNP loci by Bioinformatics was formed.272 blood samples were genotyped by SNPlex technique.The sensitivity of the method was detected and the genotyping results were verified by sequencing.The forensic application values of 48 X-SNP loci were evaluated through genetic polymorphic investigation.Results:Allele frequencies and genetic polymorphic parameters about 48 X-SNP loci were got.The coincidence rate of of genotying and sequencing was 99.17%. Statistical analysis about heterozygosity,polymorphism information content,the probability of discrimination power and the probability of exclusion showed that the 48 X-SNP loci were highly polymorphic except rs6527549.Conclusion:The X-SNP loci were polymorphic and suitable for forensic biology application.Being highly specific,highly accurate,good compatible,middle-throughout on loci and high-throughout on samples,SNPlex technique meet the needs of forensic biological detection.

The Influence on Prostaglandin E2 in Treatment for Knee Osteoarthritis by Electroacupuncture Combined with Herbal Steaming Washing Therapy

Wed, 30 Mar 2011 14:42:23 +0000

Purpose:It discusses the objectiveness of the assessment standard for the effectiveness of treatment for knee osteoarthritis by electroacupuncture combined with herbal steaming washing therapy through observation of the variation of Prostaglandin E2 in the treatment.Approach:The randomized approach and comparative approach are employed in the clinical research,which observes 60 medical cases of knee osteoarthritis(one side),covering the groups adopting electroacupuncture combined with herbal steaming washing therapy (treatment group) and foreign medicine combined with herbal steaming washing therapy (comparative group) with 30 cases in each group.After two courses of treatment,the variation of Prostaglandin E2 in the joint fluid of two groups with knee osteoarthritis,the comparison of the simplified McGill ache assessments before and after the treatment,and the variation of the standard assessment of the effectiveness(JOA) will be observed.Result:1 There is striking differences(P＜0.05) of Prostaglandin E2 in the joint fluid of the two groups before and after treatment,while there is little difference between the treatment group and comparative group(P＞0.05).2 There is striking differences(P＜0.05) between the two groups in the simplified McGill ache assessment and the assessment standard(JOA) of the effectiveness of knee osteoarthritis before and after treatment,while there is little difference between the treatment group and comparative group(P＞0.05).3 The effectiveness of the two groups is as follows:90%for the treatment group and 96.7%for the comparative group.There is little difference between the treatment group and comparative group(P＞0.05).Conclusion:1 It is proved that the Prostaglandin E2 is adopted as an indicator for the treatment of knee osteoarthritis.The electroacupuncture combined with herbal steaming washing therapy decreases the content of Prostaglandin E2 in the in the joint fluid of knee osteoarthritis so as to impair the harm to the knee joint,improve the clinical condition and relieve the sickness.2 Both the electroacupuncture combined with herbal steaming washing therapy with foreign medicine combined with herbal steaming washing therapy decrease the simplified McGill ache assessment,and increase the assessment standard(JOA) of the effectiveness of the treatment for knee osteoarthritis,which proves that it is a very efficient method by means of the combination of the electroacupuncture combined with herbal steaming washing therapy or foreign medicine combined with herbal steaming washing therapy.3 There is no striking adverse interaction related with this therapy after the treatment of the electroacupuncture combined with herbal steaming washing therapy.It is the effective combinattion of all the advantages to treat knee osteoarthritis with security and efficiency and without side effect.Therefore,it is worthwhile to put it into practice in clinic.

Randomized Controlled Trials of Promoting Blood Circulation and Removing Stasis Therapy on Insulin-resistance-related Diseases: A Systematic Review

Wed, 30 Mar 2011 14:42:22 +0000

Objective:To analyze the effects and possible adverse effects of promoting blood circulation and removing stasis traditional Chinese medicine prescription\'s treatment on insulin-resistance-related diseases,taking insulin sensitivity index(ISI),fasting plasma glucose(FPG),blood pressure,blood lipid and so on as observation indexes.Methods:A computer-based online search of China national knowledge internet (CNKI) d atabase and VIP database was undertaken to identify the articles about the treatment for cases with insulin resistance between January 1994 and February 2008, and January 1989 and February 2008.Randomized controlled trials(RCTs) and quasi-randomized controlled trials(quasi-RCTs) of patients with insulin resistance were included.The quality of included therapy was evaluated and meta-analysis was performed by RevMan 4.2 software.Results:Thirteen RCTs or quasi-RCTs involving 860 patients were included.It was found that some Chinese medicinal formulae of promoting blood circulation and removing stasis might be effective for diseases related with insulin resistance to some extent.Firstly,they could reduce fasting plasma glucose.Meta-analysis based on 9 included studies(subgroupⅠ) suggested that the effects of lowering fasting blood glucose in treatment group(Chinese medicinal formulae of promoting blood circulation and removing stasis group) were superior to those in the control group with statistical significance(WMD -0.76,95%CI -1.31 to -0.22).Meta-analysis based on other 4 included studies(subgroupⅡ) showed the similar result in the effects of lowering fasting blood glucose,but with no statistical significance(WMD -0.34,95%CI -1.10 to 0.42).Secondly,they could reduce fasting insulin.Meta-analysis based on 8 included studies(subgroupⅠ) suggested that the effects of lowering fasting insulin in treatment group were superior to those in the control group with statistical significance(SMD -1.41,95%CI -2.43 to -0.38).Meta-analysis based on other 4 included studies (subgroupⅡ) showed the similar result in the effects of lowering fasting insulin,but with no statistical significance(SMD -0.74,95%CI -1.63 to 0.16).Thirdly,they could improve insulin sensitivity index.Meta-analysis based on 7 included studies(subgroupⅠ) showed that the treatment group was more effective to elevate ISI compared with the control group with statistical significance(WMD 0.86,95%CI 0.45 to 1.26). Meta-analysis based on other 4 included studies(subgroupⅡ) told the similar result in the effects of improving ISI,but with no statistical significance(WMD 0.72,95%CI -0.03 to 1.48).Fourthly,they could decrease both systolic pressure and diastolic pressure.Meta-analysis based on 6 included studies(subgroupⅠ) suggested that the effects of lowering both systolic pressure and diastolic pressure in treatment group were superior to those in the control group with statistical significance,respectively is(WMD -0.63,95%CI -1.12 to -0.14) and(WMD -0.33,95%CI -0.62 to -0.04).Fifthly,they could reduce total cholesterol and triglyceride.Meta-analysis based on 4 included studies(subgroupⅠ) showed that the treatment group was more effective to reduce both total cholesterol and triglyceride compared with the control group with statistical significance,respectively is(WMD -0.68,95%CI -1.09 to -0.28) and(WMD -0.44, 95%CI -0.61 to -0.26).Meta-analysis based on other 4 included studies(subgroupⅡ) also told the similar result in the effects of reducing total cholesterol and triglyceride. Both the differences had statistical significance as well,respectively is(WMD -0.59, 95%CI -1.16 to -0.01) and(WMD -0.15,95%CI -0.23 to -0.06).Sixthly,they could increase high density lipoprotein cholesterol(HDL-c).Meta-analysis based on 4 included studies(subgroupⅠ) showed that the treatment group was more effective to increase HDL-C compared with the control group,but with no statistical significance (WMD 0.11,95%CI 0.00 to 0.22).Meta-analysis based on other 3 included studies (subgroupⅡ) told the similar result in the effects of increasing HDL-c with statistical significance(WMD 0.30,95%CI 0.03 to 0.57).Seventhly,they could decrease low density lipoprotein cholesterol(LDL-c).Meta-analysis based on 3 included studies (subgroupⅠ) and other 3 included studies(subgroupⅡ) both showed that the treatment group was more effective to decrease LDL-c compared with the control group.Both the differences had statistical significance,respectively is(WMD -0.65,95%CI -0.97 to -0.32) and(WMD -0.59,95%CI -1.06 to -0.13).No obvious side effects or adverse reactions were noted.Conclusions:Promoting blood circulation and removing stasis therapy might be effective for diseases related with insulin resistance to some extent.The evidence is not strong enough because of poor quality of the included studies.Therefore,more high quality randomized controlled trials are required.

Arsenic Down-regulates the Expression of Camk4, an Important Gene Related to Cerebellar LTD in Mice

Wed, 30 Mar 2011 14:42:20 +0000

Objective: To elucidate the molecular mechanism of arsenic (As) on motor learning and memory.Methods: concentration of As in cerebellar tissue of mice exposed to 1 ppm and 4 ppm As_2O_3 subchronically was determined by ICP-MS, neurobehavioral changes associated with memory was examined by the Morris Water Maze test, and the critical gene expression profiles related to the Creb-dependent phase of cerebellar long-term depression (LTD) were analyzed by GeneChip.Results: Our results showed the increased level of As concentration in cerebellar tissue of the exposed mice in a dose-response manner and the downregulated expression of Ca2+/calmodulin dependent protein kinase IV (Camk4), a very important regulator in the LTD pathway. We further analyzed the influence of As on cerebellum Camk4 expression by Western blot. The quantity of Camk4 band in the group exposed to 4 ppm As_2O_3 significantly decreased compared to the control group, agreeing well with the gene microarray result. It is strongly suggested that As accumulated in cerebellum of mice chronically exposed to As and the deposited As impeded the Camk4 expression. Therefore, the Camk4 may be target of As-induced neurotoxicity. Furthermore, the intervention of antioxidants taurine or vitamin C did not prevent Camk4 from downregulation, indicating that the downregulation of Camk4 expression by As may be via a oxidation- inde- pendent way.

The Study on the Deletion of Exons and Protein Expression of Rap1GAP, and Its Relationship with the Expression of MMP2 and MMP9 in Invasive Breast Cancer

Wed, 30 Mar 2011 14:42:19 +0000

Objective: To explore the deletion of Rap1GAP exons of E11 and E19, protein expression of Rap1GAP, MMP2, MMP9 in invasive breast cancer, and the correlation of these indicators in invasive breast cancer for understanding the molecular mechanisms of breast cancer.Breast cancer, a malignant tumor of breast ductal epithelial cells, is a common malignancy of women. In the world, there are about one million women are diagnosed with breast cancer each year, and breast cancer incidence increase by 5% -20% per year. In China, the incidence of breast cancer ranked second only to cervical cancer. The occurrence and progression of tumor is a multi-factors, multi-stages process and is based on a variety of proto-oncogene activation and tumor suppressor gene inactivation. Rap1GAP is a tumor suppressor gene and is closely related with the occurrence and progression of a variety of tumors.Rap1GAP( Rap1 GTPase-activating protein 1) is a Rap1-specific GTP- activating protein, regulating the activity of Rap1 from the GTP binding active form to the GDP binding inactive form. It regulates cell biological functions such as growth, productivity, adhesion and so on. Our group’s Preliminary studies have shown that: Rap1GAP inhibited Hela cells proliferation, migration and invasion; And Rap1GAP inhibited Hela cells migration and invasion by inhibiting the activities of MMP2 and MMP9. Cervical cancer and breast cancer are two major killers of female diseases, a variety of genes changes are same in both cancer tissues. In this paper, we studied the deletion of Rap1GAP exons of E11 and E19,protein expression of Rap1GAP, MMP2, MMP9 in invasive breast cancer, and analyzed the correlation of these indicators in invasive breast cancer.Methods: 53 cases surgical breast tissue specimens were from Dalian Central Hospital Pathological Centers, including 10 cases of normal breast tissues, 17 cases of breast lobular hyperplasia tissues and 26 cases of invasive ductal carcinoma. All specimens were diagnosed by pathology. DNA was extracted form pathohistological sections, PCR method used to check the homozygous deletion of Rap1GAP exon E11 and exon E19. Extraction DNA, PCR method used to measure the homozygous deletion of Rap1GAP exon E11, exon E19. The expression of Rap1GAP, MMP2, MMP9 in breast tissue slices were investigated by S-P method of immuno- histochemistry. the correlation of these indicators in breast lobular hyperplasia tissues and invasive ductal carcinoma were analyzed.Results:1. The deletion rates of Rap1GAP exon E11 homozygous were: 10 % (1/10), 11.76 % (2/17), 19.23 % (5/26) in normal breast tissues, breast lobular hyperplasia tissues and invasive ductal carcinoma tissues respectively. The deletion rates of Rap1GAP exon E19 homozygousthe were: 20 % (2/10), 11.76 % (2/17), 19.23 % (5/26) in normal breast tissues, breast lobular hyperplasia tissues and invasive ductal carcinoma tissues respectively. There were no significantly difference in deletion rates of exon E11 and E19 among the three groups (p>0.05).2. The expression rates of Rap1GAP were: 80 % (8/10), 52.94 % (9/17), 30.77% (8/26) in normal breast tissues, breast lobular hyperplasia tissues and invasive ductal carcinoma tissues respectively. Rap1GAP expression was lower in invasive ductal carcinoma tissues than that in breast lobular hyperplasia tissues (p0.05)4. The expression rates of MMP9 were: 10 % (1/10), 23.53 % (4/17), 57.69% (14/26) in normal breast tissues, breast lobular hyperplasia tissues and invasive ductal carcinoma tissues respectively. among the three types of breast tissues, MMP9 expression was higher in invasive ductal carcinoma tissues than that in breast lobular hyperplasia tissues (p0.05).5.2 There was negative correlation between the positive rate of Rap1GAP and MMP2 expression in breast lobular hyperplasia tissues (p0.05)and invasive ductal carcinoma tissues (p>0.05).Conclusions:1. The deletion rates of Rap1GAP exon E11 and E19 have no correlation in invasive ductal carcinoma tissues, breast lobular hyperplasia tissues and invasive ductal breast cancer.2. The expression rates of Rap1GAP protein in invasive ductal carcinoma were lower than that in breast lobular hyperplasia and in normal breast tissue. Its expression in breast lobular hyperplasia was lower than that in normal breast tissues.3. The expression rates of MMP2, MMP9 protein was higher in invasive ductal carcinoma than that in the breast lobular hyperplasia and in normal breast tissue. Its expression in breast lobular hyperplasia is higher than that in normal tissue.4. In the breast lobular hyperplasia and invasive ductal carcinoma tissue, the expression of Rap1GAP was down-regulation, but the expression of MMP2, MMP9 was up-regulation and negative correlation were observed between Rap1GAP expression and MMP2/MMP9.5. The major cause of down-regulation of Rap1GAP may not relay on gene deletion.

The Effect of P2X_7 Receptor on the Expression of CD44 and Its Function in P388D1 Cells

Wed, 30 Mar 2011 14:42:18 +0000

Objective To study the functional expression of P2X7 receptor in P388D1 and its association with CD44 and cancer cells metastasis.Methods Analysis of P2X7R expression by RNA extraction and revers transcriptasepolymerase chain reaction (RT-PCR),western blotting and analysis of P2X7R function by the bleb formation upon agonist stimulation was observed under microscope. Extracellular adenosine triphosphate(ATP) is P2X7R agonist .Analysis of P388D1 cell CD44 expression after incubated for up to 30 minutes in NaCl or KCl medium at 37°C with ATP by Flow CytometryMoreover,KN-62, the most potent inhibitor for the P2X7R were added 15 minutes before the addition of ATP then analysis of P388D1 cell CD44 expression by Flow Cytometry.Results P2X7 receptor expression could be detected in P388D1 at mRNA and protein level. The result of beble formation suggest the P388D1 cells express functional P2X7 receptors.Extracellular ATP causes fluorescence intensity of CD44 attenuating.KN62 Inhibited ATP-induced causes fluorescence intensity of CD44 attenuating by P2X7R. trypan blue staining results of the less number of the cells in bottom chamber which were stained with trypan blue showed that the cells passed through the filter and attached to the bottom chamber. The number of P388D1 cells was decreased compared without the P2X7R actived by ATP.Conclusion Base on our study, it is indicated that P2X7R can descrease expression of CD44 in P388D1.It will open a new vision for the clinic treatment of tumor.

A Study of Microsatellite Instability and Loss of Heterozygosity in Cervical Cancer and Cervical Intraepithelial Neoplasia

Wed, 30 Mar 2011 14:42:16 +0000

Objective:1.To explore the relationship of loss of heterozygosity(LOH) and microsatellite instability(MSI) in the development of cervical cancer and cervical intraepithelial neoplasia.2.Explore the mismatch repair genes and the fragile histidine traid genes in the development of cervical cancer,thus provide the basis of the early detection and treatment in cervical cancer.Methods:MSIand LOH of 6 microsatellite markers D2S123、D4S43、D4S125、D3S1297、D3S4103、D3S1298 were detected in 60 cervical cancers and 50 cases of cervical intraepithelial neoplasia(CIN) and self normal cervix.Results: Compare the PCR products of tumor tissues and corresponding normal tissue DNA at the same time.If a particular allele in tumor tissue strip with non-tumor tissue compared with the corresponding article completely disappear or relative density of more than 50%reduction,the result is the LOH;if the tumor tissue compared with non-tumor tissue strip happened translocation or increased,the result is the MSI.1.The relationship of LOH and cervical cancer:Compared 60 cases of invasive cervical organization and its corresponding normal tissue,the positive rates were gradually increased with the degree of malignancy of cervical cancer. Except D4S125,significant differences were noted between FIGO stage of cervical cancer,between the well,the moderately and the poorly differentiated cervical carcinomas and between the invasive cervical cancer with lymphnode metastasis and those without lymphnode metastasis in regard to LOH(P＜0.05).2.The relationship of MSI and cervical cancer:Compared 60 cases of invasive cervical organization and its corresponding normal tissue,the positive rates were gradually increased with the degree of malignancy of cervical cancer. On D2S123、D3S4103、D3S1298,significant differences were noted betweenⅢ、Ⅳstage andⅠ、Ⅱstage of cervical cancer,between the well,the moderately and the poorly differentiated cervical carcinomas and between the invasive cervical cancer with lymphnode metastasis and those without lymphnode metastasis in regard to MSI(P＜0.05).3.The relationship of LOH and CIN:Except D4S43 and D4S125,significant differences were noted betweenⅠ、Ⅱstage andⅢstage of 50 cases of CIN (P＜0.05).4.The relationship of MSI and CIN:On D2S123、D3S4103 and D3S1298, significant differences were noted betweenⅠ、Ⅱstage andⅢstage of 50 cases of CIN(P＜0.05).5.The relationship of the mismatch repair genes and CIN and cervical cancer: At two loci D2S123 and D3S1298 with mismatch repair gene-related,the positive rate of MSI and LOH increase with its increased level,significant differences were noted between FIGO stage of cervical cancer,between the well,the moderately and the poorly differentiated cervical carcinomas and between the invasive cervical cancer with lymphnode metastasis and those without lymphnode metastasis in regard to LOH(P＜0.05).6.The relationship of the fragile histidine traid genes and CIN and cervical cancer:At one loci D3S4103 with the fragile histidine traid gene-related,the positive rate of MSI and LOH increase with its increased level,significant differences were noted between FIGO stage of cervical cancer,between the well,the moderately and the poorly differentiated cervical carcinomas and between the invasive cervical cancer with lymphnode metastasis and those without lymphnode metastasis in regard to LOH(P＜0.05). 1.6 loci with high LOH and MSI prompt that tumor-suppressor gene with the development of cervical cancer near these sites.2.Statistical significance was noted between LOH,MSI and clinical stage, pathological grade in cervical cancer,prompt that detected LOH and MSI in these regions can be used as important indicators with tumor malignant degree and prognosis.3.Statistical significance was noted between LOH,MSI and pathological grade in CIN,prompt that LOH and MSI was a later period event in the occurrence and development of CIN.4.Mutations of mismatch repair gene are an important reason for in tumor happened,its detection contribute to prognosis and early diagnosis of cervical cancer.5.The fragile histidine traid genes occurs mutation in many kinds of tumor,and we find it occurs on early lesions of cervix,so that it is an early case.

The Effect of Thalidomide on Neointimal Hyperplasia after Balloon Injury in Rat Carotid Artery

Wed, 30 Mar 2011 14:42:14 +0000

Objective: The effects of local delivery thalidomide on inhibiting neointimal hyperplasia and VEGFmRNA expression after balloon injury in rat carotid artery and possible mechanisms were researched in the present paper.Methods: 48 male Sprague-Dawley rats were randomly divided into sham operation group (group A), alone operation group (group B) and Thalidomide group (group C). The carotid arteries of group B and group C were injured by conventional Percutaneous transluminal coronary angioplasty (PTCA) balloon catheter, group C were treated by local delivery thalidomide and group B without delivery thalidomide, The arteries of group A were not injured. 7 days and 14 days after balloon injury, rats were sacrificed. Serum concentrations of vascular endothelial growth factor (VEGF) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). HE staining was performed to observe neointima area, lumen area. Macrophage infiltration and local expressions of VEGF were measured by immunohistochemical analyses. Real time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to examine the expression of VEGFmRNA.Results:①Rat models of balloon injury and thalidomide local delivery by porous balloon were successfully completed. 7 days and 14 days after balloon injury ,the levels of VEGF and TNF-αwere more significantly increased in group B than that in group C (VEGF:4.82±0.17pg/ml vs 0.98±0.1pg/mL on day 7, P0.01;6.3±0.16pg/ml vs 1.03±0.09pg/mL on day 14, P0.05) and in lumen area of group A, B and C (B to A:0.33+0.03mm~2 vs 0.39±0.05mm~2,P>0.05; C to B: 0.39±0.04mm~2 vs 0.33+0.03 mm~2,P>0.05; C to A: 0.39±0.04mm~2 vs 0.39±0.05mm~2, P>0.05) 7 days after balloon injury. However, thalidomide administration resulted in a significant reduction of neointimal formation 14 days after balloon injury (0.07±0.01mm~2 in group C vs 0.12±0.04mm~2 in group B, P0. 05).③VEGFmRNA expression levels were no statistic meaning in group A, B and C on 7 days (B to A: 1.04±0.09 vs 0.97±0.14, P>0.05;C to B: 0.98±0.1 vs 1.04±0.09, P >0.05; C to A: 0.98±0.1 vs 0.97±0.14, P>0.05). VEGFmRNA expression of injured arteries in Group B on 14 days was significantly increased compared with that in Group C and A (6.3±0.16 vs 1.03±0.09, P 0. 05).④There were no differences of VEGF and macrophage staining among 3 groups on 7 days, the mean optical density values were respectively 0.1131±0.0112, 0.1034±0.0120, 0.1200±0.0136(P>0.05). VEGF staining was significantly increased in the nucleus and the cytoplasm of group B 14 days after balloon injury, which was attenuated in group C, the mean optical density values were 0.1784 + 0.0112 and 0.1234 + 0.0122(P<0.01). Tissue macrophage infiltration was significantly reduced in the group C compared with group B 14 days after balloon injury and the mean optical density values were 0.1084 + 0.0157 and 0.1837 + 0.0144 (P<0.01).Conclusions: The result shows that thalidomide using local drug delivery significantly reduced the levels of VEGF and TNF-αin serum, decreased macrophage infiltration and local protein content of VEGF and VEGFmRNA expression, inhibited neointimal hyperplasia and vascular restenosis after balloon injury in rat carotid artery, which suggests an essential role for VEGF in the pathogenesis of restenosis. This might provided a novel method for prevention and therapy of restenosis.

Investigation of Inhibiting A549 Cell Proliferation Mediated with RNA Interference Targeting to Gene Her2/neu

Wed, 30 Mar 2011 14:42:13 +0000

Lung cancer is the first cause of cancer death around the world.The cure rate is lower than 15%despite some developments in chemotherapeutic agents HER-2/neu gene also called erbB2,is a member of the epidermal growth factor receptor (EGFR)family and encodes a 185 kDa protein with tyrosine kinas activity and shows partial homology to the other epidermal growth factor receptors.The HER2/neu oncogene is over expressed in 30%of Non small cell lung cancer(NSCLC),and its signaling pathways play an important roles in tumor occurring,cancer cell growth, differentiation,metastasis,anti-apoptosis and resistance to chemotherapeutic agents.RNA interference(RNAi) is the phenomenon of sequence-specific,posttranscriptional gene silencing(PTGS) triggered by double-stranded RNA(dsRNA), which exists in extensive organisms such as Caenorhabditis elegans,plants, mammalian.Now RNAi has developed into a novel gene-blocking tool and used to knockdown specific gene expression efficiently in various species.Many great achievements have been made by its application includes screening new genes, functional genomics and gene therapy.However,there is elusive about the effects of depressing the expression of HER2/neu by RNAi on lung carcinoma cell line A549 cell proliferation.AimTo construct RNA interference vector to down-regulate HER2/neu gene and study the siRNA against HER2/neu effect on the cell cycle,cell proliferation of lung carcinoma cell line A549 in vitro.Methods1.Cell culture:lung carcinoma cell line A549 were cultured at 37℃/5%CO_2 in DMEM medium supplemented with 100IU/ml penicillin,10μg/ml streptomycin,and 10%heat-inactivated fetal calf serum(FCS).2.Construction of recombinant siRNA expression plasmids:①Choose appropriate 19bp sequence on the transcript of target gene and blast to eliminate any target sequences with significant homology to other coding sequences.The oligonucleotides were synthesized by Shanghai Bioasia corporation.②Annealing of each pair of oligonucleotides③Double digestion of vector by endonuclease④Ligation of annealed oligonucleotide inserts to linearized vector.⑤Transformation of DH5α⑥Identification of recombinant plasmids by sequencing.3.Cell Transfection:①Optimizing the transfection condition according to manufacture\'s introductions,choosing the best ratio of FuGene6 to plasmid DNA.②Plate the lung carcinoma cell line A549 one day before the transfection experiment.When cells were 60-80%confluent,transfected cells according to the optimal condition.4.EGFP Reporter Assay:lung carcinoma cell line A549 were transfected by pEGFP-N2 and siEGFP(target to EGFP) plasmid.The transfected cell were observed by fluorescence microscope.5.Western blotting:three days after transfection,cells were washed with PBS and collected by scraping.They were lysed in ice-cold RIPA buffer and centrifuged. The supernatant was used for protein concentration determination by BCA-200 method.The proteins were resolved on 10%SDS-polyacrylamide gels,transferred onto nitrocellulose membranes,and incubated with the appropriate antibodies.The peroxidase-based detection was performed with Chemiluminescence Reagent(Pierce) according to the manufacturer\'s instructions.6.RT-PCR:Total RNA was prepared from A549 cells 48h after transfection using the Tripure reagent according to the manufacturer\'s protocol.RT-PCR amplification products were resolved by 1%agarose gel.7.Cell cycle analysis and growth rate:①A549 cells were seeded in 6.0cm dishes with DMEM medium free of serum.The cells were synchronized in cell cycle 24h later.Then the cells were transfected with psilence 3.1-HER2 or psilence 3.1-Control vector with FuGene6 reagent.The cells were trypsinized 48h after transfection and washed by PBS for three times.FCM analysis were applied to measure cell cycle;②The cells were seeded in 24 well plates and transfected with psilence 3.1-HER2 or psilence 3.1-Control vector by using FuGene 6.On different time points after transfection,MTT method were applied to measure cell growth.All samples were done in tripicate and every sample counted was repeated at least 3 times.Results1.The recombinant siRNA expression plamids were successully constructed based on pSilencer3.1H1 vector targeted to HER-2/neu.2.Plasmid-based siRNA expression could knockdown the expression of exogenous reporter genes.After being transfected with siEGFP plasmids,the EGFP overexpression lung carcinoma cell line A549 shown decreased EGFP expression rate.3.Plasmid based siRNA expression could inhibit endogenous target genes. Several repeated experiments show that psilence 3.1-HER2 transfection can specially inhibit HER-2/neu mRNA and protein expression.4.FCM results showed that cells transfected with psilence 3.1-HER2 increased accumulation of cells in G_0/G_1 phase with a decrease in the percentage of cells in S-phase;Also,MTT results showed the growth curve of cells exhibited slower proliferation after the transfection of psilence 3.1-HER2.Conclusion1.human H1 promoter driven siRNA expression plasmids could be used in lung carcinoma cell line A549 to knockdown endogenous or exogenous gene expression.2.psilence 3.1-HER2 can knockdown her2/neu gene specially and efficiently; transfection of psilence 3.1-HER2 contributes to depressing effect of A549 cell proliferation.The data described here provided a proof of concept that vector-based RNAi targeting to her2/neu gene could be used therapeutically for lung cancer.

The Study of the Gray Matter Volume and Diffusion Tensor Imaging of Brain in Early Posttraumatic Stress Disorder

Wed, 30 Mar 2011 14:42:11 +0000

Post-traumatic stress disorder (PTSD) is a kind of psychonosema that initiates from the seriously abnormal painful incidents and also a delayed or a lasting reaction from the seriously abnormal and threatening disaster. It was first seen in , which was published in 1980. With the increasing number of the trauma events come from war, social violence, natural disaster, family violence, children maltreatment and so on, the attack rate of PTSD becomes more and more high and the research in PTSD becomes more and more widespread. But many problems about its pathogenesis are not very clear till now. Nowadays many foreign countries have widely investigated the neurobiology mechanism about PTSD, and the results indicated that the limbic system in patients with PTSD exists functional disorder, such as diminished volume in hippocampus and anterior cingutate, changed FA values in cingutate and so on. However, less research was done on the structural changes of early PTSD. This study used the high resolution magnetic resonance imaging (MRI) and magnetic resonance diffusion tensor imaging (DTI), and it began with the changes of the gray matter volume, FA values and ADC values in hippocampus, anterior cingutate and posterior cingutate. This study is to investigate whether there are changes in the hippocampus, anterior cingutate and posterior cingutate in the patients with early PTSD caused by mining disaster and to investigate the possible mechanism.ExperimentⅠ: The measurement of the limbic system gray matter volume in early PTSDObjective To investigate the possible changes of hippocampus, anterior cingutate and posterior cingutate volume in patients with early PTSD. Methods 10 patients with early PTSD caused by mining disaster were involved after six months, and 10 gender, age matched healthy persons who exposed to the same mining disaster were selected as controls. The high resolution T1 weight images were gained by Magnetom trio tim 3.0T (Siemens company) supraconductive MRI scan. Then IBASPM software toolbox was used to gain the brain gray matter volume in different anatomic site. At last, covariance methods were used to analyze the data previously obtained, so as to rule out the reflection in age and sex and to compare the differences in gray matter volume of hippocampus, anterior cingutate, and posterior cingutate between the two groups. Results After eliminating the reflection in age and sex, the gray matter volume in hippocampus and anterior cingutate, posterior cingutate did not show any significant differences between two groups (p>0.05). Conclusions The results suggest that there may be no significant differences in hippocampus and anterior cingutate, posterior cingutate volume between patients with early PTSD and healthy persons who exposed the same mining disaster . ExperimentⅡ: The changes of the limbic system DTI in early posttraumatic stress disorderObjective To investigate the possible changes of the hippocampus, anterior cingutate and posterior cingutate DTI in patients with early PTSD. Methods 10 patients with PTSD caused by mining disaster were involved after six months, and 10 gender, age matched healthy persons who exposed the same mining disaster were selected as controls. The whole brain DTI data were gained by Magnetom trio tim 3.0T (Siemens company) supraconductive MRI scan. FUNTool was used to perform the restitution of the FA image and ADC image. Analysis software(Neuro 3D)from manufacturing company in workstation was used to select and measure certain site in hippocampus, anterior cingutate and posterior cingutate. The region of interest (ROI) in hippocampus was divided into three parts, including head, body and tail. The sampling volume was equal, and it was put in the ROI center. Above work was done independently by two attending physicians in Department of Radiology in Xijing Hospital. With SPSS16.0 statistical analysis software, analysis of covariance methods was used to rule out the reflection in age and sex and to compare the differences in FA and ADC values of the hippocampus, anterior cingutate and posterior cingutate between the two groups. Simple correlation coefficientγcould be used to estimate the objective reliability among DTI data gauger. Results In comparison with controls after eliminating the reflection in age and sex, the PTSD patients had diminished FA values in right posterior cingutate (p<0.05). FA values in other regions and ADC values in the hippocampus, anterior cingutate and posterior cingutate did not show any significant differences between two groups. Conclusions The results suggest that there may be morphometric changes in right posterior cingutate in patients with early PTSD. The change of FA values detected by DTI can help to determine the involvement of neural network in PTSD patients sensitively.

The Traditional Chinese Medicine Radix Astragali Induce Gene Expression and Molecular Cloning of β-defensins-2 in Rat Womb

Wed, 30 Mar 2011 14:42:09 +0000

Through Objectiving the chinese materia medica like radix astragali influence toβ-defensin-2 in rat,so as to approach the mechanism of anti-infection of chinese materia medica,for choose defensins as inductor establish the foundation of theory and experiment.[Methods]:Sixty healthy rat random were divided into six part:blank,radix astragali,sophorae,radix,scutellaria,shuang huang lian oral liquid,each group were given intragastric administration.after seventh days,according to RNA extract,we got out the RNA of womb,to operate reverse transcription polymerase chain reaction(RT-PCR) detect the expression ofβ-defensin-2.Then,to put Amplification product purified and cloned,and sequenced recombinant plasmid.[Results]:Radix astragali,radix and shuang huang lian oral liquid were higher than normal,but others were not changed clearly.recombinant plasmid sequencing result manifest,acquired clone recombinate contain rBD-2cDNA,the result display complete same as Genebank.[Conclusions]:Radix astragali,radix and shuang huang lian oral liquid can strengthen expression ofβ-defensin-2.The result tole us that we can use these Chinese materia medica to enhance organizational immunity partly,to kill effect of pathogenic microorganism.This effect maybe one anti-infectious mechanism of the traditional Chinese medicine.

Experimental Study on Amplifide Labial Gum Soft Tissues with Different Levels Palatal Connective Tissue Flap

Wed, 30 Mar 2011 13:31:56 +0000

Objectives: Compare the healing and aesthetic effect separately after the free mucosa flap from different faults transplantation and offer the theoretical basis for clinic.Materials and methods: Chooses eight 12-16 months’healthy hybrid dogs as the experimental objects. Makes the palatal hard palate zone of the eight dogs under general anesthesia and takes the connective tissue flaps of different faults as reserve. Defect the lip-side gingival of bilateral maxillary anterior tooth, transplant and suture fix the epithelial layer (group A) and connective tissues without epithelial layer (group B) of different faults. Measure the length and width of the mucosa right after the treatment, calculates and records the initial area of transplant mucosa. Execute the animal after 1 week, 2 weeks, 3 weeks, and 4 weeks, and do the clinical inspects, measure the length and width of mucosa, calculate the area, and define the shrinkage of mucosa. Do the histology inspection analysis.Results:1 General observation: Of all the 8 experimental dogs, 1 of the tissue flap exfoliated because of infection, others healed well and had no exfoliate. The edge of transplanted flaps smoothed with the original wound edge of gingival soft tissues, and the transplanted flap of epithelial layer , with rose pink color, had similar thickness with the original gum.2 Morphological observation of HE dyeing:Transplanted flaps of epithelial layer: (1) One week after the operation, the connective tissues of transplanted flaps still had clear boundary with the connective tissues of recipient bed. Many of the inflammatory cells exuded and parts of the epithelial layer degenerated and necrossed; (2)Two weeks after the operation, there was still recognizable boundary between the connective tissues of transplanted flaps and the connective tissues of recipient bed. Fibrous tissues emerged a lot, which made the tissue density of them close to the tissue density of the recipient bed. Epithelial cells of the boundary kept on growing to the surface of the transplanted flap; (3)Three weeks after the operation, quantity of the vessels, inflammatory cells and the fibroblasts in the connective tissues of transplanted flap decreased obviously. The connective tissue density of transplanted flap got close to the tissue density of the recipient bed. Boundary of the two did not obvious any more. The epithelial tissues had already covered the whole flap. The epithelial surface-irregularities get decreased and shoaling; (4) Four weeks after the operation,inflammatory cells in the connective tissues of transplanted flap could rarely be seen. The connective density of transplanted flap increased and was unanimous with surrounding organizations and boundary between the two could not be recognized. Thickness of the epithelial tissues went normal with thick parakeratosis layer on the surface.Flap of connective tissues without epithelial layer: In the microscope, healing process of connective tissues changed consistent with the trans- planted flaps of epithelial layer, while the epithelial cells grew slower than the epithelial layer.3 Morphological observation of picric acid - sirius red dyeing:By picric acid - sirius red dyeing, all tissue slices appears cardinal red under ordinary microscope, especially the big collagen fiber, and there were no diversity between groups. Under the polarized light, there was mainly typeⅠcollagen in normal gingival soft tissues. The typeⅠcollagen fibers, which appeared red or yellow, disposed tight and orderly, and showed strong double light refraction. In the transplanted flap, the typeⅠcollagen fibers reduced significantly. As the healing period lengthened, the quantity of typeⅠcollagen fibers increased gradually. At about the fourth week, quantity of typeⅠcollagen fibers in the transplanted flap got close with those in the original gingival tissue, thinner than the original ones.Conclusions:1 The different connective tissues flaps heal well, and could repair the defection of soft tissues.2 The aesthetic effect of epithelial layer transplant flap is better.3 The subepithelial connective tissue without epithelial layer flap improves the thickness of soft tissues better, at the same time, it release the wound of the tissue donor area owing to the reserved epithelial layer.

The Theoretical Research about the Humanities Curriculum and the Growth of the Value System

Wed, 30 Mar 2011 13:31:53 +0000

The college humanities curriculum as the content vector of making the educated gain human spirit and the value of demanda plays a guiding role in the process of improving the overall quailty of the educated.The research, titled " The theoretical research of the humanities curriculum and the growth of the value system " On the base of seperately analyzing varying courses of the humanities curriculum and the value function and structure which the college humanities curruculum should process, summed up the characteristics of the humanities curriculum developing performance and its influence factors, and concluded the optimization mechanism of the growth of humanities curriculum and its value system. In particular, the role of the self-criticism.In historical,the college humanities curriculum have strong characteristics of the times in the different level of productivity. Which perform on the budding as pre-humanities curriculums, experience and natural philosophy-oriented curriculum as the prelude,and the curriculum of which oriented to secular philosophy and social ethics, after impacted by polytechnic curriculums, the college humanities curriculum was again paid attention to ,with the addition of a new time characteristics and the spirit of the times. All of which reflect the changes of people\'s social values in a certain level of social productive forces.The college humanities curriculum gradually gained a certain degree of mature after some changes on the value system. This needs national culture value as the value core of the developing value system of humanities curriculum,which were added by multicultural values in its development. In addition,social power is the base of legitimacy,the balance of multipolar power values including state\'s will,social power and school power are the bases of theory, and the amalgamation of time spirit and time value is the base of space value.All of which make the humanities curriculum in the developing space stretching and innovating.On the bases of analyzing the changes in the humanities curriculum and its value system,the study summed up the characteristics of college humanities curriculum in its process.Which including practice facilitated generating of goal, questioned the value of content structure, the main dependent on value and imperceptible influences on function implementation.In addition, analyzed the influence factors of curriculum growth,including social productive forces, social systems, social and cultural environment, traditional culture, social evaluation, the rights of society controlling, university educational philosophy, curriculum ideas, curriculum research, internal academic control, the methodology,and so on..To make the humanities course content and its value system has necessary optimizition, we must find the optimal mechanism.there exist some conditions such as the correct values, correct idea of a university, logical starting point for the correct courses,and integration of science and technology factors with a reasonable point of view. Especially, we focus on the the necessity of self-criticism to college humanities courses, the contents of the scope of self-criticism,as the optimized mechanism of college humanities curruculum and its developing value system.

Epidemiologic Study on Chronic Kidney Diseases among Military Elderly Population in Dalian

Wed, 30 Mar 2011 13:31:51 +0000

Objectives:To investigate the prevalence and possible risk factors of chronic kidney diseases(CKD) among military elderly population in Dalian.Method : 576 individuals,aged 65 or above,are selected from 16 military retirement centers in Dalian. All of them were interviewed.Their morning spot urine were tested to determine albumin to creatinine ratio,and renal function was assessed. Morning spot urine dipstick of hematuria was confirmed by microscopy. The associations among demographic charac- teristics , health characteristics and indicators of kidney damage were examined.Result:Eligible data of 576 subjects were enrolled in the study.The prevalence of albuminuria was 14.6%,hematuria 1.0%, and reduced eGFR 19.8%.The prevalence of CKD Was 27.4%.The prevalence of CKD stages, respectively, were 2.1% for stage 1; 5.6% for stage 2; 18.1% for stage 3; 1.0% for stage 4;and 0.7% for stage 5.The logistic regression model showed that hyperuricemia and diabetes contributed to the development of CKD.Conclusion:The prevalence of CKD among military elderly population in Dalian is 27.4%.Hyperuricemia and diabetes are risk factors of CKD.

The Related Clinical Research between Malnutrition and Stroke-associated Pneumonia

Wed, 30 Mar 2011 13:31:50 +0000

Objective:Stroke-associated pneumonia is common complications for stroking patient and malnutrition is pervasive phenomenon for inpatient. This investigation aims at delving into relativity of risk factors between malnutrition and SAP.This study can guide guarding against stroke-asso- ciated pneumonia as well as enhancing cognition for the old malnutrition in clinic.Methods: We have done prospective analysis for initial incidence of old cerebral ischemic stroke patients in the Department of Neurology of First Affiliated Hospital of Dalian Medical University during the period of from January 1st 2008 to December 30th 2008.The investigation mainly includes following parameters:basal state(Sex, smoking history,diabetes history, hypertension history,.hyperlipidemia, heart disease),some factors of stroke-related (the block volume, category of cerebral stroke by the type standard OCSP).Within 24-hours’s an evaluation of patient’s nutritional status (including body mass index,BMI; triceps skin-fold thickness,TSF; serum albumin, sALB; mid arm muscle circumference,MAMC).All patient have been observed for 2 weeks, monitoring whether to be attacked by SAP, All patients were divided into 2 groups by whether patient went down with SAP, and incidence rate of SAP was ascertained chi square test. Through factor analysis, selected factor(р< 0.05) compared with many factors, returning to logistic analysis.Results:1、210 chosen-patients consist of 71 pneumonia and 139 no-pneumonia, in 71 Stroke-associated pneumonia patients cover 45 early-onset pneumonia patients and 26 late-onset pneumonia patients. (Percentage: 63% versus 37%.) 2、Sixty-five patients were diagnosed in malnutrition status in 210 stroke patients, percentage of sickness was 31% . There were 21(32%) protein type malnutritions, and 32(49%) protein-energy malnutrition patien- ts and 12(19%) mixed type malnutrition patients.3、Thirty-six of malnutritional patients occoured SAP. There were 22 early-onset pneumonia patients,eleven of them were protein-energy malnu- trition,protein type malnutritions were four patients and seven mixed type malnutrition patients; There were 14 late-onset pneumonia patients, three of them were protein type malnutritions, protein-energy malnutrition were nine patients and two mixed type malnutrition patients.4、Through one factor analysis,comparing pneumonia patients with no-pneumonia patients in diabetes history,smoking history,the block volume,malnutrition,we found the value of survey in Statistics.5、Through many factors analysis,incidence rate of pneumonia of stroke patients who have diabetes history is 6.715 as high as who have no diabetes history.Probability of catching pneumonia to patients who have smoking history is 5.547 times as high as who have no smoking history. Patients who have large infarct volume is 7.505 times as high as who have smaller infarct volume in sickness rate of pneumonia.Case rate of pneu- monia to malnutritional patients is 12.144 times as high as patients with a good nutritional state.Conclusions:1、Sickness rate of malnutrition especially protein-energy malnutrition is high for the old ischemic stroke Inpatients.2、Stroke-associated pneumonia has high sickness rate in malnourished Inpatients.3、Malnutrition is one of the risk factor for cerebral ischemic stroke patients.

Carotid Artery Color Doppler Ultrasound and Analysis of Related Factors with Ischemic Stroke Patients

Wed, 30 Mar 2011 13:31:49 +0000

Objective: To investigate the characteristics,plaque nature,extent and distribution of vascular stenosis of extracranial carotid atherosclerosis in ischemic cerebrovascular disease of patients, providing an objective evidence to prevent the occurrence of carotid artery ischemic stroke.Methods: cerebral infarction and transient ischemic attack (TIA) patients of the internal carotid artery system with 121 cases and 40 healthy persons Collected from October 2007 to December 2008 was hospitalized in our hospital, were divided into three groups: cerebral infarction group, TIA group and the healthy control group. cerebral infarction patients have71 cases , 53 male, 18 women , aged 46-86 years old, on average (68±12) years old; TIA patients have 50 cases, 30 male, 20 women,aged 45-81 years old, averaged (66±13) years of old; healthy control group have 40 cases,23 cases male, 17 cases women,aged 57-76 years old, on average (65±7) years old. Mainly through the dual function of color Doppler ultrasound detection of extracranial neck vessels, testing the extent of atherosclerosis, intima-media thickness and plaque distribution, and in the light of its history, CT or MRA examination and biochemical analysisResults: 71 cases of cerebral infarction in patients with plaques of extracranial carotid artery plaque were found in 60 cases, of which 32 cases were vulnerable plaque (45.1%), stable plaque were 28 cases (39.4%). 50 cases of TIA patients who were found 36 cases of plaque, including vulnerable plaque in 11 cases (22.0%), stable plaque were 25 cases (50%). Healthy control group who were found in 18 cases of plaque, vulnerable plaque has not been detected, stable plaque in 18 cases. Cerebral infarction group, TIA group of vulnerable plaque between the two groups and the control group were 40 cases of vulnerable plaque, compared to those who have a significant difference (P0.05)Conclusion: Carotid artery color Doppler ultrasound in the detection of carotid atherosclerosis has important clinical value, some patients with the same period basically the same as in line to check the results of CTA or MRA.It can be in patients with ischemic stroke as the primary means of clinical examination. Ischemic stroke and the severity of carotid atherosclerosis is closely related, and when the carotid artery of moderate and severe stenosis and IMT≥1.5mm when a higher risk of cerebral infarction. CRP is closely related with cerebral infarction, ischemic stroke and Hcy no significant correlation.

A Study on the Association of the Methylation Status of HOXA10 and PTEN Gene Promoter Region with Endometriosis

Wed, 30 Mar 2011 13:31:48 +0000

BackgroundEndometriosis(EMs) is one of the most common benign gynecologic disorders, because it has been linked to pelvic pain,multiple operations,and infertility and difficult to cure.It is well known that certain aspects of endometriosis are similar to those of malignant disease.For example,like cancer,endometriosis can be both locally and distantly metastatic;it attaches to other tissues,invades,and damages them.And it is associated with increasing risk of cancer,The ovary is the most common site for development of malignancy in endometriosis,Nowadays the true mechanism for the pathogenesis or therapy of endometriosis is unclear.More and more evidences have indicated that malignant behavior of cancer has already became important morbidity mechanism of endometriosis.Hereditary variation of oncogene and tumor suppressor genes exists in malignant disease,so does endometriosis, especially the methylation of tumor suppressor genes.A number of studys have indicated that methylation of tumor suppressor genes may play an important role in cancer development and progression.Under the majority normal circumstances, status of tumor suppressor genes doesn\'t show methylation.Convincing evidences for the significance of CpG island or promoter hypermethylation in tumor development stem from the observation that these events are associated with silencing of tumor suppressor genes,eventually it causes cancer.Studies have indicated that HOXA10 gene can decrease invasiveness of cancer cell.This manifested HOXA10 gene is probably tumor suppressor gene.PTEN gene is a universally accepted tumor suppressor gene,which has been observed in a variety of cancers,such as breast cancer,kidney cancer,bladder cancer,melanoma, glioblastoma,and lung cancer,especially the endometrial,ovarian and glioblastoma cancer.The methylation of the CpG island of HOXA10 and PTEN promoter is important mechanism for the pathogenesis of human neoplasms.And this is often associated with metastatic diseases.Aberrant promoter methylation was related to inactivation of the HOXA10 and PTEN gene in ovarian and endometrial cancer. Endometriosis is a kind of benign disease with malignant behavior,and it\'s associated with ovarian cancer.And then does hypermethylation of HOXA10 and PTEN genes exist in endometriosis?Does the methylation of the CpG islands of HOXA10 and PTEN promoter play an important role in endometriosis? Now it is not clear.So far, the study about these aspects is eliminated,especially about the methylation of PTEN gene in endometriosis.ObjectiveTo investigate the methylation status of the 5\'CpG island locating in the promoter region of HOXA10 and PTEN gene in endometriotic tissues and uterine endometrium from women with and without endometriosis in order to study the role of HOXA 10 and PTEN in the pathogenesis and development of the endometriosis.Methods1.Tissue collection:Thirty ectopic endometrium specimens and 20 eutopic endometrium specimens from 30 patients with endometriosis(including 10 cases with endometriosis of stagesⅠ-Ⅱand 20 cases with endometriosis of stagesⅢ-Ⅳ) were used as study group from Sep.2007 to July.2008 in Department of Gynaecology and Obstetrics,Zhujiang Hospital of Southern Medical University,and 25 specimens of endometrium from women without endometriosis were used as controls study in the same period.2.Methods(1) Genomic DNA were extracted from frozen specimens by the method of proteinase K digestion. (2) All samples were detected for the methylation status of the 5\'CpG island locating in HOXA10 and PTEN gene promoter region by using methylation-specific PCR technique(MSP).The primers of the 5\'CpG island locating in HOXA10 and PTEN gene promoter region were designed according to reference.Set up blank control.(3).Statistical analysis The data were analyzed by using the chi-square test and Fisher\'s exact test(n＜40 or,when any of the expected frequencies was less than 5), Statistical analysis was performed using SPSS 13.0 software.Results1.The comparison of HOXA10 gene methylation between patients with endometriosis and without endometriosis:In women with endometriosis,presence of HOXA10 promoter methylation was seen in 12 of 30 ectopic endometriums,and 7 of 20 eutopic endometriums.However,there were no methylation occurred in HOXA10 gene of endometrium from 25 women without endometriosis.There were significantly different methylation of HOXA10 gene during three groups(P＜0.05).There were no significantly different methylation of HOXA10 gene not only between in ectopic endometrium and eutopic endometrium of women with endometriosis,but also in eutopic endometrium of women with endometriosis and endometrium of women without endometriosis,While there were significantly different methylation of HOXA10 gene between ectopic endometrium of women with endometriosis and endometrium of women without endometriosis;2.The comparison of PTEN gene methylation between patients with endometriosis and without endometriosis:In women with endometriosis,presence of PTEN gene promoter methylation was seen in 7 of 30 ectopic endometriums,and 2 of 20 eutopic endometriums,.However,there were no methylation occurred in PTEN gene of endometrium from 25 women without endometriosis.There were significantly different methylation of PTEN gene during three groups(P＜0.05). There were no significantly different methylation of PTEN gene not only between in ectopic endometrium and eutopic endometrium of women with endometriosis,but also in eutopic endometrium of women with endometriosis and endometrium of women without endometriosis,While there were significantly different methylation of PTEN gene between ectopic endometrium of women with endometriosis and endometrium of women without endometriosis;3.The comparison of HOXA10 gene methylation between patients with endometriosis inⅠ-Ⅱstage andⅢ-Ⅳstage:In women with endometriosis,presence of HOXA10 promoter methylation was seen in 11 of 20 ectopic endometriums of patients inⅢ-Ⅳstage,and1 of 10 patients inⅠ-Ⅱstage.There were significiantly different methylation of HOXA10 gene in ectopic endometrium between two group patients with endometriosis(inⅠ-Ⅱstage andⅢ-Ⅳstage)(P＜0.05);4.The comparison of PTEN gene methylation between patients with endometriosis inⅠ-Ⅱstage andⅢ-Ⅳstage:In women with endometriosis,presence of PTEN promoter methylation was seen in 6 of 20 ectopic endometriums of patients inⅢ-Ⅳstage,and1 of 10 patients inⅠ-Ⅱstage..There were no significantly different methylation of PTEN gene in ectopic endometrium between two group patients with endometriosis(inⅠ-Ⅱstage andⅢ-Ⅳstage)(P＞0.05).Conclusion1.The methylation of 5\'CpG island in the promoter region of HOXA10 and PTEN gene may be involved in occurence of endometriosis;2.The methylation of 5\'CpG island in the promoter region of HOXA10 gene may be correlated with the clinical stage of endometriosis and possibly play an important role in the development of endometriosis;3.There may be same biological characteristics about methylation between ectopic endometrium and eutopic endometrium in women with endometriosis.

Angiogenesis and Innervation of Tissue-engineered Bone Implanted with Vascular Bundle and Sensory Nerve Bundle: A Preliminary Morphological Study

Wed, 30 Mar 2011 13:31:46 +0000

BackgroundThe reconstruction of large bone segments remains a significant clinical problem in the case of extensive bone loss due to pathological events such as trauma, inflammation,and surgical treatment of tumors.Present therapeutic approaches include bone graft transplants(autologous,homologous or heterologous grafts), implants of different biomaterials or bone transport methods(Ilizarov technique),but none has proven to be fully satisfactory.Bone tissue engineering has the potential to become a real alternative to autologous bone grafts,but before this could happen it is necessary to solve the problem of homogenously supplying oxygen and nutrient to cells within a large scaffold.Sufficient blood-supply has the initial and decisive impact.Without efficient blood-supply,the tissue-engineered bone would be limited at repair the thin and small bone defects.The research of the distribution and function of nervous system in bone,has offered theoretical foundation and experience for the fact that the neural factor is incorporated to the construction of the tissue-engineered bone.In vivo studies,it maybe useful to test the feasibility and efficacy of using different approaches to facilitate the connection of the implant to the vascular network or innervation of the host.In this study,combining the tissue-engineering method and microsurgery skill, we implant the vascular bundle,sensory nerve bundle separately into the tissue-engineered bone to repair a great segmental femoral defect in rabbit,and observe the relationship between the osteogenesis effects and the reconstruction of vascular network or innervation in the tissue-engineered bone at early time.ObjectivesTo initially investigate the reconstruction of vascular network or innervation in the tissue-engineered bone with vascular bundle and nerve bundle implanted in vivo, and explore the mechanism of which the two methods affects osteogenesis.Materials and Methods1.42 New Zealand rabbits randomly assigned to 3 Groups were used for the experiment.Autologous bone marrow was harvested from the iliac crest of rabbits by needle aspiration and bone marrow mesenchymal stem cells(BMSCs) were proliferated and induction culture.These bone marrow-derived cells were seeded on beta-tricalcium phosphate(β-TCP).A stainless steel mini-plate was placed in front of the left femur.A reciprocating saw was then used to resect a 1.5-cm length segment of central diaphysis under saline irrigation.Tissue-engineered bone was imbedded into the defect at this location.A length of femoral vascular bundle was implanted to the side groove of tissue-engineered bone and fixed with thread(Group B,n=14). Similarly,an appropriate length of greater saphenous nerve was isolated,cut and then implanted to the side groove of tissue-engineered bone(Group C,n=14).Neither the vascular bundle graft nor the nerve graft was implanted into Group A(n=14).2.Four animals in each Group were sacrificed at 4、8 and 12 weeks after the implantations.Bone formation throughout the defects was assessed radiographically and evaluated by a radiologic score.Postmortem general and histology observation were also undertaken to examine bone formation.4%paraformaldehyde were used for the fixation of bone defects area and 10%EDTA were used for decalcification. The specimens was processed for routine paraffin imbedded to obtain serial paraffin sections(5μm)and the sections were then used to make HE and Masson sections to observe the osteogenesis of the tissue-engineered bone.Some of these sections were used for immunostaining or silver staining.3.The immunocytochemistry of vascular endothelial cell marker,CD34. Quantitative image analysis of the number of new vessels immunostained by CD34 was performed.Statistical software was used for statistical analysis.3×3 factorial design ANOVA was used to analyze the mean of each Group.When differences were statistically significant,the LSD method was applied for multiple comparisons.4.Observed angiogenesis by ink perfusion.After 12 weeks we selected two animals in each group undertaken ink perfusion,and prepared frozen sections of tissue engineeried bone to observe the situation angiogenesis.5.The immunohistochemistry using antibodies directed against specific neuronal markers,neurofilament 200(NF200) and synaptophysin(SY).Quantitative image analysis of the integrated optical density(IOD) of immunostainings for these markers was performed.Statistical software was used for statistical analysis.3×3 factorial design ANOVA was used to analyze the mean of each Group.When differences were statistically significant,the LSD method was applied for multiple comparisons.6.Observed the nerve reconstruction by silver staining.Gave each group samples conventional silver staining at different time points to observe nerve reconstruction of tissue engineering bone.Results1.Radiologic scoreThe radiologic scores of all the defects were increased in all groups at three time points post-implantation(P＜0.05).No significant difference between Group B and Group C in each time points(P＞0.05),but the scores in these two Groups were significantly higher than in Group A at 4、8 and 12 weeks.2.Histological observationThe histology observation of specimens in 3 Groups at 3 time points after operation.Calcification and degradation ofβ-TCP was found in all Groups at 4、8 and 12 week after operation.In Group B and Group C,material was divide into mini-particle by new bone formation,the degree of ossification around the material was higher;massive material was not degradation in Group C,and the degree of ossification around the material was lower.In all 3 Groups,new bone formation occurred around the vessel,the major way of bone formation is enchondral ossification.The extent of neovascularization and enchondral ossification in Group B and Group C were higher than Group A observed under Microscope.3.Immunohistochemistry stain results of CD34We observed the new vessels immunostained by CD34 in each group,and the total number of these new vessels was quantified.With the time prolong,the number of new vessels in each group were gradually decreased,the difference has statistical significance(F=222.578,P=0.000).The effects of sensory nerve bundle or vascular bundle implantation in the number of new vessels have statistical significance (F=1189.181,P=0.000).Factorial design ANOVA showed time and group have interaction(F=2.964,P=0.038).At 4、8 and 12 weeks,the number of new vessels in all 3 group have statistical significance(P＜0.05);In each group,the number of new vessels at all time point have statistical significance(P＜0.05).4.Ink perfusion observationsAfter 12 weeks,blood vessels structure can be seen clearly in vascular bundle implantation group material and there are a number of mature vessels with thick morphology,which are near adjacent osteoblasts.The internal blood vessels in nerve bundles implanted group with mild morphology are relatively mature,whose number is less than that of the vascular bundles implanted group.There are a small number of less mature internal blood vessels with general morphology in bone tissue engineering materials group.5.Immunohistochemistry stain results of SY and NF200With the time prolong,the expression of SY in each Group were gradually decreased,the difference has statistical signifcance(F=130.722,P=0.000).The effects of sensory nerve bundle or vascular bundle implantation in the expression of SY have statistical significance(F=103.523,P=0.000).Factorial design ANOVA showed time and group have interaction(F=4.867,P=0.004).At 4 and 8 weeks,the IOD of SY in all 3 group have statistical significance(P＜0.05);At 12 weeks,no significant difference was seen between Group B and Group C(P＞0.05),but they were obviously higher than in Group A(P＜0.05).In Group A,the IOD of SY at each time point have statistical significance(P＜0.05);In Group B and Group C,no significant difference was seen between 8 and 12 weeks(P＞0.05),but they were obviously lower than 4 weeks(P＜0.05).Small amounts of expression initially occurred at the edge of newly generated bone and periosteum and then decreased over time.With the time prolong,the expression of NF200 in each group were gradually decreased,the difference has statistical significance(F=152.848,P=0.000).The effects of sensory nerve bundle or vascular bundle implantation in the expression of NF200 have statistical significance(F=146.814,P=0.000).Factorial design ANOVA showed time and group have interaction(F=5.575,P=0.002).At 4 weeks,the IOD of SY in all 3 group have statistical significance(P＜0.05);At 8 and 12 weeks,no significant difference was seen between Group B and Group C(P＞0.05),but they were obviously higher than in Group A(P＜0.05).In each group,the IOD of SY at all time point have statistical significance(P＜0.05).The distribution of NF200 was similar with SY.Small amounts of expression initially occurred at the edge of newly generated bone and periosteum and then decreased over time.6.The results of silver stainingSilver-staining results showed that in four weeks the nerve fibers appear outside in each group of tissue-engineered bone and nerve fiber-like structures with poor form is only visible in the tissue gap inside the material.At 8 weeks,a large number of silver staining nerve fibers can be seen in the gap of new bone tissue and there are more in sensory nerve bundle implantation group and the vascular bundle implantation group than other groups.The staining nerve fibers with clear structures are staggering with each other.At 12 weeks,there are nerve fibers with good structure inside bone tissues and clear silver staining fibers are around the blood vessels.Conclusions1.In the early stage,both tissue-engineering bone implanted with vascular bundle and that implanted with sensory nerve bundle can enhance the repairing of large segmental defects in vivo,but there is no significant difference between two groups.2.In the early stage,both tissue-engineering bone implanted with vascular bundle and that implanted with sensory nerve bundle can obviously promote the reconstruction of blood supply in tissue-engineered bone,and the former group is better than the latter.3.In the early stage,both tissue-engineering bone implanted with vascular bundle and that implanted with sensory nerve bundle can obviously improve the reconstruction of innervation in tissue-engineered bone,but there is no significant difference between two groups.4.Angiogenesis and innervation in tissue-engineered bone decreased at different content in all groups during the early stage,suggesting that the early phase(4～8 weeks) maybe essential to the reconstruction of blood supply and innervation.5.The reconstruction of blood supply and innervation play an important role in the osteogenesis of tissue-engineered bone and the reconstruction of large bony defects.

Radix Astragali Normalizes the Gene Expression of Angiotensin-Converting Enzyme 2 on the Kidney in Rat Model of Metabolic Syndrome

Wed, 30 Mar 2011 13:31:45 +0000

Objective Angiotensin-converting enzyme2(ACE2) is an important peptide in renin-angiotensin system(RAS) and becoming a new target for hypertension and diabetes. At the same time, RAS have a close relationship with metabolic syndrome(MS). The present study was designed to investigate the renal function, morphology and the gene expression of ACE2 and angiotensin-converting enzyme(ACE) on kidney of rat model with metabolic syndrome, and to evaluate the effect of Radix Astragali, a kind nature herb, on them.Methods Sixty adult male Wistar rats were grouped randomly as follows: normal control group (n=10); fructose-induced MS group (n=10); MS+valsartan 20mg /(kg·d) group (valsartan group, n=10); MS+Radix Astragali 1.5g /(kg·d)group (low dose group, n=10);MS+Radix Astragali 3.0g/(kg·d)group (middle dose group,n=10) and MS+Radix Astragali 6.0g /(kg·d)group (high dose group,n=10). After 6 weeks of treatment, the metabolism indexes included SBP, visceral fat/weight ratio, TG,FFA and ISI were achieved to confirm the models establishing and to evaluate the effect of Radix Astragal on them, all rats were sacrificed and kidneys were harvested for following studies: Serum creatinine(Scr) and blood urea nitrogen(BUN) in all groups were evaluated with an automatic chemisty analyzer. HE staining was performed to observe the renal morphological changes. Real time PCR and immunohistochemistry were used to judge the changes of ACE and ACE2 in kidney separately. Moreover, AngiotensinⅡ(AngⅡ) of kidney was measured with radioimmunoassay.Results①There were significant deviation of SBP and others metabolic indexes between MS group and normal control group[SBP(144.5±3.3mmHg vs 109.3±5.0mmHg, P=0.000), visceral fat/weight ratio (0.044±0.001 vs 0.037±0.002, P=0.000), TG (1.74±0.10mmol/L vs 1.18±0.19mmol/L, P=0.000), FFA(457.6±49.1μmol/L vs 345.0±65.9μmol/L,P=0.000) increased obviously,ISI was decreased(-5.26±0.16 vs - 4.48±0.06, P=0.000)]. Compared with the normal control group, AngⅡof kidney tissue in MS group was increased statistically(20.53±1.87pg/mg vs 13.38±1.38pg/mg,P=0.000). The expression of ACE mRNA in MS group was up regulated of 76% than that in normal control group, while ACE2mRNA was down regulated of 58% (1.76±0.12 vs 1.00±0.13, P=0.000; 0.42±0.07 vs 1.00±0.07, P=0.000).②Both valsartan and Radix Astragali improved the metabolic indexes[SBP(122.1±4.6mmHg,136.0±3.90mmHg, 127.5±3.6mmHg, 116.3±4.4mmHg) vs (144.5±3.3mmHg), P=0.000, P=0.000,P=0.000,P=0.000. Visceral fat/weight ratio(0.041±0.001, 0.043±0.001, 0.040±0.001, 0.038±0.002) vs (0.044±0.001), P=0.000, P=0.441, P=0.000, P=0.000. TG(1.52±0.11mmol/L, 1.63±0.13mmol/L, 1.47±0.09mmol/L, 1.27±1.00mmol/L) vs (1.74±0.10mmol/L),P=0.004, P=0.762, P=0.000, P=0.000. FFA(406.6±41.9μmol/L, 429.1±29.0μmol/L, 400.1±30.6μmol/L, 365.1±27.5μmol/L) vs (457.6±49.1μmol/L), P=0.043, P=1.000, P=0.013, P=0.000. ISI(-4.97±0.13,-5.13±0.10,-4.81±0.11,-4.64±0.12) vs (-5.26±0.16),P=0.000, P=0.222, P=0.000, P=0.000]. The level of AngⅡof kidney tissue was found to be higher in valsartan group compared with that in MS group(27.47±2.50pg/mg vs 20.53±1.87pg/mg,P=0.000), while decreased in Radix Astragali group (Radix Astragali low dose group: 17.94±1.63pg/mg, Radix Astragal middle dose group:15.90±1.75pg/mg, Radix Astragal high dose group:13.96±1.36pg/mg) vs (MS group: 20.53±1.87pg/mg), P=0.031,P=0.000,P=0.000.③Both Radix Astragali and Valsartan improved the level of ACE2mRNA gene(Valsartan group: 0.70±0.08, Radix Astragal low dose group: 0.57±0.09, Radix Astragal middle dose group: 0.67±0.08, Radix Astragal high dose group: 0.84±0.10 vs MS group: 0.42±0.07, P=0.000,P=0.002,P=0.000 and P=0.000 respectively) and protein content of ACE2 (Valsartan group: 0.29±0.03, Radix Astragal low dose group: 0.22±0.03, Radix Astragal middle dose group: 0.27±0.02, Radix Astragal high dose group: 0.38±0.04 vs MS group:0.18±0.02, P=0.000, P=0.020,P=0.000 and P=0.000 respectively). These two medicines also could lower the level of expression of ACEmRNA (Valsartan group:1.36±0.1,Radix Astraga low dose group:1.53±0.13, Radix Astraga middle dose group: 1.33±0.1, Radix Astraga high dose group: 1.17±0.09 vs MS group: 1.76±0.12,P=0.000, P=0.001, P=0.000 and P=0.000 respectively).④There was good linear regression correlation among AngⅡand metabolic indices as a function of ACE2mRNA. But multiple regressions found that when these indicators existed at the same time, due to the interaction among them, only SBP and the ISI possessed linear regression relations with the ACE2.⑤Finally, there were no difference of Scr and BUN within these groups but renal morphological study showed the improvement in Valsartanand Radix Astragali groups compared with MS control group.Conclusions Intrarenal RAS disorder was confirmed in MS, which included morphological change, the balance between ACE and ACE2 and the level of AngⅡin renal tissue. Radix Astragali as a herb medicine has the similar capability with Valsatan torecuperate this balance, improve the metabolic indexes and renal morphological changes and there for to protect the kidney function of MS.

The Study of the Distortion Rate of Direct Digital Radiography in Measurement of Preformed Metal Posts

Wed, 30 Mar 2011 13:31:42 +0000

Preformed metal post and resin core are widely applied in the restoration of residual crowns and roots, which can simplify the process of restoration, reduce the incidence of dental root fracture and ensure adequate retention for the prosthesis in dental practice.The proportion between the length of the post in the root and the root length plays an important role in the retention of post and core and in the avoidance of dental root fracture. Direct digital radiography helps ensure a perfect restoration by making the post in the optimal position of the root.However, the images may be larger or smaller than the actual objects, which disturbs the precise placement of preformed metal posts, since direct digital radiography system can be influenced by many factors. In this study, the lengths of the preformed metal posts in the digital images were measured with software. Through comparing the measured length with the actual value, the distortion rate was calculated, which can provide valuable information to improve the clinical application of driect digital radiography.The result is that the distortion rate of direct digital radiography in placement of preformed metal posts is high, and counter measures should be taken to minimize the distortion rate to reach the need for clinical applications. MRI is one of the most valuable diagnostic tools for the examination of the head and neck regions in radiology and diagnostic sciences. However, in patients with metallic dental devices, artifacts due to misregistration at MR imaging can obscure normal anatomy or pathologic conditions and degrade the assessment quality of regions of interest near the devices.The metallic materials widely used in dental practice include Nickel- chromium alloy, Cobalt-chromium alloy, Titanium, gold alloy, etc. The popular fixed prothesises consist of crown, post and core, fixed bridge and so on. This study was attempted to evaluate the artifacts caused by metallic materials of the fixed prothesis in MR imaging. Crowns and post-and-cores made of different metals, fixed prothesises with various units made of two types of metallic materials most popularly used in dental practice were imaged by means of 1.5T MRI apparatus in specific conditions.This investigation could provide valuable information which conforms to the practice more than that of the previous studies concerning the artifacts caused by metallic dental devices in current use.The main results are as follows:1. With the use of T1-WI GRE pulse sequence, the artifacts caused by metallic dental devices were dependent on the types of metals of metallic devices. High noble metal alloys produce small artifacts, however, Cobalt-chromium alloys produce large artifacts.2. With the use of T1-WI GRE pulse sequence, the artifacts caused by metallic dental devices were dependent on the size, on the shape, on the position in the oral cavity and on the amounts of the metals of metallic devices.3. In the selection of the materials of fixed prothesis, in terms of the generation of the artifacts, preformed Titanium post resin core could be chosen to restore residual roots and crowns as one of the optimal materials, however, the effect of the fixed prothesises covering the post-and-core on MRI should be taken into account in dental practice.

Minimally Invasive Surgical Treatment of Isolated Posterior Cruciate Ligament Tibial Bony Avulsion Injuries: Retrospective Analysis of 23 Cases

Wed, 30 Mar 2011 13:31:40 +0000

Posterior cruciate ligament(PCL) arises from the lateral surface of the medial femoral condyle and extends posterolaterally to the posterior tibia 10mm below the joint line. It bears the weight of 85～100% retroposition strength and is considerded to be the key factor providing stability to the posterior directed aspect of the knee. The fibers of the PCL are usually reported as two separate bundles: the anterolateral(AL) and the posteromedial(PM) bundles. The distinct insertion sites of the two bundles on the tibia and femur are approximately equal in size. The ALbundle is two times larger in cross-sectional area than the PM bundle. During passive flexion and extension of the knee the AL bundle is more taught in flexion and lax in extension, conversely, the PM bundle is more taught in extension and lax in flexion. The distance between the femur and the tibia and the activity centre change unceasingly during flexion and extension of the knee. According to the different duty, the fibers twist correspondingly, making PCL adjust length and tension, so as to provide stability to the knee in all flexion angle. The intensity of PCL is 1.5～2 times higher than that of ACL, which hint that the PCL injuris mostly caused by high intensity, high opposability sports injuries and traffic accident. The most common trauma mechanism of PCL injury in sports is hyperflexion injuries which it more easily resulted in isolated PCL injuries. When posteriorly directed force applied to the proximal tibia, the tibia suddenly subluxates posteriorly and the posterior edge of the tibia platform stikes on the hanging wall of the intercondylar fossa, which can cut PCL off. Because of the substance of PCL tapering from the femur to the tibia, the PCL bony avulsion of tibia insertion is more common than that of femur insertion. The diagnosis of PCL injury depends on the detailed initial injury history collection and physical examination, combined with X ray, CT and MRI imageological examination. The knee′s position, weight loading state, strength laod, direct or indirect injury, patient′s momentum, and the limb′s position after injury are all important. The most accurate tests for detecting PCL injury include: posterior drawer tests, Lachman tests, quadriceps active tests , reverse pivot shift tests and Dial tests. Stress radiographs has diagnostic value on PCL tibial bony avulsion, especially when posterior subluxation of the tibial plateau and microscleres were found on the posterior edge of the tibia, higher than the joint line. If no fracture signs were found on the X ray, magnetic resonance imaging(MRI) should be executed, which has been shown to be extremely effective in confirming the diagnosis of PCL injury. If the diagnosis of PCL injury were missed, the treatment would be delayed.The treatment of PCL injuries may be the most disputed topic of knee surgery. The main reason is the deficient understanding of the natural history of PCL injuries. Traditionally most experts suggest isolated PCL injuries have good outcomes with a conservative treatment, such as bed rest, plaster or orthosis fixation. Recently more and more studies support early surgical treatment on PCL tibial bony avulsion and other associated ligament injuries. Because the proprioceptive sense lies in the femur insertion and the substance is complete, good healing capability and long-term effects can be gained after reduction. The most appropriate timing of surgical treatment is 1～2 weeks after injury when the intraarticular painful reaction has fade away and all the move range and partial strengh of the knee have recovered. The traditional surgical fixation of the avulsed PCL is usually performed through an open posterior approach with the patient in the prone position. It needs reveal the nerves and blood vessels behind the joint. The surgical trauma is serious, the operation is compicated, and the recovery postoperation is slow. Recently more and more surgeons tried to fix the cruciate ligaments bony avulsion through the arthroscope. The suture of ACL tibial bony avulsion has received good effects and was applied generally. But the PCL tibial insertion site lies in the tibial grooves and associates with the posterior articular capsule tightly, which makes the reduction extremely difficult after displacement. It often leads to insufficient repositon and PCL looseness. We treated 23 cases of PCL tibial bony avulsion via an open posteromedial approach through the space between the medial head of gastrocnemius and semitendinosus in reverse L shaped incisions, getting out of the way of all important nerves and blood vessels, revealing the PCL tibial insertion site. After accurate reduction, fix the bony avulsion firmly with screw, adsorbable screw or wire lock. Plaster cast fixed the knee in 30°-flexion position after operation. Quadriceps isometric contraction training should be taken the next day. The plaster fixation were removed after 6 weeks and performed joint active monement and CPM functional exercise. Began to gradually weigh-loading walk from 8th week. All the cases gained bone union after 2.5～6 months(average 4 months). The joint function greatly improved. No compications such as joint infection, non-union, fracture displacement and injury of nerves or blood vessels. We concluded that the posteromedial approach has the following advantage: convenient, reliable,and minimally invasive. The knee′s stability can be regained.The restricted exploration of the knee joint is the disadvantage through this approach. It should be accurately judged that the injury of PCL is isolated or complex before operation. So the overall and careful physical examination and MRI examination are quite necessary.

Preparation and in Vitro Assay of Microbubbles Targeting Solid Carcinoma by KDR with Biotin-avidin System

Wed, 30 Mar 2011 13:08:58 +0000

Purposes:The diagnosis of Tumor in early stage is the issue urgently need to be settled in imageology.As the molecular mechanism of tumor angiogenesis to be understood gradually,the progressive development of vascular functional molecular markers which based on the heterogeneity of tumor blood vessels,and applied to the diagnosis and treatment of tumor through molecular imaging.KDR as the most important vascular endothelial growth factor(VEGF) main receptor,highly expressed in endothelial cells in tumor angiogenesis,plays a key role in the early stages of tumor angiogenesis.We used KDR which expressed specifically on tumor vascular endothelial cells as the specific target,through the bridging of biotinavidin and signal multi-amplification,conjugded the small peptide K237 which has a high degree affinity of KDR to liposome microbubbles.To build a new kind of targeted microbubbles and identify the physical and chemical properties and biological activity,observe the specific imaging effect of microbubbles targeted to tumor angiogenesis.With a view to establishing a new type of ultrasound molecular imaging technology for early diagnosis of tumor.The biotin - avidin bridge system in construction of the targeted microbubble may overcome the shortcomings of the existing contrast agent targeting poor sensitivity and unstability,have significance to the diagnosis of other diseases. 1.Preparation of targeted microbubbles(MB_(K237))1.1 biotinylated liposome microbubbles(MB_B) Preparation1.1.1 DPPC,PEG,DSPE-PEG2000-Biotin and other lipid materials at a certain percentage of the amount dissolved in distilled water,at the same time give perfluoropropane(C3F8) gas,acoustic wave oscillation to form a milky white liquid, 4℃refrigerator hierarchical standing,abandoned follow fluid,injected with an equal volume of pure water to purify microbubble repeatedly to remove free liposomes.1.1.2 Microbubble solution at 1:10000 dilution ratio,Coulter particle size analyzer (MutisizerⅡ,USA) determinate the particle size,microsphere concentration,particle size distribution.1.2 Preparation of biotin - avidin bridge-mediated targeted liposomes microbubble with small peptide(MB_(K237))1.2.1 Corporation Hangzhou synthetic fluorescent-labeled peptide of 12 biotinylated peptides(FITC-Biotin-K237),the molecular weight of 2523.6 D,amino acid sequences for fitc-C6-His-Thr-Met-Tyr-Tyr-His-His-Tyr - Gln-His-His-Leu-Lys (Biotin) ~([10]).K237 is a peptide,Lei Hetian etal people isolated from phage display peptide library.Screening methods are:use KDR as the target,the use of affinity selection of phage peptide library technology,through ongoing reduce the concentration of KDR,improved the strength of washing fluid and competitive elution,to obtain strong binding of phage clones.From the two screening results, randomly selected 40 phage clones was amplified,using ELISA method to identify their binding activity,selected with the highest affinity phage clones sequenced, according to sequencing results of the chemical preparation of small peptide elements.1.2.2 Preparation of targeted microbubbles(MB_(K237))①Avidin-Biotin-Microbubbles(Av-MB_B) Preparation and detection: MB_B purified then shaked into milky white suspension,0.3ml of each suspension (about 10~8 microbubbles) as a sample,5 groups and 3 samples each group.respectively add 0μg,2μg,6μg,10μg,30μg fluorescent-labeled avidin (FITC-avidin) into each sample,after 30min incubation.Purified twice,flow cytometry detect the rate of avidin of microbubbles,evaluate the optimum dose.MB(0.3ml) and Av- MB_B(0.3ml) were deal with adding an equivalent FITC-Avidin for 30min incubation,full washing,the fluorescence microscope to observe two fluorescence effection of the microbubble.②.Targeted Microbubble Preparation and Detection: MB_B in the suspension,10~8/ml microbubbles adding 6μg no fluorescent avidin, incubated 30min,purifiy twice,shaken to the liquid,one sample is 0.3ml,7 groups and 3 samples in each group,add respectively 0μg,30μg,40μg,50μg,60μg, 70μg,l00μg of FITC-Biotin-K237 into each sample,flow cytometry to detect the rate of a small peptide bubble brought,to evaluate the optimal dose ratio.Kurt observed MB_(k237) of physical properties.Light microscopy and fluorescence microscopy in general the appearance of targeted microbubbles.2.Targted Microbubbles(MB_(k237)) in vitro assay2.1 Cell culture and Immunohistochemistry2.1.1 LOVO colorectal cancer cells,human umbilical vein endothelial cells(HUVE) and human colon cancer LS174T cells from logarithmic phase,0.25%trypsin digestion and inoculated in 6-well plates(3×105 / hole),plus 10%newborn calf 1640 serum-wide liquid-pui,5%CO2 incubator incubated until the cells in 80%of saturation.2.1.2 PBS washed 3 times,4%paraformaldehyde fixed,KDR conventional immunohistochemical staining.2.2 Targeted in vitro experiments:2.2.1 In vitro binding assay:①Three cells(KDR expression strong positive LOVO cells,positive HUVE cells, and KDR expression negative LS174T cells) were incubated with MB_(k237) for 30min, after washing observed under a light microscope and fluorescence microscope, analysing the effect of the combination of the three cell lines with targeted microbubbles.②Three types of microbubbles(targeted microbubbles MB_(k237),simply K237 modified liposome microbubblesK237-MB_B,control microbubble MB) incubated with KDR expression strong positive LOVO cells for 30min,after washing observed under light microscope and fluorescent microscope,analysing the effect of the combination of the three microbubbles targeted with LOVO cells.2.2.2 Blocking experiment: 10μg,50μg of K237 to deal with LOVO cells for 1h,incubated these treated LOVO cells with targeted microbubbles for 30min,then observed after washing,ananlysing the effect of MB_(K237)combined with teated LOVO cells.2.2.3 Mav = 6μg,Mk237 =(50μg,70μg,90ug,100μg) preparation targeted microbubble in four doses programs,LOVO cells were incubated with the four MB_(k237) for 30min,then observed after washing,annalysing the MB_(k237) in different doses the effect of targeted combination.2.2.4 Four speed(5ml/h,20ml/h,50ml/h,99ml/h) controlled by pump flushing microinjection,washing rosette structure formatted by LOVO cells and MB_(k237) at these four speed,then observed the changes in rosette structure under light microscope.3.Results Standard3.1 Immunohistochemicstry:Immunohistochemicstry was performed according to the protocol of the kit.The positive cell was determined by the appearances of light brown granules in cytoplasm or cell membrane.Semi-quantitative integral method used to determination[9],the number of positive cells≤5%,0 score;6%-25%,1 score;26%-50%,2 score;51%-75%,3 score;＞75%,4 score.The score of stain:non-specific staining for 0 score,a yellow staining intensity for 1 score,light brown for 2 score,brown for 3 score.To multipliy the two scores were denifinited as flow:0-1,negative(－),2 - 4,positive(+),more than 5 score,strongly positive(++).3.2 Rosette formation rate:Random field of vision from 10 the total number of cells,each cell-binding five or more microbubbles as a positive rosette formation,rosette formation rate calculation.4.Statistical analysisSPSS 13.0 package used for statistical analysis.The measurement data of the whole experiment was present with percentage.The Games Howell test was used to examine the normality of the value\' distribution.A value of P＜0.05 was considered statistically significant.5.Results5.1 Preparation of targeted microbubbles MB_(k237)5.1.1 Biotinylated liposome contrast agents(MB_B):MB_B appearance was white, about the concentration of microbubbles(2-6.3)×10~9 / mL,the average diameter (2.0 - 8.3)μm.Diameter of＜5μm microbubble was not less than 94.6%.200×optical microscope observation shows that microbubbles were well- distributed,no aggregation phenomenon,the appearance of a single spherical microbubble was well. MBb has a certain stability,4℃refrigerator can be stored for one month or more, and the form and concentration of microbubbles remain basically unchanged.5.1.2 Avidin - biotin - microbubble(Av-MB_B):①Under fluoroscope:two microbubbles adding equivalent of FITC-Avdin were washed fully,MB shell without fluorescence,FITC-Avdin-MB_B shell with a bright green fluorescent.②Screening adaptive dose:The bingding rate of FITC-avidin in 5 groups of microbubbles has correlation with avidin dosage,the binding rate gradually increase with the avidin dosage increased until they reach saturation.5 groups microbubble binding rate:χ~2=12.767,v=4,P=0.012＜0.05,there were statistically significant in 5 groups.Microbubble combination of avidin as a dose dependent manner,with the accession to avidin dose increase,combined with the rate gradually increased. Games-Howell multiple comparison results,0μg,2μg group were significantly different(P value of 0.000),6μg,10μg,30μg the\' combination of the rate between the each two groups were no statistically significant(P=0.781,0.882,0.793). Demomtrate that Av- MB_B systems,for about 10~7 MB_B,when the M_(Av)＜6μg when, with the avidin dose increase,combined with a gradual increase in rates,and rising faster binding rate curve;when the M_(Av)≥6μg,combined with a view to curve into the platform,increasing the dose of avidin,binding rate curve was no longer a significant upward trend.5.1.3 Targeted microbubbles(MB_(K237)) Preparation:①targeting the physical properties of microbubbles:MB_(k237) milky appearance was targeted,microbubbles concentration of about(3-8.4)×10~8/mL,the average diameter(2.6～8.0)μm.Diameter of＜5μm microbubble of not less than 90.3%.200 optical microscope observation shows that microbubbles diameter distribution,no aggregation phenomenon,the appearance of a single spherical microbubbles. Fluorescence microscope,targeted microbubble shell inspired bright green fluorescence.②The bingding rate of FITC-Biotin-K237 in 7 groups of microbubbles has correlation with FITC-Biotin-K237 dosage,the binding rate gradually increase with the FITC-Biotin-K237 dosage increased until they reach saturation.MB_(K237) binding rate of peptide results:M_(AV)=6μg,M_(k237)=(0μg,10μg,20μg,30μg,40μg,50μg,70μg, 100μg)/10~8 microbubbles under the ratio of the dose in each group microbubble Connect the small peptides have significantly different rates(χ~2=18.113,v=6,P =0.006＜0.05),it concluded that the 7 groups were significantly different;further multiple comparisons,among group 6,7,8 the multiple Comparison the differences were not statistically(P＜0.05),that when the M_(k237)≥50μg when connecting curve into the plateau.KDR adding high affinity of small peptide M_(K237)=50μg,can make 83.4600%±1.58218%percent of the microbuubbles\'surface be marked fluorescent signals. 5.2 Identification of the results of in vitro5.2.1 Cell CultureLOVO cell growth strong,good shape,showed a typical fusiform,both ends of thin, uniform and transparent cytoplasm,with little change in time.HUVECs can be seen after vaccination 2h adherent,24h after the completely adherent,the cells were short spindle or polygonal,with close,like pebbles or "cobblestone"-like for a typical endothelial cell morphology.24h completely LS174T cell-adherent,spindle or polygonal cells,showing uneven distribution of clusters is the typical characteristic of the cells5.2.2 Immunohistochemical stainingLOVO cells was KDR strongly positive expression,positive positioning in the membrane material;HUVECs were positive expression,the positive material located in the cell membrane and cytoplasm;LS174T cells were KDR negative expression.5.2.3 Targeted contrast agent in vitro to identify:5.2.3.1 In vitro binding assay:①Under a light microscope:around the LOVO cells we can see the targeted microbubble around the cells and formed the rosettes formation;under fluorescence microscopy the microbubbles issued bright green fluorescent,rosette formation rate up to 90.52%;around human umbilical vein endothelial cells(HUVE) the positive rosette formation rate of up to 53.46%;a small number micro-bubble adhesion around LS174T cells in the control group can be seen,rosette formation rate is 5.57%.②targeted microbubbles targeted LOVO cells with the combination of positive rosette test,rosette formation rate of 87.62%;small peptide - microbubble (K237-Mbs) were incubated with celis can be seen a little microbubble adhesion, rosette formation rate of 1.43%,relative microbubble targeting was significantly reduced;the surface of non-modified gaps biotinylated microbubbles were incubated with LOVO cells,rosette formation assay was negative.5.2.3.2 Blocking experiments,10μg closed group of target cells can be seen around a small number of targeted microbubble adhesion,50μg closed group of target cells around the basic non-targeted microbubble adhesion,rosette formation rate for the negative.5.2.3.3.Four doses of the program were targeted microbubble preparation,the effect of targeting among the for groups had no significant statistical difference(P＞0.05).5.2.3.4.Erosion rate of water flow in the next four LOVO cells and targeted microbubbles combined with the formation of rosettes formation structure gradually reduced.ConclusionTargeted microbubbles ultrasound contrast agent that carried peptide K237 be of high-affinity with KDR were constructed successfully in this study.The targeted microbubbles can made specific binding with KDR positive expression cells(LOVO cells,HUVEC_s) and had a certain stability,while no-specific binding with the KDR negative expression LS174T cells.There was hope for the realization of specific molecular imaging of tumor angiogenesis(such as:breast cancer) by targeting KDR, and laid the foundation for the next step experiments of targeted ultrasound molecular imaging in breast cancer transplanted in nude mice model

Clinical Efficacy of Huangqizhimushenqi Decoction for the 3rd Stage Diabetic Nephropathy with Qi and Yin Vacuity

Wed, 30 Mar 2011 13:08:57 +0000

Objectives:To observe the effect of Huangqizhimushenqi Decoction in treating microalbuminuria and clinical symptom of the 3rd stage diabetic nephropathy(DN) with qi and yin vacuity.So as to explore the therapeutic action and potential mechanism of Huangqizhimushenqi Decoction,which finally conduct as a satisfactory preparation to therapy 3rd stage diabetic nephropathy,to extend and utilize,to achieve determinate society and economic returns.Materials and Methods:62 patients from the Fuzhou General Hospital of Nanjing Mulitary Command with the 3rd stage of diabetic nephropathy with qi and yin vacuity(Male 38,Female 24;average age 56.1 years,average disease course 12.9 years) have been chosen into this clinical research.These 62 patients were divided into treatment group and contrast group at random,treatment group was 32 cases and contrast group was also 30 cases.Patients of treatment group took the Huangqizhimushenqi Decoction,adding modern medicine basical therapy;Patients of contrast group simply took the modern medicine basical therapy and Western medicine Lotensin only.The therapeutic course was 12 weeks for both two groups. Changes of clinical symptoms,microalbuminuria,blood glucose,blood viscosity,were observed and compared.Results:There was significal difference(P＜0.05) between treating group and contrast group in effectiveness of traditional Chinese medical syndrome.The clinical symptoms,as well as blood viscosity of the treating group were all significantly improved after treatment (P＜0.05 or P＜0.01),but in the contrast group improvement only showed in some of the projects above.The urine microalbuminuria/creatatine of both groups declined after cure without prominent discrepancy(P＞0.05).There was no prominent difference in blood glucose from beginning to end(P＜0.05).The both cure methods made no influence in renal function and blood pressure.No harmful response to all patients took place in the observation.Conclusion:The therapeutic effect of Huangqizhimushenqi Decoction for the 3rd stage of diabetic nephropathy with qi and yin vacuity was much better than that of modern medicine basical therapeutic in effectiveness of traditional Chinese medical syndrome.Qiyaodihuang Decoction could effectly lighten clinical symptom increasing patients\' activity quality;and treat microalbuminuria the same as Western medicine Lotensin;Qiyaodihuang Decoction also could effectly lighten blood viscosity.

The Research of the Impact of Comprehensive Nutrition Interventions to the Parts of Nutrition Indicators and KAP of the 18-25 Years Old Crowd

Wed, 30 Mar 2011 13:08:56 +0000

Objective:By the nutrition education to the 18-25 year-old volunteer groups that are lack of VA,and through the experimental observation of these groups\' different levels of VA and Zn complementarity,research the dietary intake of VA,Zn and nutrient metabolism levels before and after the comprehensive intervention,find the problems and provide useful suggestions,in order to make a further discussion of the appropriate intake parameter of VA,Zn joint complementarity of these groups and make a more farther support to the scientific nutrition education.Methods:After nutrition investigating the nutrition status and the knowledge-attitude-behavior (KAP) of the 18-25 year-old medicos in Lanzhou region,number the volunteers who are lack of VA and Zn according to their sex and the sequence of their participation, divide them into four groups with randomicity,this was conditioned in the way that the men and women were in the same number in each group.They were as follows: comparison group,single-Zn supplement group,single-VA supplement group,the joint VA and Zn supplement group.Through two ways,one was the observation(That were mostly depended on the nutrition education),and the other was integrated interventions (Combined nutrition education with joint VA and Zn supplement),make the VA,Zn levels of the volunteers stayed on a stable status at the end point of the observation (Considered synthetically the VA,Zn concentration in the peripheral blood,the level of serum RBP,dark adaptation time,AKP vitality,self-symptoms) for some time,for 5-7 days in each week during the investigation we surveyed the intakes of the dietary VA,Zn, and related nutrient of the volunteers with methods of 24h dietary recall,billing and weighting.Through the issuance of pamphlets and hold lectures and other forms of nutrition education to make the students to understand the importance of vitamins and minerals,and the harmfulness of lacks or excessive to their body and how to add nutrients through diet and reasonable in order to change bad eating habits(such as choosy food,eclipse,etc.),a reasonable diet.Before and after the intervention,we investigated the nutrition status of diet of the volunteers and analyze their nutrition knowledge-attitude-behavior.Results:1,After the integrated intervention,the volunteers formed some more reasonable diet;The energy intake of the meals in their breakfast,lunch,supper and sock in every day has increased than before;The volunteers\'intakes of the VA,Zn and vitamin C have increased evidently(P＜0.05).2,After the integrated intervention,the males\' intake parameter were increased excepted that of energy and fattiness,further more,the intake parameter of Ca,Zn,VA,VC were increased than before markedly(P＜0.05).The energy intake of the males in the breakfast, lunch,supper and sock were increased;The intake parameter of the females about the energy,carbohydrate,Fe,Zn,VA,VC were increased,thereinto the intake parameter of Zn,VA,VC was changed markedly(P＜0.05);And the others were decreased,thereinto the intake of VB2 was changed significantly than before(P＜0.05).3,After the intervention the concentration of Ca,Cu,Fe,Mg in the hole blood of the volunteers that were been investigated were increased markedly(P＜0.05),while the concentration of Zn was decreased;The lack rate of Ca,Fe,Zn and Mg was each decreased by 22.07%,10.8%,1.88%,1.88%,but the lack rate of Cu was increased by 7.04%.4,After the intervention,the RBC、HGB、HCT、MCV and RDW-SD were increased significantly(P＜0.05),while the MCH and MCHC were significantly decreased (P＜0.05).After the intervention,the number of the person whose RBC,HGB,HCT, MCV,RDW-SD were in the normal range were significantly increased,thereinto the number of them whose HCT,MCV,MCHC and RDW-SD were in the normal range were increased as 13.41%、20.67%、17.32%and 40.79%;Before and after the intervention,the prevalence rate of anemia was 3.91%and 2.81%(P＞0.05).5,Before and after the intervention,the IgA levels of the males and females were not changed significantly(P＞0.05).After the intervention,the IgG level of the males were decreased significantly(P＜0.05),and the IgM levels of the males and females were increased slightly,but there was no statistically significant difference(P＞0.05).After the intervention the complement C3 level of the males and females were significantly lower (P＜0.05);The ALP level of the males and females were decreased,thereinto the changes of the females were significantly different(P＜0.05),and the males had no significant changes(P＞0.05).6,Before the nutrition education,the crowd\'s knowledge score was 42.27±8.29,attitude score was 14.29±1.93,and behavioral score was 16.39±2.17;After the nutrition education,the three kinds of scores rose to 46.52±6.78,15.20±0.98 and 18.39±2.17. After receiving the nutrition education the crowd tends to get nutritional knowledges through more and wider range of ways;the crowd\'s correct eating behaviors increased as 15.75 percent;their demand for nutrition knowledge became more diversified,thereinto their demand for knowledge on the prevention of malnutrition increased as 20.75 percent. According to the education this crowd had an obverse change on the dietary knowledge, faith and behavior.Conclusion:After a comprehensive nutrition intervention,we found some nutrition problems here about the medical college students aged 18-25 in the region of Lanzhou, that they were mainly take animal meat food,and were inadequate about the intake of fruits and vegetables,and having single type of food diet habits,these problems have been ameliorated after the intervention,the needs of the food composition were sufficient, and the food varieties were all ready.The intervention scheme of nutrition education joined complementarity intervention to this crowd,has had a positive action to the volunteers\' series of parameters of the peripheral blood cells and the normalization of the distribution of the microelement levels in whole blood.The flexible forms of nutrition education to the crowd have had a good effect.As well as raising the level of the nutrition knowledge,we aught to make greater efforts to promote the formation of healthy eating behavior.

Development of Dengue Virus Serotype-2 NS1 Antigen Capture ELISA and Its Application in Serotyping of Dengue Virus Infectious

Wed, 30 Mar 2011 13:08:54 +0000

The mosquito-borne dengue disease is caused by infections of the dengue virus (DENV). It is a major public health problem that affects over 100 tropical and subtropical countries. Moreover, the geographical territory affected by this disease has increased markedly since the 1970s; contributing factors may include a lack of control over the mosquito vector, global warming, viral evolution, and increased ease of global travel. The dengue virus has four known serotypes (DENV1, DENV2. DENV3, and DENV4). An infection with any of these serotypes can cause dengue fever, dengue hemorrhagic fever (DHF), or dengue shock syndrome (DSS). Subsequent heterologous infections may increase the risk of the developing DHF or DSS. Since protection against heterologous serotype infections is only partial and transient, people that live in epidemic areas are at risk for contracting an average of four DENV infections in their lives. Thus, the cocirculation of various DENV serotypes in a community is the most common risk factor associated with the emergence of the severe forms of disease (DHF or DSS). Given the current absence of a vaccine that is effective against all four DENV serotypes, early diagnosis is the key to reducing the morbidity and mortality of DHF or DSS. Furthermore, symptoms of DENV infections are insufficiently specific for accurate clinical differentiation from other acute febrile illnesses, especially in areas where multiple tropical diseases such as malaria, yellow fever, West Nile disease, and St Louis encephalitis are endemic. Case identification has become increasingly important in the differential diagnosis of the acute febrile illnesses and hemorrhagic fever of unknown flavivirus infections during the early symptomatic phase. Currently, molecular techniques (reverse transcription-PCR[RT-PCR]) have widely replaced traditional virus isolation methods in the early detection of DENV infections and differentiation of the four DENV serotypes and other related flavivirus infections. Although molecular detection provides a promising sensitivity rate and rapid diagnosis, the molecular approach is costly, as it requires specialized laboratory equipment and experienced technicians; these are notable limitations in many developing countries where dengue is endemic.An \'ideal\' simple, cost-effective, rapid, and accurate diagnostic tool is necessary to overcome the main disadvantage of current molecular techniques. Viral antigen detection methods that are convenient and cost-effective have been successfully used in various infectious diseases for the diagnosis and monitoring of disease activity. DENV contains three structural (C, prM, and E) and seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5). Among these proteins, NS1 is a relatively conserved 45- to 50-kDa glycoprotein that is highly expressed in infected cells in both membrane-associated and -secreted forms. Although the precise function of the NS1 is unknown, it represents an interesting target antigen for diagnosis . Recently, two DENV NS1 antigen capture immunoassays have become commercially available for the early diagnosis of dengue virus. The two NS1 antigen capture assays are suitable sensitive and provide a rapid and reliable diagnosis of acute primary dengue disease and acute secondary dengue disease. However, one limitation of these assays is that they are unable to distinguish between the DENV serotypes, as the antibodies used in the assay is directed against cross-reactive antigenic determinants shared by all four DENV serotypes and other flaviviruses. NS1 has been identified as being a both group-specific and type-specific determinant; thus, the production of epitope-specific monoclonal antibodies (MAbs) holds potential for developing either group-specific or type-specific NS1 antigen assays. In a previously study, we prepared a panel of MAbs that recognized epitopes specific to DENV1 NS1 and successfully developed a DENV1 NS1 antigen capture ELISA. This DENV1-specific NS1 antigen assay has proved to be a valuable tool for the early detection and rapid identification of DENV1 infections. In the present study, we prepared a panel of MAbs that recognized epitopes specific to DENV2 NS1 and described the development of a DENV2-specific NS1 antigen capture ELISA.Three parts are included in this research.Part one: Preparation and characterization of monoclonal antibodies against DENV2 NS1 proteinBALB/c mice were immunized by recombinant DENV2 NS1(rDENV2-NS1) and inactivated DENV2. Then the splenocytes of the immunized mice were fused with myeloma cells to produce hybridoma cell line, which could secrete anti-DENV2 NS1 protein antibodies. A total of 37 hybridoma cell lines that stably produced MAbs were initially established on the basis of their strong positive reactivity with both rDENV2-NS1 protein and DENV2-infected cell lysates in the ELISA. The immunoglobulin isotype determinations revealed that the 37 MAbs comprised IgG2a (n=1). IgG2b (n=3). and IgG1 (n=33). Western blot analysis with rDENV2-NS1 protein and DENV2-infected cell culture supernatants revealed that all MAbs reacted to the same immunoreactive protein band with an approximate molecular weight of 45 kDa. The serotype specificity of the MAbs was further identified by testing their reactivity to the four dengue serotypes in the IFA and the rDENV2-NS1-based ELISA. Of the 37 MAbs. 20 MAbs specifically reacted to DENV2 with no cross reactivity to the other three dengue serotypes in either the ELISA or the IFA assays. The remaining 17 MAbs had cross reactivity to the four dengue serotypes in different cross-reactivity patterns. Next, the distinct binding epitopes of the 20 DENV2 NS1-specific MAbs were determined by competition ELISA using rDENV2-NS1 as the immobilized antigen. The results showed that the 20 MAbs bound to at least four different epitopes on the DENV2 NS1 protein. These obtained antibodies will provide a potential value for early diagnosis of DENV infection.Part two: Establishment of the antigen capture ELISA for detection DENV2 NS1 protein. Among the 20 MAbs, each was paired according to their abilities to recognize distinct epitopes on NS1.Then, a sandwich ELISA was performed to select the optimal capture-detector MAb pair. Pairing between eight MAbs in epitope group III and nine MAbs in epitope group IV displayed high sensitivity in DENV2 NS1 protein detection. Ultimately, the pairing of MAb M6 as a solid-phase immobilized capture antibody and MAb M14 as a labeled detecting antibody gave the highest combination of sensitivity and specificity in the detection of DENV2-specific NS1 in serial dilutions of DENV2-infected cell culture supernatants and a panel of sera from healthy humans. The minimal amount of rDENV2-NS1 detection with this assay was approximately 3ng/ml. The linear portion of the standard curve ranged from 10 to 100 ng/ml and could be used to estimate the NS1 levels in clinical samples. Furthermore, the DENV2 NS1 ELISA is specific to DENV2 and has no cross-reactivity with either other serotypes of DV or other closely related members of the flavivirus family (Japanese encephalitis and Yellow fever) when the virus-infected cell culture was used. All these results demonstrated that the established test is sensitive and specific which may lay the basis for providing a novel laboratory diagnosis tool for dengue virus infection.Part three: The preliminary application study of the DENV2 NS1 antigen capture ELISA for detection DENV2 NS1 protein in sera specimens.We compared the sensitivities of the DENV2 NS1 ELISA with the Panbio NS1 ELISA in the detection of NS1 in the DENV2- infected culture supernatants with serial dilutions. Based on the cutoff values of the two assays, the highest dilution of detectable NS1 was 1:4096 (5.8 PFU/0.1mL) in the DENV2 NS1 ELISA, while the highest dilution of detectable NS1 was 1:512 (46.8 FU/0.1mL) in the Panbio NS1 ELISA. Thus, the DENV2 NS1 ELISA demonstrated an approximately 8-fold higher sensitivity than the Panbio NS1 ELISA. Serum specimens from 504 healthy humans were analyzed to establish the background signals of the DENV2 NS1 ELISA before further clinical evaluation. We found that no false positive results were obtained with any of the samples from the 504 sera, indicating the specificity of the DENV2 NS1 ELISA was 100% (0 of 504). To evaluate the accuracy of the DENV2 NS1 ELISA for the detection of NS1 from clinical serum samples, we assessed a total of 30 acute-phase serum specimens from DENV2-infected patients with laboratory-confirmed. Among these sera, 10 were confirmed by both DENV2 isolation and RT-PCR results and 20 were confirmed by both RT-PCR results and Panbio Dengue IgM or IgG capture ELISAs. According to World Health Organization criteria, 17 sera were serologically classified as acute primary infections and 13 sera classified as secondary infections. Based on a sample dilution of 1:10. the DENV2 NS1 ELISA detected 25 of the 30 acute-phase serum specimens. The overall sensitivity of DENV2 NS1 antigen detection with respect to virus isolation and/or RT-PCR was 83.3% (25 of 30). These results were consistent with the results from the Panbio NS1 ELISA when the assays were performed at a 1:10 dilution in parallel: the sensitivities for both assays were 83.3% (25 of 30). of which 5 did not have a detectable NS1 in both the assays. Both NS1 ELISA detected 88.2 % (15/17) of the acute primary and 76.9% (10/13) of the acute secondary infections. There was no statistically significant difference in NS1 detection rate between acute primary infections and acute secondary infections (p = 0.628). To further evaluate the sensitivities of both NS1 detection assays, serial dilutions of positive sera were analyzed. From a total of 25 sera detected NS1 antigen positive in both the assays, only 19 samples had sufficient residual volume to allow us to further compare the sensitivity of both NS1 detection assays. Since the DENV2 NS1 assay displayed a marked sensitivity in the DENV2-infected culture supematants as described above, it was not unexpected that DENV2 NS1 remained detectable even when patient\'s serum was at a 1:10,000 dilution.DENV2 NS1 ELISA was also evaluated on a panel of clinical serum samples from patients infected with DENV1 or other flaviviruses and non-flaviviruses. We analyzed a total of 301 acute-phase serum samples that had previously tested positive for NS1 by the DENV1 NS1 antigen capture ELISA. Additionally, we tested a total of 132 acute-phase serum samples, comprising 50 samples from 50 cases of hemorrhagic fever with Hantan virus infections. 13 samples from 13 cases of Japanese encephalitis. 49 samples from 49 cases of measles, and 20 samples from 18 cases of leptospirosis. We found that none of these samples were positive for NS1 by the DENV2 NS1 ELISA. These findings suggested that the serotype-specific MAb-based NS1 antigen-capture ELISA might be a valuable tool for early diagnosis and serotyping of DV infections.In conclusion,1. A panel of MAbs specific to the DENV2 NS1 was successfully obtained. This may provide a potential source for studying the function of NS1 and early diagnosis of dengue virus infections.2. A MAb-based antigen capture ELISA for detection circulating NS1 protein was successfully established. This assay was displayed higher sensitivity than commercially available dengue NS1 antigen-capture ELISA. The high detection sensitivity makes it suitable for the early detection of DENV infections.3. A DENV2-specific MAb-based antigen-capture ELISA has been developed. It has high detection specificity to DENV2 without cross-reactions with either other three DENV serotypes or other closely related members of the flavivirus. The characteristic will be helpful in rapid serotyping and differential diagnosis of dengue virus infections.

Study on the Qualification and Evaluation System of Dual-Qualification Nursing Teachers

Wed, 30 Mar 2011 13:08:53 +0000

BACKGROUNDThe construction and evaluation of nursing teaching team has been the hot spot at home and abroad, the number of nursing department has increased from 18 to more than 100 with higher nursing education scale continuing to enlarge, this offered good chance for nursing development, but this also resulted in lack of full-time nursing teachers. The performance of dual-qualification can not only increase the chance of clinical practice of full-time nursing teacher, but also increase the opportunity of teaching, and then solve the problem of the lack of nursing teachers better. According to the standard of dual-teacher provided by national ministry of education, dual-qualification nursing teachers means the teachers who not only have rich teaching practice but also have rich clinical practice can engage in teaching in class and clinical nursing work and instruct students in clinical practice. The dual role can integrate teaching in class and clinical practice, embody the integrity of curriculum fully and effectively cultivate senior practical talents. The file from ministry of education pointed out that the system of evaluation and appointment of dual-qualification nursing teachers should be established singly and supported by policy. At present many colleges established the standard about qualification and evaluation individually according to their actual condition. Own to the different standard, the results among colleges made a great difference. Study on the qualification and evaluation system of dual-qualification nursing teachers can make the guide for choice and evaluation of dual-qualification nursing teachers, and make itdevelop with healthy and normally.OBJECTIVE1. Constructing selection standard can provide dual-qualification nursing teachers withevidents, and solve the lack of nursing teachers in school.2.Constructing evalution method can check dual-qualification nursing teachersregularly, and then improve teaching quality.3.The performance of dual-qualification make hospital and school close combination,and improve the quality of the whole nursing team.METHODSUsing the Delphi method, a total of 23 experts in different nursing field were consulted. Then using the analytic hierarchy process, relevant weights were developed. These experts come from four medical universities, five hospitals in Guangzhou、Chongqing、Xian、Shanghai. All experts have high technical posts. Professor is 10 persons (43.48%), associate professor is 13 persons(56.52%); undergraduate is 11 persons(47.83%), higher than graduate is 12(52.17%); working years of these experts range from 14 to 40; the mean is 27.17±8.138(SD); experts from education field is 8 persons, clinical nursing experts is 9 persons and nursing management experts is 6 persons; average age is 46.09±7.543.The questionnaire of expert consultation, which was based on consultation and retrospective literature, was designed according to the result of the pilot study. The questionnaire involves 6 main factors and 64 items: selection criteria、training method、evaluation、certification method、time and department. After three rounds of expert investigation, I analyze the feedback opinions from experts, and using analytic hierarchy process, relevant weights were developed, and then construct the system of dual-qualification nursing teachers. Based on this, three questionnaires can be evaluated dual-qualification nursing teachers from nurses、students and their selves. The next is to test the reliability and validity.In the study, describing statistics was used to analyze demographic constitution of experts. Mean and coefficient of variance was used to describe the experts\' opinion of the importance of the item; and coefficient of Kendall\'s W, percentage of questionnaire received and the authority degree of experts were used to examine the reliability and representation of expert consultation. In the three questionnaires, the internal consistency of questionnaire was examined by calculating Cronbach alpha; construct validity was evaluated by factor analysis and correlation analysis. The data was analyzed by using statistical package SPSS13.0 and Excel 2003.RESULTS1. Reliability and Consistency of the Expert ConsultationExpert is the focus for Delphi method. The results are measured by representativeness、enthusiam、authority、coordination of experts. The experts come from four medical university and five hospitals in Guangzhou Xian Chongqing and Shanghai. The percentage of questionnaire received from the three rounds of expert consultation was more than 80%, beyond to the standard of 50%. The Kendall\'s W is over 0.4, 0.4-0.5 is received normally. In addition, authority degree of experts is beyond 0.8. Therefore, based on practice and theory, the expert consultation had high reliability.2. Filtrate Index and Set Weight of ItemUsing Likert 5-scale method to describe the experts\' opinion of the importance of the item, the mean beyond 3.5 and the coefficient of variance less than 0.25 were used to filtrate index. Conclusion could be drawn according to the statistics above. Mean of item had been used to calculate the Satty sign. According to the analytic hierarchy process, next was to establish the multi-ranks judge matrix. And then the weight of item was identified. The value of RI showed that evaluation index system of certificating dual-qualification nursing teacher had logical consistency. By using principle of probability multiplication, weight of each item could be set up. Based on the evaluation index system of certificating, dual-qualification nursing teacher and weight of item set up. We established the assessment model.3. Examination of Evaluation Index System of CertificationBased on the evaluation system, three questionnaires are formed, nurses、students and their selves respectively. The Cronbach a is 0.903、0.922、0.934, the expert consultation had high reliability. It was found that in correlation analysis value of item vs. its superior grade item had higher correlation coefficient than value of items vs. other grade item. In nurses evaluation questionnaires, the factor analysis result showed that four factors were picked up which proved the role and function. The cumulative extraction sum of squared loadings were 69.051%; in students evaluation questionnaires, the factor analysis result showed that two factors were picked up which proved the role and function. The cumulative extraction sum of squared loadings were 66.241%, these data suggest that configuration of questionnaire was accordant to the result of factor analysis in the main. The questionnaire had a better reliability and construct validity.CONCLUSIONSAfter three rounds of expert investigation and statistics analysis, the dual-qualification nursing teacher evaluation index system was established and validated. Finally, we discussed the significance of establishing of the evaluation index system.1. Construct the qualification and evaluation system of dual-qualificationSelection condition includes 8 aspects: professional certificate of nursing、educational background、professional title、working time、teaching time in clinical、teaching essays、computer and English. Training methods involved lecture、classroom observations、trial teaching and examination. Evaluation included 5 aspects 35 items, the second grade is professional attitude、clinical nursing level、nursing teaching level、nursing science level and interpersonal relationship. The certification department is nursing school or personnel department in medical university. The certification method is individual application、recommendation by hospital and evaluation by experts. The certification time is 3 years.2. Foundation and Examination of the Assessment Model of CertificationBased on the Delphi method and analytic hierarchy process, weight of item ofevaluation system was calculated. Using such two scientific method, logicality andaccuracy of the study would be enhanced. By using assessment model, we could certificate the competency of dual-qualification and evaluate their role and function. According to three questionnaires, we concluded that evaluation index system had both rationality and feasibility, and the assessment model had a better reliability and validity, it provided a set of standard evaluation pattern.3. Significance of the establishment of the Evaluation Index SystemThe aim is to cultivate clinical nursing expert who can adapt to the changing medical underground and engaged in nursing practice based on nursing theory and nursing science. The duty of the teacher is to lead and guide students, cultivate students\' capable and teach them learn how to study、be a person、cooperation and survive, so enhance the selection and evaluation is very important. Though dual-qualification team construction is performed many years in a lot of university, there are many problems such as concept uncertain、selection standard not unified and so on. The significance of qualification and evaluation system of dual-qualification nursing teachers: (1)selection condition can make the dual-qualification teacher selected by evidence. (2) Dual-qualification teachers from clinic solve the lack of nursing teachers in school and increase the communication between clinical nursing teachers and full-time nursing teachers, and increase teaching quality. (3) The performance of dual-qualification make hospital and school close combination, and improve the quality of the whole nursing team. (4) It\'s role is to guide and balance for the development of dual-qualification, and ensure the nursing construction develop healthy. So The performance of dual-qualification is a win-win method for clinical nursing and nursing teaching.

The Study of Correlation between Expression of VEGF-C/D in TAMs and Lymphatic Metastasis in Breast Cancer

Wed, 30 Mar 2011 13:08:51 +0000

Objective To analyze the relationship between the expression of vascular endothelial growth-C/D(VEGF-C/D) in tumer-assoiciated macrophages(TAMs) and lymphatic metastasis in breast cancer.To provide new channel for diagnosis,treat and prognosis of breast cancer.Methods CD_(68)、VEGF-C、VEGF-D and D_(2-40) were detected by immunohistochemistry technique SP in 53 cases of breast cancer、30 cases of benign lesion and 20 cases of normal breast tissue.TAMs and microlymphatic vessel density (MLVD) were marked by CD_(68) and D_(2-40) antibody respectively;The double staining was also used to determine VEGF-C and VEGF-D expression in TAMs.Results 1.D_(2-40) expression in breast cancer(1) The MLVD level in breast cancer(17.79±4.02) was much higher than in benign lesion(6.43±2.42) and normal tissue(2.00±1.03)(P＜0.01);The MLVD level in peritumoral region(17.75±3.99) was much higher than in intratumoral region(8.68±3.38)(P＜0.01);The MLVD level in lymph node metastasis group(20.71±1.81) was higher than in no lymph node metastasis group(13.68±2.23)(P＜0.01).(2) The MLVD in breast cancer had no correlation compared with age、clinical stage and the expression of ER、PR、Cerb-2(P＞0.05,respectively);but there was statistical difference between histological gradeⅠ、ⅡandⅢ(P<0.05),the expression of MLVD in gradeⅢ(20.76±2.22) was much higher thanⅠ(14.76±2.75) andⅡ(17.84±4.30).2.CD_(68) expression in TAMs of breast cancer(1) The positive expression rate of CD_(68) in TAMs were 58.50%(31/53) in breast cancer、13.33%(4/30) in benign lesion and 15.00%(3/20) in normal tissue,there was statistical difference between them(x~2=21.89,P＜0.01);The positive expression rate of CD_(68) in TAMs were 74.20%(23/31) in lymph metastasis,36.36%(8/22) in no lymph metastasis,there was significant difference between them(x~2=7.59,P＜0.01).(2) The CD_(68) positive expression rate in TAMs of breast cancer in histological grade(Ⅰ、ⅡandⅢ) were 35.29%(6/17)、63.16(12/19) and 76.47(13/17) respectively, there were statistical difference between them(P＜0.05);It had no correlation compared with age、clinical stage and the expression of ER、PR、Cerb-2(P＞0.05,respectively).(3) The CD_(68) positive MLVD level(18.81±3.80) was higher than in CD_(68) negative (16.36±3.96)(P＜0.05);The CD_(68) expression in TAMs was related to the level of MLVD (r_s=0.418,P=0.002),That is,with the expression of CD_(68) in TAMs enhanced,the level of MLVD also enhanced.3.The expression of VEGF-C and VEGF-D in breast cancer(1) The positive expression rate of VEGF-C and VEGF-D respectively were 52.83% (28/53) and 49.06%(26/53) in breast cancer、10.00%(3/30) and 13.33%(4/30) in benign lesion、both were 10.0%(2/20) in normal tissue,there were statistical difference between them(P＜0.01,respectively);The positive expression rate of VEGF-C and VEGF-D both were 70.97%(22/31) in lymph node metastasis group,27.27%(6/22) and 18.18%(4/22) respectively in no lymph node metastasis group,there were significant difference between them(P＜0.01,respectively).(2) The expression of VEGF-C and VEGF-D in breast cancer had no correlation compared with age,clinical stage and the expression of ER,PR(P＞0.05,respectively);but there were statistical difference between histological grade(P＜0.05,respectively);In addition,The expression of VEGF-C were significantly related to the expression of Cerb-2(3) The VEGF-C and VEGF-D positive MLVD level were(19.25±3.20) and(19.58±3.09) respectively,VEGF-C and VEGF-D negative MLVD level were(16.16±4.26) and (16.07±4.10) respectively,there were statistical difference between them(P＜0.01, respectively);The VEGF-C and VEGF-D expression were related to the level of MLVD (r_s=0.413,P=0.002 and r_s=0.426,P=0.001),That is,with the expression of VEGF-C and VEGF-D in breast cancer enhanced,the level of MLVD also enhanced.4.VEGF-C and VEGF-D expression in TAMs of breast cancer(1) The double immunohistochemical staining showed that VEGF-C and VEGF-D positive expression in TAMs respectively were(17.15±4.17) and(14.05±3.32) of breast cancer,which occupied total number of TAMs(97.95±12.40) about 11.5%-21.4%and 9.0%-17.7%,the mean of them were 17.3%and 14.2%.(2) VEGF-C and VEGF-D expression in TAMs of breat cancer were both related to the number of TAMs(r_s =0.490 and r_s=0.464)(P＜0.05,respectively). Conclusions 1.TAMs may play a major role in peritumoral lymphatic neoangiogenesis and lymphatic metastasis,which may be used as a new target in diagnosis and treating of breast cancer.2.The new-born lymphatic vessels were found in breast cancer.The microlymphatic vessels density(MLVD) level in the peritumoral region is much higher than intratumoral region,it may be an important pathway of nodal metastasis in breast cancer.The MLVD is closely related to lymphatic metastasis,it could be used as a tumoral metastasis index in breast cancer.3.VEGF-C and VEGF-D participate in lymphangiogenesis and promote the lymphatic metastasis of breast cancer.4.The positive expression rate of CD_(68) in TAMs have high positive expression rate of VEGF-C and VEGF-D,it indicates that turner cells and TAMs both secrete VEGF-C、VEGF-D in breat cancer,TAMs、VEGF-C and VEGF-D are speculated to have synergistic effect in growth、invasion and metastasis process of breast cancer.

The Effect of Guiling Xiaoyi Decoction Drug Serum on the Morphology and the Secreting of IL-6 IL-8 of Eutopic Stromal-cell in Endometriosis

Wed, 30 Mar 2011 13:08:49 +0000

Endometriosis is a common intricate gynecological disease defined as endometriotic tissues growing outside the normal uterine cavity. This benign disease has confused clinicians for a long time, which behaves like malignant disease. This paper puts emphasis on the study of the theoretical foundation about the treatment by Guiling xiaoyi Decoction. We initially establish the in-vitro cultural method of purified stromal-cell, which provides an experimental model for the survey of the effect of Guiling xiaoyi Decoction on endometriosis and its mechanism of action.1、The study of the theoretical basis of the treatment of Guiling xiaoyi decoction for dometriosisThere is no term corresponding to endometriosis in traditional Chinese medical . The cardinal symptoms of endometriosis correlate with those of algomenorrhea, abdominal mass and infertility in traditional Chinese medicine. The disease is due to malfunction of the dominating reproduction of the kidney, overacting in sexual intercourse during menstruation and postpartum, and then leading to injury the Chong and Ren meridians and failure of the functions of storing and discharging of the uterus. The kidney\'s function being not sufficient is the radical factor of occurrence of the disease. Blood stasis is the pivot of occurrence and development of the disease. So the treatment for endometriosis should be based upon the concept of reinforcing warming kidney and dispersing blood stasis. Guizhi fuling pill originates from jinkui yaolue. Zhongjing Zhang ,the author of this book,designed this prescription to mildly remove mass in the abdomen,disperse blood stasis in order to prevent the abortion which was due to the abdominal mass endangering the fetus.If the prescription was utilized mainly to remove blood stasis and disperse the mass in the abdomen,there might be some shortage of the strength of the drugs in the prescription. On one hand, more drugs having the property of promoting blood circulation and resolving blood stasis should be added to the prescription. On the other hand, the prescription should be modified in accordance with syndrome differentiation. So Guiling xiaoyi Decoction was designed which combines the Guizhi fuling pill with the herbs Xianlingpi to strengthen warming kidney, and Xuejie to help dispersing blood stasis. The above mentioned notion forms the theoretical foundation of our vitro experimental study of the treatment of Chinese materia medica of warming kidney and dispersing blood stasis for endometriosis.2、In-vitro culture method of stromal-cell established initiallyWe collected endometria from patients who suffered from endometriosis and recived operation from April in 2007 to October in 2008. Stromal-cells were isolated from it and went down to the future subculture,the proliferative condition were observed by microscope,and purity coefficient of cell was characterized with immunohisto- chemistry method.As a result, the proliferative conditions of stromal-cells cultured in-vitro mainly with the method of grit separation were good. During observation, cells were all simple interstitial ones, went down to 4-5 generations and could survive 25-30 days. It can be show from the result of immunohistochemistry that the purity coefficient achieve percent 95 above.It can be seen from the result that under our experiment condition the stromal- cell cultured in-vitro has satisfactory ability of vegetation. The stromal-cells isolated from both endometriosis and non-endometriosis can be successfully cultered ( P>0.05). The menstrual cycle has an influence on the stromal-cell’s survival rate and the ability of growth (P<0.05).An effective in-vitro cultured cell model can be provided to reveal pathogenesy of endometriosis and study the pharmacodynamics.3、The effect of Guiling xiaoyi decoction on appearance of stromal-cells and IL-6、IL-8 from cell culture supernatant fluidWe made serums of different dosage drugs of Guiling xiaoyi Decoction, interfered the stromal-cells cultured in-vitro which were extracted from the parents suffering endometriosis with the serums, then observe their morphologic change by the inverted microscope, and collected supernatant liquid of cell culture at the time of 24h、48h、72h,respectively detected the concentration of IL-6、IL-8 by the method of ELISA.From morphology observation, we can initially confirm that the serums of Guiling xiaoyi Decoction can efficaciously suppress the growth of stromal-cell cultured in-vitro,and this suppress depends on the concentration of the drug serums. Besides, Guiling xiaoyi Decoction can cut down externalization of IL-6 and IL-8 from the cells cultured in-vitro obviously. The observations in this paper strongly suggest that adjusting of immunization, restraining inflammatory topo-reaction of endometrial, abating adherence at pelvic cavity, reducing the invasion and proliferation capacity of ectopic cells and inhibiting the growth of neovascularization might be the effects of Guiling xiaoyi Decoction to control mobility and development of endometriosis.

Development and Application of Cricothyriod Membrane Puncturing Device

Wed, 30 Mar 2011 13:08:47 +0000

Upper respiratory tract obstruction can be easily caused by some diseases in peacetime and war time, such as multiple complex maxillofacial trauma, inhalation of respiratory tract burns, acute laryngeal obstruction, etc.. In general, the effective therapeutic methods, include artificial ventilation, tracheal intubation, tracheotomy and establishment of intravenous access, can not be carried by specialist in time, an extreme decrease in the amount of oxygen in the body will lead the patients to loss of consciousness or death. Therefore, in the future battlefield, if the life guard and other soldiers can treat the wounded in correct and fast methods before professional medical care, the effectiveness of saving the life can be significantly improved.In this paper, a new cricothyroid membrane puncturing device was developed. It could aid the catheterization and ventilation by puncturing cricothyroid membrane and feeding high-frequency oxygen, which can effectively improve the cure rate of the patients with oxygen barrier of the upper respiratory tract, such as complex maxillofacial trauma, inhalation of respiratory tract burns, acute laryngeal obstruction, laryngeal neoplasms, tetanus cramps, throat surgery, tracheal infusion of drugs, etc. Based on the technologies of four clinic and first-aid, the medical and nursing staffs of primary military hospitals could quickly master the puncturing technology by watching video and training on the human body. The operator can the technology for emergency ventilation.Cricothyroid membrane puncturing device is suitable for first-aid of wounded person with obstacle of the upper respiratory tract in peacetime and war time. The device is portable, quick and easy to operating, minimal invasion and effective. Clinical application of new puncturing technologies, which is hopeful to become the fifth first-aid technique of military medical training and self-help saving, possesses brood prospects.

Research of EGFR/PTEN Expression and Related Apoptosis and Invasion by in Human Glioma

Wed, 30 Mar 2011 13:08:46 +0000

Gliomas are the most common malignant disease,over the past years,even though on the up in clinical glioma has been a series of active treatment measures,but the plastic stromal tumors in patients with the mortality rate remains high,so gliomas are still the refractory disease in field of neurosurgery,the study of its etiology, pathogenesis,biology characteristics as well as new means of effective treatment has become a hot topic in neurosurgery.Much current research has confirmed that the incidence of tumor development is a gene involved in steps in the evolution of complex process.Therefore,glioma is essentially a multi-gene disorders,which are happened because of the activation of oncogenes and tumor suppressor inactivation of genes cause cell cycle through the line changes.The abnormal signal transduction pathways,the life-cycle extension,as well as defects in apoptosis and so on,and thus give rise to out-of-control cell proliferation and malignant transformation.With the medical technology of molecular biology,the gene therapy of glioma study has been made exciting progress,and has the transition to clinical research stage.However,in glioma gene therapy of animal experiments and clinical studies will still encounter problems relating to the validity of questions,such as how to effectively impede the gene therapy to overcome some barriers and exert the greatest extent possible gene therapy treatment of the role of biology became a major research topic.Some protein growth factors and their receptors have been identified as protp-oncogene.Epidermal growth factor and its receptor have been most concerned. Human EGFR gene located in chromosome 7p11-13,and many of tumor cells have over-expression of EGFR.EGFR is a proto-oncogene c-erB-1(HER-1)expression of the product,encoded by anⅠ-type transmembrane protein tyrosine kinase growth factor receptor,the molecular weight of about 170kD generally expressed in human epidermal cells and stromal cells,the conversion information in the membrane play an important role,and is highly expressed in a variety of human malignancies.Signal transduction of it mediated have many effects,including proliferation,migration,cell differentiation and the stability of the environment,and is closely related to cell regeneration and the occurrence,development of malignant tumors.PTEN gene has been cloned is the first of a phosphatase activity of tumor suppressor genes,play a important role in regulating cell growth and apoptosis,cell signal transduction and tumor cell invasion,metastasis,etc.At present,we consider the role of PTEN tumor suppressor is a few ways to achieve the adoption as following: (1)Inhibited cell movement and infiltration through the phosphorylation of FAK.(2) Prevent cell growth and promote apoptosis by PIP3 dephosphorylated.(3)Inhibits cell growth and differentiation by inhibited cell MAPK signal transduction pathway.In recent years,tissue microarray developed as a morphology-based molecular biology techniques.It can be sliced up on a high-throughput access to histology,gene and protein expression information.It can simultaneously detect a tumor gene expression at different stages of the situation at the same time.It is also low cost,easy to operate.Application of tissue microarray can be used for gene expression detection, diagnosis and treatment to verify the specificity of target gene expression,detection and optimization of probes and antibodies,drug screening and disease diagnosis and so on.ObjectiveTo explore EGFR,PTEN,MMP-2/9,Caspase-9 and Bcl-2 expression in human gliomas and its relation with the degree of glioma maligancy.MethodsIn order to further elucidate human glioma EGFR and PTEN gene in the regulation of glioma proliferation and invasive growth and downstream gene expression,the subject of the application of tissue microarray analysis of 60 cases of glioma specimens have been detected.Further application of immunohistochemical method in the analysis of the expression level of EGFR,PTEN,MMP-2 / 9, Caspase-9 and Bcl-2 and the correlation between the degree of malignant glioma the contact have been made.ResultsThe expression of EGFR,MMP-2,MMP-9,BCL-2 is the plasma-positive.And different levels of expression between glioma have differences.With the increased level of its expression positive rate and the degree of expression was followed by increases.Expression levels of the EGFR signaling pathway glioma important members such as EGFR,MMP-2,MMP-9,BCL-2 by immunohistochemical analysis showed that:the above indicators at glioma positive rate and expression level of tumor with the degree of malignancy increases at low level and between malignant glioma significant statistical difference.Expression of PTEN and Caspase-9 at different levels between glioma also vary,but with the increased level of its expression positive rate and expression of positive degree is followed by reduced. Immunohistochemical analysis also showed:PTEN and Caspase-9 in glioma positive rate and expression level increased with the degree of malignant tumor is reduced,at low level and between malignant glioma significant statistical difference. Correlation analysis also found that EGFR,PTEN,MMP-2,MMP-9,Caspase-9, BCL-2 exists between the relationship significant.EGFR,PTEN,MMP-2/9, Caspase-9 and Bcl-2 between these factors and related statistically significant.Conclusion:That indicating that each of these indicators probably exist the interaction or synergy,but also implies that the above-mentioned factors have important effect in the malignant glioma.EGFR and PTEN gene has played an important role in the occurrence and development of glioma,can be used as a candidate gene for gene therapy.

Effects of β_2 Agonist Salbutamol on the Uptake of Sevoflurane and Bispectral Index Variations in Elderly Paients with Chronic Obstructive Pulmonary Disease

Wed, 30 Mar 2011 13:08:41 +0000

PartⅠEffects ofβ_2 agonist Salbutamol Aerosol on the Uptake of Sevoflurane in Elderly Patients with COPDObjective:To investigate the effects ofβ_2 agonist salbutamol aerosol on the uptake of sevoflurane in elderly patients with chronic obstructive pulmonary disease(COPD).Methods:A randomized,placebo-controlled and double-blinded trial with two groups (experiment and control group) was designed.With approval from the ethics committee in Zhong Shan Hospital,Shanghai,China,and with informed consents,26 patients were recruited to the trial and randomly allocated to salbutamol group(n= 13) or placebo group(n= 13).Eligible patients were elderly patients with ASA physical statusⅠ～Ⅱ,undergoing general surgery,with a body mass index(BMI) between 18 and 30 kg·m~(-2),well-defined clinical diagnosis of COPD and with no evidence of malignant hyperthermia,cardiac dysfunction and renal insufficiency.All subjects were fasting for 10 hours and received no premedication.Routine monitoring (consists of a three-lead ECG,pulse oximetry,noninvasive blood pressure and expired gas analysis) was instituted and 16 G right internal jugular cannula were cannulated. Left radial artery was cannulated under local anesthesia to measure direct arterial blood pressure.And bispectral index(BIS) monitoring was initiated prior to induction. All subjects were received inhaled salbutamol aerosol 200μg according to manufacturers\' recommendations and 500 mL Ringer\'s lactate solution 30 minutes before induction of anesthesia.Rapid sequence induction of anesthesia was fentanyl 3.0μg·kg~(-1),followed by propofol 1.5～2.0 mg·kg~(-1) and rocuronium 0.9 mg·kg~(-1) was administered to facilitate endotracheal intubation.When the trachea was intubated, intermittent positive pressure ventilation(IPPV) was applied with tidal volume of 8～10 mL·kg~(-1),respiratory rate 10～12 breaths per minute,inspiratory to expiratory time ratio of 1:2 to achieve pressure of end-tidal carbon dixoide(EtCO_2) 30～35 mmHg via a non-rebreathing circuit.After stable hemodynamics was maintained for 5 minutes,fresh gas flow was set to 2 L·min~(-1) with 2%sevoflurane in admixture,then HR,invasive arterial blood pressure(IABP),S_PO_2,EtCO_2,BIS,minimum alveolar concentration(MAC),concentration of inhaled sevoflurane(F_I) and concentration of end-tidal(F_E) were recorded at 1,2,3,4,5,7,10,15 min.The Ppeak and Pplat were also measured in 1,5,10 min since the start of successful endotracheal intubation.We expose the two groups after completion of whole trial and analyzed significance among the related variables.Results:There were no statistical significance in height,age,body weight,BMI, baseline HR,baseline MAP,FEV_1,FVC and arterial blood-gas parameters between two groups.The value of HR and MAP reduced with the increasing MAC of sevoflurane over time in each group,while no statistical significant variations were observed in the values of HR and MAP between two groups in respective time point (P＞0.05).There were no statistical significance in inhaled anesthetic concentration captured by side stream sampling from Y port was higher between experiment group and control group,but the inhaled anesthetic concentration of sevoflurane increased steadily over time.In comparison with group placebo,the expired sevoflurane concentration markedly lower in 2 min[(0.382±0.170)%,P＜0.05]、3 min [(0.512±0.204)%,P＜0.05],4 min[(0.653±0.224)%,P＜0.05],5 min [(0.806±0.198)%,P＜0.05]in experiment group.Peak airway pressure and mean airway pressure in experiment salbutamol were striking lowerer than control group (24.7±2.9 cmH_2O,20.1±2.7 cmH_2O,P＜0.05).The value of BIS was kept between 38 and 60 throughout the period in spite of increasing MAC over time,there were no differences between groups in each time point.Conclusion:The rate of alveolar concentration of sevoflurane was accelerated within 15 min in elderly patients with COPD after the administration of inhaled salbutamol aerosol(200μg) 30 minutes before the rapid-sequence induction of general anesthesia, its plausible mechanism may be that salbutamol nebulization partly ameliorated airway spasticity and attenuated airway resistance.While clinical significance was limited because its uptake was very short.Variations of BIS values failed to achieve an original wish since it was not sensitive for dose-response of inhaled anesthetics in the brain. PartⅡEffect of salbutamol aerosol on bispectral index value and arterial carbon dioxide in chronic obstructive pulmonary disease patients with typeⅡrespiratory failureObjective:Aged patients with chronic obstructive pulmonary disease(COPD) in accompany with carbon dioxide(CO_2) accumulation usually occur cognition dysfunction.Up-to-date,it is poorly understood that the effect of salbutamol aerosol on bispectral index and arterial carbon dioxide in COPD patients with CO_2 accumulation.This article was focussed on investigating the effect of salbutamol aerosol on bispectral index(BIS) and pressure of arterial carbon dioxide(PaCO_2)) in elderly patients with COPD in accompany with chronic CO_2 accumulation.Methods:In this prospective,randomized controlled study,12 COPD patients with clinically well-identified typeⅡrespiratory failure under bi-level positive airway pressure(BiPAP) ventilation in respiratory intensive care ward were recruited.The patients were randomly divided into experimental group(group E) and placebo group (group P).Heart rate(HR),mean arterial pressure(MAP),pulse oxygen saturation (SpO_2),BIS and parameters of arterial blood gas analysis(PaO_2,PaCO_2,pH) were recorded in 0 min and after inhaled salbutamol 15 min and 30 min respectively.At the same time,patients\' mental state were assessed by the modified Observer\'s Assessment of Alertness/Sedation scale(OAA/S).Results:7 patients in group E and 5 patients in group P were included.In comparison with baseline value(100.5±9.48 mmHg) in group E,MAP slightly decreased in 15 min (94±9.42 mmHg,P＜0.01)and 30 min(92.43±12.73 mmHg,P＜0.05).In group P, MAP mildly dropped in 15 min(90±4.3 mmHg,P＜0.05) and 30 min(95.6±7.33 mmHg,P＜0.05).While there was no statistical significance between two groups about MAP and HR variability.PaCO_2 profoundly decreased in group E(72.14±17.3 mmHg,P＜0.01;67.14±15.4 mmHg,P＜0.05) and slightly decreased in group P (70.4±14.43 mmHg,P＜0.05;64.2±10.73 mmHg,P＜0.05).BIS value was slightly increased in two groups(group E:80.14±6.3,P＞0.05 81.14±9.24,P＞0.05;group P: 81.6±8.38,P＞0.05,82.8±7.98,P＞0.05) after the administration of inhaled salbutamol.The efficiency of non-invasive ventilation is increased after the doctor\'s guidance. Conclusion:The level of PaCO_2 lowered after the administration of inhaled salbutamol via ventilator metered dosage inhaler(MDI),and this effect is more obvious in 15 min than 30 min.Brief changes of PaCO_2 level may not result in significant altertions of BIS values in COPD patients with long-term CO_2 accumulation.

Study of Relationship between Traditional Chinese Medicine Syndrome Differentiation-Type of Chronic Renal Insufficiency and Mineral Bone Disorder

Wed, 30 Mar 2011 12:14:36 +0000

Objective:To make TCM Syndrome Differentiation(TCM-SD) for chronic renal insufficiency(CRI) patients and determine some index relative to Mineral&Bone disorder of these patients.And then,the prevalence of different TCM-SDTs of CRI patients and its\' correlation with those index mentioned above were study.By doing this, it may provide reliable basis for making TCD differentiation objectively and treating exactly.Methods:40 CRI patients have been divided into 5 types by TCM differentiation standard, which were Syndrome of Qi deficiency of spleen and kidney(QSK),Syndrome of Yang deficiency of spleen and kidney(YSK),Syndrome of deficiency of both Qi and Yin(QY),Syndrome of Yin deficiency of liver and kidney(YLK),Syndrome of deficiency of both Yin and Yang(YY).While for control group,there were 25 healthy persons enrolled.And then,serum Ca,P,Alkaline phosphatase(AP),parathyroid hormone(PTH),bone density of this two groups have been measured and compared.In addition,correlation of TCM-SDT and those index mentioned above have also been studied.Results:1.The incidence rate ofQSK,YSK,QY,YLK and YYwas 32%,30%,18%,15%and 5%respectively.2.The serum creatinine concentration of YSK,QY,QSK and YLK was 524.62±134.89 umol/L,725.62±170.14umol/L,688.33±82.39umol/L,846.00±177.85umol/L, 1346.50±99.70umol/L respectively.And that of QSK was significantly lower than other 4 types.3.Compared to control group,serum Ca and P of CRI patients were significantly abnormal.And the concentration of serum Ca of QSK,QY,YLK and YSK were 1.94±0.25mmol/L,1.92±0.38mmol/L 1.89±0.19mmol/L and 1.67±0.34mmol/L respectively.While for serum P,the figures of YSK,YLK,QY and QSK were 2.47±0.519mmol/L,2.40±0.65mmol/L,2.09±0.3 lmmol/L and 1.97±0.50mmol/L respectively.There also existed significantly difference between QSK and YSK patients(P＜0.01).4.The concentration of serum AP of YSK,QY,QSK and YLK was 99.00±33.35U/L, 91.43±13.72U/L,80.92±22.31U/L and 69.33±24.48U/L respectively.And concentration of YSK and QY were significantly higher than control group(P＜0.05),there also existed significantly difference between YSK and YLK patients(P＜0.05).5.As for concentration of serum iPTH,the figure of YLK,QY,YSK and YY was 434.67±191.86pg/ml,415.29±270.74pg/ml,340.67±270.74pg/ml and 199.93±121.23 pg/ml respectively which were all significantly higher than control group (QSP vs control group,P＜0.05;other types vs control group,P＜0.01).In addition, there also existed significantly difference between QSP and YLK patients(P＜0.05).6.The figure of bone density of QSK,YSK,YLK and YY was 0.32±1.04,-1.09±1.144, -1.52±2.07 and -2.04±1.86 respectively.And that of YSK,YLK and YY were significantly lower than QSK and control group(P＜0.01).Conclusion:1.The level of serum creatinine of QSK patient was significantly lower than other TCD types,so QSK may be the major TCM type at early stage of CRI.And then, the TCD type may develope from QSK,YSK/YLK,QY to YY graduallay.2.Abnormality of Ca & P metabolism and secondary hyperparathyroidism which occurred in QSD patients suggested the importance and necessity of correcting these abnormalities at early stage and/or QSD of CRI patients.3.Bone densitometry of YSK,YLK,QY patients were obviously reduced, demonstrating osteoporosis occurred in these patients.At the same time,OBD has been found to be associated with TCM-SDT.4.iPTH and OBD could be looked as markers for making TCM-SDT and judging its turnover of CRI patients.

The Expression and Clinical Significance of Integrinανβ3 and Claudin-1 in Cervical Squamous Carcinoma

Wed, 30 Mar 2011 12:14:34 +0000

Objective:Cervical cancer is the second malignant diseases of women in the world, each year more than 280,000 patients died , of which 80% occured in developing countries [1]. Study shows that there is only a small part of the HPV-infected cervical lesions will develop into invasive cervical cancer. A number of other oncogenes, tumor suppressor genes and micro- environmental factors also play an important role in the progress of cervical cancer. The aim of this study was to determine the expression and clinical significance of integrinανβ3, tight junction protein claudin-1 in cervical squamous carcinoma of cervix.Methods: We examined integrinανβ3 and tight junction protein claudin-1 expression in 30 cases of cervical squamous cancer by immunohistoche-mistry SP method. All the tissue paraffin blocks were filed in Dalian Central Hospital Pathology diagnostic squamous cancer from January 2003 to December 2007: 20 cases of well-differentiated group, 10 cases of poorly differentiated group. SPSS10.0 was used to analysis the data.Results:1.The positive expression rates of integrinανβ3 of well-diffe- rentiated and poorly differentiated groups in squamous cell carcinoma of cervix were 55% (11/20) and 80%(8/10) separately. The expressive inten- sity of integrinανβ3 in poorly differentiated groups was significantly higher than in well-differentiated group (P <0.05).2.The positive expression rates of claudin-1 protein of well- differentiated and poorly differentiated groups in squamous cell carcinoma of cervix were 80%(16/20)、20%(2/10). The expressive intensity of claudin-1 protein in well-differentiated group was significantly higher than in poorly differentiated groups (P <0.05). Conclusion:The expression of integrinανβ3 is negative correlative with the pathologic degree of differentiation in squamous cell carcinoma of the cervix. With the descending of pathologic degree of differentiation, the expression level of integrinανβ3 is up-regulation, so integrinανβ3 in patients with cervical squamous cell carcinoma has certain clinical sign- ificance in prognosis.The expression of claudin-1 protein is positive correlative with the pathologic degree of differentiation in squamous cell carcinoma of the cervix. With the descending of pathologic degree of differentiation, the expression level of claudin-1 protein is down-regulation, so claudin-1 protein in patients with cervical squamous cell carcinoma has certain clinical significance in prognosis.

Expression of Urinary Soluble Fms-like Tyrosine Kinase-1 with Preeclampsia

Wed, 30 Mar 2011 12:14:33 +0000

Objective: Pre-eclampsia is a specific pathologic process of gestation period.Although amount of reseach have done about it,the etiology and pathogenesis of pre-eclampsia remains unclear.Clinical diagnostic methods are monitoring of blood pressure, analysis of urine protein, test of serum uric acid,but the specificity and sensitivity are not optimal.Update,the study showed that the imbalance between angiogenesis factor and antiangiogenic agent is the important reason.Among them,it focuses on soluble fms-like tyrosine kinase-1 (sFlt-1)generated by placenta.sFlt-1 is inhibit receptor of vascular endothelial growth factor (VEGF). It can inhibit the physiologic action of VEGF and placental growth factor (PlGF) by combined them.It was recently shown that concentration of sFlt-1 in placenta and blood of patients is high.So far the study is concentrated on the variety of sFlt-1,VEGF and PlGF in serum.And it is found that there exists singnifi– cant difference of sFlt-1 in serum between pre-eclampsia and normal pregnant.Meanwhile sFlt-1 indentify the degree of the disease. However,in our experiment, we sought to the possibly change of sFlt-1 in urine and the action of it.At the same time, in order to find out an noninvasive method,an experimental diagnostic mark with high specificity and sensitivity,we compared sFlt-1 with other clinical indexes.Methods:Gathering the serum of 36 patients in pre-eclampsia without complications and using the drugs affecting the vacular activity, severe pre-eclampsia ,non severe pre-eclampsia and normal pregnant are 16、13and15 respectively.sFlt-1 is measured by ELISA,urine protein by biuret colorimetry, serum uric acid by phosphotungstic acid. Results: (1)In severe pre-eclampsia,non severe pre-eclampsia and normal pregnant groups, the quanlity of urine protein is 2.88、1.31 and 0.37 respectively, the quanlity of uric acid in serum is 380、312 and 209umol/L respectively (P=0.004),There exists singnificantly difference(P<0.05). (2)the quanlity of sFlt-1 is 2986.25、2189.38 and 1231.60mg/ml respectively(P=0.000).There exists singnificantly difference (P<0.05). (3)Among them,there exist correlation ship.The r between urine protein and uric acid,urine protein and sFlt-1,uric acid and sFlt-1 is 0.582 and 0.325 respectively (P<0.05).(4)From the ROC curve,we can see that the specificity and sensitivity of sFlt-1 is the highest.Conclusion: (1) sFlt-1 plays a important role in the creation of pre- eclampsia.And sFlt-1 indentify the degree of the disease.(2)Evaluation of sFlt-1 in urine is an noninvasive and quick meathod.The specificity and sensitivity of sFlt-1 are the much highest than another to identify the degree of severe pre-eclampsia.(3) sFlt-1 in casual urine has positive singnificance while larger than 2676mg/ml.

The Role of Oxidative Stress in Copper 8-quinolinolate-induced DNA Damage in HepG2 Cells

Wed, 30 Mar 2011 12:14:31 +0000

Objective:Copper 8-quinolinolate(Copper 8-quinolinolate,CuQ) is ex- tensively used as a fungicide in many countries and was used as a fungicide for controlling the diseases of freckle, scab, and black spot in vegetables and fruits.In addition,CuQ is approved for use as a wood preservative for control of sapstain, mold and decay in unfinished wood and wood products. Treated wood materials may be used in contact with fruit, vegetables and other foodstuffs in an area such as greenhouses and greenhouse items, produce picking boxes, mushroom trays and vegetables stakes.The carcinogenic risk of PAT to humans is classified in Group3 by the International Agency for Research on Cancer (IARC), since the evidence of carcinogenicity is inadequate in humans and in experimental animals. CuQ has been reported to dose-dependently increased the reverse mutation of S. typhimurium TA97, TA100, TA102 in assay with rat liver S9, and positive response of S9-activated in S. typhimurium TA100 with Ames test and in Bacillus subtilis with rec assay CuQ-induced hepatotoxicity has been ob- served in experimental animals. Therefore, we explore the DNA damage effects of CuQ in Human hepatoma cell line (HepG2) cells and the reaction mechanisms. HepG2 cells were selected as test system, which was orig- inally derived from a human hepatoblastoma. HepG2 cell line retains many of the functions of normal liver cells and expresses the activities of several phases I and II xenobiotic metabolizing enzymes. It was considered to be more suitable for reflecting the metabolism of xenobiotics in the human body better than other mammalian cell lines.The aim of this study was to assess the DNA damage effects of CuQ and to elucidate the reaction mechanisms by use of human hepatoma HepG2 cells. Thus it may provide some information for the safety assessment to humans on CuQ.Methods: HepG2 cells were selected as test system. DNA-strand breaks in HepG2 cells were evaluated by single cell gel electrophoresis assay (SCGE). To elucidate the DNA damage mechanism in HepG2 cells, Catalase (CAT) activity was determined and the level of lipid peroxidation was determined by measurement of thiobarbituric acid-reactive substances (TBARS). We measured the effect of CuQ on NF-κB by western blotting analysis. 8-hydroxyderoxyguanosine (8-OHdG), which is a reliable marker for oxidative DNA damage, was also measured by immunoperoxidase stain- ing analysis. We used the O-phthalaldehyde (OPT) to monitor the levels of GSH. DL-buthionine sulphoximine (BSO) were used to modulate the level of GSH in HepG2 cells, and evaluate the effects of GSH on CuQ-indudced DNA damage.Results:Exposure to CuQ caused significant increase of DNA migra- tion in comet assay at all tested concentrations (0.5μM-4μM) in HepG2 cells,suggesting that CuQ caused DNA damage in HepG2 cells. CuQ signi- ficantly decreased levels of glutathione (GSH) and the activity of catalase (CAT) in HepG2 cells and increased levels of lipid peroxidation product thiob-arbituric acid reactive substances (TBARS). Moreover, CuQ signi- ficantly increased accumulation of the p65 subunit of NF-κB into nucleas, the formation of 8-hydroxydeoxyguanosine (8-OHdG). The intracellular GSH level was modulated by pre-treatment with buthionine(S,R) sulf- oximine (BSO),a specific GSH synthesis inhibitor, it was found that depletion of GSH in HepG2 cells treated with BSO dramatically increased the susceptibility of HepG2 cells to CuQ-induced DNA damage.Conclusion: CuQ exerts DNA damage by oxidative stress in HepG2 cells; GSH, as a main intracellular antioxidant, is responsible for cellular defense against CuQ-induced DNA damage.

The Effect of Different Initial Warfarin Dosages on the Time of Target International Normalized Ratio in Patients with Nonvalvular Atrial Fibrillation or Atrial Flutter

Wed, 30 Mar 2011 12:14:29 +0000

Objective: To investigate the effect of different initial warfarin dosages on the time of target international normalized ratio (INR) and the safety during dose adjustment in patients with nonvalvular atrial fibrillation (NVAF) or atrial flutter (AFL).Methods: 85 patients with NVAF or AFL who had the indication of anticoagulation therapy were randomly assigned to three groups according to different initial warfarin dosages, 3.125 mg/d (Group 1), 3.125mg/d after 5 mg/d for tow days (Group 2) and 5 mg/d (Group 3). Baseline data collected included demographic characteristics (age, sex, height, weight), conjoined medication, assident diseases, and the INR before taking warfarin. Warfarin should be administered at 16:00 daily except for the patients following an AF radiofrequency current catheter ablation (RFCA) who should take warfarin come back ward. Low-molecular-weight heparin (LMWH 5000～7500U every 12 hours by subcutaneous injection) can be restarted 2-4 hours postoperatively along with warfarin and the combination continued for 4 to 7 days until the INR increase to the desired range (>1.6). If patients are considered to be at high risk of postoperative bleeding, LMWH can be delayed for 24 hours or longer. The INR was measured on the 3rd、4th、5th、7th and 9th day before 10:00 am, the dose of warfarin was adjusted according to INR, until INR was stabilized between 1.8 and 3.0 for at least one week at the same dose of warfarin. When the INR is above the therapeutic range 3.0 but less than 5.0, the patient has not developed clinically significant bleeding, the next dose of warfarin can be omitted and resumed (at a lower dose) when the INR approaches the desired range (0.05). Compared with group 1 and group2, the INR change in group3 was more quickly, the ratios of INR within target range on day 4, 5and day 7 were the highest (P0.05). The mean time achieving the target INR was 8.2±2.4 days, 7.6±3.1 days and 6.2±2.1 days and reach a stabilized target INR was 15.5±2.8, 14.8±3.1 and 13.0±2.0 respectively in Group 1, 2 and 3. Compared with group 1 and group 2, the mean time achieving and reaching the stabilized target INR in group 3 was the shortest (P0.05). There was no significant difference on the incidence of exorbitant INR in three groups. There were not thromboembolism and important hemorrhage complication in 3 groups (P>0.05).Conclusions: In Chinese patients with NVAF or AFL who had the indication of anticoagulation therapy,an initial warfarin dosage of 5 mg/d treatment may reach the stabilized INR range quickly, safety and efficiently without increasing the bleeding complication.It’s not necessary to measure INR in the first 3 days. If INR reaches target range at 4th day,warfarin should be decreased to 2.5～3.125mg according to INR, and INR should be mearsured every other day. If INR is lower than target, INR should be measured at the 5th day and the 7th day until the therapeutic range has been reached, and then warfarin should be decreased to 2.5～3.75mg according to INR. It’s indicated that the maintenance dose of warfarin is higher if INR not yet reach target range at the 4th day. It’s not ought to decease dose of warfarin after INR reach target range. Once the INR becomes stable, the frequency of testing can be reduced to intervals as long as 4 weeks.

The Study on Aβ_(1-42) Oligomer Preparation

Wed, 30 Mar 2011 12:14:27 +0000

Alzheimer\'s disease(AD) is the most common form of dementia among aged population,which causes memory impairment and cognitive dysfunction and characterized by presence of extracellular fibrillar Aβin amyloid plaques,intraneuronal neurofibrillary tangles.At present,42-amino acid form of Aβsoluble oligomers (amyloidβ-protein) has been recognized as the pivotal impact factor in Alzheimer\'s disease.Abundant evidences suggest that the soluble Aβ_(1-42) oligomer has become the common target in the treatment of Alzheimer\'s disease.Here,the chemically synthetic Aβ_(1-42) oligomer has been assemblied in vitro.The resulted oligomer has been observed and assessed via SDS-PAGE,Western,ELISA and immunou blot assays.Our results showed that the profiles of this kind of oligomer appeared as a ladder in the molecular weight ranges from 4-170 kD,which were tetramer,hexamer and other higher moleculndar weight aggregations.In addition,there were still a little bit of monomer. The Aβ_(1-42) oligomer is stable in coating buffer and can be detected effectively in indirect ELISA.Immobilized Aβ_(1-42) oligomer on nitrocellulose filter could be recognized by monoclonal antibody 6E10 in immunoblot assays.Furthermore,the arrayed Aβ_(1-42) oligomer dots could be applied in fast and high throughput antibody detection and screenings.In conclusion,our results may provide important tools in Aβdetection,basic and therapeutic research in AD.

The Expression and Its Significance of CD147, MMP-9 and VEGF in Renal Cell Carcinoma

Wed, 30 Mar 2011 12:14:26 +0000

Objiective To study the expressions of CD147, MMP-9 and VEGF in renal cell carcinoma, and their correlation with tumor growth, tumor invasion and metastasis. Methods The expression of CD147, MMP-9 and VEGF were detected immunohistochemically SP method in tissue from the carcinoma (RCC , n=107) and normal renal tissue (n=10). The relationships between the expression and clinical pathology were analyzed, and correlation among CD147, MMP-9 and VEGF was studied. Results Expressions of CD147, MMP-9 and VEGF in RCC were significantly higher than that of normal renal tissue (P0.05), but related to clinical staging (P0.05). Conclusion CD147, MMP-9 and VEGF correlate significantly with aggressiveness and invasion in RCC, suggesting of expression of CD147, MMP-9 and VEGF may be biological indexes in detecting tumor growth, invasion and metastasis of RCC.

Study on the Response of Osteoblasts to Overuse and Disuse in Vitro

Wed, 30 Mar 2011 12:14:25 +0000

The balance of bone resorption and bone formation keeps the bone mass level, while many factors including the mechanical loading can break it. Normally the balance is kept, even though the physical loading input on bone changes in a range due to the change of our activity. However, it is broken down when our activity and the loading exerted on bone and its cells are greatly changed, such as the obvious weight gain/loss, long-term bed rest, space flight. These phenomena were consistent with the hypothesis put forword by Burger in 1999 that overuse will profit bone formation, while disuse will profit bone resorption, from the point of signaling and nutrients supply among cells in bone. However, if it is true in cellular level remains unknown. In this study, the aim of our investigation is to make clear if the overuse increases bone formation and disuse increases bone resorption in vitro cellular level.We used Wistar primary cultured osteoblasts and studied its differentiation as bone formation index and the ratio of secreted RANKL and OPG as bone resorption index. Stepwise increased and decreased laminar shear stress was used to mimic the overuse and disuse. The flow chamber was to provide controlled shear stress. The main work and conclusions were as follows:Flow chamber was designed and used in our study. It provided laminar flowshear stress which can be adjusted via the flow rate. The assembly and disassembly were very easy. About 35 mL of flow fluid was used in our flow system.The primary Wistar rat’s osteoblasts were cultured from the calvaria of new-born rats by the tissue piece culture method. The osteoblasts were identified by the morphology and Von Kossa staining. The third passage of osteoblasts cultured on glass slides was used in mechanical experiments after they reached confluence.Six processes of shear stress were designed here to validate the effect of overuse and disuse on the gene expression of osteoprotegerin ( OPG ) and osteoclast differentiation facto(rRANKL)and alkaline phosphatase(ALP)and mineralization. The results confirmed that the stepwise increased shear stress inhibited bone resorption; while the stepwise decreased shear stress gave rise to adverse response. The stepwise increased shear stress promoted the mineralization, while 30-20-10-5 dyn/cm2 stepwise decreased shear stress inhibited it, the other two stepwise decreased shear stresses promoted the mineralization. Our results confirmed that overuse can profit bone formation by increasing bone formation and inhibiting bone resorption; while disuse can profit bone resorption at least by increasing bone resorption.

The Extraction, Purification and Biologic Activity of Ostreidae Polysaccharides

Wed, 30 Mar 2011 12:14:22 +0000

Oysters polysaccharide was extracted from oysters. In the article, we discussed the process of oysters polysaccharide extraction systematically and then analysed the purified polysaccharides’s strcture , the biological activity in vivo and in vitro .We hope to provide polysaccharide reference for the further study of oysters medicinal value and oyster products.Crude polysacharides were isolated from Ostreidae by a method so called“water extraction followed by ethanol precipitation”, Single factor test was used to analyze the influence of each factor, such as extraction times, solid-liquid ratio, and temperature. The optimal conditions for extraction and isolation of Ostreidae polysacharides (OPS) were studied by L933orthogonal experiment design.The results of orthogonal test on OPS showed that the optimal conditions for extraction of polysacharides from Ostreidae were as follows: temperature-90℃, solid-liquid ratio-1:80, extraction running times-3 hours.The polysacharide content of Ostreidae was determined by phenol-sulfuriic acid method. Finally, the content {percentage) of total sugar was determined to be 72.43%.After extraction, the free protein and other impurities in the crude extract were wiped out by Sevage method. After the process of dialyzing, the obtained polysaccharides were further purified by DEAE-52 column chromatography and Sephadex G-100 chromatography,OPS-II and OPS-II b were obtained. By using UV spectrum, it was found that there were no absorption peaks of protein and nucleic acids at 280 nm and 260nm. OPS-II b was tested by HPLC for purity and the result showed that OPS-IIb had a characteristic absorption peak, so the composition and structure of OPS-II b could be analyyzed.The physicochemistrical properties of OPS-IIb were preliminarily analyzed by HPLC, gas chromatography (GC), infrared spectroscopy (IR), thin layer chromaotography (TLC) and other Physical and chemical analysis methods, The results indicated that OPS-II b had typical absorption peaks,it is a component of glucose.The results of antioxidation test in vtro showed that OPS played a good role in the antioxidation process. OPS was obviously able to scavenge super oxide free radical in the concentration of 3.0mg/mL and the scavenging rate was 43.84%; while in the concentration of 4.0 mg/mL inhibition rate of hydroperoxide-induced hemolysis of red blood cells in mice and hydroxy free radical rate were 58.9% and 65.69%,respectively;the inhibition rate of liver lipid peroxidation reach 62.57%. Experiments in vitro showed that OPS could enhance macrophage phagocytosis, increase immune enzymatic activity of mice and increase their spleen indeces.Immune function test results in mice showed that the oyster 3mg/ml group can effectively increase the normal mouse spleen cells in Pakistan and carbon leaching dissection vitality, and 6mg/ml, 9mg/ml group had no significant effect.

Electrophysiology Characteristic of CIPM and the Influence to the Duration of Mechanical Ventilation

Wed, 30 Mar 2011 12:14:20 +0000

Objective:To understand the incidence of neurological critical illness polyneuromyopathy(CIPM) in a specific group of people,to explain the electrophysiologic character of critical illness polyneuromyopathy,to increase in the diagnosis of the disease based on electrophysiological and deepen our understanding of the nature.Observating the influence of CIPM to the duration of mechanical ventilation,and explore the reasons for its impact.Methods:To utilize Denmark Keypoint 4 electromyogram instrument to make electroneurophysiology detection on twenty-seven subjects with respiratory failure who were treated with mechanical ventilation over 1 week and had difficulty in weaning from the ventilator in intensive care unit,13 male,14 female,age(69.1±16.2) years old.Electroneurophysiological examination including physical side of the median nerve,ulnar nerve,posterior tibial nerve and common peroneal nerve of nerve conduction;anterior tibial muscle,quadriceps,and deltoid abductor digiti needle EMG polar;median nerve of the F wave;ulnar nerve of the repetitive nerve stimulation(RNS);limb sympathetic skin response(SSR);under the limbs evoked potential(SEP);blink reflex(BR).To record mechanical ventilation time of CIPM and non-CIPM,at the same time to compare their detection of the phrenic nerve conduction.Results:In 27 cases of patients with nerve conduction and electromyography testing 13 were diagnosed CIPM.The incidence is about 53%(13/27),in which one subtype of critical illness polyneuropathy(CIP) in patients with 10 cases and critical illness myopathy(CIM) in patients with three cases.10 cases were detected in patients with CIP had 40 neurons,each case has two or more nerve compound muscle action potential(CMAP) and sensory nerve action potential(SNAP) amplitude lower end of motor nerve latency(DML) or less normal,and all normal nerve conduction velocity;At the same time testing the 40 muscles,25 neurogenic damage prompted. CIM detected three cases of the 12 patients with nerve,SNAP amplitude and nerve conduction velocity(NCV) were normal,eight of which reduce nerve CMAP amplitude;6 of 12 muscle show myogenic damage,mainly in the proximal muscle damage.In CIPM patients sympathetic skin response(SSR) did not lead to positive waveform.The occurrence of F wave in 3 cases of patients with CIP and 2 cases of patients with CIM reduce,and conduction velocity were normal.CIPM all patients with RNS,SEP and BR testing were normal.The patients with CIPM compound action potential amplitude of the diaphragm(diaphragmatic compound muscle action potential,DCMAP) reduced than non-CIPM was significantly(0.23±0.11 vs 0.35±0.13,P = 0.018),the difference was statistically significant.Their distal motor latency phrenic nerve(phrenic distal motor latencies,PDML) had no significant difference (7.67±1.27 vs 7.39±1.23,P = 0.562).Comparison of CIP and non-CIP group of patients with the phrenic nerve conduction in patients DCMAP was statistically significant difference(0.22±0.21 vs 0.34±0.12,P = 0.024),while no difference PDML in two groups(7.82±1.41 vs 7.35±1.13,P = 0.347).The median duration of mechanical ventilation(quartile interval) in CIPM patients were 30.0(22.5) d,and was significantly longer(U=20.0,P＜0.01) than non-CIPM 14.5(8.5) d.Analysis of the subgroup of patients with CIP group the median duration of mechanical ventilation and quartile range of 30.0(19.0) d,more than non-CIP patients 15.0(10.5) d was significantly longer(U= 30.0,P＜0.01).Conclusion:CIPM is the more common neurological complications in patients with respiratory failure mechanical ventilation over 1 week in ICU.Neural electrophysiological testing is an effective means of CIPM.In CIPM conventional electrophysiological examination of patients is mainly expressed in compound muscle action potential and(or) sensory nerve action potential amplitude reduction,while the normal NCV.All CIPM patients have abnormal SSR and there are no central nervous system and neuromuscular junction damage.Diaphragm and(or) phrenic nerve damage in patients with CIPM,which might be the direct cause of the time offiine.

Effect of Expression of CD50 in Macrophage on SIRS Induced by SAP with Combined Treatment of Traditional Chinese and Western Medicine

Tue, 29 Mar 2011 15:31:51 +0000

Background and objective: Acute pancreatitis (AP) is a common surgical disease, whose clinical pathological change is complex, with various responses to treatment. Light edematous pancreatitis is a self-limiting disease with fine responses to treatment and prognosis, while severe acute pancreatitis (SAP) is disease with acute onset and rapid progress, which can lead systemic inflammatory response syndrome (SIRS)at early stage. Although there is rapid development in the treatment of SAP, the mortality rate is as high as 10 ~ 30%. SIRS is considered the main reason leading to death. A large number of studies have confirmed that the phagocytosis ability of macrophage in clearing apoptotic neutrophil decrease in SIRS, resulting in the release of a great number of inflammatory mediators increased systemic inflammatory response. CD50 molecule on the cell membrane of macrophages plays an important regulatory role in the clearance of apoptosis neutrophil, laying the foundation for macrophage identifying, removing apoptosis neutrophils. Therefore, the study of macrophage expression of CD50 in SIRS induced by SAP and its expression in different treatment is significant for treating SAP-induced SIRS and reducing the mortality rate. This experiment is to study the effect of traditional Chinese medicine Yinchen Chengqi Decoction, Dexamethasone and combination of the two drugs on expression CD50 protein in macrophage in SIRS induced by SAP rat model.Methods: SD rat SAP / SIRS model [3] was produced by injecting 1.5% sodium deoxycholate retrogradly into pancreatic duct through duodenal papilla. 75 SD rats were randomly divided into sham operation group (SO), severe acute pancreatitis SIRS model group(SIRS/SAP),Yinchen Chengqitang Decoction treatment group(YCCQT), Dexamethasone treatment group(DEX), traditional Chinese associates with western medicine treatment group (ZXY), dose of the Yinchen Chengqi Decoction was 1ml/100g (ig),while dose of Dexamethasone was 1mg/100g (intraperitoneal injection), the treatment group in two hours and after producing the model with a re-delivery in 12 hours. Model samples were collected after 24h for detection.Pathological changes and pathology score of pancreat were observed with microscope in each group; the positive rate of CD50 in macrophages was detected by flow cytometry; Use western blotting to detect differences of macrophage CD50 protein expression level between each group; the differences of levels of CD50 protein were detected by immunofluorescence.Results:1.Pathological changes of pancreas in each group: (1)compared with SO group, pathological changes in SI Group were significantly obvious (edema: 3.50±0.21 vs 0.290±0.05, inflammation: 2.90±0.15 vs 0, hemorrhage: 2.10±0.18 vs 0, necrosis: 2.80±0.24 vs 0, P <0.01); (2)compared with the SI group,various improvement in treatment groups can be seen with statistical significance (P <0.05), particularly in ZX group (edema: 3.00±0.14 vs 3.50±0.21, P <0.05, inflammation: 1.40±0.23 vs 2.90±0.15, hemorrhage: 0.40±0.15 vs 2.10±0.18, necrosis: 1.40±0.12 vs 2.80±0.24, P <0.01).2.Expression of macrophage CD50 positive rate in Flow cytometry:(1)Compared with SO group, CD50-positive macrophages in SI group was significantly decreased(12.32±8.82% vs 36.52±12.61%, P <0.01);(2) Compared with SI group, macrophages CD50-positive rate slightly increased in YC Group (12.47±4.36% vs 12.32±8.82%, P <0.05), while DE Group(26.98±3.72% vs 12.32±8.82%, P <0.05) and ZX group(35.35±5.74% vs 12.32±8.82%, P <0.01) were significantly increased;(3)Compared within treatment groups: the CD50-positive rate was significantly increased in ZX group than inYC group and DE group (35.35±5.74% vs 12.47±4.36% vs 26.98±3.72%, P <0.05).3.Western Blotting Detection of macrophage CD50 protein expression:(1)Compared with SO group, SI group macrophage expression of CD50 were significantly different (0.47±0.09 vs 0.76±0.15, P <0.01);(2)Compared with the SI group: YC group macrophage CD50 expression levels were significantly different (0.47±0.09 vs 0.55±0.08, P <0.05), DE group macrophage CD50 expression levels were significantly different (0.47±0.09 vs 0.56±0.11, P <0.05), while the ZX group macrophage expression of CD50 were significantly different (0.47±0.09 vs 0.71±0.13, P <0.01);(3)To compare within treatment groups: macrophage CD50 expression levels in ZX group were significantly different (0.71±0.13 vs 0.56±0.11,0.71±0.13 vs 0.55±0.08, P <0.05 )comparing with DE groups and YC group; DE group compared with the YC group, macrophage expression of CD50 significantly different (0.56±0.11 vs 0.55±0.08, P<0.05).4.AU value of macrophage CD50 protein expression detected by Immuno- fluorescence:(1)Compared with SO group, expression of macrophage CD50 in SI group were significantly different (235.44±21.35 vs 354.72±28.46, P <0.01);(2)Compared with SI group: macrophage CD50 expression in DE group were significantly different (235.44±21.35 vs 275.43±30.27, P <0.05), macrophage CD50 expression in YC group are different (235.44±21.35 vs 267.39±25.36, P <0.05), while the macrophage expression of CD50 in ZX group were significantly different (235.44±21.35 vs 305.52±31.14, P <0.01);(3)Compared within treatment groups: macrophage CD50 expression in ZX group were significantly higher than YC group and DE group (305.52±31.14 vs 267.39±25.36,305.52±31.14 vs 275.43±30.27, P <0.05 ); compared with the YC group, macrophage CD50 expression in DE group were higher (275.43±30.27 vs 267.39±25.36, P <0.05).Conclusions:Compared with SO group, SI group has more serious pathological changes in the pancreas.The treatment group have varying degrees of improvement, with the most obvious effect in ZX group, implying that Yinchen Chengqi Decoction alone or Dexamethasone alone has less significant effect on pathological changes of rat pancreas than joint Application of Western and Chinese Medicine. Expression of macrophage CD50 in SI Group significantly decreased, the treatment groups increased in varying degrees, in which Chinese and Western Medicine has better effect on expression of macrophage CD50 protein than the single treatment group, suggesting that the SAP / SIRS model of the membrane of macrophages may be changed so that the CD50 in the surface of macrophage decreased significantly, affecting the apoptosis of macrophages and neutrophils adhesion and cell-to-cell signal transduction, resulting in decreasing phagocytic function of macrophages removing apoptosis neutrophil , and further rise to a large number of neutrophils accumulated in the body to release excessive inflammatory response to increase inflammatory mediators, so that a deterioration in conditions. Therefore, traditional Chinese and western medicine may be effective way to increase the capacity of macrophage phagocytosis and become treatment of SAP caused by SIRS.

Studies on Autophagic Cell Death Induced by TIPs in Human Hepatoma Cells HepG2

Tue, 29 Mar 2011 15:31:49 +0000

AIM:To investigate the mechanism of proliferation inhibition of TIP-6 (7-(4-methoxyphenyl)-5,Sa-diphenyl-1,2,3,7,8,Sa-hexahydroimidazo[1,2-a]pyridine) in human hepatoma cells HepG2 and human normal hepatocyte L02.METHODS:After treatment with varying concentration of TIP-6, Typan Blue Assay and MTT assay was used to determine the proliferation rate treated-cells.The IC50 of cells was figured out.The morphology of cells was observed by the phase contrast microscope.The cell cycle was determined by Flow cytometry(FCM) analysis in treated-cells.Autophagy and autophagic cell death were detected after acridine orange(AO) staining under fluorescent microscopy.The submicrostructure was observed by transmission electron microscope.Apoptosis was analyzed by Annexin V/7-AAD,DAPI staining and DNA agarose gel electrophoresis assays.The expression of nuclear factor-kappa B(NF-κB) was detected by cellular immunochemistry.RESULTS:We found that TIP-6 can induce different cell line cytotoxity. While TIP-6 causes 50%inhibition in HepgG2 at 138.6μmol/ml for 24 hrs,88.6μmol/ml for 48 hrs,52.5μmol/ml for 72 hrs,same inhibition in L02 was caused at 142.5μtmol/ml for 24 hrs,93.3μmol/ml for 48 hrs,59.1μmol/ml for 72 hrs.The result based on Typan Blue Assay shown that TIP-6 can suppress the proliferation of HepG2 cells in a dose- and time- dependent manner.TIP-6 has stronger inhibition in HepG2 than in L02 cells.After treatment with varying concentration of TIP-6 for 24, 48,72 hrs,more vacuoles in cells were found by phase contrast microscope.FCM showed that cells were blocked in G2/M phase.Annexin V/7-AAD and DAPI staining and DNA agarose gel electrophoresis assays indicated that TIP-6 can\'t inhibit proliferation followed by inducing apoptosis in cells.The of AO staining and transmission electron microscope showed that TIP-6 can cause autophagic cell death in HepG2 cells.Moreover,we found that the expression level of NF-κB increased by TIP-6 treatment.CONCLUSION:TIP-6 inhibited cell proliferation,as cells were blocked in G2/M phase.TIP-6 can inhibit proliferation of cells by causing autophagic cell death in HepG2 and L02 cells.The NF-κB pathway may be involved in these processes.

Clinical Study on the Repair of Chronic Skin Ulcer with Tissue-Engineered Acellular Dermal Substitute

Tue, 29 Mar 2011 15:31:47 +0000

Background The chronic ulcer of skin is a common disease, caused by local or systemic factors su ch as High blood sugar,acral ischemia, Neuropathy,Immune abnormalities and infection. people has paid much attention to it because of its seriousity and increasing incidence. Now, the conventional treatment includes autugraft, allograft and xenograft. But such three treatments have their respective defects. The source of autologous skin is very limited and might make new wound surface. Allograft and xenograft cound cause immunological rejection and spread diseases, so their application is restricted. Tissue engineering as a newly emerging science develop rapidly. Construction of tissue engineering skins provides a new method in treatment of the chronic ulcer of skin. Tissue engineering dermal substitutes offer a notable advantage,guaranteeing a greter clinical success in the procedures of wound healing. It cound take up good external appearance after wound healing. Many dermal substitutesa are in development or have been identified by FDA, such as Alloderm produced by Lifecell in 1995 have been applied to clinic widely. But Reports indicated that acellular dermal matrix cound cause immunological rejection and potentially dangerous infectious diseases, it sources are limited. Cell-free synthetic dermal matrix at the lack of extracellular matrix (ECM) components of acellular dermal matrix such as elastic fibers, proteoglycans and glycoproteins, and bovine collagen type I may stimulate the immune response, resulting in reduced graft survival rate. A new type of tissue- engineered acellular dermal substitute has been developed in Tissue Engineering Center of the Fourth Military Medical University. The greatest advantage of tissue-engineered acellular dermal substitute is that the structure of the extracellular matrix is close to natural structure, and its histocompatibilit is better than natural acellular dermal.Objectiv The purpose of the clinical trial is to investigating the clinical effects of tissue-engineered acellular dermal substitute in repairing of chronic skin ulcer, and finding a practical and feasible method of promoting the healing of chronic skin ulcer.Methods 24 patients with chronic skin ulcer in Out-patient clinic and ward were selected and enrolled in this study. After surgical debridement, ulcers (n=30) were randomly divided into experimental group and control group, tissue-engineered acellular dermal substitute was applied directly on the wound in experimental group(n=15) and vaseline dressing was applied on the wound in control group (n=15) respectively.All the patients were conventionally dressed and fixed. We Used Photoshop 8.0 image processing software for calculating ulcer area in each group. Analyzing statistically the healing time in each group (day),calculating and analyzing statistically the percentages of healing ulcer area in each group (%) at 2,4 and 6 weeks after operation. Observing changes in the course of treatment such as local wound inflammatory response, immunological rejection, evaluating qualityof ulcer healing after ulcer healing. We used SPSS 11.0 software for analysing difference between the two groups. Analysis of variance (ANOVA) or T test was applied for measurement date statistics ( x±s).Result The wound healing time in test group(28.13±4.42 days) was significantly shorter than that in control group(41.07±6.31days).The percentages of healing ulcer area in test group was Significantly larger that in control group. There was statistical significant diference between the two groups. There was no evidence of inflammatory response and immunological rejection in test group.Conclusion The chronic ulcer of skin could be repaired by tissue-engineered acellular dermal substitute in promoing the early healing, shortening the ill course and reducing its complications. It is practical and feasible for clinical application of tissue-engineered acellular dermal substitute.

Study of Presurgical Orthopedics in Infants with Unilateral Cleft Lip and Palate

Tue, 29 Mar 2011 15:31:45 +0000

Congenital cleft lip and palate is one kind of most common developmental malformation in oral and maxillofacial region. The incidence rate demonstrates varieties for different ethnicity and areas. In China, the rate is 1.8:1000 (14.01% of total defect rate), secondary only to that of cerebromedullary tube malformation. The etiology of cleft lips; clefts of the lip and alveolus; clefts of the lip, alveolus, and palate; and isolated cleft palates not associated with a specific syndrome is still an important subject of research and so far has not fully been understood. Genetic as well as environmental factors play an important role in the development of a cleft. It might be important to learn about etiological connection between prevalence rate and characteristics of cleft.Because of the limited factors by the social economy and medical condition, the treatments of cleft lip and palate were mostly subjected to single surgery except for partly team approach, such as presurgical orthopedics. A great deal of research reports have revealed that the benefits of presurgical orthopedics were more evident than that of single surgery. In this study, we modified Grason’s model of presurgical orthopedics, that is, presurgical nasolveolar molding (PNAM). On one hand, we designed two nose-supported scaffolds which were put two sides of nasal columella top to support the nasal tip. On the other hand, we momentarily gathered images from the three Dimensional Sensing System (3DSS) to evaluate the treatment effect, especially for those infants. There were two parts in the study.Part 1. Simultaneous lip-nose repair following presurgical nasoalveolar molding for infants with unilateralcleft Iip and palateObjective To evaluate the outcome of presurgical nasolveolar molding (PNAM) therapy in the threatment of patients with cleft lip and palate. To apply a new way to evaluate the outcome of PNAM therapy in the threatment of patients with cleft lip and palate. Methods 10 cases with cleft lip and palate underwent simultaneous lip and nose repair following presurgical nasoalveolar molding.The nasoalveolar molding included gap-closing of alveolar cleft and nose-stent until the age of approximately 3-5 months when the surgery was performed. Student’S t test was used for data analysis with SPSS15.0 software package. Results After nonsurgical nose-alveolar molding in 10 infants with cleft lip and palate about 2-3months, the width of the cleft lips in all cases was significantly narrowed (P<0.01), the soft tissue of both sides of the clefts grew and expanded. The deformity of the columella, the dome,and the wide alar of the nose was significantly reduced following nonsurgical nasoalveolar molding (P <0.01). The width of the alveolar cleft became significantly narrower in these patients (P<0.01). Conclusion In order to obtain satisfactory lip-nose configuration and uninterrupted alveolar bone for unilateral complete cleft lip and alveolus, the presurgical nasoalveolar molding and early simultaneous lip-nose repair are useful in cleft lip and palate team approach.Part 2. Research and analysis of infant cleft lip and palate in north of Shaanxi province.Objective To detect prevalence rate and characteristics of infants cleft by case survey. Methods All 30 patients were newborn infants from hospitals of north of Shaanxi province. The research contents included cleft classification, sex ratio, infected malaria during pregnancy and cleft family history. Results There were 25 unilateral cleft lip (83.67%) and 5 bilateral cleft lip (16.67%), including 24 cleft palate(80.00%). In the group of unilateral cleft lip, the number of left upper lip cleft (16 cases) was more than that of right upper lip cleft (14 cases). The ratio was 1.14. The classification of cleft lip showed that three degree cleft (25 cases, 83.33%) was more than two degree cleft (4 cases, 13.33%) and one degree cleft (1 cases, 3.33%). Three mo thers had medical history of malaria. Most patients (23 cases, 76.67%) were from rural areas, and their parents were mainly illiterate. Conclusion The occurrence of cleft lip and palate in this area might be relevant to family condition, nutrition level an, infection and inheritance during pregnancy, etc.

Expression and Clinical Value of Gastric Cancer-associated Antigen MG7 and COX-2 in Gastric Precancerous Lesions

Tue, 29 Mar 2011 15:31:43 +0000

The development of gastric cancer is a multi-step and multi-factor involved process. The transformation of normal gastric mucosal cells into malignant cells can not be finished at one time, and it may take many steps. Intestinal metaplasia and dysplasia in gastric mucosa are the common steps in the development of gastric cancer, which are known as gastric precancerous lesions.MG7, a monoclonal antibody against gastric cancer was firstly established in our institute, by immunizing the BALB/C mice directly with a poorly-differentiated gastric adenocarcinoma cell line MKN-46-9. The antigen recognized by the antibody is different from other gastrointestinal tumor markers as reported. It has been considered as a promising biomarker in gastric cancer screening because of its high specificity. Prelimary studies had shown that the level of MG7-Ag expression was valuable in predicting and diagnosis of gastric cancer. Cyclooxygenase (COX) is a rate-limiting enzyme controlling the conversion of arachidonic acid to prostaglandinH2. The two major types of COXs are COX-1 and COX-2. The role of COX-2 in inflammation and cancer has been studied intensively. Previous studies revealed that the expression of COX-2 was closely related to the development of gastric cancer, and our laboratory further discovered COX-2 had a positive correlation with MG7-Ag in gastric cancer. However, the relationship between expression of COX-2 and MG7-Ag in gastric precancerous lesions remained unclear.In this study, we investigated the gastric cancer-associated antigen MG7 expression in 334 gastric precancerous lesions and the correlation with expression of COX-2 by immunohistochemical analysis. In addition, a one-year follow-up of 107 patients was conducted to explore the clinical value of combined detection of MG7-Ag and COX-2 in prediction of advances in gastric precancerous lesions.Objectives:To explore the significance of detection of MG7-antigen and COX-2 in gastric precancerous lesions and the correlation between the two molecules. To explore the clinical value of combined detection of MG7-Ag and COX-2 in prediction of advances in gastric precancerous lesions.Material and methods:1.Samples collection501 gastric mucosal biopsies were collected from patients with gastric precancerous lesions(334), superifcial gastritis(59), atrophic gastritis(67), gastric cancer(41) from the Fourth Military Medical University, Tianjing Medical University and Shandong Medical University. 107 patients were followed up for more than one year. According to the dynamic histology, 107 cases were divided into two groups including advanced group(intestinal metaplasia→dysplasia, dysplasia→gastric cancer) and inadvanced group(intestinal metaplasia→intestinal metaplasia, dysplasia→dysplasia).2.Methods1) MG7 hybridoma cells were defrosted and cultured for MG7-Ab preparation.2) Immunohistochemistry was performed by using mouse monoclonal antibody to MG7-Ag and COX-2.Results1. Expression of MG7 antigen and COX-2 in precancerous gastric mucosal tissues.MG7-Ag was mainly located in the cell membrane and cytoplasm. COX-2 was mainly stained in the cell membrane and cytoplasm too.The expression of MG7-Ag and COX-2 was not significantly correlated with age and gender of patients with different precancerous gastric mucosal tissues (P>0.05). The positive rates of MG7-Ag and COX-2 expression were increased gradually with the development of gastric cancer lesions, and the difference was significant (P0.1).Conclusions:We successfully detected the gastric cancer-associated antigen MG7 in gastric precancerous lesions and its correlation with expression of COX-2. In our study, from intestinal metaplasia, low-moderate grade dysplasia to high grade dysplasia, expression of MG7-Ag and COX-2 was increased gradually. The positive rates of MG7 antigen and COX-2 expression in superficial gastritis,atrophic gastritis, intestinal metaplasia, dysplasia and gastric cancer also increased gradually. It indicated that MG7 antigen and COX-2 could be used as predictive factors of cancer development from gastric precancerous lesions. There is a positive correlation between the expression of MG7 antigen and COX-2 in gastric precancerous.Combined detection of MG7-Ag and COX-2 in followed-up patients, double positive staining in gastric precancerous lesions had an increased risk of precancerous progression over 22 times, which implys its great value of prediction of early gastric cancer from precancerous lesions.

Hemodynamic Change of Hepatic Cirrhosis Rat after Orthotopic Liver Transplantation

Tue, 29 Mar 2011 15:31:41 +0000

Aim As a common and frequently-occurring disease, portal hypertension possesses specific regional and systematical hemodynamic characteristic. Final stage benign liver diseases are indication of clinical liver transplantation. Only liver transplantation can cure PHT. There are regional and systematical hemodynamic changes during and after liver transplantation. Study on the hemodynamic changes after liver transplantation in PHT rats can identify the hemodynamic characters before and after liver transplantation. On this foundation, we can investigate the hemodynamic changes in cirrhosis rat after orthotopic liver transplantation. We can investigate the therapic effect and can improve the survival rate of liver transplantation. This study is helpful to enlarge the indication of clinical liver transplantation.Method①100 male SD rats were randomly divided into 5 groups. One group was the control group; The others, as the experiment groups, were investigated after 2,4,7,10 weeks injection with CCl_4 respectively.②The 1st group included 50 male SD rats, as the recipients, which were duplicated the PHT rats model with injected CCl_4 10 weeks. The 2nd group included 30 male SD rats, as the donators, which were bought and fed 3 weeks before liver transplantation. The two groups were paired up together in accordance with their weight. The method of two sleeves anastomoses was performed. 3) Electromagnetic blood flow meter was used to detect the portal venous flow. The ileocolic vein, inferior vena cava and abdominal aorta were pricked respectively, and were connect to electrophysiolograph and pressure transducer, so that the value of portal venous pressure, inferior vena cava pressure and mean arterial blood pressure could be determined.Results①During the process of cirrhosis, the hepatocyte underwent four phases including degeneration, necrosis, fibrosis and pseudolobule proliferation. The value of mean arterial pressure, portal venous pressure,portal venous flow, portal vascular resistance and splanchnic vascular resistance in the rats with carbon tetrachloride injecting 2 weeks were different to it in the control group rats(P<0.05). But injection for 2 and 4 weeks, there were no significant deviation. After injection for 4 and 7 weeks, there were significant deviation in mean arterial blood pressure, portal venous pressure, portal venous flow and portal vascular resistance(P<0.05). But after injection for 7 and 10 weeks, there were no significant deviation, except portal vascular resistance(P<0.05).②After liver transplantation had completed, the value of mean arterial blood pressure and portal venous flow of recipient rose(P<0.05), but the value of portal venous pressure and portal vascular resistance decreased(P<0.05). Conclusions During the duplication of cirrhosis rat model, the portal vascular resistance plays an important role in the forming of portal hypertension. It also affected the hemodynamic changes. The vasoactive substance is important to the portal hypertension forming. The values of portal venous pressure, portal vascular resistance and portal vein flow are decreased after liver transplantation in short term, but the long term effect still need to be studied.

Studies on the Cytotoxicity of Silver Nanoparticles

Tue, 29 Mar 2011 15:31:39 +0000

Silver nanoparticle is clinically used as antibacterial agent because of its good antibacterial activity. However, it is found that nanoAg will also interact with the cell matrix, and subsequently cause the worry that concern to the potential harm of nanoAg products.In order to investigate the cytotocity of nanoAg, the influence of nanoAg on the cell adhesion state, membrane fluidity and integrity of Human umbilical vein endothelial cells (HUVEC) and Human umbilical artery smooth muscle cells (HUASMC) was studied in vitro by means of transmission electron microscopy (TEM), X-ray diffraction (XRD), LDH assay, and fluorescence polarization method. In additional, Raman spectroscopy and UV- Spectrophotometry were used to study the influence of nanoAg on the conformation of protein of both cells, and HSA as well.The results are as follow: (1) Interacting with cells, silver nanoparticles trend to aggregate as clusters and deposit on cell membrane without any change on its chemical state. (2) When the concentration of nanoAg ranged from 0.0039-0.5 mg/mL, no significant acute death of both cells are found, which implies that nanoAg has no acute cytotoxicity. However, nanoAg shows certain cytotoxicity after several hours later, which was verified by following aspects: the adhesion state of both cells became abnormal, the membrane fluidity of the cells decreased, and the integrity of cell membrane was damaged. The effect trends to be non-linear vs. the concentration of nanoAg. (3) Interacting with silver nanoparticles, the comformation of proteins on both cells is changed, which may subsecquently influence the cell proliferation. Therefore, the interaction between silver nanoparticles and proteins may be the molecular mechanism of the cytotoxicity of nanoAg.

The Effect of the Intervention with Unfractionated Heparin (UFH) or Low Molecular Weight Heparin (LMWH) on Fluid-resuscitated Sepsis Rats

Tue, 29 Mar 2011 15:31:38 +0000

Sepsis has become a huge disease burden in today\'s world.Although human have got a deep insight into the pathogenesis of sepsis,there are few effective remedies yet.Since 1990\'s,anticoagulants have aroused a comprehensive interest in the field of sepsis research,which is a result of the achievement that more and more knowledge about the crosstalk between inflammation and coagulation has been obtained.Unfortunately rhAPC is the only anticoagulant recommended by SSC in 2008 as a remedy to improve the survival of patients with severe sepsis.As for heparin,there have been too much debate on the issue of whether such a cheap anticoagulant has the same power as,if not more than,rhAPC to treat sepsis patients,and whether it can improve tissue oxygenation among the sepsis victims.In this article,the assessment of the sepsis rats treated by unfractionated heparin(UFH) about their 7 days\' survival after CLP operation was made in the first part, then organ damage,tissue oxygenation,and histopathology of the sepsis rats treated by UFH or LMWH at the time point of 36 hours after CLP operations were implored in the second part.Section 1 The Effect of unfractionated heparin(UFH) with 3 kinds of dosage on the survival of the sepsis rats during the 7 days after CLP operation.Objective:To evaluate the effect of different dosage of UFH on the 7 days\' survival of the rats after CLP operations.Methods:24 S-D rats were equally allocated into 4 groups at random.Each group received an unique remedy at the time of 12 hours after CLP operations.These remedies comprise a baseline fluid resuscitation(BFR) and three kinds of UFH interventions with different dosage.During the 7 days after CLP operation,all rats\' physical activity and the number of deaths were observed and recorded at 12 hour intervals.All rats underwent autopsy within 8 hours after death.Results:At the time of 144 hours after CLP operation the numbers of survivors of group A,B,C,D are 0,0,2,4 respectively.Statistical analysis of survival rate during 120 hours after CLP operation shows no difference among all groups(P＜0.05),but Log-rank test shows statistical difference of survival curve between two pairs of groups,i.e.group A and D,group B and D.Conclusion:Compared with the BFR intervention and the UFH(300IU/KgBW/d) intervention,the intervention with UFH(1500IU/KgBW/d) can significantly improve the survival of sepsis rats during the 7 days after CLP operations.This benefit may have a positive relevance with the dosage of UFH.Section 2 The influence of the intervention with unfractionated heparin or low molecular weight heparin on the changes of organ function,tissue oxygenation,and histology of the sepsis rats at the time of 36 hours after CLP operations.Objective:To evaluate the influence of the intervention with unfractionated heparin or low molecular weight heparin on the changes of organ function,tissue oxygenation,and histology of the rats at the time of 36 hours after CLP operations.Methods:24 S-D rats were equally allocated into 4 groups at random.Each group received an unique remedy at the time of 12 hours after CLP operations.These remedies comprise a BFR,an intervention with UFH(250IU/KgBW/4h) and an intervention with LMWH(250AXaIU/KgBW/4h).At the time of 36 hours after CLP operations,the parameters of organ function and tissue oxygenation of the sepsis rats were monitored. Afterwards,the rats were killed and fresh liver,kidney,and lung sections were harvested and fixed in 10%buffered formalin.Results:1.Compared with group E,the other 3 groups show significant increase in the value of ALT, BUN,Scr,Lac,Pa-vO_2,O_2ER and significant decrease in the value of ScvO_2. (P＜0.05)2.Compared with group F,group G shows significant increase in the value of PaO_2, Pa-vO_2,,O_2ER and significant decrease in the value of ScvO_2.,Lac.(P＜0.05)3.Group H shows no statistical difference with group F in the value of ALT,BUN,Scr, PaO_2,Lac,PH,Pa-vO_2,O_2ER,ScvO_2.(P＜0.05)Conclusion:1.The rats will develop into the phase of severe sepsis at the time of 36 hours after CLP operations,manifesting organ damage and poor tissue perfusion.2.Compared with the intervention of solitary BFR,the UFH(250IU/KgBW/4h) intervention combined with BFR can significantly increase oxygen extraction ratio(O_2ER) of sepsis rats with a lower level of anaerobic metabolism.3.At the time of 36 hours after CLP operations,among the sepsis rats undergoing the combined intervention of UFH(250IU/KgBW/4h) and BFR,O_2ER manifests a positive relevance with the anticoagulative intensity of UFH.4 No evidence demonstrates that the LMWH intervention(250AXaIU/KgBW,SC,q4h) combined with BFR is superior to the solitary BFR intervention to increase O_2ER of and to alleviate organ damage of sepsis rats at the time of 36 hours after CLP operations.5 Compared with the combined intervention of LMWH(250AXaIU/KgBW,SC,q4h) and BFR,the combined intervention of UFH(250IU/KgBW/4h) and BFR can significantly increase oxygen extraction ratio(O_2ER) of sepsis rats at the time of 36 hours after CLP operations.

Significance of Thromboelastogram in Coagulation of Patients with Severe Traumatic Brain Injury

Tue, 29 Mar 2011 15:31:17 +0000

Objectiv To investigate parameters and figures of thrombelastogram of coagulation in patients with severe traumatic brain injury(STBI).Methods Three groups were studied:(A)patients with severe isolated traumatic brain injury(n=31).They had no operations.CTs measured when patients were just in hospital supported that they were with severe isolated traumatic brain injury.(B) patients with general trauma without TBI(n=6).(C) healthy volunteers(n=18).TEG parameters of A and B groups were measured in 24 hours,72 hours and a week.And C group as control group only measured one time.The parameters contained R(time of reaction)、K(coagulation time)、Ma(maximum amplitude of thrombus)、α(solidification horn)、APTT(activated partial thromboplastin time)、TT(thrombin time)、PT(prothrombin time)、FIB(fibrinogen) and PLT(platelet count).Results The results revealed the hypercoagulabllity was in the patients with severe isolated TBI in 24 hours and the hypocoagulability was in 72 hours.It was statistically significant.It was different between A group and B group and C group.1.here are hypercoagulability in 24hours,but the state of low coagulation in 72hours, and almost normal in a week in A group.2.It is different betweenA and B groups in 3 days but consistent with eath other in a week.Conclusions1.There are obvious coagulation abnormalities in early stage of patients with severe traumatic brain injury.2.We could find a process of coagulation abnormalities of patients with severe traumatic brain injury is from hypercoagulability in 24 hours to the state of low coagulation in 72 hours and then returned to normal in a week.3.TEG Comparison of A group and B group shows that coagulopathy is specific and regularity.4.There is correlativity between Prognosis of the patients with severe isolated TBI and GCS scores.

Study of Complement and Regulatory T Cells of the Patients with Immuno-related Pancytopenia

Tue, 29 Mar 2011 15:31:15 +0000

Objective:To investigate the bone marrow complement levels,the quantity and function of regulatory T cells in the IRP,then expore the mechanism of hematopoietic cells destruction and significance of regulatory T cells in the pathogenesis of IRP.Methods:Fifty-nine untreated IRP patients,65 recovered IRP patients and 24 healthy donors as controls were enrolled in this study.The bone marrow levels of CH50,C3, C4,C5b-9,IL-2,TGF-βwere tested with ELISA.The auto-antibodies on bone marrow hematopoietic cells(BMHC)and the quantities of regulatory T cells(CD4~+CD25~+/CD4~+ cells,CD4~+CD25~+CD127~-/CD4~+ cells)were examined with flow cytometric assay.The expressions of FoxP3 and Galectin-10 mRNA in BMMNC were measured with semiquantitive RT-PCR.Results:1.The level of C5b-9 in bone marrow of untreated IRP patients(119.82±53.95)was significiantly higher than that of recovered IRP patients(100.74±33.42)or normal controls(93.86±28.81)(p＜0.05),and there was no significant difference between recovered IRP patients and normal controls(p＞0.05);The levels of CH50 in bone marrow of untreated or recovered IRP patients(33.29±11.51,30.77±10.34)was significiantly higher than that of normal controls(24.09±6.37)(p＜0.05),and there was no statistic significance between untreated IRP patients and recovered IRP patients (p＞0.05);The level of C3 in bone marrow of untreated or recovered IRP patients (4.90±2.19,4.95±3.47)was significiantly lower than that of normal controls (6.98±5.56)(p＜0.05),and there was no statistic significance between untreated IRP patients and recovered IRP patients(p＞0.05);The bone marrow level of C4 among three groups was no statistic significance(p＞0.05);The level of complement in peripheral blood was consistent with in bone marrow.Bone marrow CH50 level of IRP patients was negatively correlated with C3 level(r=-0.303,p=0.007)and positively correlated with C5b-9 level(r=0.241,p=0.003).IgG autoantibody accounted for 55.17%,IgM 54.31%,IgG+IgM 31.90%in all patients,BMHC-IgM positive group accounted for 86.21%in all patients.The bone marrow level of C5b-9 in IRP patients with BMHC-IgM(117.59±55.69)was higher than that in IRP patients without BMHC-IgM(99.15±26.15)(p＜0.05).The positive rate of CD34~+-IgG or CD34~+-IgM of IRP patients was positively correlated with their C5b-9 levels(r=0.593 p=0.000,r=0.326 p=0.049).The percentage of reticulocyte(r=0.421,p=0.000)and the serum level of indirect bilirubin of IRP patients were positively correlated with their CH50 levels(r=0.230,p=0.032).After treatment,the percentage of reticulocyte reduced to normal(p=0.038)and the quantity of leukocyte increased to normal(p=0.002).The bone marrow level of C3 was higher than that of before treatment(p=0.003);and the positive rate of CD34~+-IgG or CD34~+-IgM was lower than that of before treatment(p=0.016,p=0.022).2.The bone marrow levels of IL-2 in untreated or recovered IRP patients(5.64±1.70, 6.19±2.53)was significiantly lower than that in normal controls(7.91±3.71)(p＜0.05); The bone marrow levels of TGF-βin untreated or recovered IRP patients(1.79±0.67, 1.86±0.77)was significiantly lower than that in normal controls(2.48±0.94)(p＜0.05), and there was no statistic significance between untreated IRP patients and recovered IRP patients(p＞0.05);The quantity of CD4~+CD25~+/CD4~+ cells in untreated IRP patients(22.46±9.47)was significiantly lower than that in normal controls (30.59±8.58)(p＜0.05);The quantity of CD4~+CD25~+CD127~-/CD4~+ cells in untreated IRP patients(7.18±2.72)was significiantly lower than that in normal controls (10.44±3.24)(p＜0.05),and there was no statistic significance between recovered IRP patients and normal controls(p＞0.05).The relative mRNA expressions of FoxP3 and Galectin-10 were 0.34±0.25,0.69±0.51,0.82±0.65 and 0.66±0.11,0.74±0.11, 0.76±0.09 in three groups.The expression of untreated IRP patients was significiantly lower than that of recovered IRP patients or normal controls(p＜0.05).Conclusions:The destruction mechanism of hematopoietic cells of IRP patients especially with BMHC-IgM have some relationship with auto-antibody activating complement.The regulatory T cells play important role in the pathogenesis of IRP.

Study of Hypomethylation of CAGE, MAGE-A1 and MAGE-A3 Promoter in Gastric Carcinoma

Tue, 29 Mar 2011 14:35:27 +0000

Objective:DNA methylation,as an important epigenetic modification,plays an important role in tumorigenesis of gastric cancer.Aberrant DNA methylation,such as the hypermethylation of tumor suppressor genes and the hypomethylation of oncogene,is recognized as the main mechanisms of tumor suppressor gene inactivation and oncogene activation.Therefore,it is important to study aberrant DNA methylation for tumor prevention and treatment.CAGE,MAGE-A1 and MAGE-A3 are included in cancer/testis antigen genes.The hypomethylation of the genes promoter has been reported in some tumor tissues and is related to the occurrence and development of tumor.The objectives of the study were to reveal the relationship between the hypomethylation of CAGE,MAGE-A1,MAGE-A3 promoter CpG island and clinicopathological features and to explore the role of the hypomethylation of CAGE, MAGE-A1,MAGE-A3 promoter CpG island in earlier and development of gastric carcinoma,in order to search new molecular markers for cancer detection.Materials and Methods:1.The endoscopic biopsy specimens were collected randomly from 25 normal control,32 premalignant lesions and 30 gastric carcinoma in Department of Gastroenterology.The specimens were divided into three groups by pathological results including normal control,premalignant lesions and gastric carcinoma.25 cases of normal control included 14 males and 11 females,average age 52 years old. 32 cases of premalignant lesions included 20 males and 12 females,average age of 56 years old.30 cases of gastric carcinoma included 18 males and 12 females,average age of 58 years old.Gastric carcinoma group,according to different histological types, were divided into 3 groups including adenocarcinoma,signet-ring cell carcinoma and undifferentiated carcinoma;according to different differentiation degree,were divided into 3 groups including well-differentiated,moderately-differentiated and poorly-differentiated;according to different lymphatic metastasis,were divided into 2 groups including lymphatic metastasis positive and lymphatic metastasis negative; according to clinical stages,was divided into 2 groups includingⅠ+ⅡandⅢ+Ⅳ.2.DNA was extracted from all tissues and then treated by Sulfite.The hypomethylation status of CAGE,MAGE-A1 and MAGE-A3 promoter was detected by MSP. Results:1.The hypomethylation of the CAGE,MAGE-A1 and MAGE-A3 promoter 1.1 The positive rate of hypomethylation of CAGE promoter was 4.0%(1/25), 37.5%(12/32) and 80.0%(24/30) in normal control,premalignant lesions and gastric carcinoma respectively.There was significant difference among three groups (P＜0.05).1.2 The positive rate of hypomethylation of MAGE-A1 promoter was 0.0%(0/25), 21.9%(7/32) and 60.0%(18/30) in normal control,premalignant lesions and gastric carcinoma respectively.There was significant difference among three groups (P＜0.05).1.3 The positive rate of hypomethylation of MAGE-A3 promoter was 8%(2/25), 12.5%(4/32) and 46.7%(14/30) in normal control,premalignant lesions and gastric carcinoma respectively.The positive rate of hypomethylation in normal mucosa and premalignant lesions was significantly lower than that of the gastric carcinoma tissues (P＜0.05),but the difference between the normal mucosa and the premalignant lesions was not statistically significant(P＞0.05).1.4 Among the 30 gastric carcinoma tissues,more than one gene promoter hypomethylation were found in 27 eases;more than two genes promoter hypomethylation were found in 23 cases;three genes promoter hypomethylation were found in 7 cases.Therefore,the combined hypomethylation analysis of the genes promoter may enhance the efficiency of detecting the hypomethylation.2.The relationship between the hypomethylation of CAGE,MAGE-A1,MAGE-A3 promoter and the clinicopathologic features.2.1 The relationship between the hypomethylation of the genes promoter and the differentiation:The positive rate of hypomethylation of CAGE promoter in well-differentiated, moderately-differentiated and poorly-differentiated groups was 62.5%, 81.8%and 90.9%respectively.No statistical significance was observed among three groups(P＞0.05).The positive rate of hypomethylation of MAGE-A1 promoter was 25.0%,63.6%and 81.8%respectively.The positive rate of hypomethylation of MAGE-A3 promoter was 12.5%,45.5%and 72.7%respectively.There was significant difference among three groups.(P＜0.05)2.2 The relationship between the hypomethylation of the genes promoter and the lymphatic metastasis:The positive rate of hypomethylation of CAGE promoter in the lymphatic metastasis positive and negative groups was 94.7%and 54.5%respectively. There was significant difference between two groups.The positive rate of hypomethylation of MAGE-A1 promoter was 68.4%and 45.5%respectively.No significant difference was observed(P＞0.05).The positive rate of hypomethylation of MAGE-A3 promoter was 63.2%and 18.8%respectively.There was significant difference between two groups.(P＜0.05).2.3 The relationship between the hypomethylation of the genes promoter and the TNM stage:The positive rate of hypomethylation of CAGE promoter inⅠ+ⅡandⅢ+Ⅳgroups was 58.3%and 94.4%respectively.The positive rate of hypomethylation of MAGE-A1 promoter inⅠ+Ⅱgroup andⅢ+Ⅳgroup was 25.0% and 83.3%respectively.There was significant difference betweenⅠ+ⅡandⅢ+Ⅳgroups.The positive rate of hypomethylation of MAGE-A3 was 33.3%and 55.6% respectively.No significant difference was observed(P＞0.05).2.4 No statistical significance was observed between the positive rate of hypomethylation of CAGE,MAGE-A1,MAGE-A3 promoter and age,sex and tumor histological type(P＞0.05).Conclusions:1.The hypomethylation of CAGE,MAGE-A1 and MAGE-A3 promoter was significantly increased from normal mucosa,premalignant lesion to gastric carcinoma. The hypomethylation of CAGE,MAGE-A1 and MAGE-A3 promoter may occur in the early stage of carcinogenesis and plays a crucial role in the occurrence and development of gastric cancer.2.Hypomethylation of CAGE promoter was significantly correlated with lymphatic metastasis and TNM stage;Hypomethylation of MAGE-A1 promoter was significantly correlated with the differentiation and TNM stage;Hypomethylation of MAGE-A3 promoter was significantly correlated with the differentiation and lymphatic metastasis.3.The combined hypomethylation analysis of CAGE,MAGE-A1 and MAGE-A3 promoter may enhance the efficiency of detecting the hypomethylation.

Study of Activation of Akt1 in Gastric Adenocarcinoma and Moudulation in Human Gastric Cancer Cells SGC7901

Tue, 29 Mar 2011 14:35:14 +0000

Objective:Gastric cancer is the most common malignant tumor in digestive system,which doesn\'t have special symptom in early stage,and most of which is advanced stage when discovered,patients will die in a short time.Although there are many methods to treat it,such as radiotherapy,chemotherapy and so on,except early gastric cancer can be healing well by surgery,others are not satisfied.In recent years, With the development of molecular biology and molecular genetic,it has been found a new therapy,that is to say gene therapy,and it has been a focus to study.Akt is the key gene in PI3K/Akt survival signal pathway,which regulate the cell growth,survival,proliferation,invasion and apoptosis through phosphorylate downstream substrate,and abnormal activation of Akt closely relates to the genesis and development of tumor.Methods:The present study focused on the abnormal expression of Akt signal pathway in gastric adenocacinoma,and RNA interference technology was used to detect the effect of malignant phenotype of gastric cancer when it is abnormally actived.The study was divided into 3 parts.In the first part,using tissue microarray and immunohistochemistry to detect the expression of Akt1 and the members in the signal pathway,found the abnormal expression of MMP2,Ki-67,Bcl-2 and PTEN.In the second part,RNAi technology was used to observe its inhibitory effect on the growth of human gastric cancer cells SGC7901.SiRNA targeting Akt1 was transfected into SGC7901 cells mediated by oligofectamine.Akt1 expression were detected by realtime PCR and western blotting technology after transfection.The expression of other main members in Akt1 patheway were also studied The phenotypic change of SGC7901 cells including proliferation,apoptosis and invasive ability was studied by MTT assay,annexin V staining,flowcytometry and Matrigel 3D growth experiment.In the third part,subcutaneous SGC7901 gastric cancer model was established in nude mice.The mixture of oligofectamine and siRNA was injected into the tumors and the tumor volumes were measured.And the expression of Akt1 and other members in Akt1 pathway were studied by immunohistochemistry after resected. Apoptosis in tumors were detected by TUNEL method..Results:The first part:The expression of Akt1,MMP2,Ki-67,Bcl-2 in gastric carcinoma were upregulated,and,high expression was observed in poor differentiation. gastric carcinoma.However,the expression of PTEN was reversed.The second part:The expression of Akt1 was obviously knocked down after transfection,and the expression of other members were also decreased.The result of flowcytometry revealed that apoptosis cells increased,cell cycle was arrested in G0/G1phase.MTT assay indicated that cell proliferation rate was inhibited obviously,matrigel experiment and transwell assay showed that cell invasive ability was attenuated,cell immigration decreased dectected by scrach assay.The third part:The tumors in mice treated with siRNA targeting Akt1 grew slowly The expression of Akt1 and other members were decreased detected by immunohistochemistry.Meanwhile,cell apoptosis was increased.Conclusion:1.Abnormal expression of Akt1 and other members in the pathway lead to cell proliferation and invasion,block the apoptosis of cell.2.Using RNAi technology,siRNA targeting Akt1 mediated by oligofectamine efficiently knocks down the expression of Akt1,inhibits activation of Akt1 pathway, results in decrease of cell proliferation activity and invasive ability,arrests cell cycle and induces apoptosis.3.The established subcutaneous SGC7901 gastric cancer models in nude mice are treated with siRNA targeting Akt1.The tumor growth is inhibited and cell apoptosis is induced.4.Akt1 can be the candidate genes for gene therapy of gastric cancer.RNAi is a new gene silencing technology with high efficiency,which will have potential application prospects in clinics.

Study on the Effect of Demethylzeylasteral to Bladder Cancer Cell Line T-24

Tue, 29 Mar 2011 14:35:07 +0000

Background:Bladder cancer is the most common malignant tumor in urology system. the postoperative recurrence rate is very high,while retaining the bladder after surgery for about 50%-70%of patients with postoperative recurrence.Urology by scholars at home and abroad over the years to explore,discover Intravesical chemotherapy drugs regularly can be effective in reducing and delaying the recurrence of bladder tumor,improve the survival rate of patients.Therefore,the use of chemical treatment before and after surgery,as well as the use of advanced bladder cancer chemotherapy are the current treatment of bladder cancer treatment and the focus of the study.Choose the appropriate chemotherapy is the current treatment of bladder cancer hot spots.In recent years,the role of anti-tumor Tripterygium wilfordii has been more and more attention,but because of the complex components of Tripterygium wilfordii people eager to be separated from the monomer composition effective to find an ideal anti-tumor monomer.The topics on the latest extract of Tripterygium wilfordii demethylzeylasteral(Dzy,T-96) on the role of bladder cancer study.Part OneStudy on the toxic effect of demethylzeylasteral to bladder cancer cell line T-24Objective:Investigate the toxic effect of demethylzeylasteral to bladder cancer cell line T-24.Select certain concentration of demethylzeylasteral for the next studies.Methods:Cytoactivaty of T-24 cells were measured after treated with varies concentrations of THP and demethylzeylasteral with the method of MTT.The index of cytotoxicity(IC) was calculated.Results:IC of THP and demethylzeylasteral were increased with the increase of the concentration.A " flat phase" turns up after a certain concentration both in the group of THP(lug/ml).Conclusion:Take the pharmacokinetics account,two different concentrations of each drug were selected for the next studies.Part TwoStudy on the cytotoxicity of demethylzeylasteral to bladder cancer cell line T-24Objective:Investigate the dose-effect and time-effect connection of THP and demethylzeylasteral to bladder cancer cell line T-24.Whether there is a significant difference between the cytotoxicity of THP and demethylzeylasteral.Methods: Cytoactivity of T-24 cells were measured after treated with varies concentrations index of cytotoxicity(IC) was calculated,too.Results:IC of THP and demethylzeylasteral were increased with the increase of treated time.There is a significant difference between IC of the lower and higher concentration group of THP and demethylzeylasteral on the third day(lower group:0.624±0.035 vs 0.563±0.032,P=0.015;higher group:0.792±0.026 vs 0.672±0.028,P＜0.001 ).There is no significant difference between IC of the lower and higher concentration group of THP and demethylzeylasteral on the first and second day(Lower group:the first day:0.152±0.063 vs 0.127±0.055,P=0.456,the second day 0.504±0.031 vs 0.507±0.042,P=0.879;Higher group:the first day:0.266±0.068 vs 0.231±0.057, P=0.256,the second day 0.585±0.053 vs 0.594±0.039,P=0.660).Conclusion: Whether the lower or higher concentration groups of THP and demethylzeylasteral have a similar cytotoxicity to bladder cancer cell line T-24.Part ThreeThrough Annexin V and PI double staining,using flow cytometry to study the way of demethylzeylasteral effect on bladder cancer ceU line T-24Objective:study the way of demethylzeylasteral effect on bladder cancer cell line T-24(to kill cells or induce apoptosis),explore the possible mechanisms of demethylzeylasteral effect on tumor cells.Methods:Through Annexin V and PI double staining After treated with THP and demethylzeylasteral.Then,screened by flow cytometry to distinguish among them were killed or apoptosis of cells.Result: Demethylzeylasteral:Lower group:Necrotic cells8.29%±1.03,Apoptotic cells 14.37%±2.50;Higher group:Necrotic cells 27.935%±3.80,Apoptotic cells 55.105%±9.75;THP:Lower group:Necrotic cells 64.41%±15.53,Apoptotic cells 2.16%±1.91;Higher group:Necrotic cells 96.25%±2.28,Apoptotic cells 2.81%±2.49 Conclusion:demethylzeylasteral mainly by inducing cell apoptosis pathway effect on bladder cancer cell line T-24.Part FourStudy on the relationship of demethylzeylasteral with bax and caspase-3 gene in inducing bladder cancer cell line T-24 apoptosisObjective:study on the relationship of demethylzeylasteral with bax and caspase-3 gene in inducing bladder cancer cell line T-24 apoptosis.Methods:Use Immunohistochemical method,detect qualitatively the expression of bax and caspase-3,after treated with demethylzeylasteral.Compare with treated with THP and nothing.Result:bax and caspase-3 expression was positive in T-24 cell after treated with demethylzeylasteral.Bax and caspase-3 expression were positive after treated with THP.Conclusion:demethylzeylasteral induce T-24 cell apoptosis, Activate bax and caspase-3 gene expression may be possible mechanisms.

The Application of Fluorescence in Situ Hybridization in the Diagnosis of Urothelial Carcinoma

Tue, 29 Mar 2011 14:35:03 +0000

Objective:To establish the experimental platform of Fluorescence in situ hybridization(FISH) in detecting urothelial carcinoma.Detect the chromosome abnormalities in the urine cells of patients with urothelial carcinoma.To discuss the value of FISH as a non-invasive diagnosis method for urothelial carcinoma.Methods:Urine samples of both suspected urothelial carcinoma patients and healthy controls were collected,urine cytology and FISH were synchronously checked.FISH were used to detect the abnormalities of chromosome 3,7,17 and 9p21(p160.100 suspected patients of urothelial carcinoma and 20 healthy controls were enrolled in this experiment.Data of the healthy group were collected to establish the threshold and patients of carcinoma were diagnosed by this threshold.Through the comparison and statistical analysis between the results of the experiments and pathology,we concluded the sensitivity and specificity of the two methods,as well the difference between high grade and low grade carcinomas.Results:Urine samples of patients with urothelial carcinoma have obviously increased chromosome abnomilities compared with the control group.After the threshold were established,results of FISH and urine cytology were compared.The sensitivity and specificity of urine cytology were 71%and 80%respectively,the sensitivity and specificity of FISH were 88%and 80%respectively,the sensitivity of FISH is higher than urine cytology.As for the low grade urothelial carcinoma,the sensitivity of urine cytology and FISH were 49%and 78%respectively.And for high grade urothelial carcinoma,the sensitivity of urine cytology and FISH were 88%and 96% respectively.The receiver operating characteristic(ROC) curve was drawn using the data of sensitivity and specificity,the areas under the curve were 0.758 and 0.842 for urine cytology and FISH.Conclusions:FISH is a valuable and non-invasive experiment for patients who is suspected to have urothelial carcinoma.The overall sensitivity of FISH is higher than urine cytology, the specificities are the same.Especially for the low grade urothelial carcinomas, while the sensitivity of urine cytology is low,FISH has a much higher sensitivity than urine cytology.With this opportunity,FISH can be a good supplementary.Among the many urine markers used for clinical use,FISH is the one that has both good sensitivity and specificity.Moreover,it\'s an non-invasive process.FISH has considerable value in the diagnosis and follow-up for urothelial carcinoma.

The Expression of TSLC1 in Bladder Transitional Cell Carcinoma

Tue, 29 Mar 2011 14:35:01 +0000

Objective:To master the preparation technology of monoclonal antibody against TSLCl,after prepqration of monoclonal antibody agaist TSLCl,we apply immunohis-tochemistry dyeing technology with the sample we have chosen,whih is simple and cheap.taken together,studying the relationship between the occuring and developing of BTCC and the abnormal expression of TSLC1 protein.Methods:At first,we immunize Balb/c female mice,and preparation of monoclonal antibody agaist TSLC1 by spleen cell and myeloma cell fusion technology.We use ELISA to determine the potency.Meantime,we choose 65 BTCC pathology wax objects,40 of 65 wax objects are low malignant tumor,the others are high malignant tumor.Classified by clinical staging,the 25 wax objects are Ta-1 tumors,the 20 wax objects are T2 tumors,the 20 wax objects are T3-4 tumors.And we also choose 20 normal mucous membrane of urinary bladder.Then we start to determine the expression of TSLC1 protein with immunohistochemistry technology(sp method).The pathology doctors help us to analyze the dyeing states.they determine three states:negative(-)、weakly positive(+)、strongly positive(++).We also use IPP6.0 to quantitative analysis with the positive expression cells.At last,we use SPSS13.0 to analyze the TSLC1 protein expression results of different clinical staging and different grading.Results:We use ELISA to detect serum TSLC1 polyclonal antibody,when dilution is 1:8000,the OD value is 1.355,much higher than the negative group(OD=0.545),it means good effect on mice immune.After cell fusion,we get twenety-two positive clones.Strictly limited dilution to clone,finally retain six hybridoma cell lines,which have the stability of a high titer of McAb secretion.they are AD6,BC10,CB7,CE8,DC5,DF4,respectively.Then we determine antibody subtype,the result reals:AD6 and CE8 belong to IgGl;BC10 and DF4 belong to IgG2a;CB7 and DC5 belong to IgG3.Check AD6 cells cultured for amplification,10~7 cells injected in mice to produce ascites to prepare IgG1,After purification,of the post-test titer was 1:100000.The results of immunohistochemistry technology are the TSLC1 protein expression in cell plasma membrane.High malignant BTCC samples,the weakly positive(+) is only 12%(3/25),most others are negative(22/25);Low malignant BTCC samples,the weakly positive is 75%(30/40),the others are negative(10/40),the difference of two groups has statistical meaning (P<0.05.what is more,with all normal mucous membrane of urinary bladder,the TSLC1 protein express in all samples,and they are strongly positive(++),therefore,the normal mucous membrane of urinary bladder is obviously different from the BTCC samples.We also find that the TSLC1 protein expression has relationship with the clinical staging,the weakly positive of Ta-1 is 92%(23/25),the weakly positive of T2 is 60%(12/20),the weakly positive of T3-4 is 10%(2/20),the differences of three groups also have statistical meaning(P<0.0167).Meantime,we use IPP6.0 to quantitatively analyze them,in the grading,the PI of low malignant samples is 68.55±10.50%,the PI of high malignant samples is 20.65±5.52%;PI and pathological grading of BTCC show a negative correlation(r= -0.905,P=0.000).in clinical staging,PI of the Ta-1 samples is 82.15±5.12%,PI of the T2 samples is 60.55±10.25%, PI of the T3-4 samples is 18.25±6.58%,PI and clinical staging of BTCC also show a negative correlation(r=-0.932,P=0.000).Conclusion:We mastered the preparation technology of monoclonal antibodies against TSLC1,and which may be used to detect the biological behavior of BTCC,the staining results are satisfactory.By immunohistochemistry,we consider the low TSLC1 protein expression or null is associated with the happening and developing of BTCC,and the positive expression of TSLC1 protein(PI) in BTCC showed a negative correlation with pathological grade and clinical stage,so in determining the malignant degree of BTCC,it has important clinical value,and can also be used to evaluate the prognosis of BTCC.It is a valuable marker in the biological behavior of BTCC.

Research on Clinicopathological Features and Related CK19 Expression of Malignant Thyroid Nodules

Tue, 29 Mar 2011 14:34:58 +0000

Objective:1.To improve the diagnosis and surgical treatment of patients with thyroid nodule;2.To assese the diagnostic value of immunohistochemical staining of CK19 on thyroid nodule tissues at differentiating between benign and malignant thyroid nodules;Methods:1.Clinical and pathological characters of 2844 cases with thyroid nodule which underwent operation in Tianjin Medical University General Hospital during 1999-2008 were retrospectively analyzed to improve the accuriety of differentiat diagnose of benign and malignant thyroid nodule,and to improve the treatment.2.The expression of CK19 were examined immunohistochemically in 40 cases of nodular goiter and 40 cases of papillay thyroid carcinoma.Results:1.2226(78.27%) cases were diagnosed as nodular goiter,300(10.55%) as thyroid malignant tumor,258(86.0%) cases as papillary thyroid carcinoma of thyroid. The ratio of benign to malignant is 8.48:1 and male to female is 1:3.38.Malignant thyroid nodules from benign lesions combined accounted for 34.33%(103/300),the combined 85 cases of nodular goiter,nodular goiter with malignant transformation rate of 3.68%(85/2311).2.The average age of malignant cases is signifcantly younger than that of benign cases(P＜0.05).Follicular thyroid cancer was the minimum age (34.43 years) in malignant cases,papillary carcinoma(44.47 years),medullary carcinoma(47.4 years) and undifferentiated carcinoma(56.87 years) followed by increased,those groups were significant differences(p＜0.05).3.1 year incidence of malignant nodules surgery rate(43%) higher than benign nodules(34.63%).From 2000 to 2008,the course is less than 1 year of operation the number of cases(142→390) and surgical cases each year a number of malignant nodules(25→56) gradually increased,a positive correlation(p＜0.05).However,no correlation between the proportion of that(70.65%→70.40%,12.44%→10.11%).4.The major reason of treatment is finding of neck masses,malignancy group were significantly more than the benign group in emergence of preoperative hoarseness,cough,neck lymph nodes, hyperthyroidism,abnormal thyroid antibodies,the second surgery(P＜0.05).5.The reoperation rate of malignant thyroid nodules is significantly higher than that of benign nodules.6.in malignant group the number of nodules,the ratio of solid nodules,the number of enlarged lymphonodes are significantly higher than that in benign gruop,differentiated thyroid cancer is less than 4cm in size-based,accounting for 39.22%(91/232),and other malignant nodules greater than 4cm in diameter significantly larger number of cases,the two groups was statistically differences (P＜0.05).7.Differentiated thyroid cancer were more in the middle size(2cm-4cm between),and malignant nodules larger more than.8.Unilateral lobectomy plus isthmusectomy is the major procedure in malignant nodules cases(25%),and the rest from more to less for Unilateral lobectomy,local mass excision,the entire thyroid gland or near-total resection,subtotal lobectomy,glandular leaves some or subtotal, etc.2003 for the sector,compared before and after 5 years less than the affected gland excision leaf,Unilateral lobectomy plus isthmusectomy and near total or total thyroidectomys were have significant difference(p＜0.05).9.The positive rate of CK19 histoimmunochemical detection in malignant thyroid was significantly higher than that of benign ones(P＜0.01).Conclusions:1.Surgical treatment of t malignant nodules cases accounted for 10.55 percent,more women than men(1:3.38),in the majority of papillary carcinoma.More than half of the malignant transformation from benign disease,Malignant transformation rate of nodular goiter was 3.68 percent.2.High incidence of follicular thyroid cancer in 20 to 40 years old,papillary carcinoma and medullary carcinoma of the high incidence of age group 30 to 50 years old,the other of the high incidence of thyroid malignancy in the age of 60 years old.3.Although the main manifestations of malignant nodules as found in the neck tumor,but which is often accompanied by hoarseness,cough,neck lymph nodes,hyperthyroidism,abnormal thyroid antibodies etc.Ultrasonography indicate single,solid mass is also risk factors.Thus treatment of malignant nodules was significantly higher than that of benign nodules.4.Data in this group prompted the incidence of thyroid nodules may increase,but it did not increase the proportion of malignant.5.Malignant nodules higher reoperation rate showed that preoperative diagnosis of the disease is still difficult.6.Thyroid ultrasonography with high specificity and diagnostic accuracy,should be as routine examination.ECT, MIBI,thyroid function,antibody measurement diagnostic examination of thyroid nodules in the application of more and more.Intraoperative frozen examination of the diagnostic value is the highest,but the role of MIBI to be further assessed.7.For malignant thyroid nodules,Unilateral lobectomy plus isthmusectomy is the most used technique,“not less than lobectomy gland”More and more used in the treatment of malignant thyroid nodules.8.CK19 is a very valuable indicator in the diagnosis of pathological of papillary thyroid carcinoma and is a good marker in distinguishing papillary thyroid carcinoma and papillary hyperplasia.

The Research of MicroRNA-20a Expression in Colorectal Carcinoma and Its Pathoclinical Relevance

Tue, 29 Mar 2011 14:34:52 +0000

Objective To detect the miR-20a expression in the colorectal carcinoma and normal colorectal tissue,and analyze the differences of miR-20a expression between the colorectal carcinoma and normal colorectal tissue,the clinical stages and the degree of differentiation.To investigate the mechanism of miR-20a in the process of the tissue malignant transformation.Method Collect 46 specimens of the colorectal carcinoma after operation,and detect the expression of miR-20a in the carcinoma tissue(T) and the normal colorectal tissue(N) by fluorescence in situ hybridization.Statistically analyze the relevance between the miR-20a expression and the pathoclinical characteristics.Results 1.The positive rate of miR-20a expression in colorectal carcinoma is 63.04%,and that of colorectal normal tissue is 23.91%.There is significant difference between them by the statistical analysis(P＜0.01).2.The positive rate of miR-20a expression in colorectal carcinoma of Dukes stage A+B,C and D period are 53.85%,78.57%and 66.67%,respectively.There is no statistical difference among the Dukes\' staging groups of colorectal carcinoma(P＞0.05).3.The positive rate of miR-20a expression in the well differentiated, moderately differentiated and poorly differentiated colorectal carcinoma tissues are 66.67%and 77.27%and 44.44%,respectively.There is no statistical difference among the each differentiation group of colorectal carcinoma(P＞0.05).Conclusions 1.The miR-20a expression in the colorectal carcinoma is higher than that of the colorectal normal tissue.It points to that in the colorectal carcinoma, the miR-20a with high expression negatively regulate E2F-1 expression,and it leads to the loss of E2F-1-induced apoptosis and then the carcinoma continues to progress.2.There is no statistical difference among the miR-20a expression of Dukes staging groups and each differentiation group of colorectal carcinoma.It prompts that miR-20a may be an important regulatory factor in the process of tumorigenesis,while it is nothing to do with the progress and tumor differentiation.

The Significance to Detect Twist、P53、E-cadherin Gene in Esophagus Carcinoma

Tue, 29 Mar 2011 14:09:18 +0000

Objective:To detect the expression level of Twist in esophageal cancer,adjacent and normal tissues.At the same time,P53,E-cadherin protein are detected together, further to explore the relationship of Twist and P53,E-cdherin protein with clinic-pathological indicators,and its function and mechanism at the diagnosis and prognosis of malignant tumors.Methods:30 cases of surgical resection specimens in patients of esophageal cancer with pathological diagnosis,which contains 1 case of adenocarcinoma,29 cases of squamous cell carcinoma;Esophageal cancer and adjacent tissues were collected in each of patients,adjacent tissues from the cancer area at the edge of 3-5 cm between the organization,simultaneously to select 10 cases normal esophageal tissues of innocent disease of esophagus in the same period of hospitalization as compared group,specimens were placed in 10%neutral formalin fixed and buried in paraffin, 4μm serial sections standby;All specimens were examined respectively Twist,P53 and E-cadherin protein by SABC immunohistochemical method,and the experimental results were dealed by spss15.0 statistical data analysis package.Results:Twist protein expression in esophageal cancer tissue was significantly higher than adjacent and normal esophageal tissues,adjacent tissues and normal tissues were not significantly different in expression levels;at different depth of tumor invasion,lymph node metastasis,TNM staging and tumor differentiation degree,the expression of Twist protein was significantly different;Expression of P53 and E-cadherin protein in esophageal carcinoma was significantly higher than adjacent tissues and normal esophageal tissues;and at different depth of invasion, lymph node metastasis,TNM staging and varying degrees of tumor differentiation, P53 and E-cadherin protein expression has a significant difference;Twist expression level of esophageal carcinoma has obvious correlation with the existence of P53 and E-cadherin.Conclusion:Twist may be the biological parameter of esophageal diagnosis,as well as determine the course of disease in patients with esophageal disease-specific indicator of prognosis.P53 protein,E-cadherin plays an important role in the process of esophageal cancer growth,invasion and metastasis,Twist probably inhibits apoptosis and promotes tumor cell formation by regulating pathway of ARF/MDM_2/P53,meanwhile impacts expression of E-cadherin to promote situation of EMT,and participates in the occurrence and metastasis of esophageal cancer.

CCL21-CD40L Fusion Gene Augment Antitumor Activity

Tue, 29 Mar 2011 14:09:16 +0000

Objective:Dendritic cell is the most functional antigen presenting cells in vivo,it can capture,processing,presenting tumor antigen to T cells,initiating immune response. However,DC vaccine is hard to develop because of its long processing,short survival time and difficulty in mimicing its natural maturation.We construct eukaryotic expression vector carried CCL21-linker-exCD40L fusion gene,test its chemotactic function,and investigate its anti-tumor activity.Methods:Search the gene coding sequence of mouse CCL21 and CD40L in GeneBank, and determine the amplified region.The total mRNA was extracted from mouse spleen using trizol,and the aimed segment were amplified from total mRNA by the method of reverse transcription polymerase chain reaction.CCL21-linker-exCD40L fusion gene were constructed by overlap PCR connecting CCL21 and exCD40L,then it were inserted into pMD18-T for seqencing.Eukaryotic expression plasmid pcDNA3.1 was used to carried the CCL21-linker-exCD40L fusion gene after confirmation of sequencing.We also cloned CCL21 and CCL21sig-CD40L into pcDNA3.1 as comparision.The recombinant plasmid were respectively transfected into Chinese Hamster Ovary(CHO) cells and wsestern blot was used to identify the fusion protein.The chemotatic function of the expressed product was detected by chemotaxis assay and the anti-tumor activity of it was tested with vivo transfection.Result:Gene sequencing,gel electrophoresis of PCR product and restrictive digestion proved the successful construction of pcDNA3.1/CCL21,pcDNA3.1/CCL21sig-exCD40L and pcDNA3.1/CCL21-linker-exCD40L,and its expression was conformed by western blot, which is 14KD,30KD,43KD respectively.We test that the fusion protein had a significant chmotactic function to DCs using Millicell-Cabinet Method,which the cell numbers passing through the film is 14.95 times of blank control every high power microscope visual field. We construct colon cancer model in Balb/c mouse with C26 cell line,and after tumor orthotoic injection of plasmid carrying fusion gene,the expressed product in vivo act anti-tumor function.Conclusion:The recombinant murine CCL21-linker-exCD40L,CCL21 and CCL21sig-exCD40L eukaryotic expression plasmid was successfully constructed,and they all can be expressed by eukaryotic cell after transfection.The expression product of CCL21-1inker-exCD40L fusion gene in tumor cells had a remarkable function to DCs.After tumor orthotopic injection,it can inhibit tumor growth and prolong the mouse survival time, which is more effective than all the control.

Effect Observation of Granulocyte Macrophage Colony-stimulating Factor Combined with Allogeneic Tumor Cellular Immunotherapy for Advanced Cancer

Tue, 29 Mar 2011 14:09:12 +0000

ObjectiveThe aim of this trial was to evaluate the feasibility,toxicity,immunologic,and clinical activity of GM-CSF secreting allogeneic cancer cell vaccine in advanced-stage patients.Methods and materialPatients:Patients were enrolled at our hospital,after institutional review board approval and written informed consent was obtainded,Including melanoma(n=11), ovarian cancer(n=10),breast cancer(n=10),renal cell carcinoma(n=15), gastric/colorectal carcinoma(n=14).All the patients were diagnosed stage IV,and stoped chemo- or radiotherapy at least one month.Patients were required to have a Karnofsky performance status of 60 to 80.Vaccine preparation:Cell line which have the same pathology with patients\' tumor were used as the allogeneic tumor cells.Each injection contains inactivated tumor cell more than 1×10~7 and GM-CSF-secreting cell which can secret GM-CSF more than lug/10~6/24h.Vaccines administration: Vaccines were administered subcutaeous every week for a total six vaccinations.The vaccines were administered on the upper 1/3 inside of upper arms or thighs on a rotated.DTH were administered intradermal at lower 1/3 inside of forearms on the first and fifth vaccinations.Immune responses evaluation:1.Collected 10ml peripheral venous blood at pre- and protreatment,check the percentage of CD4+ T cell,CD8+ T cell,CD4+/CD25+/CD127-T cell,CD3-/CD16+56+/CD45+ cell by using flow cytometry,The levels of interferon-gamma,interleukin-4,transforming growth factor beta-1 were evaluated by using the ELISA.ResultsSafty:all the patients completed the treatments.Vaccines treatment were generally well tolerance.the most common vaccine-related adverse events were self-limited injection site reactions,erythema induration(94%),pruritus(60%),vesicular(4%) and pain or soreness at the injection site(94%).All the injection site reactions were grade I orⅡin severity.Seven patients reported fever,lower than 39℃.one patients had headache and dizziness,but got better without treatment.No clear evidences of vaccine-associated adverse events.Clinical responses:One patient dead.No patients achieved a partial or complete tumour responses.Four achieved mixed responses (MR),with regression of at least one tumour mass.40 patients maintained stable disease for at least 4 weeks following vaccine initiation.16 patients progressed, among them seven had liver metastasis,two had brain metastasis,and 1 had lung metastasis.To 30/6/2008 8 patients dead results in diseases progression,among them 6 had liver metastasis,one had brain metastasis.Median overall survival was 5.5 months,four patients were alive more than 1 years.Immune responses:Analysis of immune responses to vaccine was hampered by the polyvalent natural of the whole tumor cell used as the immunogen.Thus,immune responses to vaccination were assessed by measurement of vaccines injections site induration,and DTH reactions of intradermal injection of allogeneic tumour cell.DTH were positive in 44 of 50 patients,3 of 16 in progressed patients,34 of 40 in stable disease patients.DTH reactions were seen in stable disease patients more than in progressed patients(χ~2 =22.54,P <0.005).DTH responses level and injection site reaction level exist linear correlation(R~2=0.594,P<0.001).Findings,patients immune system was activated at pro-treatment by test the changes of lymphcytes substype and related cytokines.TGF-β_1 significantly decreased in melanoma and renal cell carcinoma patients at pro-treatment(p<0.05). CD8+T cell significantly increased in melanoma,ovarian cancer,renal cell carcinoma patients at pro-treatment(P <0.05).Treg cell significantly decreased in all patients at pro-treatment.IFN-γonly observed increased tendency,no statistical significance at pro- and pre-treatment.Other cytokines and lymphocytes substypes no clear tendency. CD8+T cell significantly increased and Treg cell decreased in stable disease patients at pro-treatment(P <0.001,P <0.001).Treg cell was observed significantly decreased only in progressed patients(P <0.05).ConclusionsThis non-patient-specific cancer immunotherapy has a favorable safety profiles and is immunologically active.Continued clinical tial is warranted

Correlation of STAT3 Expression to Cisplatin Resistance in Head and Neck Squamous Cell Carcinoma

Tue, 29 Mar 2011 14:09:10 +0000

ObjectiveSignal transducers and activators of transcription 3(STAT3) is an important member of family of signal transducers and activators of transcription.Many tumor-derived cell lines as well as samples from human cancer have been reported to express high levels of constitutively active Stat3 protein,for example head and neck squamous cell carcinoma(HNSCC),breast cancer,prostate cancer,multiple myeloma and so on.Constitutively active Stat3 has important roles in several biological responses such as tumor prolifertion、differentiation、apoptosis and migration.Besides its classic functions,STAT3 oncogenic pathway may be involved in drug resistance. However,its role in cisplatin resistance of HNSCC is unclear.This study is to explicit the expression levels of STAT3 in HNSCC,and explore the correlation of the STAT3 oncogenic pathway to cisplatin resistance.Methods1.Twenty-eight cases of HNSCC pathologically confirmed after resection were obtained as surgical samples from patients treated at the head and neck neoplasms department of Tian jin cancer hospital,including 8 cases of laryngeal carcinoma,7 cases nasopharyngeal cancer,5 cases tongue carcinoma,5 cases gingival carcinoma,1 case carcinoma of the floor of mouth,1 case carcinoma of upper esophagus,1 case maxillary sinus carcinoma.All the specimens were divided into two portion,the sensitivity to cisplatin of one portion was detected by collagen gel droplet embedded culture-drug sensitivity test(CD-DST),another was embedded by paraffin and tested the expression levels of STAT3 by immunohistochemistry. Besides,17 cases of normal squamous epithelium were collected for the control.Cell line of tongue squamous cell carcinoma was recoveried,and creep plates of induction culture cell were collected for immunocytochemistry.2.The sensitivity to cisplatin of 28 cases of HNSCC was detected by CD-DST. 3.Labelled streptavidin biotin method was used to detect the expression levels of STAT3 protein in the cisplatin resistant group、cisplatin sensitive group、normal squamous epitheliums and Tb3.1 cells.4.After Tb3.1 cells were treated by AG490 which was one inhibitor of the STAT3 oncogenic pathway for 24 hours,the sensitivity to cisplatin of the AG490 treatment group and the control group were detected by CD-DST.5.SPSS13.0 statistical software was used for the data analysis.The expression levels of STAT3 in HNSCC and its correlation to cisplatin resistance were analysed by x~2 test.The difference of cisplatin sensitivity between the AG490 treatment group and the control group was analysed by t test.Results1.The chemosensitivity was successfully evaluated by CD-DST for 25 of 28 patients.The efficacy rate was 89.3%(25/28).Eight cases were assessed as sensitive,and sevevteen as resistant.The response rate was 32.0%.2.The expression levels of STAT3 protein were elevated in HNSCC tumor samples as compared with levels in normal squamous epithelium(x~2=13.137, P＜0.005).In the cisplatin resistant group,4 cases have higher expression of STAT3,12 cases have moderate expression of STAT3,1 case has weak expression.While in cisplatin sensitive group,1 case has higher expression of STAT3,2 cases have moderate expression of STAT3,5 cases have weak expression(x~2=9.616,P=0.008).3.STAT3 protein is positively expressed in the cytoplasm of Tb3.1 cetls,with brown granulations.4.The cell survial rate of Tb3.1 cells was(96.59±10.01)%after treated by DMSO,and(95.22±7.16)%by AG490 respectively.There was no statistical difference as compared with the blank control group.The Tb3.1 cells were sensitive to cisplatin,and the cell survial rate was(22.69±2.79)%.There was significantly different cisplatin sensitivity between the AG490 treatment group(8.94±1.06)%and the DMSO treatment group(21.19±2.74)% (P=0.000). Conclusions1.STAT3 protein was positively expressed in HNSCC.The expression levels of STAT3 were elevated in HNSCC tumor samples as compared with levels in normal squamous epithelium.2.There was a positive correlation between the expression levels of STAT3 protein and cisplatin resistance in HNSCC.3.STAT3 protein was positively expressed in Tb3.1 cells.4.The sensitivity to cispaltin of Tb3.1 cell could be improved after the STAT3 oncogenic pathway was blocked by AG490.

Radiosensitivity Enhancement by Combined Treatment of COX-2 Inhibitor and EGFR Inhibitor on A549 Human Lung Adenocarcinoma

Tue, 29 Mar 2011 14:09:04 +0000

Objective:To characterize the radiation-enhancing effects and underlying mechanisms of combined treatment with celecoxib,a cyclooxygenase-2 selective inhibitor,and erlotinib,an epidermal growth factor receptor(EGFR) tyrosine kinase inhibitor,in human lung adenocarcinoma A549 cell lines.Methods:Human lung adenocarcinoma A549 cell lines was cultured in vitro.The subjects were divided into 8 groups:control group(O group),celecoxib group(C group), erlotinib group(E group),celecoxib + erlotinib group(C + E group),radiation group (R group),radiation + celecoxib group(R + C group),radiation + erlotinib group(R + E group),radiation + celecoxib + erlotinib group(R + C + E group).The antiproliferative activity of celecoxib and erlotinib were determined by MTT analysis, and the IC_(50) and IC_(20) were caculated,respectively.The 20%inhibitory concentrations (IC_(20)) were needed for the next subjects.Clonogenic radiation survival assays after treatments with celecoxib and erlotinib with or without radiation were done on A549 cell lines.The combining effect was quantified by calculating the SF(Surviving Fraction).Alterations in apoptosis and cell cycle observed by Flow cytometry were measured and Western blot for Akt and phosphorylated Akt were conducted to identify the mechanisms underlying radiation-enhancing effects of celecoxib and erlotinib.Results:1.The effect of the dose-dependent growth inhibition was observed by the MTT analysis in human lung adenocarcinoma A549 cell lines.And the IC_(50) and IC_(20) were(50.13±2.41 )μmol/L and(40.32±1.26)μmol/L for celecoxib and (12.21±0.83)μmol/L and(5.15±0.14)μmol/L for erlotinib,respectively.2.Clonogenic radiation survival assays showed that the Dq、Do、SF_2 of(R+C+ E) group was lower than those in(R + C) group or(R + E) group.The SER of(R+ C+E) group was 2.217,the SER of(R+C) group was 1.299,the SER of(R+E) group was 1.503.The radiation-enhancing effects of drug combination were bigger than those of single drug.And the drug combination could enhance the radiosensitivity more significantly.3.The Flow cytometry showed that both celecoxib and erlotinib could induce G_1/G_0 phase arrest,and the reduction of S phase combined treatment with radiation, especially in(R+C+E) group.Both celecoxib and erlotinib could enhance radiation-induced apoptosis,and the apoptosis was highest in drug combination. So drug combined treatments could enhance radiation-induced apoptosis more significantly.4.Western blot showed that the expression of Akt protein had no difference in any group;both celecoxib and erlotinib could inhibit the expression of pAkt and radiation can enhance the expression of pAkt.In(R + C) group or(R + E) group, the expression of pAkt was lower than R group,and the expression of pAkt was lowest in(R+C+E) group.Conclusions:Both celecoxib and erlotinib could enhance the radiosensitivity,and the drug combination could enhance it more significantly.The mechanisms seemed to involve the of reduction of radiation-insensitive S phase and synergistic enhancement of radiation-induced apoptosis,possibly inhibiting PI3K/Akt signal transduction pathway was one of the mechanisms of radiosensitivity enhancement by combined treatment of celecoxib and erlotinib.

Prognostic Factors for Hepatocellular Carcinoma and Comparisons of Prognostic and Therapeutic Guiding Values of 8 Staging Systems in a Chinese Cohort

Tue, 29 Mar 2011 14:09:01 +0000

Background:Hepatocellular carcinoma(HCC) is a major health problem worldwide. Clinical staging of cancers is important for guiding therapeutic interventions and assessing prognosis.Although several staging systems have been developed in the last few years,there is still no worldwide consensus on which is the preferred prognostic staging system for HCC,and only the Barcelona Clinic Liver Cancer(BCLC) staging system embodies therapeutic schedule.Furthermore,few studies were based on the Chinese patients,who account for more than a half of the worldwide cases.Therefore, these staging systems still need to be validated in the Chinese HCC patients.Objective:The aims of this study were to identify independent prognostic factors for HCC,to validate 8 staging systems in Chinese HCC patients and to compare both the prognostic and the therapeutic guiding values of these systems for HCC patients in China.Methods:We retrospectively studied 861consecutive HCC patients without prior anticancer therapy who were admitted in Liver Cancer Institute of Zhongshan Hospital between January 2001 and December 2002.Demographic,laboratory,and tumor characteristics and performance status were determined at diagnosis and before therapy.Factors determining survival were analyzed by univariate and multivariate analysis using the Kaplan-Meier method and Cox proportional hazard regression models.Tumor-Node-Metastasis(TNM)、Okuda、BCLC、Cancer of the Liver Italian Program(CLIP)、Japan Integrated Staging score(JIS)、Chinese University Prognostic Index score(CUPI)、GRoupe d\'Etude et de Traitement du Carcinoma Heatocellulaire (GRETCH) and Chinese Scoring System evaluated before treatment were applied. Each system was entered in to the Cox regression model to calculate the LR x~2,AIC and Harrell\'s C values.The area under the ROC curve was calculated to evaluate the accuracy of prediction of death at 1-,3-,5-year for each staging system.Survival curves for the patients treated with different therapies in each stage category according to the discussed eight staging systems were also calculated using the Kaplan-Meier method and compared by log rank test.Results:At the end of follow-up,496 patients(57.61%) were deceased.A total of 114 patients(13.24%) had withdrawn,including 91 patients(10.57%) being followed up with less than 3 years,and 108 patients(12.54%) less than 5 years.Overall median survival was 38±2.84 months,with estimated 1-,3-,and 5- year survival rates as 74.64±1.52%,51.27±1.78%,and 40.08±1.78%,respectively.By multivariate Cox regression analysis,tumor size,tumor numbers,vascular invention,extra-hepatic invention,cirrhotic node size,ascites,alkaline phosphatase(AKP),HBe antigen (HBeAg),were independent prognostic factors for overall survival.By Kaplan-Meier analysis,all the 8 systems showed prognostic abilities.However,only the JIS,Chinese and BCLC systems showed significant difference among survival rates for each subgroup.With the Cox regression model,BCLC showed the best homogeneity(LR x ~2=158.19) and informative explanatory ability(AIC=5049.75),followed by the Chinese staging system.Chinese staging system showed the best discriminatory ability and monotonicity of gradients than the other systems(Area under the ROC curve:1-year:0.7848;3-year:0.7492;5-year:0.7152,Harrell\'s C=0.6877),followed by the BCLC and JIS staging systems.On the other hand,only the Okuda,BCLC, CLIP and Chinese systems showed the ability to decide which patients may not benefit from resection.There were no significant survival differences between the surgery group and the TACE/TAE group for patients in Okuda stageⅡ,BCLC stage C,CLIP scores more than 3,and Chinese stageⅢA,while resection may benefit for patients with the earlier stages. Conclusion:The BCLC,JIS and Chinese staging systems revealed best prospective ability,and the Okuda,BCLC,CLIP and Chinese systems revealed good therapeutic guiding ability.Only the Chinese and the BCLC staging systems were good at both prospecting prognosis and deciding preferred treatment for patients.However,the BCLC treatment schedule is not validated by this study.

Investigation of Molecular Mechanism of Vasculogenic Mimicry in Malignant Tumor from Muscle

Tue, 29 Mar 2011 14:08:56 +0000

Aims:To observe and identify VM existence in malignant tumor from muscle,and indicate the clinical and pathological significance of VM through a retrospective study with human samples.Based on these,we will to investigate the role of VM and the molecular mechanism involving in VM formation.Methods:1.Forty-five leiomyosarcoma cases and forty-six rhabdomyosarcoma cases undergoing surgery in Tianjin Medical University from 1~(th)/1,1982 to 30~(th)/12,2002 were collected in this study.The clinicopathologic information of samples is complete. HE staining and CD34/PAS immunohistochemical double staining were performed to observe the existence and distribution of VM,and to explore the clinicopathologic significance of VM and the relationship between prognosis and VM.2.Tissue microarray and immunohistochemical staining were applied to detect the expression of FAK,cathD and MMP-2.Staining index(SI) was assessed to investigate the different expression between VM-positive group and VM-negative group,and to study the relationship between them and VM.3.Bwteen January 2006 and December 2008,twenty leiomyosarcoma cases and thirty rhabdomyosarcoma cases with fresh tissue were collected from Tianjin Medical University.Real-time PCR was applied to detect the mRNA expression of FAK, CathD and MMP-2,and to investigate the different expression between VM-positive group and VM-negative group.Results:1.VM exists in human leiomyosarcoma and rhabdomyosarcoma.In HE staining slides,there were VM,formed by tumor cells but not endothelial cells,in which red blood cells presented;near these structures,there were no hemorrhage,necrosis and inflammatory cells infiltratin.In CD34/PAS double staining slides,VM was tumor cells formed channel with CD34-negative,there was PAS-positive material separate tumor cells and red cells.2.Statistical results showed that 55.6%(25/45) of leiomyosarcoma had VM,and 56.5%(26/46) of rhabdomyosarcoma had VM.VM is close-related with tumor sizes, metastasis and survival status(P＜0.05),however,unrelated with the age and gender of patient,location and recurrence.Kaplan-Meier survival analysis indicated that the prognosis of patients with VM was worse than the prognosis of those without VM (P＜0.05).3.IHC results shows that the expressions of FAK,CathD and MMP-2 in VM-positive group were statistically significant higher compared with those in VM-negative group.The expression of FAK is close-related with the expression of MMP-2 in rhabdomyosarcoma and leiomyosarcoma.4.Real-time PCR results showed that the mRNA expressions of FAK,CathD and MMP-2 in VM-positive group were statistically significant higher compared with those in VM-negative group in leiomyosarcoma and rhabdomyosarcoma.Conclusion:1.VM exists in malignant tumor from muscle.Under appropriate matrix microenvironment conditions,remodeling of the extracellular matrix(ECM) and the connection of the VM channel and host blood vessels to acquire a blood supply from the host tissue.2.VM is close-related with tumor sizes,metastasis and survival status.Tumors with VM are likely to spread via blood vessels in the early stage,and the patients faced an inefficient curative effect and a poor prognosis.3.FAK,CathD and MMP-2 expressed higher in VM-positive group than VM-negative group.This shows that the three molecules may participate in VM formation by degradating base-membrane and extracellular matrix in malignant tumor from muscle.4.The expression of FAK was closely correlated to the expression of MMP-2.This shows that can enhance invasion and metastasis ability of tumor cells and contribute to the formation of VM together.

The Trend Analysis of Clinical Incidence, Diagnosis and Treatment of Prostate Cancer in Tianjin

Tue, 29 Mar 2011 12:37:34 +0000

Objectives:By analysing the data of hospitalized patients with prostate cancer in Tianjin,to learn the status and changing trends of clinical incidence,diagnosis and theraty of prostate cancer.To improve the diagnosis and treatment level of prostate cancer in Tianjin.Methods:Supported by the prostate cancer database of Tianjin and Cooperation Group of Prostate Cancer Diagnosis and Treatment,we retrospectively analyse the clinical data of 1536 prostate cancer cases in fourteen hospitals of Tianjin from 1980 to 2008.We analyse the thrands of patients age,clinical strage,tumor grade, diagnosis and therapy methods.Results:The incidence of prostate cancer in 1981 and 2000 in Tianjin are about 1.07 per 100,000 men and 4.09 per 100,000 men.1536 cases of prostate cancer are diagnosed and hospitalized from 1980 to 2008,namely 49.41±57.65 cases diagnosed per year.The 60～and 70～age segments are principal parts(76%).The diagnose rate of CT,MRI and ECT is 59.8%,82.8%and 64.1%,respectively.The diagnose rate of DRE+PSA,DRE+TRUS,PSA+TRUS and DRE+PSA+TRUS is 90.0%,86.3%,97.1%和98.6%,respectively.The proportion of cases in(A+B)stage increases from 12.3% to 20.3%.(C+D)stage decreases from 87.7%to 79.7%(x~2=8.525 p=0.004).The proportion of bilateral orchiectomy in Endocrine therapy is higher than 50%.The use of androgen increases from 8.6%to 86.2%.Recently,the clinical use rate of radical prostatectomy,Seed brachytherapy,bisphosphonates,argon-helium cryoablation are 16.2%,11.1%,15.2%and 6.0%.Conclusions:The incidence of prostate cancer in Tianjin are on a level from 1980 to 1988,but in the late 1980s it shows increasing trend.Prevalence of age is still 60～,70～-based,but there is a gradual increase in the average age of the trend. Serum PSA testing has been basically popularized in clinical use,and the clinical detection rate of prostate cancer has greatly enhanced.Preoperative TRUS-guided prostate biopsy in the diagnosis of prostate cancer has been generally carried out in clinical.And CT,MRI and ECT can give significant help for diagnosis and treatment of prostate cancer.Now bilateral orchiectomy,maximal androgen blockade play important roles in hormonal therapy of prostate cancer.Radical prostatectomy increases recently with the increase of early staged cases.And some new treatments have been used in clinical,such as Seed brachytherapy,radionuclide therapy, bisphosphonates,radiofrequency thermal ablation(RFTA)and argon-helium cryoablation,However,fewer cases.Patients of prostate cancer with late stage and poorly differentiated grade has high proportion in all prostate cancer patients n Tianjin,which is in line with the national conditions of our country,so the most urget and important thing is to improve the early diagnosis for prostate cancer.

Comparation of Quality of Life in Different Types of Urinary Diversion after Radical Cystectomy

Tue, 29 Mar 2011 12:37:05 +0000

Objectives:Using bladder cancer specific measurement FACT—BL to evaluate quality of life of patients who underwent the operation of Studer\'s ileal neobladder, ileal conduit or cutaneous ureterostomy,copare the difference of general quality of life and bladder cancer related quality of life among the three types of urinary diversion and to analsys the cases of this differences in order to improve the quality of life by providing basis of quality of life for patients who underwent radical cystectomy and want to choice a type of urinary diversion.Material and methods:In this study we collected the patients who have diagnosed with bladder cancer,underwent radical cystectomy and accept urinary diversion of Studer\'s ileal neobladder,ileal conduit or cutaneous ureterostomy in our department during Dec 2006 and May 2008,choice the suitable cases,then collect the information of the patients such as name,sex,age,operation date,type of operation, type of pathology,pathological stage,address and telephone number.Test the quality of life using bladder cancer specific measurement FACT—BL by inviting the patients to our department or mail them questionnaires or phone them.compare the quality of life of patients among the three types of urinary diversion in different time after the operation.All descriptive data are reported as the mean±standard deviation(SD).The differences in the mean values between the ileal neobladderand the cutaneous diversion patients were analysed by Indipendent Samples T test,The data regarding the percentage were analysed by x~2 test.Significance was defined at the 5%level.The statistic analysis was completed by SPSS-13.1,tables and picyures were made by Excel 2003.Results:The HRQOL of the three groups patients were gradually rised after the operation,within the 12 mouths after the operation the difference among the three groups was not significant(P＞0.05) and from the 12 mouths after the operation the Studer\'s ileal neobladder group had a higher HRQOL then ileal conduit and cutaneous ureterostomy,the difference was significant(P＜0.05).the difference in physical well being,social and family well being,emotional well being and FACT-G was not significant(P＞0.05),but the neobladder group had a batter HRQOL in functional well being and FACT-BL,the difference was significant(P＜0.05).Conelusions:In our study the neobladder group had a batter HRQOL then the other two groups.So when choice a type urinary diversion,the Studer\'s ileal neobladder should be considerd first if the patients can tolerate.The sample of this study was small and the time of following up was short,so further study are necessary to investigate the influence of different types of urinary diversion in HRQOL.

An Experimental Study of Effects of Argon-helium Cryotherapy on Angiogenesis in Prostate Cancer

Tue, 29 Mar 2011 12:37:01 +0000

ObjectiveThe purpose of this study was to explore the effect of Argon-Helium Cryosurgery(AHCS) on the mice of bearing RM-1 prostate cancer through detecting angiogenesis and cell proliferation in prostate cancer of mice,To analyze the effects of cryotherapy on VEGF、Ki67 protein expression and microvesel density of prostate cancer,and provide an experimental evidence on cryoablation comprehensive treatment for advanced prostate cancer.Materials and MethodsA hundred mice experimental models with RM-1 prostate cancer were established,experimental mice were randomized into two group:the AHCS therapy group(group A,n=50),the tumor-bearing control group(group B,n=50).after the AHCS therapy,Peripheral tumor tissues of 50 mice were obtained from group A at 1d,3d,5d,7d,14d.at the same time,the tumor-bearing control group B(not do the AHCS therapy),tumor tissues of 50 mice were obtained from group B at 1d,3d,5d, 7d.14d.MVD,VEGF,Ki67 protein expression were detected by immunohistochemical staining at different time after the freezing.Results1.Prostate cancer cell has stable biology characteristics,and the incadence of cancer was 100%.2.MVD、VEGF and Ki67 protein expressions in freezing group were decreased.The lowest value was detected at the third day after freezing.The value increased slowly after that.There was no change in MVD and VEGF,Ki67 protein expressions at different time in control group(P＞0.05),There were significant change in MVD,VEGF,Ki67 protein expressions at different time in freezing group(P＜0.05). Correlation analysis showed suggest a positive correlation between VEGF protein expression and MVD(r=0.8793,P＜0.01) suggest a positive correlation between ki67 protein expression and MVD(r=0.7614,P＜0.01).suggest a positive correlation between VEGF protein expression and ki67(r=0.6921,P＜0.01).Conclusion After argon-helium cryoablative treatment for prostate cancer,local tumor MVD, VEGF,Ki67 was reduced at the first day,the lowest value was detected at the third day after freezing,then increase,in the dynamic process of change,MVD,VEGF, Ki67was positively correlated.And cryoablation joint anti-angiogenesis is an effective means that may be integrated in the treatment of advanced prostate cancer.

Expression of Coxsackie and Adenovirus Receptor in Non-small Cell Lung Carcinoma and Its Significance

Tue, 29 Mar 2011 12:36:43 +0000

Objective:To study the expression of coxsackie and adenovirus receptor(CAR) mRNA and protein in lung normal tissue,paraneoplastic tissue and carcinoma tissue, analyzing their correlation with carcinogenesis and development,as well as their different expression in different clinicopathologic parameter,Confirming the role of CAR in lung cancer formation especially adenocarcinoma,and demonstrating the clinical significance of CAR as a new therapeutics target.Method:Collecting fresh lung cancer 32 samples,as well as its corresponding paraneoplastic tissue and normal tissue,put into liquid nitrogen as soon as removed from the patient\'s body,-80℃followed.Simultaneously the clinical data of the patients was collected for the results analysis.The expression of CAR protein was determined by IHC and Western blot and CAR mRNA was detected by RT-PCR in lung norm tissue,and paraneoplastic tissue carcinoma tissue respectively.Results:1.CAR protein expression in lung normal tissue,para-carcinoma tissue and carcinoma tissue.Using immuhistochemical method,19 lung cacinoma tissue were positive in the 32 lung cancer cases:14 adenocarcima(6-bronchiolo-aleolar carcinoma BAC ) and 5 squamous cell carcinoma..By Western blot assay,a band of about 46 kd was detected in in lung normal tissue, paraneoplastic tissue and carcinoma tissue,with the expression value 0.902±0.038, 0.944±0.042,1.08±0.052.Carcinoma tissue were statistically higher than lung normal tissue and paraneoplastic tissue(P=0.09,P=0.027).2.CAR mRNA expression in lung normal tissue,para-carcinoma tissue and carcinoma tissue.The expression value of CAR mRNA in lung normal tissue,paraneoplastic tissue and carcinoma tissue were 1.000±0.012,1.048±0.035,1.282±0.072.And Carcinoma tissue were significantly higher than lung normal tissue and paraneoplastic tissue,the difference having significance(P=0.022,P=0.007).3.There was a statistically positive correlation between expression of CAR mRNA and protein(r=0.448,P=0.026).By McNemar method,We calculated the value ofк=0.652,P=0.011,P＜0.05.The two has high anastomosis. 4.The correlation of CAR expression with different clinicopathologic parameters. There was no statistical significance in other clinicopathologic parameters(P＞0.05), including gender,age,smoke or not,tumor size,with or without lymph node metastasis and TNM stage.The expression level of CAR has significant difference between squamous cell carcinoma and adenocarcinoma(P=0.012),with 7 BAC all expressing higher level CAR than non-BAC(P=0.029).But significance disappear when we excluded the BCA from adenocarcinoma(P=0.125).Conclusion:1.Carcinoma tissue were statistically higher than lung normal tissue and paraneoplastic tissue;CAR expression in lung adenocarcinama was higher than squamous cell.These data predicted that CAR expression might be necessary for the formation of human lung cancer,particularly for the development of adenocarcinoma.2.The expression level of CAR has significant difference among different pathological types but no statistical significance in other clinicopathologic parameters. 7-BAC all expressed high level CAR than non-BAC.This might thoroughly demonstrate the role of CAR in the formation of BAC.But need more research.3.CAR mRNA and protein has significantly positive correlation.When the expression of CARmRNA rises,the CAR protein increases.4.If CAR is a promoting factor for human lung cancer,we might develop the antibody-directed therapy as EGFR TKI did in human lung cancer.If CAR is not a promoting factor of human lung cancer,the elevated expression of CAR in lung cancer will be more favorable to the gene therapy of human lung cancer.

Silencing of E2F3 Expression in Bladder Cancer Cell Line 5637 Using RNAi with AAV Virus Expressing Vector

Tue, 29 Mar 2011 12:36:39 +0000

Bladder cancer is one of the most common urological malignant diseases,most of bladder cancer is transitional-cell carcinoma.The most important features of urothelial cancers of the bladder are metachronous or synchronous multifocal occurrence with high frequeney.Operation has been the main measure in treating bladder cancer,but after operation bladder cancer easily recurs and the recurrence cancer often increases its progression.For this reason the use of intravesical drugs such as chemieal drug and immune revulsive have been the most important part of treating.However these therapeutie methods we used cannot control the Progression and recurrence of bladder cancer and at the same time the side effects of drugs and drug resistence in Patients remain the tough problem.For more than twenty years research gene therapy has gradually become mature and is accepting as extremely promising approach to treating cancer after regular treatment such as operation, chemotherapy and radiotherapy.The discovery that RNAi works in mammlian cells has sparked intense investigation into its role in normal mammlian cell function,its use as a tool to understand or screen for genes functioning in cellular pathways in healthy and diseased cells and animals,and its potential for therapeutic gene silencing.RNA interference(RNAi) is an evolutionary conserved mechanism for silencing gene expression.In primitive organisms,RNAi Protects the genome from viruses and other insertable genetic elements and regulates gene expression during development。The antisense(guide) strand of short double-stranded RNAs is incorporated into an RNA-induced silencing complex that can either suppress protein expression or direct degradation of messenger RNAs that contain homologous sequence(s).RNAi may provide an important new therapeutic scheme for treating illnesses especially for cancer.On the condition of high safety、efficiency vector,RNAi may process in cells favorably Recently Adeno-associated virus vector is researched extensively in gene therapy, because of its broad host、low immunogenicity、high safety and stability,AAVs is considered as one of new generation virus vectors.Hitherto;AAV vector has been used for treating tumor in different animal models,the results show that it is a potential vector.The E2F transeription factors control expression of genes that are essential for cell proliferation,including key components of both the DNA-replication and cell cycle control machinery.E2F3 is sufficient to induce quiescent cells to enter S phase. The overexpression of E2F3 in human bladder cancer cells is a common phenomenon, now many scholar think that The tumorigenesis and development of tumor has intimate relationship with the overexpression of E2F3,knock-down of the expression of E2F3 may restrain development of bladder cancer.Objective:In this article we want to know if AAV can be used effectively in RNAi of Bladder cancer cell line and wheher it can take siRNA into cells.To answer these questions we did the following jobs:we have developed effective RNAi AAVs correponding to E2F3 gene transfected the AAVs into human bladder cancer cell line 5637,assessed the E2F3 mRNA and protein levels in 5637 cells.Methods:AAV 293 cells were co-transfected using pAAV-siRNA-E2F3,pRC and pHelper to package by electroporation rAAV-siRNA-E2F3.The cells were under Chloroform-NaCl sediment-Chloroform and ice alcohol to dissociate,purify and concentrate rAAV.Viral particle of purified were assayed by AVSachTM ELISA.Use SDS-PAGE Coomassie brilliant blue staining to observe capsid protein of rAAV.The rAAV-siRNA-E2F3 without endotoxin was extracted successfully and then was transfected into 5637 cells respectively.rAAV-siRNA-E2F3 were transfected into human bladder cancer cell lines 5637 and the transfection efficiency can be acquired by observing green fluorescence of cells.RT-PCT was used to determine if rAAV-siRNA-E2F3 can downregulate the expression of E2F3 mRNA.The silencing effect can also be detected by Western blot in protein levels.Results:Under AVSachTM ELISA mensuration,the MOI of rAAV is 5.6×10~(13)v.p/ mL.Under SDS-PAGE and Coomassie brilliant blue staining,we could find three notable viral capsid protein strap of AAV.Green fluorescence can be seen at 30 min. Relative to control,rAAV-siRNA-E2F3 induced 69.84%(P＜0.01) inhibition of E2f3 mRNA treatment in 5637 cells.Comparing with control,E2F3 protein levels of 5637 cell significantly reduced and inhibition rate is 83.27%(P＜0.01)Conelusion:5637 cells can be transfected efficiently by AAV.The rAAV-siRNA-E2F3 was a better transfection reagent for RNAi in 5637 cells.The sequence specific siRNA showed a block effect in downregulation of E2F3 gene expression,Silencing E2F3 may be developed into a potential tool for bladder gene therapy.

Transcriptional Regulation of PTPRO Gene by E2F1 during PC12 Differentiation after NGF Stimulation

Tue, 29 Mar 2011 12:36:34 +0000

PTPRO is a member of the receptor-type protein tyrosine phosphatases(RPTPs) family, which is involved in podocyte function,axon pathfinding and tumor suppression.The expression of PTPRO varies under different conditions.For example,in kidney,PTPRO is expressed on the apical surface of the podocyte early in development and is present on the apical surface of podocyte foot processes in the mature phenotype;in brain,maximal expression of PTPRO iscoincident with mid to late gestation and axonogenesis.It is reported that Receptor-type protein tyrosine phosphatases(RPTPs) are required for appropriate growth of axons during nervous system development in Drosophila.In the vertebrate,typeⅡa RPTPs [protein tyrosine phosphatase(PTP) and LAR(leukocyte common-antigenrelated)]and the typeⅢRPTP have been implicated in the regulation of axon growth,However,people know little about the regulation mechanisms for PTPRO expression and its function in neuronal differentiation.To clarify the regulation of PTPRO gene transcription during PC12 neuronal differentiation after stimulated by NGF,PCR(Real-time PCR) and Westem bloting was used to examine the up-regulation of PTPRO gene in the differentiation of PC12 cell line.By applying a series of transient transfection and relative luciferase activity assays,we found that transcription factor E2F1 promoted the activities of the PTPRO promoter evidently.The results of real-time PCR and Western bloting confirmed the effects of E2F1 in promoting the PTPRO gene expression.Additionally,by using ChIP,we showed that E2F1 bound to PTPRO promoter in vivo.Our data demonstrated that the transcription factor E2F1 was involved in the transcriptional regulation of PTPRO gene.PTPRO was identified as a new target of E2F1. This work provides further insight into the regulation mechanisms of PTPRO gene and its function in neuronal diffenentiation.

The Relationship between Anatomic Measurements of Pharynx and Other Factors in Patients with Obstructive Sleep Apnea-hypopnea Syndrome

Tue, 29 Mar 2011 12:36:32 +0000

Objective To investigate the morphosis of pharynx in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) during sleeping.And to study its relationships with apnea-hypopnea index(AHI),body mass index(BMI),breathing control,and so on.Methods 35 OSAHS patients(with an apnea-hypopnea index,AHI≥5),27 men and 8 women,were studied.And they were subdivided into two groups by AHI:mild group included 14 patients with 5＜AHI≤20,and moderate-severe group included 21 patients with AHI＞20.Polysomnography,Breathing control and CT of the upper airway during sleep periods were performed in both groups at supine position without oxygen.All parameters were compared between the two groups.Results In sleeping without Oxygen,data comparative results were as follows:1.Tongue area,width and height of soft palate,and Genioglossal(GG) width in AHI>20 group were larger than those in 5＜AHI≤20(P<＜0.05),and the former had the smaller minimum area of retropalatal region(P＜0.05).2.P0.1 showed lower in AHI＞20 than 5＜AHI≤20 group,and there were significantly statistical differences between them(P＜0.05).At the same time, Correlation analysis showed that P0.1 was negatively correlated with GG width (r=-0.574,P＜0.05) for patients with AHI＞20 group.And this relationship was only associated with AHI＞20 group.3.There were obviously statistical significance in the minimum area of RP,tongue length and tongue area between different BMI(P＜0.05).With the increasing of BMI, the measurement parameters of the soft palate width,soft palate height,soft palate area,tongue height were increasing,too.But there were not obviously statistical significance.And we found that there were significant positive correlation between AHI and BMI for obese patients(r=0.608,P＜0.001).Conclusions1.Patients with AHI＞20 have more narrowed pharyngeal cavity than 5＜AHI＜20, suggesting that there are some differences of pharyngeal morphology for differences AHI in OSAHS patients during sleeping. 2.PO.l was negatively correlated with GG width(r=-0.574,P＜0.05) for patients with AHI＞20 group.That is to say,with the increasing of AHI,the compensation activity of GG increases.But the impulse to the cerebral cortex is disappearance when GG width is at a certain extent,then the respiratory driver drops even more.3.Patients with different BMI had the differences of pharyngeal morphology and the mechanism for OSAHS in sleeping.With the increase of BMI,the severity of OSAHS is increase,too.AHI is associated with BMI.BMI can indirectly reflect on the degree of upper airway obstruction and the development of disease in OSAHS.

Research of Linear and Nonlinear Methods of Heart Rate Variability for Assessment of Automatic Nervous System during General Anesthesia

Tue, 29 Mar 2011 11:09:11 +0000

Objectives:Heart rate variability(HRV) refers to minor differences between cardiac cycle.HRV contains a lot of information about the cardiovascular regulation, while subtracting and analyzing of the information of HRV can offer a quantitative evaluation of the tenseness and the balance of the sympathetic and vagus nerve activities and their effect on cardiovascular motion.Along with the development of anesthesia,the analysis Of HRV has been the-focus of attention.Anesthesia drugs, procedures and surgical stimulations can significantly affect the functions of autonomic nerve system,followed by evident HRV changes.In recent years,the methods used for the analysis of HRV are mainly time-domain analysis and frequency-domain analysis.But the characteristics of HRV are nonlinear,while the nonlinear analysis is.seldom used.The purpose of this study is to explore the evaluation of cardiac ANS by using the linear and nonlinear analysis under the general anesthesia.Methods:Twenty-four patients scheduled for elective oral and maxillofacial surgery under general anesthesia were studied with mean age 43±5.4 years old and mean weight 56±6.1 kg,ASAⅠ～Ⅱ.Patients with cardiac.disease,diabetes, hypertension or psychoneurosis were excluded.All patients were given intramuscular injection with atropine 0.01 mg/kg,Midazolam0.05～0.1 mg/kg in 30 minutes before the operation.After entering the operation room,ECG,SpO_2,BP were monitored,and the electrocardiosignal was collected by home-built equipment mading by soundcard. The signal was manipulated and calculated by Matlab.Patients were induced by Midazolam 0.1mg/kg,Etomidate 0.3mg/kg,Fentanyl 3～5ug/kg.Anesthesia was maintained by propofol 50ug/kg·min,fentanyl 5ng/kg,vecuronicum 0.08mg/kg.The HRV signals were collected in 5 minutes at four time points,before induction(To), induction(T_1),operation(T_3 ) and analepsia(T_4).Both HR and MAP were recorded at the same time of HRV.Result:1、The hemodynamics features of patients:HR and MAP increased in response to tracheal intubation comparing to other time points. 2、The results of Time-domain analysis:In contrast with T_0,the value of SDNN decreased at T_2,but increased at T_1,T_3;And rMSSD and triangular index decreased at T_1,T_2,but increased at T_3 significantly.PNNso decreased at all time points after indication.3、The results of Frequency-analysis:The value of TP and VLF increased at T_1 and T_3,decreasing at T_2 in contrast with T_0;The values of HF and LF decreased at T_1 and T_2,but increased at T_3.Meanwhile the value of LF/HF increased at.T_1 and T_2,but decreased at T_3.4、The result of nonlinear analysis:The values of Complexity and Entropy decreased statistically,but recovered after analepsia.In the Lorenz plots analysis,the value of VAI decreased at T_1 and T_2,but the value of VLI increased significantly at T_1 and T_3,decreasing at T_2。The index of DFA increased at T_2,but no significant change was observed at other time points.5、There are different degrees of relevance among the various indicators of HRV, exactly,SDNN and TP,SDNN and VLI,SDNN and triangle index,VAI and rMSSD, HF and rMSSD,VAI and HF,VAI and PNN50,complexity and ApEn,scale index and ApEn,scale index and complexity.SDNN,TP,VLI,and triangle index can reflect the activity of autonomic nervous system;VAI,HF,rMSSD and PNN50 can reflect the vagus nerve on heart rate control in correlation analysis.Conclusion:The time-domain,frequency-domain and nonlinear analysis of HRV can reflect the activity of autonomic nervous system function in different degrees.The nonlinear analysis method is more sensitive than linear analysis.Individual differences are larger in frequency-domain analysis,and the data do not follow normal distribution,this phenomenon may be related to the analysis method.The nonlinear analysis can reflect the activity of autonomic nervous system function stably and sensitively.There are different degrees of relevance among the various indicators of HRV.

Clinical Study of Sinusitis with Tracheal Intubation

Tue, 29 Mar 2011 11:09:08 +0000

Objective:To compare the incidence rate and outcome of sinusitis in patients who had undergone nasotracheal,orotracheal intubation and incision of trachea for mechanical ventilation.Methods:A total of 87 patients who had undergone mechanical ventilation for COPD, pulmonary interstitial fibrosis,severe pneumonia,bronchial asthma and coronary-artery bypass surgery were classified into three groups:the nasotracheal group,orotracheal group and incision of trachea group.The nasotracheal group also was separated according to the size of nasotracheal tubes:diameter 7.0 mm and diameter 7.5mm.Sinus computed tomography(CT) scans were performed before removing intubation,five days and ten days postoperatively.All patients had no history of previous sinusitis through radiographic evidence.Nasal discharge brush specimen,culture species,temperature,chest X-ray,and blood leucocytes were recorded.Results:The overall incidence rate of sinusitis in nasotracheal group and orotracheal group were 55.07%.Patients in nasotracheal group had higher incidence rate of sinusitis than that in orotracheal group(x~2=8.179,P＜0.01) and incision of trachea group(x~2=6.613,P＜0.05)respectively.Using nasotracheal tubes of 7.0mm diameter had a lower incidence rate of sinusitis than 7.5mm diameter(x~2=4.080,P＜0.05). Clinical evidences showed that mechanical ventilation associated sinusitis always happened in tubes inserted side of sinuses which most affected in maxillary sinus.Conclusion:The incidence rate of sinusitis in patients had undergone nasotracheal intubation was statistically significant higher than oral intubation and incision of trachea groups.Factors which may contribute to the development of sinusitis included the use of nasotracheal tubes of large diameter,traumatic intubation,blocking sinuses, prolonged of mechanical ventilation.There was no evidence showed that sinusitis could increase the occurrence of ventilator associated pneumonia.

Reorganization of Sleep Patterns after Individual Surgical Treatment for OSAHS

Tue, 29 Mar 2011 11:09:07 +0000

ObjectiveTo investigate the character of sleep architecture for obstructive sleep apnea hypopnea syndrome(OSAHS)and give them individual surgical treatment according to their multiple obstructive parts.And to investigate the change of the sleep parameter,the reorganization of sleep architecture,the drowsiness and the reduce of the saturation of blood oxygen after the treatment.MethodsWe chose 40 moderate and severe patients who were finally diagnosed with OSAHS by polysomnography(PSG),electro-fibro laryngoscope and M(u|¨)ller test and all of them were consistent with the 2002 Hangzhou criteria.And we chose 40 primary snoring as the control group.All patients were performed individual surgical treatment after preoperative treatment with continuous positive airway pressure treatment(CPAP) 5-7 days.They filled the Epworth sleepiness scale with the special guidance.And they were given different surgical treatment according to their different obstructive parts.After 6 months of the surgery,they refilled the ESS, rechecked PSG and compared it to the previous results.Results1.Compared with the control group,the AHI,BMI,neck circumference, LSaO_2,MSaO_2,MAI,ESS,the superficial sleep time,the deep sleep time and the rapid eye movement of OSAHS are significantly different(P＜0.01).2.In the 40 cases(severe 30 and moderate 10)after the individual surgical treatment,healed 17.5%,utility 65%,improved 17.5%,no improved 0.3.After the operation,there is no significant change in BMI(P＞0.1),ESS was reduced from(14.60±3.70) to(8.32±2.22)(P＜0.01),and BMI was correlated with AHI(r=0.886).There is also an negative correlation between BMI and LSaO_2% (r=-0.742).We rechecked the PSG and found that AHI was reduced from (50.12±17.04) to(13.61±7.30).The lowest saturation of blood oxygen(LSaO_2) was increased from(70.02±9.66)%to(84.63±4.90)%,the Mean saturation of blood oxygen(MSaO_2) was increased from(92.03±2.95)%to(95.59±1.67)%(P＜0.01),the sleep efficiency was increased from(79.60±6.23) to(85.74±6.23).At the same time, the sleep architecture were effectively changed,the percent of S1 phase in total sleep time(TST) was reduced from(16.94±4.51)%to(10.86±5.64)%,the percent of S2 phase in total sleep time was reduced from(58.12±6.65)%to(50.69±7.13)%,the percent of S3+S4 phase in total sleep time was increased from(9.73±5.81)%to (17.51±8.21)%,the percent of REM phase was increased from(15.44±4.95)%to (21.37±5.31)%(P＜0.01).The Micro-arousal index was reduced from(38.42±19.32) to (14.31±8.88)(P＜0.01)and MAI was correlated with AHI(r=0.125).There is also an negative correlation between MAI and ESS(r=-0.735).Conclusions1.In the patients with moderate and severe OSAHS,the sleeping architecture is indiscriminate,the sleep gragmentation and the sleep cycle is insufficient,the superficial sleep time was extended,the deep sleep time and the rapid eye movement was decurtated or destitute,and the sleepiness is conspicuous aggravated.2.Individual surgical treatment for patients with moderate,severe OSAHS,the saturation of blood oxygen was significantly improved,the AHI and MAI were reduced,the percent of S1 and S2 phase were reduced,the percent of S3+S4 and REM were improved,the sleep architecture was also changed,and the sleep efficiency improved.So the individual surgical treatment is an effective method,and has a significant practical importance.

Study of Complement 9 Expression in the Laser-induced Choroidal Neovascularization and Cobra Venom Factor Intervention in Mice

Tue, 29 Mar 2011 11:09:06 +0000

ObjectiveAge-related macular degeneration(AMD) is the most common cause of low vision and blindness in the elderly worldwide persons,choroidal neovascularization (CNV) is the leading cause of blindness.The aim of theses experiments is to observe the components of Drusen- membrane attack complex(MAC)expression in the laser-induced choroidal neovascularization in mice and to study the formation of MAC playing the role in the development of CNV.In this study we prewent CNV development with cobra venom factor(CVF) by intraperitoneal injection.Further explain the immunological mechanisms of AMD and explore a effective methord to inhibite complement activity,provide a theoretical basis and new methods.Method1.Thirty six 6-8 weeks C57BL/6J mice,male and female unlimited,they are all healthy and no eye disease checked by ophthalmoscope and slit lamp.The mice divide into three groups by random,group one as control group(n=12),group two,CVF treated:C57BL/6 mice(n=12) are treated i.p.with 0.1μg/g of CVF 2 days before laser photocoagulation and every day after laser treatment,group three,C57BL/6 mice(n=12) are only treated by laser photocoagulation.2.Induction of CNV in mice:the mice are anesthetized by the 10%chloral hydrate(0.3ml/100g),mydriasis with Tropine amide compound,surface anesthesia, drops 1%methyl cellulose amount in conjunctival sac of experimental eye,place the slide as a contact lens,the krypton red laser(50μm spot size;0.1s duration;120 mw, Wavelength 647nm),four laser spots are placed in each eye close to the optic nerve. The bubbles generated after photocoagulation can be regarded as the symbol of breakage of bruch\'s membrane,checked the laser spots by ophthalmoscope.3.At the 1,3,5,7day,the mice are anesthetized by the 10%chloral hydrate.Put out the eyes taking the Frozen section to the SABC-FITC histochemical stain and observe the positive stain under Fluorescence microscope.Each eye select two laser regional integrity retina structure by SABC-FITC histochemical stain and HE stain.Each section selects 5 fields by random(×200 times),figure the positive stain in each field.The HE stain is to observate the morphological changes in retina under the light microscope.4.Statistics method:The result put into the spss13.0 statistic software.after normality test and homogeneity test for variance.The only experimental group or only time group data use one way ANOVA,the different experimental group and time groups dates use two way ANOVA,then compare the each group using SNK test,p＜0.05 has the statistic meaning.Results:1.HE stain display that in the control group,the mice retina has the regular layers, the photoreceptor cells are regular,the nuclear stains deep violet,the boundary is sharp,retinal pigment epithelium(RPE) cell nuclear is light violet,ellipse,clear and stain uniformity.Two group mice:One day after retinal photocoagulation,we can find that retinal tissue have edema,Bruch membrane and RPE layer are significantly damaged,the outer and inner nuclear layers structure of retina are floc chaos;three days after retinal photocoagulation,choriocapillaries is obviously congestive,RPE cell are hyperplasia,but don\'t find neovascularization;seven days after retinal photocoagulation,a small amount of CNV are formed in facula area,mild retinal detachment and subretinal scarring lesions,cells arrange irregularly in scar lesions, injuried RPE decompose and break,photoreceptors fracture and some disappear,outer nuclear layer disappeare,inner nuclear layer disorder.Three group mice:one day after retinal photocoagulation,retinal edema is lighter than group two;three days after retinal photocoagulation retinal tissue remains edema,congestion of choriocapillaries are lighter than guoup two;seven days after retinal photocoagulation don\'t find CNV.2.SABC-FITC histochemical stain appears that under different treatment groups and at different time point groups,with the two way ANOVA.The mice C9 IOD(integrated optical density)in SABC-FITC histochemical stain are differernts,and these difference between different treatment groups are significant at the 0.05 level analyzed by statistics software spss13.0.(p=0.000).So do the different time points.We compare each group with SNK test,except the group one and group three, the difference among other groups all has significant statistic meaning(P=0.000). Under the only treatment group,we use one way ANOVA,besides the group one and group three(P_1=1.000,P_2=1.000),then we use SNK test shows besides group two between one day,three day and five day(P=0.993,P=0.924,P=0.917),the mice C9 IOD in SABC-FITC histochemical stain are differents among other groups,and these difference between different treatment groups are significant at the 0.05 level analyzed by statistics software(P＜0.05).Under the only time factor,we use one way ANOVA,at every time period,the mice C9 IOD in SABC-FITC histochemical stain are significant differents at the 0.05 level(P=0.000),then we use SNK test shows that besides every time period between the group one and group three(P= 0.995,P=0.983,P=1.000,P=0.977),the mice C9 IOD in SABC-FITC histochemical stain between other groups are significant differents at the 0.05 level(P＜0.05).Conclusion:Our reseach sets up the model of laser-induced choroidal neovascularization in mice.We then focused our attention on the formation and deposition of the MAC in CNV lesions,we observed that the neovascular complex stained very strongly for C9 in complement-sufficient C57BL/6 mice with SABC-FITC histochemical stain.In contrast,C57BL/6 mice depleted of complement with CVF,laser spots did not stain for C9 and did not develop CNV,we demonstrated that complement was essential for the development of CNV in C57BL/6 mice after laser photocoagulation.These results showed a correlation between the presence of complement,MAC deposition,and choroidal new vessel formation following laser photocoagulation.

The Expression of Neuritin mRNA and Protein in the Visual Cortex of Normal and Monocular form Deprived Rats

Tue, 29 Mar 2011 11:09:04 +0000

ObjectiveTo detect the expression of Neuritin mRNA and protein in the visual cortex of normal and monocular form deprived rats by SYBRgreenⅠreal-time fluorescent polymerase chain reaction and immunohistochemistry stain methods.To explore the associatiion Neuritin with visual experience and ages,and influence by Abnormal vision.To provide the molecular mechanism of the visual plasticity.Methods1.Fifty healthy Wistar rats were divided into 10 groups:5 in every group,in which 1-6 groups for normal group(N),7-10 groups for monocular deprivation group (MD).The eyelids of MD groups were sutured right eyelid at 14 days.Rats were housed in alike environment,the time-point for normal group is postnatal 7、14、21、28、45、90 days.The time-point for MD group is postnatal 21、28、45、90 days.The rats were killed at the design time-point,the procedur is anesthesia、decapitation、get the brain、extract total RNA、detect the quality and density of total RNA、reverse transcription reaction、common PCR reaction、SYBRgreenⅠreal-time fluorescent polymerase chain reaction,then apply computer self-software compared the sample with standard preparation,and combine with references to obtain the Neuritin mRNA quantity in rats\' visual cortex.Analyzed and compared the Neuritin mRNA quantity,experiment data was indicated with mean±standard deviation((?)±s). Analyzed data with SPSS16.0,adopted t-test in normal and MD groups,ANOVA in the different stages of same group.2.the rats\' groups and animal model the same to the part one.The procedur is anesthesia、perfusion,fixation and opening cranium,cut the posterior brain coronally, putted it in fixative solution for 24-48h,then imbedded with paraffin routinely.Sliced the prepared tissue in 4μm,carried out routine HE stain and immunohistochemistry stain.Observed the expression of Neuritin in visual cortex under the light microscope.Pictures were Collectted with Olympus micro-image system,integrated optical density(IOD) were determined with image analysis system.Analyzed and compared the IOD,experiment data was in dicated with(?)±s.Analyzed data with SPSS16.0,adopted t-test in normal and MD groups,ANOVA in the different stage of same group.Results1.(1)To analysis the standard curve:from the solubility curve we know that Neuritin and GAPDH amplification single PCR production in any density ranges, without primer dimer and hairpin;From the amplification curve and standard curve we know that there is a well linear correlation,there could obtain exact quantitation.(2) The expression of Neuritin mRNA in normal rat visual cortex: before open the eyes there is a small quantity expression,P7 was 0.152±0.011,then increased gradually,P14 was 0.234±0.052,P21 was 0.527±0.047,then reached its climax value in P28,it was 0.846±0.067,then decreased slightly and maintained stabiy in adults,P45 was 0.445±0.020,P90 was 0.410±0.017.Significant deviation existed among different ages with one-factor analysis of variance(F=177.52, P＜0.01).3.The MD groups:the expression is obviously less than normal groups,at MD21 the quantitation was 0.412±0.015(t_(21)=5.262,P＜0.01)、MD28 was 0.501±0.019(t_(28)=11.223,P＜0.00)、MD45 was 0.381±0.030(t_(45)=3.970,P＜0.01) and MD90 was 0.384±0.015(t_(90)=2.492,P＜0.05)。2.(1).HE stain results:The neurons of visual cortex in rats distributed in layers,from superficial to the deep layer was molecular layer,external granular layer,external pyramidal layer,internal granular layer,internal pyramidal layer and polymorphous cell layer,the neurons in every layer lined in order.There was not obviously abnormality in the visual cortex of monocular deprivation group.(2).The expression of Neuritin in rat visual cortex in different stages:In light microscope, Neuritin positive reaction product was yellow granulous,main located between neurons.Neuron cell body,blood vessel and callosum were not labeled.The Neuritin positive granula were seen in every lays of visual cortex.The expression of Neuritin increased developmentally in the normal groups.At P7 the IOD was 6.48±0.62、P14 the IOD was14.56±2.37,at P21 it increased to23.36±4.28,at p28 reach to the peaking the IOD was 67.94±5.98,then decreased P45 was 26.85±2.33, maintained at higher level.The expression of Neuritin in visual cortex of monocular deprivation rats decreased,the IOD at MD21 was18.17±1.42(t=5.571,P＜0.05), 30.92±2.17 at MD28(t=6.802,P＜0.01),19.33±2.08at P45(t=5.379,P＜0.01).Conclusion1.The expression of Neuritin mRNA and protein in any ages of rats visual cortex,as well the both to show the synchronism.To suppose may that the Neuritin gene to participate in developing visual system,and play an important roles in the growth and maturation of visual cortical neuron.2.The age factor effect the expression of Neuritin mRNA and protein,to accord with the wave change of the visual development critical.3.Visual experience regulats the expression of Neuritin mRNA and protein in development stage of visual cortex,to suppose may that affect Synapse development and maturation in the visual cortical neuron.In order to provide theory for the pathogenetic of amblyopia.

The Analysis of Dental Model on Normal Occlusion in Han Race of Chinese Adult

Tue, 29 Mar 2011 11:09:02 +0000

Objectives This study is aim to establish the norms of dental model on normal occlusion in Han race of Chinese adult,reveal the characteristics of dental model, compare with different sexual and racial population and provides the reference for diagnoses in orthodontics.Methods 42 cases of normal occlusion dental model of Han race of Chinese adult(20 males and 22 females) were selected.The width of crown,the width and length of the dental arch,the basal arch width and length were measured.The Bolton index,Pont index,and Howes value were figured out.Correlation analyses of some norms were calculated.The measurements were analyzed with statistic methods and compared with that of different groups,meanwhile,the value of Bolton index and Pont index in clinic were eval uated.Desgined a program Han race of Chinese to output customized dental arch with Matlab 7.0.Result 1.The norms of dental model on normal occlusion in China adults are established.2.The Bolton index,Pont index,and Howes value of Han race of Chinese adult are established.3.Establish the system of customized dental arch in Han race of Chinese adult.Conclusion 1.The crown,dental arch and basal arch are different in sex.2.The Pont index is different in sex.Racial difference should be paid more attention.3.Statistically significant correlations between the crowns width of front teeth in maxillary and mandibular could be established,so do the crowns width of the whole teeth.The crowns width of front teeth or the whole teeth in maxillary and mandibular is highly correlated with the dental arch.The crowns width of front teeth or the middle and back dental arch in maxillary are markedly correlated with basal arch.4.The system of customized dental arch can instruct the clinical therapy.

Effects of Anti-TLR4 and Pyrrolidine Dithioearbamate on Lipopolysaccharide-stimulated Human Periodontal Ligament Cells

Tue, 29 Mar 2011 11:09:01 +0000

ObjectiveThe purpopse was to investigate the effects of anti-TLR4 and PDTC,a NF-κB inhibitor,on lipopolysaccharide-mediated nuclear translocation of NF-κB and cytokine IL-1βand IL-8 expression in human periodontal ligament cells,and study the mechanism of TLR4-NF-κB signaling pathway in periodontal injure.MethodsHuman periodontal ligament cells were prepared by using explant culture technique.Collecting of the 3-6 passage cells in logarithmic growth phase were used for the following experiment.There were four groups in this study.Control group: cells were treated with Dulbecco\'s modified Eagle\'s medium(DMEM) containing 1% FBS.LPS group:cells were treated with DMEM containing 1%FBS and 10μg/ml LPS.TLR4 group:cells were incubated with anti-TLR4 for 30min before DMEM containing 1%FBS and 10μg/ml LPS was added.PDTC goup:cells were incubated with PDTC for 30min before DMEM containing 1%FBS and 10μg/ml LPS was added.Cells of all groups were cultured at 37℃in 5%CO2 for 24 hours.Then the amounts of interleukin 1βand interleukin 8 secreted into the culture medium were determined by an enzyme-linked immunosorbent assay(ELISA) respectively.TLR4 and nuclear translocation of NF-κB in the cells was evaluated by indirect immunofluorescence.Results1.The amounts of IL-1βand IL-8 secreted into the culture medium decreased when cells were incubated with anti-TLR4 for 30 minutes,which had statistic difference compared to cells without anti-TLR4(P＜0.05).2.PDTC suppressed cell proliferation,significantly different from the control (P＜0.05).The effects of 1000μmol/ml and 200μmol/ml PDTC were more significant. 50μmol/ml PDTC and above incubated cells for 24 hours promoted apoptosis.3.The secretion of IL-1βand IL-8 was not inhibited by 10μmol/ml PDTC,and, more pronounced inhibition of 100μmol/ml and 1000μmol/ml PDTC showed there was significant difference from the control(P＜0.05). 4.Cytoplasmic staining by TLR4 antibody was obvious and it had no significant difference before and after LPS stimulation.No TLR4 antibody staining could be observed with anti-TLR4 treatment.The NF-κB antibody staining showed strong green fluorescence signals in the nuclei with weak cyroplasmic staining after LPS stimulated.The anti-TLR4 and PDTC treatment strongly reduced the intensity of nuclear staining.Conclusions1.TLR4 exsits on human periodontal ligament cells surface and can recognize lipopolysaccharide.2.PDTC can suppress human periodontal ligament cells proliferation and induce apoptosis.3.Nuclear translocation of NF-κB and IL-1βand IL-8 expression in LPS-stimulated human periodontal ligament cells can be inhibited by anti-TLR4 and PDTC,which indicates that TLR4- NF-κB signaling pathway play a significant role in periodontal injure.Blocking the pathway can be a new treatment with periodontal disease.

Retrospective Analysis of 185 Inpatients with Drug Eruption

Mon, 28 Mar 2011 11:44:28 +0000

ObjectiveTo investigate the clinical feature of inpatients with drug eruption,the relationship between causative agents and clinical types,the impact factors of severity and systemic damage,provide the prevention and treatment of drug eruption.MethodsA retrospective analysis was carried out on the clinical data of 185 inpatients in Tianjin Medical University General Hospital nearly 7 years,from Jan 2002 to Dec 2008,to analysis the sex,age,latent period,causative agents,types,systemic damage, laboratory examination,therapy of drug eruption with excel 2003 software.The difference of systemic damage and severity between the four age groups(0-20、21-40、41-60、61-),were compared.Statistical analyses are performed with SPSS 15.0 software.Results185 patients,85 males and 100 females,with drug eruption were enrolled with an age range of 4 years to 89 years,mean(39.16±20.58years).The largest percentage of total number was from 21 to 30(22.70%).The longest mean incubation period was anti-epilepsy drugs in all drugs(16.78±9.45 days) and Steven-Johnson syndrome in all clinical types(13.87±10.51).The drugs most often responsible for the drug eruption were antibiotics(penicillins 12.4%,cephalosporins 12.97%,etc.),followed by the antipyretic analgesics(23.78%),Chinese patent medicine(9.19%),anti-epilepsy drugs(6.49%).The commonest clinical types were erythema multiforme(35.14%), urticaria(23.24%),morbilliform-scarlatiniform exanthesis(20.54%).The commonest type is urticaria in furaxone-induced drug eruption and fixed drug eruption in sulfonamide-induced skin rash.6 patients with fixed drug eruption are all male and there was significant difference in male and female.37 patients were concomitant with the liver damage,the commonest causative agents were antipyretic analgesics, cephalosporins,carbamazepine.The incidence rates were higer in younger(0-20) and elder(61-) than the other groups(p＜0.05).Among them there were 29 cases of severe drug eruption.Administering corticosteroid early and sufficiently and preventing & curing complication actively was the key to cure severe drug eruption.The severe drug eruption\'s mean course was longer than the others(p＜0.05).Conclusions1.There were no difference between men and women.2.The most frequent causes were antibiotics(mainly penicillins and cephalosporins),followed by the antipyretic analgesics,Chinese patent medicine,anti-epilepsy drugs.3.The commonest clinical types were erythema multiforme,urticaria,morbilliform- scarlatiniform exanthesis. The commonest type is urticaria in furaxone-induced drug eruption and fixed drug eruption in sulfonamide-induced skin rash.The commonest drug leading to severe drug eruption was antiepileptics.The fixed drug eruption was very common in male.4.Different drug and different types had regularity of the incubation period.5.The commonest drug leading to systemic damage were antipyretic analgesics, cephalosporins,antiepileptics and systemic damage mainly including liver,kidney, hemopoietic system.6.Age had little effect on the severity of drug eruption,but had difference on systemic damage.7.Administering corticosteroid early and sufficiently and preventing＆curing complication actively was the key to cure severe drug eruption.

Study on the Establishment and Application of the PI Forecasting Model for Radiation Induced Lung Injury

Mon, 28 Mar 2011 11:44:27 +0000

Objective:By the analysis of the clinical data,radiotherapy data and prognosis data,and serum cytokines detection in NSCLC patients,picking out the factors associated with radiation induced lung injury.And establish a PI forecasting model using to predict radiation induced lung injury.Methods:120 NSCLC patients from the radiotherapy department in Tianjin Cancer Hospital,was recorded from 2008.3.18 to 2008.8.5.And the follow up time is to 2009.3.18.These factors were designed in a table.1.General clinical data.Including:age,sex,KPS score,smoking or not,primary disease in lung,diagnosis time,primary symptom,the tumor location,pathology types,clinical staging,radiotherapy method,treatment Taxinol using,pulmonary function index and so on.Blood sample were collected at the time of before radiation,in radiation at 30-40Gy,and 3 months after radiation.The cytokines such as IL-1,IL-6,TGF-β1,ACE,were detected.By the analysis of the radiotherapy program and DVH graph,get the date associated with,the V5、V10、V15、V20、MLD、PTV MLD,and the total dose and fraction.According to the CTC AE 3.0,RTOG acute lung pneumonia criteria,Grade 2 or worse lung injury was used as end point.Results:1.The development of the radiation lung therapy.There were 120 cases,the follow-up time was to Month 18th,2009.The middle time was 10 months,from 8month to 12month.we had a evaluation at 3month and 6 months.There were 39 cases to happen the injury.Among the total,18 cases is grade 1,12cases is grade 2,7cases is grade 3,2cases was grade 4.There were 21 cases≥2 cases.The incidence rate was 17.5%, and there were two cases died for other reasons. The statistics analysis show that,chemotherapy,Taxinol using,lung function IL-6、TGF-β1,V5,V20,V10,were associated with injury.PI forest model:PI=h(t,x)=h0(t)exp(1.138X1+2.833X2+1.025X3)When V5≥51%,the risk increase 1.12 times by the 1%increase.When V20≥25%,the risk increase 1.999 times by the 1%increase.The IL-6 6ng/ml, and P=0.040.the risk increase 2.049 times by the 1ng/ml increase.Conclusions:1、radiotherapy lung injury was developed by all kinds of factors.Taxinol using chemotherapy and the lung function was important before radiotherapy.V5 was more important in the early injury.and the V20 was important in the late injury.when we designing a program,we should pay attention to V5,V20 and MLD.And the special character in different patients should be individed.PI foresting model should be proofed at the following research.

The Clinical Study of Computed Tomographic Laser Mammography

Mon, 28 Mar 2011 11:44:25 +0000

SectionⅠComputed Tomographic Laser Mammography and Images features in Normal BreastsObjective:The objective of this study was to investigate the scan technique of CTLM(computed tomographic laser mammography) and analysis the images features in normal breasts.Materials and Methods:The study involved 150 female Volunteers(20-69 years of age;median age:42years) underwent CTLM scan from September 2007 to June 2008.Result:The breasts diameter of 150 women were from 52mm-126mm.The slices of these breasts is from 9-50 and 1mm,2mm and 4mm slice thickness were be set.The normal vessel were shown areas of high absorption like ribbons(53 cases),branches(66 cases), stars(11cases),cones or cylinders(39 cases).9 cases shown deep vessels.Conclusion:Our study indicated that the skill of CTLM was the foundation of quality images.It was important for technicians to chose the suitable scan conditions according the size and shape of the breasts. The images features in normal breasts were regular and characteristic. SectionⅡComputed tomographic laser mammography, (CTLM),as an Adjunct to Mammography in Patients with Dense BreastsPurpose:The objective of this study was to evaluate the utility of CTLM,(computed tomographic laser mammography),as an adjunct to mammography in women with dense breast tissue(Breast Imaging Reporting and Data System[BI-RADS]density category 3 or 4).Materials and Methods:The study involved 155 women scheduled for biopsy or surgery from January 2008 to August 2008. Breasts density was evaluated initially with Mammography. Parenchymal patterns of"heterogeneously dense"(BI-RADS Grade 3:51 to 75%fibroglandular) and "extremely dense",(BI-RADS Grade 4:＞75% fibroglandular),were used to classify breast density.CTLM examination was performed subsequently to mammography on patients with dense breasts.We then compared the findings of mammography alone,CTLM alone,and CTLM adjunct to mammography(mammography+CTLM), with pathology.Sensitivity,specificity and positive predictive values were calculated by using standard methods.Between-group differences were evaluated with the x~2 test for categorical variables.Result:155 women were classified by mammography into 74 heterogeneously dense breasts and 81 extremely dense breasts. Pathologic analysis revealed 79 malignant and 76 benign breast lesions. Angiogenesis was observed significantly more often in malignant than in benign lesions(79.75%VS.31.57%x2=36.550 p=0.000),Among extremely dense breasts the sensitivity of mammography alone,CTLM alone,and mammography+CTLM was respectively,35.13%、78.37%、81.57%.The sensitivity of mammography VS mammography+CTLM was 35.13%VS.81.57%(x~2=13.071 p=0.000).The specificity of mammography alone,CTLM alone,mammography+CTLM was 89.19%、72.97%、78.13%.Among heterogeneously dense breasts the sensitivity of mammography alone,CTLM alone,mammography+CTLM was 66.67%、80.95%、95.34%.The sensitivity of mammography VS mammography+CTLM was 66.67%VS 95.34%(x~2=11.131 p=0.001). The Specificity of mammography alone,CTLM alone, mammography+CTLM was 87.18%、64.10%、55.26%.The sensitivity of CTLM in heterogeneously dense breasts VS extremely dense breasts was (78.37%vs 80.95%x~2=0.446 p=0.504).The specificity in these cases was 72.97%vs 64.10%x~2=0.0001 p=1.0)Conclusion:CTLM provides information about angiogenesis.It does not appear to be affected by breast density and when used as an adjunct to mammography in heterogeneously and extremely dense breasts,increases diagnostic sensitivity CTLM could successfully distinguish malignant from benign lesions of dense breast.This study suggests that in clinical practice,using CTLM as an adjunct to mammography may help solve the dense breast problem.

Research of the Relationship between the Concentration about ADMA、eNOS、NO with Acute Cerebral Infarction

Mon, 28 Mar 2011 11:44:24 +0000

Purpose:Ischemic cerebrovascular disease has high morbidity and mortality both at home and abroad,damage in endothelial function of cardiovascular and cerebrovascul -ar diseases(CCD) be considered as a widespread phenomenon,with the developing of its research,the treatments are divided into improving the lack of regional blood supply gradually and protect the ischemic nerve cells,with in these the nitric oxide (NO) which be produced by vascular endothelial cells is an inportant intrinsic agent in regulate cerebral blood flow and protect nerve cells.Under physiological condition ,NO is released sustainly,L-arginine as its substrate and mainly by endothelial nitric oxide synthase(eNOS) catalytic generated.ADMA is the analogues of L-arginine, and it can compete with eNOS active site to inhibit the activity of NOS,thus will to reduce the generation of NO,resulting in endothelial dysfunction.This study through exam the levels of ADMA in plasma and NO,eNOS levels in serum about patients of who suffered with cerebral infarction combined with or not hypertension.In order to explore the pathogenesis of CCD that through ADMA induced,on the other hand,to explore the neuroprotective effect of eNOS.All of these will be benefit to cerebral infarction in clinical intervent early and drug treatment.Moreover,it will providing theoretical guidance and objective basis to cerebral infarction.Methods:Select fifty cases who suffered with acute cerebral infarction but normote -nsive,sixty cases of acute cerebral infarction with hypertension,and fifty cases of healthy controls,three groups of age,sex matched.Patients who presented with in one-week after the onset,and collected venous blood after fasting at least twelve hours,then examed the levels of ADMA,NO and eNOS each group.ADMA was examed by HPLC,NO was examed by nitrate reductase method,eNOS was examed by enzyme-linked ELISA method.Value to mean±standard deviation of that result, all statistical analyses were performed with the statistical software SPSS11.5.Results:1.Acute cerebral infarction with normal blood pressure group plasma ADMA level was(0.73±0.16μmol/L),cerebral infarction with hypertension plasma ADMA level was(1.03±0.22umol/L),the healthy control group,plasma ADMA level was 0.47±0.16umol/L,there have statistically significant difference among there groups(p＜0.05).Moreover,ADMA and the level of hypertension were positive -ly correlated(p＜0.01),there were non-related to age,TC,TG,smoking,drinking and other factors.2.Cases of infarction patients with hypertension whose eNOS level was(47.61±20.2 1U/ml),significantly lower than the normal blood pressure group which eNOS level was(55.70±18.96U/ml)(p＜0.05),and both lower than the normal control group,its eNOS levels in serum(66.54±24.13U/ml)(P＜0.05).3.NO level in Infarction combined with hypertension group was(55.28±10.78umol /L),the group cerebral infartion but normal blood pressure whose serum NO level was(60.20±14.03umol/L),there were not significant difference between the two groups(p＞0.05),but were lower than NO level of the control group(70.27±6.78 umol/L),the difference has statistical significance(p＜0.05).4.Determination of high blood pressure(HBP) classification at all levels with hypert -ension in patients with cerebral infarction inspection indicators:patients of HBP classl whose plasma ADMA level of 0.74±0.23umol/L,were significantly lower than class 2 and 3 patients(whose ADMA levels to(0.96±0.17 umol/L) and(1.05±0.11 umol/L),and between the later two groups was not significant different (p＞0.05);eNOS levels among three HBP groups show:classⅠwas of(51.27±16.76 umol/L),class 2 group was 49.43±19.22umol/L,both they were higher than class 3 group(45.45±14.10 umol/L),and have significant differences(p＜0.05);three groups of patients with serum NO concentrations were(53.16±11.48 umol/L), (50.87±16.03 umol/L) and(46.02±16.82 umol/L),but there were no significant differences(p＞0.05).5.After the OCSP classification founded that level of ADMA in both TACI and PACI were higher than LACI;eNOS level in TACI and POCI were lower than LACI group; and the NO level in TACI was lower than LACI.The differences were significant.6.Pearson-correlation analysis showed:normal blood pressure group cerebral ADMA levels with eNOS were negative correlation(r=-0.501,p＜0.05);cerebral infarction combined with hypertension group ADMA levels with eNOS were also negatively correlated(r=-0.717,p＜0.05).ADMA and NO levels were negatively correlated between two groups,correlation coefficients were -0.484 and -0.312. 7.Respectively,the traditional risk factors for cerebral infarction such as gender,age, hypertension,diabetes,smoking,drinking,etc.and ADMA to do one-factor analysis, statistic standard is p＜0.05,it will be removed if p＞0.05,and factors of p＜0.05 were performed the multiple stepwise Logistic regression analysis found that in the control of other traditional risk factors of ischemic cerebrovascular disease,elevated plasma ADMA levels was still a independent risk factor.Conclusion:The levels of ADMA in plasma were higher significantly in acute cerebral infarction,but levels of eNOS in serum were lower obviously,indicated that vascular endothelial dysfunction was prevalence in cerebral infarction.There have significant difference in the levels of ADMA and eNOS which in the two groups—CI combined with hypertension and CI but normotension group,the finding suggesting that there have correlation between the hypertension with levels of ADMA and eNOS, and there were correlation with the degree of hypertension.On the other hand,based on OCSP classification,ADMA may be close correlated with cerebral infarction area. ADMA was negatively correlated with eNOS,eNOS as a key enzyme in produce NO, so the quantity and biological effects of NO will be regulated directly by eNOS,thus, we think that the influence of ADMA to vascular endothelial function through the eNOS/NO pathway.Controlling for other factors,the multi-factor analysis showed that ADMA may serve as an independent risk factor for cerebral infarction,it can be used as predict factor for stroke.As a result,ADMA will be as a treatment target of CI and may expe -cted to reduce its morbidity and mortality.

Long-term Therapeutics Effects of Varying Treatment Choices in Patients with ST Elevation Acute Myocardial Infarction

Mon, 28 Mar 2011 11:44:22 +0000

Objective:We sought to compare the effect of different treatments on the long-term outcome of acute ST elevation myocardial infarction(STEMI) patients,as well as to evaluate the impact of varying time from symptom onset to reperfusion on long-term prognosis.Moreover,we want to identify the independent predictive factors of STEMI patient in developing MACE.Methods:From January 2004 to July 2007,432 patients treated in the inpatient department of Zhongshan Hospital affiliated to Fudan University were reviewed.(1)According to treatment methods,432 patients were classified into the conservative group(n=115),the immediate PCI group(n=141) and the late PCI group(n=176).The general conditions of the patients in three groups and the PCI information of the immediate and the late PCI group were collected and compared.The primary end-point was the happening of all-cause death,the combined end-point was MACE, including all-cause death,recurrent infarction and the target vessel revascularization. Kaplan-Meier analysis and log rank test were used for univariate analyses of factors predictive of survival and MACE free survival,and multivariate analysis was carried out using the Cox proportional hazards model to determine the variable relevant to survival and MACE free survival.(2) The patients who received immediate PCI was divided into the≤6h group(n=64) and＞6h group(n=77) according to the symptom-onset to balloon time.The end-point was MACE,as it said in part one.The method was the same as part one.The patients who received late PCI was divided into 12h＜t≤1w group(n=50),1w＜t≤2w group(n=75),and t＞2w group(n=51).The end-point was MACE,as it said in part one. The method was the same as part one.Results:(1)The general characteristics of patients were well-balanced in three groups except that age,stroke history,new arrhythmia,cardiac shock,killip class,LVEF, LDL-C and the use of tirofiban and LMWH showed statistically significant difference(P＜0.05).Collateral circulation,the maximal narrowness of IRA,the mean length of stents,and the number of stents were found to be significantly different between the immediate PCI and the late PCI group,and the rest of the PCI information were similar across two groups.After a median follow-up of 30.1 months, treatment choice is associated with mortality(43.2%vs.3.7%vs.7.2%) and were an independent predictor of late survival(P=0.001)by Cox regression.The survival of the immediate PCI or the late PCI group is better than the conservative group(immediate PCI:HR=0.220,P=0.003;late PCI:HR=0.319,P=0.002),though the two PCI groups had no significant impact on the late survival(P＞0.05). Multivariate analysis also revealed that age,stroke history,EF,killip class,the standard use of Clopidogrel were significantly associated with the survival of patients(P＜0.05).Treatment choice is also associated with the occurrence of MACE(45.9%vs.7.4%vs.16.2%) and were an independent predictor of late MACE free survival(P=0.003) by Cox regression.Patients treated with immediate or late PCI had equal MACE free survivals but the immediate group had a better trend(HR=0.490,P=0.062),both of which compared favourably with those in the conservative group(immediate PCI:HR=0.282,P=0.001;late PCI:HR=0.575,P =0.045).Multivariate analysis also revealed that age,stroke history,hyperlipidemia, killip class and the standard use of Clopidogrel were significantly associated with the MACE free survival of patients(P＜0.05).(2) The general characteristics,PCI information of patients and drug use were well-balanced in two immediate PCI groups except that LDL-c and TIMI flow grade 3 after PCI showed statistically significant difference(P＜0.05).Cox regression showed that new arrhythmia,the length of stents,that TIMI flow grade 3 after PCI were independent predictor of MACE free survival.PCI time,multiple vessels involved and the use of statins had a trend to be associated with MACE free survival.The general characteristics,PCI information of patients and drug use were well-balanced in three late PCI groups except that LDL-C,EF,shock,the maximal narrowness of IRA,the number of stents and the standard use of LMWH and Clopidogrel showed statistically significant difference(P＜0.05).Cox regression showed that hyperlipidemia, EF and that TIMI flow grade 3 after PCI were independent predictor of MACE free survival.Conclusions:1)The immediate PCI and late PCI had similar efficacy in treating STEMI patients,but were both superior to the drug treatment and treatment choice was an independent predictor of long-term prognosis 2)Time from symptom onset to balloon had a trend to be associated with MACE free survival in immediate PCI subgroup,but no significant impact on late PCI subgroup.

The Clinical Analysis of Acute Myocardial Infarction in the Elderly

Mon, 28 Mar 2011 11:44:20 +0000

Objective To analysis the risk factors,the clinical manifestation,the arrhythmia outbreak situation,the outcomes of CAG and the peak of creatine kinase of patients with acute myocardial infarction,accordingly to summarize the clinical characteristics of the elderly as a special group with acute myocardial infarction.Materials and methods Retrospective analysis of 1015 people with acute myocardial infarction who are performed primary PCI in the Second Hospital of Tianjin Medical University from Jun.2000 to Oct.2008.They are divided into two groups according to the age≥60 year old:Old age group 587 examples,male 387 examples,female 200 examples;Non-old age group 428 examples,male 377 examples,female 51 examples.Compares two groups of risk factors,the clinical manifestation,the arrythmia outbreak situation,the outcomes of CAG and the peak of creatine kinase difference,thus summarizes two group of patient\'s clinical characteristic difference.Results 1 the risk factor:The sex and the smoking history difference of the old age group and the non-old age group have statistics significance(P＜0.05). Hypertension,diabetes and the hyperlipemia and so on risk factor comparison,the differences of two groups do not have statistics significance(P＞0.05).In has in similarly the above three kind of risk factor crowd,has statistics significance compared with the old age group and the non-old age group\'s CAG result difference(P＜0.05).2 The preinfarction angina pectoris formation rate differences of the old age group and the non-old age group do not have statistics significance(P＞0.05).The typical angina pectoris\'s formation rate differences of two groups have statistics significance(P＜0.05).The heart function KILLP graduation(the KillipⅠ,the KillipⅡ, Killip≥Ⅲ) differences of the old age group and the non-old age group have statistics significance(P＜0.05).3 The atrial premature beats,atrial fibrillation,two degree above atrioventricular and premature ventricular beats formation rate comparison differences of the old age group and the non-old age group have statistics significance(P＜0.05).The difference of the ventricular tachycardia and ventricular fibrillation formation rate does not have statistics significance(P＞0.05).4 The infarction related artery distribution comparison differences of the old age group and the non-old age group do not have statistics significance(P＞0.05).The old age group and the non-old age group CAG result comparison difference has statistics significance(P＜0.05).5 Compare the difference of creatine kinase peak(CK＞upper limit 2 times、CK＞1000U/L、CK＞2000U/L) of the old age group and the non-old age group do not have statistics significance(P＞0.05).Conclusion(1) The female patients of the old age group with acute myocardial infarction compare with the non-old age group increase obviously,but the male patients are still crowd\'s majority.The old age patients with acute myocardial infarction suffer from hypertension,diabetes and hyperlipemia\'s probability does not have the difference with the non-old age group,but to compare with the non-old age group who have the similarly hazard factors,the old age group CAG results have relatively serious blood vessel pathological change.(2) The preinfarction angina pectoris formation rate of the old age group do not have the difference with the non-old age group,but after acute myocardial infarction, the typical angina pectoris\'s formation rate is low,namely the senior citizen indolence myocardial infarction formation rate is high.After acute myocardial infarction,the heart function of the senior citizen is bad.(3) The atrial premature beats,atrial fibrillation,two degree above atrioventricular and premature ventricular beats formation rates of acute myocardial infarction in the elderly are high.(4) The infarction related artery distribution of the elderly does not have difference comparing with the non-old age group,the elderly CAG results have relatively serious blood vessel pathological change.(5) The senior citizen and the non-senior citizen creatine kinase peak value does not have the difference.

Microarray-based Analysis of MicroRNA Expression in Rat Cerebral Cortex after Traumatic Brain Injury

Mon, 28 Mar 2011 11:44:19 +0000

ObjectiveUsing the microRNA microarray technology to analysis the differential expressed microRNA in traumatic brain cortex tissue of rats subjected to TBI (traumatic brain injury) at 6h,24h,48h and 72h post injury,respectively,to find candidate microRNA for microRNA interference strategy in TBI therapy.MethodsTen wistar rats were randomly assigned into 5 groups:four experiment groups (according to 6h,24h,48h and 72h post injury time points) and 1 sham operated control group.Each group has two rats.The rats of four experimental groups subjected to lateral fluid percussion injury.Sham operated rats were subjected to anesthesia and craniotomy but were not attached to the fluid percussion device. Separate the cerebral cortex tissues of experimental operated and sham operated rats, to preserve in -80℃.The total RNA of brain tissues were extracted by Trizol reagent, hybridized with the microarray probe that designed and printed to the aminoslide, after rinsing and drying,the slides were scanned with single-channel confocal laser scanning microscopy,the raw data were inputted into SAM(Significance Analysis of Microarrays) statistical analysis software,and we used the list of differential expressed microRNAs from SAM analysis to perform the hierarchical clustering using Cluster 3.0 software.Target gene prediction was made on the PicTar website. The potential physiological functions of microRNAs were generated from the bioinformatic analysis.ResultsSAM statistical software analysis(set twofold as threshold) shown that in six hours post injury group a total of 136 microRNAs expressed,of which 13 microRNAs up-regulated,14 microRNAs down-regulated;in 24 hours post injury group a total of 118 microRNAs expressed,of which 4 microRNAs up-regulated,23 microRNAs down-regulated;in 48 hours post injury group a total of 149 microRNAs expressed,of which 16 microRNAs up-regulated,11 microRNAs down-regulated;in 72 hours post injury group a total of 203 microRNAs expressed,of which 19 microRNAs up-regulated,5 microRNAs down-regulated.Of these,only miR-21 was highly expressed in the four time points after brain injury,the ratio of differentially expressed microRNAs were 2.0215,2.0401,2.0702,and 2.7910,respectively. Through the online target gene analysis tool PicTar,we found that mir-21 has 175 target genes at present.Including apoptosis related gene BCL2、CAPN3、PDCD4、TIMP3、PPP3CA and FASLG,etc.The targeted genes and the signal pathways in which they participate include:PPP3CA in LTP(long-term potentiation);FASLG, PPP3CA,RASA1,NTF3 and RASGRP1 in MAPK(mitogen-activated protein kinase) signaling pathway;PPP3CA in Wnt signaling pathway;JAG1 involved in Notch signaling pathway;PPP3CA in VEGF(vascular endothelial cells growth factor) signaling pathway;CNTFR,STAT3,SPRY1 and SPRY2 in JAK-STATs signal transduction pathway;PITX2,ACVR2A and SMAD7 in TGF-β(transforming growth factor-β) signaling pathway;BCL2,CAPN3,PPP3CA and FASLG in apoptosis.Conclusion1.MicroRNA microarray technology can be used for high-throughput screening of miRNA.2.The list of differential expressed microRNAs of the rat cerebral cortex after traumatic brain injury was generated from SAM statistical analysis and hierarchical clustering.We found that miR-21 was the only microRNA which up-regulated in all the four time points(6h,24h,48h and 72h) observed post injury,respectively,indicate that miR-21 might playing some important roles in the pathological process that occur following TBI.3.We found 175 genes were the target of miR-21 by retrieving the web-based miRNA target gene prediction tool PicTar,including the apoptosis related gene BCL2、CAPN3、PDCD4、TIMP3、PPP3CAand FASLG.4.Through the MAS(molecular annotation system) analysis,we got a preliminary understanding of the interaction between the miR-21 target genes,interaction between miR-21 encoding proteins.We presume their potential important roles in signal transduction pathway,long-term potentiation and apoptosis that participated in the pathophysiological process after traumatic brain injury.This study laid the foundation for investigating the molecular mechanism of traumatic brain injury in respect of microRNA events.5.The function of many miRNAs are not yet well understood including miR-21, thus the practical application of miRNAs have not carried out in clinical practice. The loss-of-function and gain-of-function of specific miRNA and their effect on the expression pattern of the encoding proteins of miRNA target genes will be a new direction of further experimental TBI study.

The Effect of Serum Cholesterol on the Mortality of Severe Acute Pancreatitis

Mon, 28 Mar 2011 11:01:19 +0000

Aims:To analysis risk and protective factors which can early predicting the in-hospital motality in the presence of SAP.To analysis risk and protective factors which can early predicting the in-hospital organ failure in the presence of SAP.Methods:Prediction factors for in-hospital motality were evaluated retrospectively from the clinical data obtained from 346 SAP patients of Tianjin Nankai hospital between January 1999 and December 2008,who were treated with intensive care、blood routine、blood biochemical test and even computed tomography on admission.Results:This analysis revealed that serum WBC、Cr、CRP were risk factors while serum albumin was a protective factors for in-hospital motality.The TC was a protective factor when it between 4.37mmol/Land5.23mmol/L;Further analysis showed that elevated CRP appeared to be associated with high mortatily(P<0.05) while elevated ALB associated with improved survival(P<0.05).Elevated TC was associated with reduced mortality,but the mortality rised up again when the concentration of TC was over 5.23mmol/L.Evaluated TC was accompanied by decreased CRP.Conclusions/Infections Renal insufficiency、Inflammatory reaction、hypoalbminemia were prognostic factors for in-hospital motality;Both higher levels of CRP and lower ALB levels were asoociated with SAP in-hospital mortality.Elevated TC properly could resisted Inflammation and adverse survival in patients with SAP,reduced the in-hospital mortality..

Effects of Xuebijing Injection on Tissue Factor of Monocyte and Coagulation Parameters in Septic Rats

Mon, 28 Mar 2011 11:01:17 +0000

Objectives 1.To investigate the effects of TF and PARs in rat monocytes of peripheral blood stimulated with LPS in vitro.2.To observe effects of Xuebijing injection on expression of TF,IL-6,TNF-αand PAR-1,PAR-2 in rat monocytes stimulated with LPS in vitro.3.To investigate the effects of Xuebijing injection on PAR-1,PAR-2 of monocyte and plasma TF,IL-6,TNF-αin septic rats.4.To observe the effects of Xuebijing injection on coagulation parameters and PLT in septic rats.Methods 1.The monocytes were isolated from the peripheral blood of male Wistar rats,were suspended in cell culture plates,then were stimulated by 1μg/ml LPS. Compared with APC,exposuring to the 10mg/ml Xuebijing injection for different duration(12,24,48 and 72h) the cell supernatants were harvested to determine the levels of TF,IL-6 and TNF-αby ELISA,the monocytes were denatured in cell culture plates to determine PAR-1,PAR-2 by FCM.2.The monocytes were stimulated by 1μg/ml LPS then exposuring to different concentration Xuebijing injection(1,10 and 100mg/ml) for 24h,the cell supernatants were harvested to determine the levels of TF,IL-6,TNF-αby ELISA,the monocytes were denatured in cell culture plates to determine PAR-1,PAR-2 by FCM.3.One hundred and four Wistar rats were randomly divided into four groups:normal group,sham-operation group,CLP group, and Xuebijing-treated group.Blood samples of each group were obtained from abdominal aorta after 12,24,48 and 72h of operation.PAR-1,PAR-2 of monocyte was assessed by FCM,the levels of plasma TF,IL-6 and TNF-αwas determined by ELISA, the expression of TF and IL-6 mRNA of the peripheral blood mononuclear cells was determined by RT-PCR and the PT,APTT,FIB and PLT were assessed.Results 1.In vitro,compared with normal group,PAR-1 up-regulated in LPS group (P<0.01),PAR-2 up-regulated after each time(P<0.05 or P<0.01),TF,IL-6 and TNF-αin the supernatant significantly increased after each time(P<0.01).Compared with LPS group,PAR-1 down-regulated in LPS+APC group after 24,48 and 72h(P<0.01), PAR-2 up-regulated after 24 and 48h(P<0.01),TF,IL-6 and TNF-αdecreased after each time(P<0.05 or P<0.01).Compared with LPS group,PAR-1 down-regulated in LPS+ Xuebijing injection group after 24,48 and 72h(P<0.01),PAR-2 up-regulated after each time(P<0.01),TF,IL-6 and TNF-αdecreased after each time(P<0.05 or P<0.01).2.In vitro,Compared with LPS group,PAR-1 down-regulated(P<0.01) and TF,IL-6 and TNF-αdecreased(P＜0.05 or P＜0.01) treated with 1,10 and 100mg/ml Xuebijing injection.It\'s obvious in 10mg/ml group.PAR-2 up-regulated(P＜0.01) in 10mg/ml group.3.Compared with normal group and sham-operation group,PAR-1 up-regulated in CLP group after 48 and 72h(P＜0.05 or P＜0.01),PAR-2 up-regulated after each time(P＜0.05 or P＜0.01),TF increased after 24,48 and 72h(P＜0.05 or P＜0.01),IL-6 and TNF-αincreased after each time(P＜0.05 or P＜0.01),TF and IL-6 mRNA increased after 12,24 and 48h(P＜0.05 or P＜0.01),PT shortened after 24,48 and 72h(P＜0.05 or P＜0.01),APTT extended after 12 and 48h(P＜0.01),FIB decreased after 12,24 and 48h(P＜0.05 or P＜0.01),PLT decreased after 12 and 24h(P＜0.05 or P＜0.01).Compared with CLP group,PAR-1 down-regulated in Xuebijing injection group after 12 and 24h(P＜0.05 or P＜0.01),PAR-2 up-regulated after each time(P＜0.05 or P＜0.01),TF decreased after 24,48 and 72h(P＜0.05 or P＜0.01),IL-6 decreased after 24 and 48h(P＜0.01),TNF-αdecreased after each time(P＜0.05 or P＜0.01),TF mRNA decreased after 12,24 and 48h(P＜0.05 or P＜0.01),IL-6 mRNA decreased after each time(P＜0.05 or P＜0.01),PT extended after 24 and 48h(P＜0.05 or P＜0.01),APTT shortened after 48h(P＜0.05),FIB increased after 12,24 and 48h(P＜0.01),PLT increased after 24 and 48h(P＜0.05 or P＜0.01).Conclusions 1.Xuebijing injection and APC could decrease LPS-stimulated PAR-1 expression on monocytes and TF,IL-6 as well as TNF-αrelease in supernatants.It\'s the best after 24 and 48h treated by 10mg/ml Xuebijing injection.Xuebijing injection was better than APC in the experiment 2.Xuebijing injection would obviously decreas the plasmic IL-6 and TNF-α,ameliorate the coagulation dysfunction in sepsis by reducing monocytic TF and down-regulating monocytic PAR-1.

The Effect of Different Types of Exercise on Big Mouse Bone Density, Bone Metabolism Biochemical Index and Bone Tissue Metrological Index under Neutralised Gravitation

Mon, 28 Mar 2011 10:57:33 +0000

Osteoporosis(OP) is a disease which charactorised in bone density reduction, bone micro-structural damage with increased bone brittleness therefore more prone to bone fracture.However,the academic community has valued intervention of bone movement as a positive means of development.This experiment was aimed at understanding the different forms of movement of simulated weightlessness on the tail-suspended rats and its bone mechanism.Forty 6-month-old SD rats were divided into 5 groups:a controlled group to simulate the loss of the reorganization of the restoration of simulated weightlessness + free group,simulated weightlessness + swimming group,simulated weightlessness group + running group.The tibia cancellous bone histomorphometry indicators,femur and lumbar spine bone mineral density and bone metabolism in the blood concentration of related indicators of the rats in each group were determined after slaying the rats.Experimental period was 4 weeks;suspension time for 4 weeks;movement form of swimming and treadmill training 6 days/week.Purpose:1.The establishment of an ideal simulated weightlessness in rat tail-suspension model, the model of simulated weightlessness on bone mineral density and bone metabolism in the impact of the internal control mechanism.The rear suspension is osteoporosis in the present study a better model.Therefore,to establish and select the appropriate animal model to study the pathogenesis of osteoporosis,at the same time to find effective methods of prevention and treatment.2.To study the different forms of exercise movement in rats after simulated weightlessness on bone mass,bone metabolism and bone metrology organizations such as the role of the effect of the differences between the means of intervention at the same time the choice of sports and exercise prescription to provide the theoretical basis for the formulation.3.Explore the treadmill movement patterns and swimming patterns on bone mineral density in rats.4.Explore the treadmill movement patterns and swimming patterns of biochemical markers of bone metabolism affected. 5.To explore the treadmill movement patterns and swimming patterns of the bone tissue in rats the impact of metrology.Methods:1.The establishment of animal model of tail suspension device.Rat tail will be using medical adhesive tape wrapped around the roots scattered around other parts of the bear,in the light of Chen Jie,the model of the rat tail-suspension and improved methods,a reasonable suspension;With continuous growth in rats,it is necessary to continue to mediate a high degree of tail suspension.2.Tail-suspended rat model of response to the observation,observation of sporting activities during the rat,diet,weight change,skin color of the tail.3.Biochemical markers of bone metabolism measurement.Microplate reader and Nanjing,the company completed biological reagents measured serum calcium,phosphorus ions,alkaline phosphatase,tartrate-resistant acid phosphatase4.Rat femur bone mineral density measurementThe use of dual-energy-ray scanners detect rat femur bone mass,bone mineral density5.Tibial bone tissue measurement metrology.Removed after tibial fixation with 10%formaldehyde in 2 days,2 hours and water washing,dehydration alcohol gradient(75%,80%,90%,100%all day),methyl methacrylate-embedded,50μm slices(not part of grinding,direct observation of fluorescence),glue stick and remove chip code compression for 24 hours,fine sandpaper grinding,polishing alumina(5μm),HE staining,observed under the microscope and film.Results1.indicators of bone mineral density in the femur,swimming and treadmill rats compared with the loss of significant reorganization of the latter is greater than(P＜0.05);Sports and Freedom group have increased compared to the restoration group,and the treadmill group increased more significantly(P＜0.01).Indicators of bone mineral density in the spine,swimming group,treadmill group compared with the loss of reorganization than the latter(P＜0.05);the restoration of the exercise group and the Liberal group,significantly higher than the latter(P＜0.05). 2.calcium concentrations,simulating loss of the reorganization of higher than control group(P＜0.05);exercise group lost reorganization and simulation group has decreased significantly(P＜0.05);exercise group compared with the freedom of the restoration of a significant decrease(P＜0.05);;Phosphorus concentration,the restoration of movement and freedom of group compared with the loss of the reorganization,there are significant differences(P＜0.05).ALP,lost control group compared to restructuring and a significant reduction (P＜0.05);exercise group compared with the simulated loss of reorgan-ization significantly increased(P＜0.05);treadmill group compared with the freedom to resume significant increase(P＜0.05)TRACP,missing reorganization compared with the control group increased significantly(P＜0.01);loss of reorganization with the treadmill group,free to resume a significant reduction in group(P＜0.01);swimming group and the freedom to restore group significantly increased(P＜0.01),the Running group and the freedom to run the restoration group decreased significantly(P＜0.05).3.The static parametersTrabecular bone volume(TBV(%)),significant loss of restructuring than quiet group(P＜0.01);exercise group lost significantly higher than that of recombinant (P＜0.05).Trabecular thickness(Tb.Th)lostless than the reorganization of a quiet group,but no statistical significance;treadmill group restructuring and losses,to restore freedom group significantly increased(P＜0.05).Trabecular number(Tb.N),lost less than the quiet group of recombinant(P＜0.05); swimming group and the Liberal loss of the restoration group was significantly higher than recombinant(P＜0.05);treadmill group was significantly higher than the loss of recombinant(P＜0.01);treadmill group compared with the freedom to resume significant increase(P＜0.01).Mean trabecular spacing(Tb.Sp),lost more than a quiet restructuring group(P＜0.05); the freedom to restore group,swimming group compared with the loss of the reorganization,are lower than the loss of recombinant(P＜0.05);treadmill group was significantly lower than the loss of recombinant(P＜0.01). Cortical thickness(MCT),lost significantly less than the reorganization of a quiet group(P＜0.05);running higher than the loss of the reorganization of the Running group,(P＜0.05);swimming group and the relative loss of freedom of the restoration group has increased the trend of restructuring.4.The dynamic ParametersPercentage of fluorescent-labeled perimeter(%L.Pm),higher than the control group lost recombinant(P＞0.05);relative simulated reorganization,significantly reduce the freedom of the restoration group(P＜0.05),the exercise group were lower than the loss of the reorganization,but no significant difference(P＞0.05);swimming group, treadmill group was significantly higher than that of freedom of the resumption of group(P＜0.05).Cortical bone mineral apposition rate(MAR),missing more than a quiet restructuring group(P＞0.05);the freedom and the restoration of swimming group compared with the trend of lowering the loss of the reorganization;treadmill group was significantly higher than the loss of recombinant(P＞0.05);treadmill group compared with the freedom to resume significantincrease(P＜0.01)Bone formation rate BFR/BS(%),higher than the control group lost recombinant (P＞0.05);the freedom of the restoration group was significantly lower than the loss of the reorganization,swimming group,treadmill group compared with the trend of lowering the loss of reorganization.Bone formation rate BFR/BV(%),loss of the reorganization was significantly higher than control group(P＞0.01);treadmill group was significantly lower than the loss of recombinant(P＜0.05);swimming group was significantly higher than the freedom of the resumption of group(P＜0.05).Osteoclast surface absorption(ORS),the freedom to restore group,swimming group reorganization of the relative loss of a downward trend;treadmill group was significantly lower than the loss of recombinant(P＜0.05);Swimming Group,the Running group and the freedom to run the restoration group,nosignificant difference.Conclusion:(1) In this experiment the body weight of rats in each group are increasing with aging. Suspension disposal methods can significantly inhibit the growth of body weight. Swimming exercise group observed in the vertical growth of the relative freedom of body weight recovery and treadmill Small;Help Desk movement running than swimming for the growth of body weight in rats is a suitable form of movement.(2) Allows suspension of bone formation activity,bone mass,bone structure,such as lower performance.(3) Different forms of exercise on osteoporosis to restore the role of rats significantly different results.Four weeks of exercise training on the solution of tail-suspended rats lumbar and femoral bone mineral density,bone metabolism and blood parameters tibia histomorphometry have clear targets,but better than the treadmill group swim team.(4) Different forms of movement for the role of bone metabolism are obvious differences between sites.(5) Bone mineral density and indexes have a significant correlation between bone mineral density and biochemical markers of bone metabolism and alkaline phosphatase and the ratio of acid phosphatase was significant correlation between

The Research of Asymmetric 1,3-rearrangement of Organozinc Halides and Imines and Its Application in the Preparation of Chiral Tert-Leucine

Mon, 28 Mar 2011 10:57:31 +0000

α-amino acids exist in a wide range of proteins in nature as the important material basis of life.Naturalα-amino acids are generally chirality except Glycine, many of which are precursor compounds of drugs.There were a lot of asymmetric synthesis reactions of chiralα-amino acids,and imines were often used as substrate to firstly form newα-amino acids via cyano-group\'s asymmetric addition of the C=N bond,removal of N-substituent and hydrolysis.In our preliminary work,we studied the asymmetric addition of organozinc halides with imines,but found a novel organozinc halides-induced 1,3-migration of tert-butyl from sulfur to carbon in racemic N-tert-butanesulfinyl iminoacetates.In this thesis,we tried to further study this unexpected rearrangement reaction for preparation ofα-amino acids and derivatives using enantiomerically pure imines, discuss the optimized conditions,conduct a series of substrates and propose a reasonable mechanism.Moreover,this rearrangement reaction provided a new way of preparing enantiomerically pure tert-Leucine.27 new compounds in all were synthesized,and confirmed by spectrum analysis and X-ray.

Study on Ramipril and BQ-123 Against Myocardial Ischemia/Reperfusion Injury in Vivo in Rats

Mon, 28 Mar 2011 10:57:29 +0000

Objective:To study the protection of ramipril,BQ-123 and their combination against myocardial ischemia/reperfusion(I/R) injury in vivo in anesthetized rats.Methods:Healthy male Wistar rats were divided into 5 groups randomly:①sham operated(sham) group;②I/R group;③ramipril(RAM) guoup;④BQ-123(BQ) group;⑤ramipril and BQ-123(R&B) group.All groups but not sham were subjected to 30 min ischemia/120 min reperfusion through ligation of the left anterior descending(LAD) coronary artery.Twenty four hours before ligation,ramipril(1 mg/kg) was given orally to rats in RAM and R&B groups.The same volume of normal saline was given to rats in other three groups.BQ-123(10μg/kg/min) was infused intravenously from 10 min before ligation to the end of 30 min ischemia to rats in BQ and R&B groups.The same volume of normal saline was given to other three groups. Rats in sham group was not subjects to I/R procedure.A leadⅡelectrocardiogram was monitored throughout the experiment and ventricular arrhythmia(VA) was recorded during ischemia.The activity of plasma creatine kinase(CK) and lactate dehydrogenase(LDH) was measured by spectrophotometer.Myocardial infarct size was measured by TTC staining. Changes of myocardial morphology were observed after HE staining. Apoptosis of cardiomyocyte was assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) assay and confirmed by detection of DNA ladder formation.The expression of Bcl-2 and Bax was determined by immunohistochemistry.Activity of caspase 3 in myocardium was detected by spectrophotometer.Results:1.Effects of ramipril and BQ-123 on myocardial ischemia/reperfusion (I/R) injury in rats.In I/R group,the elevation of ST-segment was dramatically increased (p＜0.001);onset of ventricular premature contraction(VPC) and ventricular tachycardia(VT) were 5.10±0.71 and 6.05±1.65 min,durations of them were 19.94±3.56 and 13.72±5.06 min;incidences of VT and ventricular fibrillation (VF) were 100%respectively;activity of plasma CK and LDH was significantly enhanced;infarct size/area at risk(IS/AAR) and IS were 36.68±4.30%and 83.74±8.73 mg;apoptosis index(AI) of cardiomyocyte was 37.72±0.87%;the ratio of Bcl-2/Bax was 0.35±0.06;activity of caspase 3 in myocardium was 5265.70±387.56 U/μg.Compared with I/R group,the parameters in RAM and BQ groups were changed dramatically.The elevation of ST-segment was decreased(p＜0.01), onset of VPC(10.27±2.63;7.77±0.87) and VT(10.57±1.32;8.74±0.96 min) were delayed,durations of VPC(9.55±1.10;7.59±1.99 min) and VT (5.63±1.45;5.38±0.80 min) were shortened,incidences of VPC,VT and VF were decreased.Activity of plasma CK and LDH was decreased.IS (46.72±9.23;41.71±7.85 mg),IS/AAR(22.65±2.30;20.43±3.79%),AI (23.94±1.81;21.41±1.67%) and caspase 3(4070.04±417.86;3894.71±374.96 U/μg) were decreased respectively.The ratio of Bcl-2/Bax(0.75±0.13; 0.84±0.14) was increased.The morphology of myocardium was improved. DNA-laddering demonstrated that apoptosis of cardiomyocyte was attenuated.2.Effects of ramipril in combination with BQ-123 on myocardial I/R injuryCompared with I/R group,the R&B group was changed dramatically.The elevation of ST-segment was decreased(p＜0.001);onset of VPC(9.17±1.58 min) and VT(8.91±1.45 min) were delayed,durations of VPC(8.91±1.45 min) and VT(4.69±0.90 min) were shortened;incidences of VT(59%) and VF (12%) decreased;activity of plasma CK and LDH was significantly degraded; IS(31.09±6.08 mg),IS/AAR(15.83±4.67%),AI(15.68±1.45%) of cardiomyocyte and caspase 3(3501.62±266.79 U/μg) were decreased, respectively;the ratio of Bcl-2/Bax(0.92±0.13) was,increased.The morphology of myocardium was improved.DNA-laddering demonstrated that cardiomyocyte apoptosis was attenuated.Compared with RAM and BQ groups,the parameters of R&B group were changed dramatically.Mean arterial blood pressure(MAP) was increased in reperfusion 30 and 120min(p＜0.001);activity of plasma CK in ischemia 30 min and LDH in reperfusion 120 min were both decreased(p＜0.001);IS, IS/AAR,AI and caspase 3 were decreased respectively.The morphology of myocardial was improved.DNA-laddering demonstrated that cardiomyocyte apoptosis was attenuated.And compared with BQ group,onset of VPC was delayed(p＜0.05);compared with RAM group,duration of VPC was shorten (p＜0.01),the ratio of Bcl-2/Bax were risen(p＜0.05). Conclusions:1.The results documented that ramipril and BQ-123 protected myocardium from I/R injury by decreasing the elevation of ischemia-induced VA,decreasing the activities of CK,LDH and caspase 3,increasing the ratio of Bcl-2/Bax associated with reduced necrosis and apoptosis of cardiomyocytes.2.It was the first time to investigate the protective effects of ramipril in combination with BQ-123 on myocardial I/R injury in vivo.The results demonstrated that combined using these two agents may protect myocardium from I/R injury.The protective effects on decreasing the activities of CK, LDH and caspase 3,decreasing infrarct size and apoptosis were better than using ramipril and BQ-123 alone.

Study of Self-Navigation Fast Spin Echo Technology at Low Field MR Scanner

Mon, 28 Mar 2011 10:57:27 +0000

Magnetic Resonance Imaging (MRI) has rapidly developed since early 80\'s of 20th century. Now it is widely used in medical diagnosis. Low-field open permanent MR scanners have become widely disseminated in many parts of the world, particularly the developing countries, as a means for low cost access to MRI technology for routine clinical examinations.Motion artifacts at low field MRI system are more severe than that at high field strength MRI due to longer scan time to improve the signal-noise-ratio. As a result, blurring and ghosting caused by patient motion may reduce anatomic detail and limit the detection of pathological findings. Multishot PROPELLER (periodically rotated overlapping parallel lines with enhanced reconstruction) approach, a self-navigation fast spin echo technique, is effective in suppression motion artifacts using segmental phase corrections and oversampling at the central k-space. Diffusion weighted imaging is an important technique for the assessment of acute ischemia. Single-shot echo planar imaging (EPI) , which is available at high field strength MRI systems with relatively strong and fast gradients, is the most common acquisition of choice for diffusion weighted imaging since it is very fast and minimizes any patient-related motion artifacts. However, EPI technology is not readily available on the lower-field open MRI systems.The main contents of this paper include:1. Implementation of self-navigation motion correction fast spin echo sequence at 0.35T open permanent MRI scanner.2. Study of image reconstruction method on self-navigation motion correction fast spin echo sequence.3. Study of diffusion weighted imaging combined with self-navigation motion correction fast spin echo sequence.

The Measurement of Femoral Anteverion and Neck-shaft Angle in Rats and the Femoral Neck of Rats Biomechancal Test

Mon, 28 Mar 2011 10:57:24 +0000

PartⅠThe measurement of femoral anteversion and neck-shaft angle in ratsObjective Through measuring the neck-shaft angle and the femoral anteversion angle, get the range of normal value of the neck-shaft angle and the feromal anteversion angle,therefore,to set the foundation for the study of proximal femoral mechanics research in the future.Methods Scanning the femoral specimens of 50 pair of Wistar rats with scanner and input the pictures into computers to measure them,work out the neck-shaft angle and the femoral anteversion angel,which will be analysed in statistics ways to gain the normal valueResults The scanning method obtains the neck-shaft angle is 123.29°±3.85°,the femoral anteversion angle is 24.86°±4.44°.The neck-shaft angel and the feromal anteversion angel of rat carry on Independent-Samples T Test,the former difference has statistics significance,left side is bigger than right side,The latter difference non-statistics significance.The neck-shaft angle and the feromal anteversion angle comparison between the female and male rat,have statistics significance.The female feromal anteversion angle is bigger than the male,but the male the neck-shaft angle is bigger than female.Conclusion The scanning method does not need any expense nearly,may be many times redundant,and result stable reliable,moreover it may simultaneously demonstrate that two-sided and the multi-lateral of the neck-shaft angle and the feromal anteversion angle,is advantageous in carries on two-sided or the multi-lateral contrasts.Scanning is a accurate and reliable method that is improvement of anatomise measurement to measure femoral anteversion and neck-shaft angle,may provide the reference value for femoral biomechanical experiment. PartⅡThe femoral neck of rats\' biomechanical testObjective Through changes the neck-shaft angle and femoral anteversion angle of rat, obtains the femoral neck failure force under the different rat neck-shaft angle and feromal anteversion angle,thus obtains the data to carry on statistics to analyze the conclusion to instruct the clinical treatment femoral neck disease.Methods First gains 80 normal growth male Wistar rats femur specimen,random average divides into 8 groups:The normal control group,the experimental group of neck-shaft angle reduces 3°,the experimental group of neck-shaft angle increases 3°, the experimental group of neck-shaft angle increases 6°,the experimental group of neck-shaft angle increases 9°,the experimental group of feromal anteversion angle increases 3°,the experimental group of feromal anteversion angle reduces 3°,the experimental group of feromal anteversion angle reduce 6°,Then fixes the embedding rat femur specimen in EnduraTEC ELF3200 biomechanical testing apparatus and carries on mechanics test obtains the data.SPSS15.0 statistical package was used for a homogeneity of variance test and a single way analysis of variance(ANOVA) with a Scheffe test.The level of statistical significance was set to 0.05.Results The control group compares with each experimental groups has the significance difference(P<0.05 ),control group\'s failure force is bigger than each experimental groups,and is bigger along with the angle change.Conclusion Biomechanics of the femoral neck tested in rat experiments show that under the anatomy position the femoral neck have the best mechanical properties:the largest failure force,the lowest femoral neck stress,If neck-shaft angel and the feromal anteversion angel increased or reduced,mechanical properties became lower,the failure force became smaller,the stress increased,and the greater the deviation from the original point of view,smaller maximum load and stress increase the more obvious.

The Influence of Multi-walled Nanotubes on Mice Liver and Urine Metabolism

Mon, 28 Mar 2011 10:57:22 +0000

Objective:With the development of nanoscience,carbon nanotubes are widely used in medicine field,accordingly,more and more attentions are paid to their security.There are many studies and reports on nanotoxicity both in home and abroad,among which the biocompatibility of carbon nanotubes is an important factor that results in toxicity to organism,thus carbon nanotubes are needed to improve their biocompatibility by modification to reduce their toxicity,and these new types of carbon nanotubes are used as drug carriers and biosensors etc.Although some pilot reports found that some modified carbon nanotubes showed no toxicity to organism in vivo,the detailed toxicology data was still lack.Our test is to investigate carbon nanotubes toxicology effect through researches on the influences of multi-walled carbon nanotubes and modified multi-walled carbon nanotubes in mice by liver injures detection and urine metabolism analysis.Methods:Prepare for MWCNT-COOH and MWCNT-PEG and determine their characteristics. Select 70 mice and divide them into seven groups as following:phosphate buffered saline group,multiwalled carbon nanotubes group(contain two different dosages:10 millgram per kilogram and 60 millgram per kilogram),polyethylene glycolylated carbon nanotubes group(contain two different dosages:10 millgram per kilogram and 60 millgram per kilogram),carboxylated multiwalled carbon nanotubes group (contain two different doses:10 millgram per kilogram and 60 millgram per kilogram),10 mice in each group.Drugs were injected i.v.into the tail caudal vein with the rate of 1mL per minute.We collected 24 hour urine of mice\'s half of each group by metabolic cages at 14 days and 59 days respectively,and executed mice of different groups to collect serums and liver tissues for whole specimens as well as pathological sections observation and for serum TB、ALT、AST、Cr as well as SOD、GSH of liver tissues determination.We applied ECF to urine sample derivatization, analyzed urine metabolism by gas-chromatography-mass spectrometry and analyzed urine metabolisms with GC-MS technique.Results:1.Liver injures:①Compared with PBS group,we found more nanotubes deposition in 60 days liver tissues than in 15days liver tissues of the rest groups,and more focus necrosises in HE stain of pathology tissue.Within the same breeding time(15days and 60days),we found more nanotubes depositions in liver tissues of different administrationed groups with a dosage of 60mg/kg than those with a dosage of 10mg/kg,HE stain of pathology tissue revealed that there were mild inflammation cell infiltration at portal vein area,spotty necrosis or small focal necrosis among a small quantity liver cells,especially in pristine group,while there were focal necrosis among a small quantity liver cells in PEG group,no obvious liver pathological changes were found in other groups.②At the breeding time of 15 days and 60 days, we found the value of AST was rised among administrationed groups compared with PBS group,which indicated liver injures.There were no significant differences of Cr among administrationed groups compared with PBS group.③At the breeding time of 15 days and 60 days,there were no significant differences of SOD and GSH among different groups with the same dosage and different dosages of the same group.2.Influence on urine metabolism:①At the breeding time of 15 days and 60 days, there were no significant differences of urine metabolisms among control group、MWCNT group、MWCNT-COOH group and MWCNT-PEG group.②At the breeding time of 15 days and 60 days,there were no significant differences of urine metabolisms between high dosage group and low dosage group of all the three administrationed groups.③There were significant differences of urine metabolisms between pristine group at the breeding time of 60 days,so did PEG group,but we also found significant differences of urine metabolisms between the control group at the breeding time of 15 days and those at the breeding time of 60 days,which indicated that such differences maybe dose no mean to our toxicology research.Conclution:1.Pristine multiwalled carbon nanotubes result in mild liver injures,and these injures can be reduced by COOH and PEG modification.3.There were no significant differences of low molecular urine metabolisms among pristine and modificated multiwalled carbon nanotubes groups.

Preliminary Studies on Muscle Engineering by Using Myogenic Differentiated Bone Marrow Derived Mesenchymal Stem Cells and Alginate

Mon, 28 Mar 2011 10:57:10 +0000

OBJECTIVESSUI is a kind of disease which commonly occurs among adult females.The symptoms increase with age,seriously affecting the physical and mental health of older women.We intend to use bone marrow derived mesenchymal stem cells(BMSCs) which is induced by myogenic differentiation and alginate gel for transurethral injection in the mouth,on the one hand to increase the urethral resistance,on another hand,the use of myogenic differentiation induced by bone marrow-derived mesenchymal stem cells to replace the relevant parts of reverse and degenerative changes in cells to repair pelvic floor muscles,fascia and intrinsic urethral sphincter the physiological functions.This experiment,acting as part of the research,focuses on the isolation and cultivation of the BMSCs.We hypothesized that BMSCs isolated from rat are induced into myogenic cells and compounded with Alginate gel to enhance the stability of Alginate gel.The way the diversification of the BMSCs and the myogenic cells in vivo is ensured.MATERIALS and METHODSPartⅠ,studies on the isolation of rat bone marrow derived mesenchymal stem cells(BMSCs) BMSCs were isolated from newborn rats,which were 1 week old,by using percoll and anchoring methods,and analyzing the surface antigens with the help of Flow Cytometer.Acridine Orange(AO) was used as a differentiating material preparing for the following step.PartⅡ,studies on the differentiation of rat bone marrow derived mesenchymal stem cells into myogenic cells The BMSCs were inducted with 10 mol/L 5-azacytidine for 24 hours.After 3 weeks,the expression of desminandα-SMA in the cells was studied with immunohistochemistry.PartⅢ,studies on the preparation of Alginate gel Mixing 2%sodium alginate with different chroma calcium chloride for volume at the rate of 1:1,5:1,10:1,analyzing the character of the Alginate gel at 4℃、room temperature and 37℃.PartⅣ,studies on the Cells compounded with Alginate gel for tissue engineering Mixing 2%sodium alginate with 1%chroma calcium chloride for volume at the rate of 5:1 made up the injectable material. Alginate gel was used as the frame of BMSCs and myogenic cells were injected into the rat(weight 200～250g) After 4 weeks and 8 weeks, the gel was analyzed according to the change of the volume of the mass under the skin and the expression of desmin andα-SMA in the cells with immunohistochemistry.RESULTSPartⅠ,studies on the isolation of rat bone marrow derived mesenchymal stem cells(BMSCs) BMSCs could be harvested effectively and safely by using percoll and anchoring methods from rats\' bones.After 2 days, BMSCs anchored and grew like spindle,and the cells were round or ellipse.After 5～10 days,BMSCs\' inosculation rate reached 90%, and the form changed into as star shape,and the rate of the growing slowed down.0.25%Trypsinprotease plus 2g/L EDTA were used to assimilate the BMSCs to impress at the rate of 1:3.We could get 3～5×10~6 cells per 10cm culture medium.After being passed for 2～3 times, BMSCs were almost pure cells.The expression of CD29 and CD105 were 89.4%and 46.0%,CD34 and CD45 were negative.Acridine Orange(AO) could make the BMSCs express green fluorescence.PartⅡ,studies on the differentiation of rat bone marrow derived mesenchymal stem cells into myogenic cells After 24 hours of induction directly with 10μmol/L 5-azacytidine and being cultivated for 3 weeks,the ceils grew into coarctation.The inducted cells were polygonal,we could see fascicle-liking shape in the cells.BMSCs,cells inducted 1 weeks and cells inducted 3 weeks were analyzed by immunohistochemistry.There were no differences among them in H&E.The desmin andα-SMA in the cells with immunohistochemistry were no color. Cells inducted for 1 weeks were cambridge blue as in desmin andα-SMA,in the same case,was brown.Cells inducted 3 weeks looked strong blue and brown.PartⅢ,studies on the preparation of Alginate gel The 2%sodium alginate was snuff color ropy.Alginate gel was no color transparent or white. The gel which was formed by mixing the volume at the rate of 1:1 with calcium chloride was stable and hard.And the gel formed by adding calcium chloride at the rate of 5:1 was a little flowing and uniform. After 24 hours,the 1%chroma calcium chloride group was suitable for the injectable material.After the calcium chloride was added at the rate of 10:1,there were more sodium alginate,and the gel could not be formed immediately.The gel became frangible and its volume was smaller than the one before.The temperature ranging from 4 to 37℃did not affect the form of the gel.After drying,the gel\'s volume reduced into 5～10%of the one before.The observation with the scan electron microscope showed that the higher the chroma of the calcium was,the less holes on the gel\'s surface there are.PartⅣ,studies on the Ceils compounded with Alginate gel for tissue engineering The scan electron microscope found the cells spread uniformly in the small interstices of the Alginate gel.The volume of the gel mass reduced 50%in 1 month,and 80%in 2 month.After 4 weeks there was envelope formed outside the gel mass.Analyzing them in the way of immunohistochemistry,that there was inflammatory reaction around the mass,and new blood vessel grew into the mass. BMSCs and myogenic cells survived,but the color of the immunohistochemistry is so feebleness,so the question,is there new born muscle come from the cells which we put in,still need us to do more research. CONCLUSIONSBMSCs could be harvested effectively and safely by using percoll and anchoring methods from rats\' bones.The BMSCs expanded rapidly in the culture concentration.After 3 weeks of induction with 5-azacytidine,BMSCs can express characteristic desmin andα-SMA.But the consistency and the affecting time must be limited.Sodiumalginate,mixed with calcium chloride of different consistency,produce the different gel.It was the higher the consistency,he harder the gel.The consistency needs to be controlled to some exten,which we need is injectable alginate gel.The BMSCs and the myogenic cells compounded with alginate gel can grow up in vivo.But there were no obvious evidence to say that the cells we put in grew into muscle.

Bronchial Mucus Transport Velocity in Patients Receiving Sevoflurane-based General Anesthesia

Sun, 27 Mar 2011 14:40:46 +0000

Objective:The objective of this study was to investigate the influence of sevoflurane(0.3MAC-1.2MAC) on bronchial mucus transport velocity(BTV) in ventilated patients during sevoflurane-based general anesthesia.Methods:Forty patients scheduled for elective general surgery were randomized to one of four groups,receiving sevoflurane(end tidal) targeted at 0.3MAC(group 1),0.6MAC(group 2),0.9MAC(group 3) and 1.2MAC(group 4) after standardised induction with propofol,fentanil and scoline.Other drugs(propofol,fentanil,vecuronium or local anesthetics through an epidural catheter et al.) were adjusted to clinical need.A drop of methylene blue dye(approximately 0.02ml) was placed onto the dorsal surface of the left main bronchus through an epidural catheter inserted into the working channel of a fiberoptic bronchoscope.BTV was measured at 5,15 and 30 min after placement of the dye by observing the movement of it using the scope.Results:At 5min and 15min after placement of the dye,the difference of BTV between group land 2,and between group 3 and 4 was not significant (P＞0.05).BTV in group 3 was significantly lower compared to group 1 and 2(P＜0.05),and BTV in group 4 was sinificantly lower too(P＜0.05). In group 1 and 2,at 5 min after placing the dye,BTV was significantly higher than that at 15min(P＜0.05),but in both group 3 and 4,the difference of BTV at 5 min and 15min was not significant(P＞0.05). Conclusions:Anesthesia with sevoflurane may significantly depress the bronchociliary clearance in patients without lung disease,but this effect is dose dependent.On one hand,when the dose of sevoflurane is low(≤0.6MAC),the reduction of BTV is not significant,whereas when the dose is high(≥0.9MAC),BTV is significantly reduced.But there is no more reduction of BTV when the dose is changed from 0.9MAC to 1.2MAC. On the other hand,when the dose is high(≥0.9MAC),sevoflurane may significantly depress BTV in a relatively short period of time.

The Evaluation of Fluid Therapy with Hydroxyethyl Starch 130/0.4 in Patients Undergoing Major Abdominal Surgery

Sun, 27 Mar 2011 14:40:45 +0000

Objective:To investigate the influence of Hydroxyethyl Starch 130/0.4 on fluid balance;metabolism of natrium;hemodynamics;APACHEⅡscores and pulmonary function after major abdominal surgery.Methods:60 patients undergoing elective major abdominal surgeries were selectd and devided into two groups,crystalloid group and Hydroxyethyl Starch(HES)group. Patients in crystalloid group were just administrated with crystalloid.That in HES group,additional 500ml HES 130/0.4 was given for consecutive 3 days.The changes of fluid and natrium balance;hemodynamics;APACHEⅡscores were recorded on the operation and post opration days.Forced vital capacity(FVC),forced expiratory capacity in first second(FEV1.0),maximal voluntary ventilation(MVV),oxygenation index(PaO_2/FiO_2),respiratory index(RI) were measured by lung function equipment and blood gas analysis pre and post opration days.Results:The positive fluid balance of crystalloid group was significantly more than that in HES group on postoperative day 1,2,3(P＜0.05).The hemodynamics of HES group was significantly better than that in crystalloid group on postoperative day 1,2(P＜0.05).The APACHEⅡscores of crystalloid group were higher than those in HES group on postoperative day 2,3(2＜0.05).FVC,FEV_(1.0)、MVV increased significantly in HES group more than those in crystalloid group on postoperative day 2,3(P＜0.05). The alteration of PaO_2/FiO_2、RI in HES group were significantly more than those in crystalloid group on postoperative day 1,2,3(P＜0.05).Conclusion:The positive fluid balance and natrium retention happened after the major abdominal surgery.The positive fluid balance and natrium retention decreased with the application of HES 130/0.4.It was also able to decrease the APACHEⅡscores,promote pneumonic function recovery and diminish lung injury.It means that the application of HES 130/0.4 can promote the recovery of the patients undergoing major abdominal surgery.

Preliminary Study on Epidemiology of Deep Vein Thrombosis of Lower Extremity of In-Patients with High Risk

Sun, 27 Mar 2011 14:40:42 +0000

Objective:To make preliminary cross-sectional study on epidemiology of deep vein thrombosis of lower extremity of in patientswith high-risk in Zhongshan hospital, including bedrid stroke patients,post-surgery bedrid patients from department of orthopaedics、gynecology、general surgery and vascular surgery and patients of surgical intensive care unit.To analyze risk factors for deep vein thrombosis of lower extremity in these patients.Methods:From Sep 2008 and Feb 2009,we performed compressed ultrasound to detect deep vein thrombosis of lower extremity of in-patients with high-risk in Zhongshan hospital,including bedrid stroke patients、post-surgery bedrid patients from department of orthopaedics、gynecology、general surgery and vascular surgery and patients of surgical intensive care unit.Incidence of DVT of the whole study group as well as each department were calculated and analyzed.We collected detailed clinical information of each recruited patient,including gender,age,history of hypertension or diabetes,family history of DVT,history of vascular surgery,bedrid time,application of hemostasis drugs or anticoagulants or NSAIDS or hormonal drugs,types of stroke,types of surgery,trauma,tumor and laboratory examination including routine blood test and blood coagulation function as well as D-Dimer array. We made statistical single-factor and then Logistic regression multiple-factor analysis of these data with SPSS16.0,to determine high risks of deep vein thrombosis of in-patients. Results:(1) Totally 372 patients were enrolled in our study with 125 men and 247 women,mean age 62±16y.Three main departments were:department of neurology: 102 in-bed stroke patients,department of orthopaedics 100 post-surgery patients and department of gynecology:111 patients with surgery or chemotherapy.(2) 38 patients were detected with DVT of lower extremity,so the incidence of whole series was 10.2%(38/372),including 19(50%) with central type,15(39.5%) with mixed type and 4(10.5%) with peripheral type.8 patients(21.1%) suffered bilateral thrombose while 30(78.9%) only had unilateral thrombose,with 19(50%) left-sided and 11(28.9%) right-sided.15(39.5%)patients presented clinical symptoms of swelling and pain of lower extremity,while the rest 23(60.5%) were asympotomatic(3) High risks of lower extremity deep vein thrombosis for the whole series were bedrid time(30 day boundary)(P=0.000),thrombin time(P=0.007) and history of vascular surgery(P=0.009).(4) The incidence of DVT of lower extremity of department of neurology,orthopaedics and gynecology were 12.7%(13/102),of 10.0%(10/100) and 6.3%(7/111) respectively.(5) High risk for DVT in post-surgery patients of department of gynecology was resection of uterus and both attachments(P=0.032), with no other statistically significant factors in each department.Conclusion:(1) In our study,the incidence of DVT of lower extremity for high-risk in-patients was 10.2%,most of whom were asymptomatic and left-sided.(2) Bedrid time(30 day boundary),thrombin time and history of vascular surgery were high risk factors with DVT.(3) The incidence of DVT from department of neurology,orthopaedics and gynecology were 12.7%(13/102),of 10.0%(10/100) and 6.3%(7/111) respectively,which may arouse enough clinical awareness to strengthen prophylactic anticoagulation.

The Effect of Yi Qi Yang Yin Fang Intervention on Glomerular Filtration Barrier in Adrimacycin-induced Nephropathy Rats

Sun, 27 Mar 2011 14:40:20 +0000

BackgroundPrimary nephrotic syndrome(PNS) is a common disease during childhood. About 80 percentage of PNS in children is minimal change nephrotic syndrome (MCNS).Although most of the patients were sensitive to steroid,some patients were prone to frequent-relapse or to be resistant to steroid treatment,even developing into renal insufficiency.In view of this,we have been seeking to new drugs and new methods which can improve effectiveness and reduce side effects of glucocorticoid and immuno-suppressive agents.Recently,traditional Chinese medicine has made great progress in treating kidney diseases.However,the suppressive effect and mechanism of TCM on MCNS have not well known yet in spite of many studies on the suppressive effect of TCM with MCNS,in particular proteinuria.In addition to the profound involvement of podocytes in the pathogenesis of glomerular diseases the sophisticated function of podocytes and the molecules associated with podocytes are highly attractive in the research of the mechanisms of proteinuria.Permselectivity of serum protein dertermined by the integrity of glomerular filtration barrier.Heparan sulfate(HS) in the glomerular basement membrane(GBM) is important in maintaining the charge-dependent permeability.It is proposed that The beta-D-endoglyeosidase heparanase should be important in the pathogenesis of proteinuria by degrading the negatively charged HS side chains with in the GBM selectivity Similarly,nephrin is one of important component of podocyte slit diaphragm,precious studies showed that it was involved in several diseases such as nephritic syndrome,and all of these diseases have close correlation with proteinuria.Therefore,in this study we used adrimacycin-induced nephropathy rats to observe the heparanase and nephrin expression and their changes in kidney in the protein and the gene level respectively,and to investigate the protective effect and mechanism of Yi Qi Yang Yin Fang on nephrotic syndrome induced by adriamycin(ARD). PARTⅠEffect of the Yi Qi Yang Yin Fang intervention on adrimacycin-induced nephropathy ratsObjective To observe the 24hr urinary protein,routine biochemical indicators of expression and the pathological changes in kidney morphology within normal and adriamycin -induced nephrotic rats,and in order to investigate the suppressive effect of Yi Qi Yang Yin Fang on nephrotic syndrome induced by Adriamycin(ADR).Methods Adriamycin(ADR)-induced nephrotic rat models were established by a single injection of 5mg/kg adriamycin via the tail vein.30 Sprague-Dawley rats were divided randomly into five groups:ADR group injected with adriamycin and the control group injected with normal saline.The 24hr urinary volume were collected at 7days,14days,21 days and 28days after adriamycin injection respectively,and all of rats were sacrificed at 28days.24hr urinary protein,serum cholesterol,serum triglyceride and renal function of rats were measured by routine biochemical method. The ANOVA test was used to analyse the results.Differences were regarded as significant when P＜0.05.All values are expressed as mean±SEM unless otherwise stated.Results(1) In ADR rats,the level of urinary protein and triglyceride and cholest -erol in serum increased significantly at 14th day(P＜0.01,0.01,0.05,respectively),at same time,albumin in serum decreased significantly(P＜0.01).The changes of these indices reached the peak level at 28th day.Under electron microscope,at 28 days after adriamycin injection,extensive fusion of foot processes was observed in ADR rats.(2) In Yi Qi Yang Yin Fang group(Z)、Glucocorticoid group(J) and Yi Qi Yang Yin Fang plus Glucocorticoid group(ZJ),urinary protein and the level of triglyceride and choles -terol in serum decreased,albumin in serum increased at 28th day compared with Model,especially ZJ group significantly(P＜0.01).Under electron microscope,inter -ventions groups dramatically reversed the loss of foot processes back to normal mor -phology,especially ZJ group significantly.Conclusion Yi Qi Yang Yin Fang attenuates foot process injury of ADR-induced kidney,especially ZJ group dramatically.PartⅡEffect of Yi Qi Yang Yin Fang intervention on haparanase and nephrin in adrimacycin-induced nephropathy ratsObjective To detect the HPA and nephrin level in kidney of every group rats,and to discuss mechanism of Yi Qi Yang Yin Fang on adrimacycin-induced nephropathy rats.Methods Real time quantitative RT-PCR was used to determine the expression of HPAmRNA and nephrin mRNA respectively in renal cortex of every group rats at 28 days after adriamycin injection.The level of HPA in urine of every group rats were detected by ELISA.The expression and distribution of nephrin in kidney were detected by immunohistochemical staining(IHC),and were quantitatively analyzed with Motie image 3.2 analysis system.Results(1) The mRNA of HPA in total cortex of kidney in model rat(4.55±0.38) increased at 28th day(P＜0.05) compared to control(1.5±0.79).In invention groups, the level of HPA mRNA in total cortex of kidney decreased respectively at 28th day compared with Model,especially Z(3.566±0.18) and ZJ(3.125±0.807) group significantly(P＜0.05).The expression of HPA in total cortex of kidney correlated significantly to the proteinuria level(r=0.708,P＜0.005).(2) No significant difference of HPA in urine was found between inventions groups and model group at 28day, while model rats had a higher HPA level in urine than control(P＜0.05).(3) The mRNA of nephrin in total cortex of kidney in rat model increased at 28th day compared to the control(P＜0.05).In invention groups,the level of nephrin mRNA in total cortex of kidney decreased respectively at 28th day compared with the Models, especially ZJ(0.91±0.01) group significantly(P=0.031＜0.05).(4) However,The expression of nephrin protein in kidney in model rats(15.1±1.42%) increased at 28th day compared to the control(23.77±4.5%,P＜0.05).In other groups,the level of nephrin in kidneys decreased respectively compared with the Models,especially in ZJ (19.9±3.3) group significantly(P=0.016＜0.05).(5) The expression of nephrin in kidney correlated negatively to the proteinuria level(r=-0.66,P=0.001)Conclusions Yi Qi Yang Yin Fang group(Z)、Glucocorticoid group(J) and Yi Qi Yang Yin Fang plus Glucocorticoid group(ZJ) can attenuate the expression of HPA in total cortex of kidney.It is suggested that these inventions groups improve proteinuria through regulation of haparanase on GBM.Proteinuria are correlated with the expression of nephrin in kidney.Yiqiyangyin herbals and/or steroid are capable of regulating the expression of nephrin in kidney.It is suggested that these inventions may decrease the proteinuria through the regulation of nephrin on SD.Conclusions1.Yi Qi Yang Yin Fang attenuates foot process injury of ADR-induced kidney, especially combined with the steroid.2.Yi Qi Yang Yin Fang attenuates the development of proteinuria in ADR-induced rats possibly through inhibiting HPA in kidney3.Yi Qi Yang Yin Fang can regulate the expression of nephrin on SD.The study illustrated that the treatment with Yang Yin Fang plus Glucocorticoid.was better than other singular intervention,and i its pharmacological mechanism was explored by biochemieal and molecular biological analysis to provide the basis for clinical application.

The Hemodynamic Changes before and after Pneumoperitoneum Laparoscopic Cholecystectomy

Sun, 27 Mar 2011 14:40:10 +0000

Objective:To observe the impact of pneumoperitoneum on hemodynamics during laparascopic cholecystectomy.Metheds:Eighteen patients(ASAⅠ-Ⅱ) scheduled for laparoscopic cholecystectomy were included in this study.A radial artery catheter was inserted before anesthesia and connected to an arterial pressure-based cardiac output(APCO) measurement device (Flotrac~(TM)/Vigileo~(TM),Edwards Lifescience).Routine perioperative monitor was also connected with patients.Patients were induced with dexamethasone 10mg,fentanyl 0.1-0.2mg,lidocaine 1mg/kg,rocuronium bromide 0.6-0.8mg/kg,propofol TCI target concentration 4ug/kg and remifentanil TCI target concentration 3ng/ml.Anesthesia was maintained with propofol TCI target concentration 2.5ug/kg,remifentanil TCI target concentration 3ng/ml and fentanyl 5ug/kg in total.If MAP or HR continuous increased(＞1.2basic value) and last more than 5 minutes,change the target concentration of remifentanil,the maximum≤4ng/ml.Hemodynamic parameters(HR、MAP、SBP、DBP、APCO) were measured at different times before and after anesthesia induction,before and after CO_2 inflation,before and after CO_2 deflation.Results:Compared with the period before peritoneal insufflation,After CO_2 insufflating 1 minute,HR,DBP increased,APCI decreased(P＜0.05).After peritoneal insufflation 5 minutes,SBP,MAP and APCI increased,DBP elevated further.MAP, SBP,DBP,APCI and HR were similar with the values that recorded after peritoneal insufflation 5 minutes and satble until exsufflation.Compared with the period pneumoperitoneum lasting,HR and DBP decreased,APCI increased(P＜0.05) after exsufflation 1 minute.When pneumoperitoneum had been removed 5 minutes,HR and APCI returned to the level before peritoneal insufflation;MAP and DBP decreased (P＜0.05),but still higher than the level before peritoneal insufflation. Conclussion:Peritoneal insufflation affected the hemodynamics significantly during laparoscopic cholecystectomy.When Peritoneal insufflation started,cardiac ouput reduced,APCI dereased,HR and DBP increased.When the pneumoperitoneum had last 5 minutes,APCI increased and returned to the level before Peritoneal insufflation; HR,SBP,DBP and MAP incresed and stay at a high level until exsufflation.Then these hemodynamic varies almost unchanged until pneumoperitoneum removed.After exsufflation,cardiac output increased significantly,as a result,APCI elevated.When pneumoperitomeun had been removed 5 minutes,these hemodynamic parameters returned to the level before Peritoneal insufflation.

Risk Factors and Prognosis for Proteinuria in Renal Transplant Recipients

Sun, 27 Mar 2011 14:40:08 +0000

Background.Proteinuria is a sensitive marker of renal dysfunction.The proteinuria in renal allograft recipients has been regarded as a sign of poor prognosis.The cause of post-transplant proteinuria include chronic rejection、glomerulonephritis、acute rejection and cyclosporine nephrotoxicity.Among them, chronic rejection is known to be most frequent.The purpose of the present study was to investigate risk factors of the persistent proteinuria and whether persistent proteinuria was associated with poor graft outcome.Methods.This study included 178 patients who had first renal transplantation in Huashan hospital.The patients were divided into two groups according to their level of urinary protein excretion.We recorded clinical and laboratory data for each patient,including gender,age at transplantation,donor status(living or cadaveric), duration of pretransplant dialysis,presence of acute rejection,systolic and diastolic blood pressure levels,immuno- suppressive treatment,graft survival and so on. Statistical analyses were performed with SPSS software.we compared categorical variables with the chi-square test;and continuous variables with one-way anova or Kruskall-Wallis;Kaplan-Meier curves were use to calculate graft survival(log-rank test).Logistic regression analysis was performed to estimate relative risks(RR) for independent variables of protenuria following renal transplantation.Cox proportional hazards regression models were used to examine the relationship of variables with survival.P values less than.05 were considered significant.Results.After a mean follow-up of 20 months,43 patients(24.16%) were found to have persistent proteinuria.Hepatitis C(P =0.009),transfusion(P =0.022),induction treatment(P=0.026),cholestero(P<0.001)and LDL(P<0.001)were associated with proteinuria.Multivariate analysis confirmed the independent value of Hepatitis C (OR =7.426;95%confidence interval[CI]1.239 to 44.493;P=0.028) to predict postransplant proteinuria.The mean percentage of decline in serum creatinine per month was persistently higher from the early stages after transplantation in the proteinuric group.Graft survival was 70%among patients who developed proteinuria and 93%in those without proteinuria(P<0.001).Cox proportional hazards regression models confirmed the independent value of protenuira(RR =8.405;95%confidence interval[CI]1.174 to 60.168;P=0.034) to predict graft survival.Conclusion.Proteinuria in renal transplant recipients was related to Hepatitis C、transfusion、Induction treatment、cholestero and LDL.Proteinuria in renal transplant recipients predicted a worse allograft survival.

The Application of Radionuclide Imaging in Mild Cognitive Impairment

Sun, 27 Mar 2011 14:40:04 +0000

Purpose:Patients with mild cognitive impairment(MCI) are at risk for developing Alzheimer disease(AD).To diagnose AD at an early stage,one must develop highly specific and sensitive tools to identify it among at-risk subjects.The purpose of this study was to evaluate and compare the ability of FDG-PET,SPECT,and structural MR imaging to predict conversion to AD in patients with MCI.Materials and Methods:Relevant studies were identified with MEDLINE from January 1990 to April 2008.Meta-analysis and meta-regression were done on the diagnostic performance data for each technique from eligible studies.We estimated and compared the weighted summary sensitivities,specificities,likelihood ratios(LRs), and summary receiver operating characteristic curves of each imaging technique.Results:Twenty-four eligible studies were included,with a total of 1112 patients. FDG-PET performed statistically better in LR+ and OR,whereas no statistical difference was found in pooled sensitivity,specificity,and LR- for each technique. No statistical difference was confirmed between SPECT and MR imaging.The Q* index estimates for FDG-PET,SPECT,and structural MR imaging were respectively 0.86,0.75,and 0.76.In meta-regression,statistical significance was found only between technique and log OR,with a regression coefficient of-0.575.Conclusion: This meta-analysis showed that FDG-PET performs slightly better than SPECT and structural MR imaging in the prediction of conversion to AD in patients with MCI; parallel performance was found between SPECT and MR imaging. Purpose:Mild cognitive impairment(MCI) represents a transitional phase between normal aging and Alzheimer\'s disease(AD).The amnestic subtype(aMCI) is the most predictive of progression to AD.The aim of this activation likelihood estimation (ALE) meta-analysis was to quantitatively analyze functional neuroimaging(PET, SPECT and fMRI) studies that estimated brain functional alteration of aMCI at rest condition and during episodic memory performance.Materials and Methods: Nineteen relevant articles from January 1998 to May 2008 were identified.Three main meta-analyses were conducted by comparing aMCI with normal control(NL) at the rest state,aMCI with NL during episodic memory performance,and aMCI alone during episodic memory performance versus the rest state.For each cluster,locations of voxels with peak probabilities were generated.ALE maps were masked and overlaid onto anatomical template using MRIcroN software.Results:The results largely conformed to previous studies and further confirmed the role of hippocampus, precuneus and posterior cingulate cortex(PCC) in aMCI patients at rest,as well as that of the frontal lobe and medial temporal lobe(MTL) structures in episodic memory.Aside from the same areas in the left cerebrum which were activated in aMCI both during encoding and retrieval,increased activation was found in the right medial frontal gyrus during retrieval task.Conclusion:These ALE findings provided the average location and extent of functionally impaired brain region of aMCI patients and generated regions of interest for the testing of new hypotheses.Purpose:To investigate the characteristics of cerebral blood perfusion in the patients with mild cognitive impairment(MCI) by comparing to normal controls and to evaluate the potential roles of SPECT in identifying subjects with MCI from normal aging.Materials and Mehtods:Tewnty patients,who were diagnosed as having MCI, and 20 normal aging subjects each underwent a ~(99)Tc~m-ECD SPECT cerebral blood perfusion imaging.We processed SPECT images with SPM5b software and performed a voxel-based statistical analysis comparing MCI patients and normal aging subjects.Results:In comparison with normal controls,subjects with MCI showed asymmetry hypoperfusion in the posterior cingulate gyrus,inferior and superior parietal lobule,cingulate gyrus,inferior and superior frontal gyrus.Conclusion:The results were consistent with previous research,which further confirmed the specific regions with decreased blood perfusion in patients with MCI.

Hair Growth Parameters of Chinese Subjects in Spring and Autumn

Sun, 27 Mar 2011 14:01:36 +0000

Objective:To assess and compare the spring and fall,the Han people hair growth parameters.Materials and Methods:1.Study:18-50-year-old Han volunteers 60 cases(30 cases among men and women).2.The past did not accept hair,perm,such as treatment of this study informed consent.3.Using PTG assessment of hair at the vertex of the first,temporal,occipital hair growth parameters(hair density,hair growth rate and the proportion of quiescent period),spring and fall,once the assessment.4.All results using Stata 7.0 statistical software at the vertex of the scalp and the temporal,occipital comparison between the hair growth parameters using analysis of variance(Residual Normality),or rank transformation test(residual non-normality),sex,Seasonal comparison using t test(homogeneity of variance) or rank sum test(variance arrhythmia),P＜0.05 indicated statistical difference.Results:1.At the top of the density of hair density were higher than the temporal and occipital(P＜0.0001),temporal and occipital there is no significant difference compared;gender comparison in the spring,male hair density is 143±17 root/cm2 than 120±11 female root/cm2(P＜0.0001),men and women in the autumn there is no significant difference in hair density(P＝0.6151);the spring season were relatively hair density of 132±18 root/ cm2,the fall to 118±15/cm2,spring hair density was significantly higher than in autumn(P＝0.0004).2.The proportion of parts of the quiescent period was no significant difference (P＝0.1925);gender relatively quiescent period in the autumn the proportion of females 22.1±7.1%higher than that of men 17.6±8.7%.; The fall season is relatively quiescent period the proportion of 19.8±8.2% higher than the spring,16.4±4.5%.3.At the top of the hair growth rate in the spring and autumn(492±65μm/day;483±31μm/day) are the three parts of the slowest,temporal and occipital no significant difference compared;male hair growth rate of 511±39μm/day in the spring than females 496±53μm/day;spring,the fall of the hair growth rate was no significant difference(P＝0.4845).Conclusion:1.The present study,vertex is three positions for the highest density of hair density in men than women,the spring than the fall of hair density;2.Quiescent period in fall is higher than the spring;3.Hair at vertex of the slowest growth rate in the spring is the growth rate of males than females.Chinese Library Classification:R758.71

437 Cases of Idiopathic IgA Nephropathy: Analysis of the Clinical Characteristics and Histopathological Lesions

Sun, 27 Mar 2011 14:01:34 +0000

【Background and Objective】IgA nephropathy is the most common form of glomerulonephritis in the world,and is also the main cause of end stage renal disease. Now it is considered to be a clinicopathological syndrome with the same immune-histological feature,characterized by deposits of IgA with or without other immune-complex in the mesangium of the glomerulus.The disease has lots of various clinical presentations and histological lesions.A renal biopsy is required for an accurate disgnosis of IgA nephropathy and the degree of the histological lesions plays an important role on prognosis and treatment of the disease.Anyway,many hospitals in our country are not able to do renal biopsies until now and little patients accept a repeat renal biopsy during the follow-up time.So clarifying the relationship between clinical presentations and renal pathological lesions of IgAN may give us important references in clincal practice.In this research we anlysis the clinical and pathological characteristics of 437 cases digonised IgA nephropathy these years in our hospital, aim to investigate the relationships with each other,find out the clinical risk factors associated with the renal pathologic lesions and to predict the degree of the active and chronic histological lesions with the clinical presentation.【Subjects and methods】437 cases with IgA nephropathy is involved in our research.The clinical and pathological data at the time of renal biopsy were analyzed. We described the clinical characteristics and common pathological lesions in IgA nephropathy,assessed whether clinical presentation(24-hour proteinuria,uria red blood cell,arterial hypertension,renal function,age and gendar) correlated with the pathological lesions(the indices of masangial proliferation,glomerular sclerosis, crescents,tubelar atrophy,interstitial infiltration and fibrosis,especially the indices of the chronic pathological lesions) using the software SPSS 16.0.【Results】(1) IgAN account for 41.46%in the patients with primary glomerular disease,the ratio of male to female was 1.06:1,the apparen peak incidence age was 20-40 years old(60.7%),the average incidence age was 34.33±12.38years old(13-76 years old),the apparent age of male and female has no difference(p＞0.05). (2) 27.7%of the patients was found out the disease through a routine health examination without any symptoms.The incidence of gross hematuria was 29.8%, more common in younger patients.The incidence of other symptoms such as edema, lumbago was 28.6%、24.0%.Proteinuria with hematuria is the most common presentation,the incidence of it is 82.4%,respectively the incidence of proteinura without hematuria or isolated hematuria without proteinura was 10.8%and 6.7%.31 cases(7.1%) performed as nephrotic syndrome.The percentage of CKDⅠ-Ⅴstage was 48.7%,27.6%,16.3%,5.1%and 2.3%.(3) The incidence of arterial hypertention is 45.7%.Respectively,the incidence of high urine-acid hemaria,high triglycerideria and high cholesteria was 30.2%, 40.0%and 17.4%.The incidence of aterial hypertention and high triglycerideria was increased by aging.And high urine-acid hemaria and high triglycerideria were more common in male than in female.(4) Pathological lesions such as masangial proliferation,glomerular sclerosis, cresents formation,tubelointerstitial lesions and renal vascular injury was common in IgA nephropathy,the incidence of them was respectively 98.2%,69.1%,33.6%, 89.5%and 35.2%.The percentage of patients with LeeⅠ、LeeⅡ、LeeⅢ、LeeⅣ、LeeⅤis 7.78%、33.9%、33.9%、19.9%、,4.6%。(5) Patients with advanced chronic pathological lesions was also accompanied with the severe active lesions and clinical presentations in which the percentage of patients with proteinuria＞1.0g/d,decreased renal function and hypertention was increased.(6) The chronic lesions and active lesions such as masangial proliferatio and interstitial infiltration were more severe in the patients with＞lg/d proteinuria.When the proteinuria was more than 2.5g/d,the incidence and the degree of cellular cresent would be increased.In the patients with gross hematuria in one month before renal biopsy or with high level of urine red blood cells at the time of renal biopsy,we could see more cellular cresent formation.As the renal function decreasing,the chronic lesions were advanced,and the cellular cresent became more common to see.And the chronic lesions was also advanced accompany with the arterial pressure elevated.The chronic lesions was more severe in male patients and older patients.(7) The high level of blood pressure,the decrease of renal function and proteinuria more than l.0g/d are independent predictors of the advanced pathological lesions.On the other hand,the high level of urine red blood cells and the proteinuria more than 2.5g/d are independent predictors of cellular cresent formation.(8) Patients with isolated hemeturia is characterized by mild clinico-pathological presentation.Patients with nephrotic syndrome have severe active lesions but relatively mild chronic lesions,61.3%of the cases was in lowest chronic level, however 41.9%cases have the cellular cresent formation.【Conclusions】IgAN is the most common glomerulonephritis in China,and is account for 41.5%in primary glomerular disease.The ratio of male to female is 1.06, the peak incidence age is 20-40 years old,27.7%of the patients have no symptoms at the onset of the disease,29.8%of the patients may have gross hameturia.There are 7.1%of the patients presented nephrotic syndrome and 51.3%of the patients with decreased renal function in our data.Patients with advanced chronic lesions were in the more severe state of illness.Logistic Multivariete analysis shows he high level of blood pressure,the decrease of renal function and proteinuria more than 1.0g/d are independent predictors of the advanced pathological lesions.It also shows the high level of urine red blood cells and the proteinuria more than 2.5g/d are independent predictors of cellular cresent formation.

The Analysis of Clinical, Pathological and Prognostic Relationship of IgA Nephropathy Presented by Nephrotic Proteinuria

Sun, 27 Mar 2011 14:01:32 +0000

Objective:To analysis the clinicopathological features and relationship of IgA nephropathy presented by nephrotic proteinuria,to investigate the influence of clinical parameters,renal pathological lesions and treatment on renal survival.Methods:The clinical and pathological data of eighty patients with IgA nephropathy presented by nephrotic proteinuria were analyzed retrospectively.Histological evaluation of patholiogical lesions was based on the histological grading and semiquantitative integral for IgAN described by Lee and Kathafuchi,acute and chronic changes were estimated separately,which was the method used by Shigemastsu.We used Kruskal-Wallis H test to investigate the relationships of the clinical and pathological features.Kaplan-Meier curves and Cox proportional hazards model were also used in the renal survival analysis.Comparisons between different treatment groups were by the chi-square method.Results:1(1) General indices:Among 80 patients,46 male and 34 female,mean age(35.58±13.3)years old.20-40 year-old age group was the most prominent.(2) Clinical characteristics:25%patients had an incentive before onste.Mean course of disease was 21.55 months.Incidence of gross hematuria,hypertension,renal decrescence was 22.5%,32.5%and 53.75%,respectively.There was no significant difference in incidence of gross hematuria between the normal renal function group and decreased renal function group.62.5 percent of the patients were with edema at onset.The level of plasma albumin was significantly higher in edema group than that in non- edema group.(3) Pathological lesions:GradeⅣwas the most prominent in those patients,holding 37.5%.Immunofluorescence:the common deposition of IgA, IgM and C3 was the most prominent in glomerular mesangial region,holding 33.75%. (4) There was no relationship between crescent with the other pathological lesions. But there was positive correlation between glomerulosclerosis with inflammatory infiltration,interstitial fibrosis,tubular atrophy and vessel lesions,in contrast. 2 The relationship between clinical and histopathological changes:(1)There was no significant difference of the distribution of pathological grade and the scores of pathological lesions in different hematuria group(P＞0.05).The scores of lesions of global glomerulosclerosis,inflammatory infiltration,interstitial fibrosis,tubular atrophy and vessel lesions in the decreased renal fuction group were higher than that in the normal renal function group,which had statistically significance(P＜0.05). Pathological grade was rising with the increase of the hypertensive grade.The scores of lesions of global glomerulosclerosis,interstitial fibrosis,tubular atrophy and vessel lesions in hypertensive group were significantly higher than that in normal hypertension group.(2) There was positive correlation between urinary protein and crescent lesions.The positive correlation of global glomerulosclerosis,inflammatory infiltration,interstitial fibrosis,tubular atrophy and vessel lesions was mean arterial pressure,the negative correlation were eGFR and hemoglobin.(3) The proteinuria with no glomerulosclerosis was higher than that with glomerulosclerosis,significantly (P=0.007).But there was no significant difference in groups of different degree of glomerulosclerosis.The higher the sclerosis scores were,the higher MAP was,the lower eGFR was,the more severe renal anemia,acute and chronic tubulointerstitial lesions and vessel lesions were.(4) The patients in GradeⅤnot only had severe chronic lesions,but also had serious acute lesions including crescents and inflammatory infiltration.(5) The degree of sclerosis(P=0.041) and inflammatory infiltration(P=0.031) was higher when there was stronger intensity of IgA in immufluorescence,as well as the pathological grade(P=0.046).3 Renal survival analysis and estimate of the prognostic factors:54 patients were followed up,follow-up period was 2-60 months.12 patients developed to renal deterioration,6 patients required dialysis or renal transplantation,2 patients died of uremia.Since renal biopsy,the survival rates were 94%at 6 months,94%at 1year,90% at 2 years,80%at 3 years,63%at 5 years.Univariate Cox regression indicated the following parameters as risk factors:proteinuria,mean arterial pressure,eGFR, hemoglobin,global glomerulosclerosis,inflammatory infiltration,interstitial fibrosis, tubular atrophy and vessel lesions.Multivariate Cox regression indicated the following parameters as the independent risk factors:proteinuria,mean arterial pressure,renal anemia and vessel lesions(OR=1.799,1.052,0.860,21.163). Kaplan-Meier curve indicated the following parameters as risk factors:mean arterial pressure≥106.7mmHg,eGFR＜60ml/min,hemoglobin＜110g/L,scores of global glomerulosclerosis＞6,severe inflammatory infiltration(scores＞2),severe interstitial fibrosis(scores＞2),moderate and severe tubular atrophy(scores＞1),vascular intima thickening and glass samples.4 Treatment:one patient didn\'t receive the regular treatment,2 patients were treated with ACRI or ARB alone,no remission.34 patients were treated with oral steroids,17 patients were treated with prednisone and immunosuppressive agents.22 patients achieve the remission in group of prednisone,with the rate of 64.7%,14 patients achieve the remission in group of therapeutic alliance,with the rate of 82.4%,there was no significant difference between two groups in remission rate.The complete remission rate in group of prednisone and therapeutic alliance were 14.7%vs 17.6%, which was with no statistically significance.Conclusion:1 The common age range in IgA nephropathy with nephrotic proteinuria was from 20 to 40 year-old.The clinicopathological characteristics were not slight.The renal survival at 1year and 5years were lower than the average level of IgA nephropathy.2 Hematuria,whether gross hematuria or not,whatever degree of microhematuria was, was not the impact factor of pathological lesions in IgA nepropathy presented by neprhotic syndrome.The urinary protein excretion increased with crescent lesions., which could reflect the acute gloermular lesions.The high MAP,the low eGFR and the moderate or severe renal anemia were the important parameters that reflected the severity of chronic lesions.3 Proteinuria,mean arterial pressure,renal anemia and vessel were the independent risk factors of renal survival.4 Mean arterial pressure≥106.7mmHg,eGFR＜60ml/min,hemoglobin＜110g/L, scores of global glomerulosclerosis＞6,severe inflammatory infiltration(scores＞2), severe interstitial fibrosis(scores＞2),moderate and severe tubular atrophy(scores＞1), vascular intima thickening and glass samples were risk factors of renal survival.5 We founded that the steroid therapy or immunosuppressive agents could decrease proteinuria effectively and protected renal function.But the difference of these two therapeutic effect remains large sample RCT researches to prove.

The Study of Clinical Characteristics and Glycyl-tRNA Synthetase Gene Mutations in Adolescent-onset Myoatrophy of Bilateral Distal Upper Extremities with Knee-jerk Hyperreflexia

Sun, 27 Mar 2011 14:01:29 +0000

Background:Adolescent—onset bilateral distal upper extremities muscular atrophy is an uncommon clinical entity,which includes Juvenile muscular atrophy of the distal upper extremity(JMADUE,also named Hirayama disease),Charcot-Marie-Tooth disease type 2D(CMT2D),and motor neuron disease(such as distal spinal muscular atrophy typeⅤ(dSMAV) and amyotrophic lateral sclerosis(ALS)).We have encountered five cases of young bilateral distal upper extremities muscular atrophy with Knee-jerk hyperreflexia.These patients are characterized by adolescent-onset bilateral weakness and myoatrophy in hand with Knee-jerk hyperreflexia, as well as slowly progressive aggravation.The clinical manifestations exhibited by these patients are different from Hirayama disease,CMT2D, and dSMAV.To make a definitely diagnosis,we carefully analyzed their clinical data and sequenced the fragment containing 10 mutations reported of the gene for glycyl-tRNA synthetase(GARS gene).Objective:To explore the association of adolescent—onset bilateral distal upper extremities muscular atrophy with GARS gene mutation and to help make a definitely diagnosis. Materials and Methods:1.A retrospective review and analysis was performed to the five patients with adolescent—onset bilateral distal upper extremities muscular atrophy,consisting of clinical characteristics,the determination of thyroid function,chest X-ray,electroneuromyography, head and cervical MRI.2.RNA was extracted by standard methods from fresh blood samples of the five patients and 15 healthy controls.RT-PCR and sequencing were performed to detect the 10 known mutations of GARS gene.Results:1.All five patients were characterized by adolescent-onset slowly progressive muscle weakness and atrophy,preferentially involving the muscles of bilateral distal upper extrimities,particularly the thenar eminence and first dorsal interossei muscles.Bilateral knee-jerks of five patients were increased.Electroneuromyography examination showed a neurogenic damage in all patients,involving the anterior horn cells of spinal cord.Conventional cervical MRI and flexed cervical MRI did not find abnormal lesions.2.GARS gene detection did not find abnormal mutations in the 10 known mutations sites.Conclusion:1.Adolescent-onset bilateral distal upper extremilties muscular atrophy with knee-jerk hyperreflexia is a rare disease of the nervous system.The clinical manifestations exhibited by these patients are different from Hirayama disease,CMT2D,and dSMAV.2.GARS gene sequencing did not find abnormal mutations in 10 known gene mutation sites.It indicates that these patients suffer from CMT2D/dSMAV with few possible.It needs DNA microarray detection to find the Candidate mutation gene furtherly.

Preliminary Analysis of SP Phenomenon, Chemotherapy Resistance and Stem Cell Related Genes Expression in Gastric Cancer Side Population Cells

Sun, 27 Mar 2011 14:01:27 +0000

Objective:To analyze the relationship among the side population phenomenon,chemotherapy resistance and ABCG2 in gastric cancer cell lines.To compare stem cell related gene expressions between SP cells and NSP cells.Methods:1.We choose gastric cell line MKN28(well differentiated) and BGC823(poor differentiated) as candidates,labeled them with Hoechst33242 and PI,then SP detection and fluorescence-activated cell sorting(FACS) were performed using flow cytometry.2.Observe morphologic differences between SP cells and NSP cells after FACS by inverted microscope,especially diameters.3.Detect SP cells percentages in BGC823 and MKN28 under different Hoechst33342 concentrations,and compare dye refluxing ability.Meanwhile,detect ABCG2 expression in both cell lines by semi-quantitative polymerase chain reaction(PCR) and Western-blot.4.Carry out chemotherapy resistant test in both cell lines by using 5-FU and cisplatin, then compare ABCG2 expression in SP cells and its counterpart by semi-quantitative PCR and fluorescent-quantitative PCR.5.Compare expression of OCT-4,NOTCH,NANOG,BMI-1,CD133,CD24 and CD44 in BGC823 SP cells and NSP cells by semi-quantitative PCR.Results:1.After FACS and plated 4 hours,BGC823 SP cells\' diameter is longer than NSP cells\'(SP=30.03,NSP=28.23,P<0.05),however the difference disappeared after cells were plated 24 hours(SP=53.48,NSP=52.01,P=0.335).2.The percentage of SP cells in BGC823 is about 1%under 5μg/ml dye,but there is no SP cells found until dye concentration is reduced to 1μg/ml in MKN28(0.4%).Results of Semi-quantitative PCR and Western-blot show ABCG2 is expressed more in BGC823 than in MKN28.3.Chemotherapy resistant test in BGC823 show SP cells grew more quickly than NSP cells in 72h-cisplatin(1μg/ml) Group and 120h-5FU(10μg/ml) Group.PCR results indicate ABCG2 is expressed more in SP cells than in NSP cells.4.Semi-quantitative PCR results show OCT-4,NOTCH,NANOG,CD24 and CD44 were expressed more in BGC823 SP cells than NSP cells.Conclusion:Side population cells in gastric cell line BGC823 possess stronger chemotherapy resistant ability,which may have some relationship with ABCG3,than its counterpart.Compare with NSP cells,SP cells are expressed more stem cell related genes,such as OCT-4,NOTCH, NANOG,CD24 and CD44.

Establishing a Citrate Pharmacokinetics Model and Its Application in RCA-CRRT

Sun, 27 Mar 2011 14:01:24 +0000

Background and Objectives:Continuous renal replacement therapy (CRRT) has, in recent years, been a means of indispensable organ support method to save critically ill patients with acute renal failure or multiple organ dysfunction. Due to a prolonged treatment course and ever-changing conditions of patients, critically patients who are often complicated with active bleeding or high risk of bleeding tendency are bound to increase the risk of bleeding when using heparin for extracorporeal circuit anticoagulation. On the other hand, the use of frequent saline flushes without anticoagulant has a lot of drawbacks such as a shorter life-span dialyzers, poor dialysis effects and high cost. So to use what anticoagulation approach is a cumbersome problem.Citrate causes anticoagulation by chelation of ionized calcium in the extracorporeal circuit, not affecting the blood coagulation system. Thus, it has the advantages of fewer bleeding complications, excellent anticoagulation effect, good biocompatibility and long life-span of dialyzers. In addition, because of chelation of ionized calcium, citrate probably can inhibit complement activation and leukocyte degranulation which may have anti-inflammatory and antioxidant synergistic therapeutic effects. Therefore, regional citrate anticoagulation is an ideal method of anticoagulation for critically ill patients with active bleeding, with high tendency of bleeding, and with hemodynamic instability as well. However, it is dangerous for critically ill patients to using regional citrate anticoagulation in CRRT. A large amount of citrate will be infused into body during anticoagulation program. In addition, critically ill patients who are present with hypotension, inadequate tissue perfusion or hypoxia and multiple organ dysfunction, particularly with abnormal liver function, probably have difficulties metabolizing citrate which easily leads to citrate accumulation and cause toxicity. The question is how about the citrate body clearance of critically ill patients with acute renal failure. What a role does the dialysis play in the clearance of citrate in the extracorporeal circuit? Will critically ill patients occur citrate accumulation during the course of RCA-CRRT? How to carry out RCA-CRRT both effectively and safely?To answer these questions, the aim of our investigation is to establish a citrate pharmacokinetics model which provides us with a detailed understanding of the interactions of citrate with plasma and of citrate fluxes in the patient and in the extracorporeal blood circuit. We intend to preliminary validate the model through the pharmacokinetics study of the population of acute renal failure and acute renal failure with multiple organs dysfunction, meanwhile, to explore the change of plasma citrate level when change the CRRT mode.MethodsOur investigation is divided into two parts:1. Establishing the comprehensive citrate kinetic model: First, analysis these citrate fluxes. The source of new citrate for anticoagulation, the body clearance and filter elimination of citrate are the three major citrate fluxes that determine the systemic citrate level during RCA. Second, define the kinetics parameters to describe all major citrate fluxes during kinetic modeling and combined them into a single pool, first order kinetic equation. Then, data from three published clinical study of citrate kinetics was applied to evaluate the model. We predict the risk of systemic citrate accumulation in patients with abnormal, impaired and absent body clearance while different CRRT protocols are being carried out. The model helps us to find out the safe and effective CRRT protocol without citrate accumulation in patients with impaired and absent body clearance.2. The preliminary application of citrate pharmacokinetics model in the CRRT: Investigate acute renal failure and acute renal failure complicated with multiple organ failure patients in Huashan Hospital during RCA-CRRT. The tri-sodium citrate solution (1020mmol/L) was infused into the arterial line prior to the blood pump at a dose of 2.5mmol/L of blood flow. Systemic citrate was repeatedly measured to calculate citrate pharmacokinetics. Different CRRT modes were tried. The first mode: CVVH (pre-dilution),Qb200ml/min,Qpre1L/h;The second mode: CVVH(pre-dilution), Qb150ml/min,Qpre 6L/h.Systemic citrate was repeatedly measured to calculate citrate pharmacokinetics. Evaluate the agreement between true citrate measurements and our predictions. The citrate fractional extraction ratio (Ecit) and plasma clearance of citrate on the filter (CLf) were measured in both CRRT modes. Analyze thedifferences between the measured and estimated Ecit and CLf.Results1. The single pool, first order citrate kinetic modeling equation is :2. There is excellent agreement between published citrate measurements and our predictions.3. Kinetic model shows that patients with different body clearance undergo RCA-HFD safely. The plasma citrate concentration of patients with normal citrate body clearance is no more than 1mM so that patients can tolerant well during RCA-CRRT. The plasma citrate concentration of patients with impaired citrate clearance is two times higher than patients with normal citrate clearance and nearly goes up to the toxic concentration. Unfortunately, systemic citrate accumulates in a short time when the body metabolism of citrate is absent.4. Theoretically, larger portion of citrate can be removed when the citrate fractional extraction ratio is elevated. The plasma citrate concentration reduces and the risk of systemic citrate accumulation declines.5. The model predicts that when Ecit＞66%, systemic steady citrate concentration will be among the safe range even in patients of impaired body metabolism of citrate. When Ecit>80%, systemic citrate accumulation is not possible even in the complete absent of body metabolism.6. The result of our citrate kinetic of patients with acute renal failure and acute renal failure complicated with multiple organs dysfunction shows there are marked differences. The plasma citrate concentration of the patients with liver dysfunction and hemodynamic instability is elevated. These two populations probably have a high risk of systemic citrate accumulation during RCA.7. Our preliminary clinical study validates the citrate kinetics model. Citrate measurements are in good agreement with predictions. There is no significant difference between our citrate measurements and predictions by the repeated measurements ANOVA test.8. Our clinical study has proved that the systemic citrate plasma concentration will be reduced by adjusting the CRRT parameters to achieve a higher Ecit.Conclusion We propose a citrate kinetic model of RCA-CRRT that help us to predict the the risk of systemic citrate accumulation and to design the CRRT protocols for the patients with impaired citrate body metabolism during RCA delivering. Our clinical study has shown that the plasma citrate concentration will be reduced by adjust the treatment mode of CRRT, indicating its guidance role in the RCA- CRRT. The result of citrate pharmacokinetics indicates that it is very necessary to investigate pharmacokinetics and metabolism of Chinese citrate in critically ill patients because there are marked differences among critically ill patients with acute renal failure in order to reduce the risk of systemic citrate accumulation in this population.

Clinical Characteristics and Features of Airway Inflammation of Difficult-to-treat-asthma in Adult

Sun, 27 Mar 2011 14:01:20 +0000

[Background]Clinically,there are approximately 5%-10%asthma patients whose asthma still may not be effectively controlled after receiving treatment regimens recommended for gradeⅣpatients by the Global Initiative for Asthma(GINA).This type of asthma is called difficult-to-treat-asthma(DTA).Although the number of patients with DTA is low,they consume about 80%of medical resources related to asthma.DTA is a type of asthma that is persistent and associated with a high risk of death.Therefore,DTA is a huge challenge for clinicians.[Objective]Patients with either DTA or Well-controlled asthma were included in this study.Clinical data concerning these two groups of patients was collected.Serum inflammatory markers and fractional exhaled nitric oxide(F_ENO) levels were measured.Differences in family history,allergen skin test and concurrent allergic disease were analyzed between patients with DTA and those with Well-controlled asthma. Differences and correlation in inflammatory markers were analyzed between the two groups.Clinical characteristics and features of airway inflammation of DTA were investigated.In addition,effects of inhaled corticosteroids(ICS) on the levels of total serum IgE(TsIgE),TNF-αand F_ENO in asthma patients were examined.[Methods]Fifty-eight sporadic cases of asthma(n=30 for DAT,n = 28 for Well-controlled asthma) from asthma specialty clinics in Zhongshan Hospital included in this study.Clinical data of patients were recorded for classification of the level of asthma control.ICS-based treatment regimens were administered according to the GINA 2006.During the 12-month follow-up,lung function was detected and the level of asthma control was evaluated.The levels of TsIgE and TNF-αwere detected at both the starting point and end point of follow-up.F_ENO was measured at end point.Differences in clinical data,treatment regimens,response to treatment(evaluated by FEV_1 and ACT scores) between patients with DAT and those with Well-controlled asthma were analyzed.Investigations on correlation among the levels of TsIgE,TNF-αand F_ENO were performed.Differences in the levels of TsIgE,TNF-αand F_ENO between DTA patients and those with Well-controlled asthma were analyzed.A P value of less than 0.05 was defined as statistically significant for differences or correlation. [Results]A total of 58 patients completed follow-up:30 patients(n = 15 for male and female,respectively) in the DTA group and 28 patients (15 men and 13 women) in the Well-controlled asthma group.Compared with the Well-controlled asthma group,the DTA group had a longer course of asthma,a greater mean age(56.4±8.9 years vs.45.6±14.3),and a higher BMI and proportion of smokers.There were no differences in onset age,family history and biparental history of asthma between the two groups.Although the total positive rate of allergen skin prick test was higher in the DTA group than in the Well-controlled asthma group,there were no significant differences in concurrent allergic disease between these two groups.Patients with DTA received treatment regimens for gradeⅣandⅤ,and all failed to response to degraded treatment regimens. In this group,during the twelve-month follow-up,14 patients suffered from more than 2 times(mean:1.33 times/person) of aggravation of asthma.In the Well-controlled asthma group,pharmaceutical treatments were completely discontinued in 3 patients among 14 patients whose treatment regimens were degraded,and during the twelve-month follow-up,3 patients suffered from aggravation of asthma.FEV_1 and ACT scores at starting point and end point were significantly lower in the DTA group than in the Well-controlled asthma group.In the DTA group, the mean daily dose of ICS at end point was comparable to that at starting point;no significant improvements in FEV_1 were seen;inconsistent with improvements of FEV_1,ACT scores at end point were significantly increased(3.53±4.82 points,P=0.000) when compared with those at starting point.TNF-αlevels at both starting point and end point were higher in the DTA group than in the Well-controlled asthma group(P＜0.05).In both groups,TNF-αlevels at end point were significantly decreased as compared with respective levels at starting point.At starting point,the DTA group had lower TsIgE levels with the differences reaching statistical significance(P＜0.05)and lower blood EOS count but without the differences reaching statistical significance(P＜0.05),as compared with the Well-controlled asthma group.TsIgE levels at end point did not differ significantly between the two groups but were substantially decreased as compared with respective levels at starting point.FeNO levels were lower in the DTA group than in the Well-controlled asthma group,with the differences reaching statistical significance(P＜0.05).In the DTA group,there was a linear correlation between FeNO and TsIgE(r=0.711,P=0.000) but no correlation between TNF-αand TsIgE;total FeNO was linearly correlated with TsIgE(r=0.597,P=0.000).[Conclusion]Compared with those with Well-controlled asthma, patients with DTA had a longer course of asthma,a greater mean age and a higher BMI.In the DTA group,ACT scores improved better than FEV_1 after treatment.Compared with those with Well-controlled asthma, patients with DTA showed higher serum TNF-αlevels but lower TsIgE levels.ICS-based treatment regimens led to significantly decreased serum levels of TNF-αand TsIgE.F_ENO levels were lower in patients with DTA than in those with Well-controlled asthma.F_ENO was positively correlated with TsIgE.

The Growth Hormone Receptor (GHR) Exon 3 Polymorphism and Its Relationship with Metabolic Profile in Chinese Obesity Children

Sun, 27 Mar 2011 13:31:28 +0000

Objective:To investigate the GHR exon 3 polymorphsim and its relationship with the metabolic profile in Chinese obesity children.Subject and Methods:409 obesity/overweight children and 206 normal weight children were recruited.Body weights and heights were measured,body mass indexes were calculated,obesity degree were evaluated using IOTF standard.Genomic DNA was extracted from their peripheral blood leukocytes,GHR exon3 gene polymorphism were determined using PCR method.Fasting glucose,insulin and lipid profile were analysed, and HOMA-IR and ISI were calculated using homostais model.All the data were analysed by SPSS 12.0 software.Result:1.GHR exon3 gene polymorphism distribution in obesity and normal control group:The frequency in obesity and overweight group of the three gene types d3/d3,f1/d3,f1/f1 is 7.3%,7.9%and 4.8%;The frequence in control group of the three gene types d3/d3,f1/d3, f1/f1 is 4.8%,17.5%,77.7%.The frequence of d3 gene in obesity group is significantly higher than that of the control group(χ~2=10.66,p=0.005).2.GHR exon3 gene polymorphism and its relationship with metabolic profile in obesity group:The bioactivity of GHRd3 isoform is about 30%higher than GHRf1,so the d3/d3 and f1/d3 genotypes were combind as d3 group.In the obesity group,144 subjects were d3 genotype,and 265 were f1/f1.There existed no statistical significant difference in age (t=-0.21,p=0.83),heigh(t=-0.20,p=0.84),weight(t=-1.73,p=0.17),and sex ratio between the two genotypes(χ~2=0.11,p=0.74).The BMI(t=-2.4,p=0.02),fasting insulin(t=-2.91, p=0.004),HOMA index(t=-5.11,p=0.00),total cholesterol(t=-2.34,p=0.02), triglyceride(t=-2.16,p=0.032) of the d3 group was significantly lower than of the f1/f1 group.The insulin senstive of the d3 group is significantly higher than that of the f1/f1 group(t=4.37,p=0.00).3.GHR exon3 gene polymorphism and its relationship with metabolic in normal control group:In the control group,46 subjects were d3 gene,and 160 were f1/f1.There existed no statistical difference in BMI(t=1.03,p=0.30),fasting insulin(t=0.25,p=0.80),HOMA index(t=-0.08,p=0.94),insulin sensitive index(t=0.59,p=0.56),total cholesterol (t=-0.23,p=0.82) and triglyceride(t=-0.20,p=0.84) between the two genotypes.Conclusion:The GHR d3 polymorphism has no significant effect on children\'s obesity development,but GHR d3 genotype can preventing obesity children from developing into metabolic syndrome via increasing insulin sensitivity. Objective:To identify the association of the IGF1 Gene SNP rs5742694G/T with adult height and body mass index(BMI) in young Chinese adults.Patients and Methods:1000 young adults including 500 male and 500 female from a college were recruited.Body heights and weights were measured,and BMI were calculated.Genomic DNA was extracted from their peripheral blood leukocytes.SNP rs5742694G/T was determined by SnaPshot.The genotype and phenotype correclation were analyzed by SPSS15.0.Results:1.SNP rs5742695 existed three genetypes,TT,TG and GG.The most frequent genetype is TT,and GG is rare both in male and female subtypes.The mean height of each genotype is,for male,TT 172.10cm,TG 172.74cm,GG 172.12cm,there existed no significant statistical difference among the three genotypes(one-way ANOVA analysis p=0.57).The mean height was TT 160.19cm,TG 159.21cm,GG 159.74cm in female group,there existed no significant statistical difference in the three genotypes(one-way ANOVA analysis p=0.73).2.The extreme discordant phenotype design was used to find the height difference between the three genotypes.100 shortest male(H≤170cm) and 111 tallest female (≥177cm) were selected,comparing their gene constituent ratio,and find there existed no statistical significant between the gene constituent composition between the shortest and tallest groups(p=0.70).92 shortest female(H≤155cm) and 108 tallest female(≥165cm) were selected,comparing their gene constituent ratio,and find there existed no statistical significant between the shortest and tallest groups(p=0.47).3.There existed no statistical significant BMI difference among the 3 genotypes (p≥0.05).There also existed no statistical significant in the geneconstituent ratio between the thinnest and fattest males and females(p≥0.05).4.There revealed no significant association among the 3 SNP genotypes and heights or BMI in both sexes using superposition linear regression model analysis.Conclusion:The IGF-1 intron 3 SNP rs5742695 G/T polymorphism showed no significant effect on the adult height and body mass index in current Chinese young people. Objective:To identify the association of the ESRαGene SNP rs2179922G/A with adult height and body mass index(BMI) in young Chinese adults.Patients and Methods:1000 young adults including 500 male and 500 female from a college were recruited.Body heights and weights were measured,and BMI were calculated.Genomic DNA was extracted from their peripheral blood leukocytes.SNP rs2179922G/A was determined by SnaPshot.The genotype and phenotype correclation were analyzed by SPSS15.0.Results:1.SNP rs2179922 exists three genotypes,TT,TG and GG.The most frequent genotypes are GG and GA,and AA is rare both in male and female subtypes.The mean height of each genotype were,for male,TT 172.22 cm,GA 172.23cm,AA 172.03 cm,there existed no significant statistical difference in the three genotypes(one-way ANOVA analysis p=0.88).The mean height was TT 160.34cm,TG 159.74cm,GG 160.30cm in female group,there exixted no significant statistical difference in height of the three genotypes(one-way ANOVA analysis p=0.43).2.The extreme discordant phenotype design was used to find the height difference between the three genotypes.100 shortest male(H≤170cm) and 111 tallest female (≥177cm) were selected,there existed no statistical significant difference in the gene constituent ratio between the shortest and tallest group(p=0.43).92 shortest female(H≤155cm) and 108 tallest female(≥165cm) were selected,there existed no statistical significant difference in the gene constituent ratio between the shortest and tallest group(p=0.37).3.There existed no statistical significant BMI difference among the 3 genotypes(male p=0.59,female p=0.29).There also existed no statistical significant difference in the gene constituent ratio between the thinest and fattest group by extreme discordant phenotype design study(p≥0.05).4.There revealed no significant association among the 3 SNP genotypes and heights or BMI in both sexes using superposition linear regression model analysis.Conclusion:The ESRαGene SNP rs2179922G/A polymorphism showed no significant infulence on adult height and body mass index in young Chinese people.

Preparation and Application of Multifunctional Polymer Drug Carriers

Sun, 27 Mar 2011 13:31:26 +0000

With the research development of nano-biotechnology,multi-functional and smart polymer microspheres have attracted more and more attentions.Especially in the field of cancer therapy,multi-functional polymer microspheres could be used as the drug carriers in the targeted drug delivery system,which has a broad potential application. Based on the research background,this thesis focused on the preparation > characterization and application of the multi-functional polymer drug carriers,which involves the following two parts.First,the magnetic thermo-sensitive micro-containers were prepared in a novel solvent-templated approach,and the products were used as the carriers to load anti-cancer drug Doxorubicin(DOX). Second,the magnetic thermo-sensitive micro-containers were further modified with the conjugation of cancer cell targeting agent folic acid.The products had response to the magnetic field、environmental temperature and recognization of cancer cells.The drug loading and in vitro releasing of these carriers were carefully investigated.And finally the multi-function drug carriers were applied in vitro cancer cell experiments to testify the targeting of the products.The results of each part are listed as follows:1)Based on the previous work,a new solvent-templated approach to the fabrication of magnetic thermo-sensitive micro-containers of Fe_3O_4 nanoparticles with poly(N-isopropylacrylamide) walls is presented.The micro-containers undergo a temperature-induced volume phase transition and present an impressive magnetic response.The micro-containers have a well-defined structure with a narrow size distribution.The wall thicknesses of the micro-containers can be controlled according to requirements.Compared to other preparation methods,the process is simple and reproducible.The magnetic saturation of these micro-containers is high enough to meet most requirements of bioapplications.To further investigate the potential application of these micro-containers,they are tested as drug carriers,with the drug loading and releasing processes carefully studied.The drug encapsulation efficiency and drug content in the carriers are pH-dependent,and the carriers have a maximal drug loading of about 50 wt%under alkaline conditions.The release of the drug from the micro-containers can be controlled by the environmental pH,temperature,and magnetic force.2)Based on the work of part one,the magnetic thermo-sensitive micro-containers were further modified by the conjugation of the targeting agent folic acid,which resulted in the multi-functional drug carriers.These products had response to magnetic field、environmental temperature and cancer cells.With well defined morphology and structure,temperature induced volume phase transition and high magnetic saturation,the products could be used to load anti-cancer drug DOX.The in vitro release of drugs was also investigated in two different conditions which were simulated physiological condition and simulated cancer cell interior condition. We found that DOX release much faster under a simulated cancer cell interior condition when compared to that under physiological condition.Finally,the multi-functional products were utilized in vitro cancer cell experiments to testify the targeting effect.The products exhibited good capability in recognizing cancer cells, which indicated that this carrier-drug complex could be used in cancer targeted drug deliver system.

Study on People Living with HIV/AIDS in Rural Area of Shanxi Province: A Prospective Cohort Study

Sun, 27 Mar 2011 13:31:24 +0000

1.Clinical response and adverse effects of Antiretroviral Therapy among HIV/AIDS patients in rural area:a prospective cohort study[Objective]The study aims to understand the effectiveness and toxicities of free antiretroviral therapy program for adult HIV-Infected people in China,so as to provide scientific evidence for better administering antiretroviral therapy(ART) and for future researches on ART.[Methods]All adult HIV-infected patients in Wenxi county of Shanxi province who started antiretroviral treatment(ART) between 01 July 2001 and 30 November 2008 and aged above 18 years were included in our cohort study.First,all patients underwent epidemiological investigation and laboratory tests at the initiation of the ART.Then,patients were followed up at months 0.5,1,2,3,6,9...after the initiation of ART,mainly to recognize adherence to the regimens,efficacy(CD4 cell counts) and drug toxicities.Data entry was using EpiData software and data analyses were performed using SPSS 11.0 software.[Results]From 01 July 2004 to 30 November 2008,a total of 188 HIV/AIDS patients were enrolled in our study.The average age of these patients was 43.9 years. Among them,61.7%were males and 38.3%females.Most of the patients were married(92.6%),under 8 years\' education level(96.3%) and farmers(99.5%).The major routes of HIV transmission were commercial blood in the middle 90th.From the initiation of ART to the end of the study,the median follow-up time period was 38.0 months(IQR:19.0—43.0months).Over the duration of the follow-up, self-reported adherence was good in 97.8%of patients(adherence rate＞90%),53.9% patients reported never missing administering any antiretroviral drug dose.The results of the generalized estimating equation showed that after adjusting other factors, patients without clinical symptoms at baseline had an lower hazard of missing drugs than those with clinical symptoms at baseline,the odds ratio(OR) was 0.18(95%CI:0.06—0.56):patients with CD4 cell count＜50cells/ul at baseline had an lower hazard of missing drugs than those with CD4 cell count＞=50cells/ul at baseline, the odds ratio(OR) was 0.59(95%CI:0.35—0.99):Patients treated with EFV/IDV had a lower hazard of missing drugs than those with NVP,the odds ratio(OR) was 0.19(95%CI:0.08-0.46).Treatment was effective in most of the patients,the CD4 cell count of patients increased after the initiation of ART.Significant increases in the median CD4 cell count were found at all time points after treatment initiation(p＜0.001).And also the percentage of patients with CD4 cell count＜200cells/μl at all time points after treatment decreased significantly(p＜0.001).According to the results of the generalized estimating equation about the short-term effect,after adjusting the other factors,CD4 cell count had an average increase of 13.55cells/ul every three months; patients＜=40 years old had an average increase of 30.17cells/ul on CD4 cell count than those＞40years old;patients treated with EFV/IDV had an average increase of 33.81 cells/ul on CD4 cell count than those with NVP;CD4 cell count at baseline had a increase of 1 cells/ul,the treated patients had an average increase of 0.68cells/ul.Until the endpoint of our study,eleven patients had stopped the treatment during the follow-up,one patient lost to follow-up,twenty-five patients died of AIDS-related diseases,one hundreds-fifty-one patients still on treatment.The product-limit estimate of survival at3、12、24 and 54 months were0.90、0.88、0.87 and0.84,respectively. According to a mutivariate Cox\'s proportional hazard model,the hazard of death was significantly different with the CD4 cell count at baseline(＞=50cells/μl versus＜50cells/μl) and the type of initial regimens(NVP versus EFV/IDV).Patients with a CD4 cell count＞=50cells/μl at baseline had a lower hazard of death than those with＜50cells/μl at baseline,the HR was 0.27(95%CI:0.17-0.63).Patients treated with EFV/IDV had a lower hazard of death than those with NVP,the hazard ratio(HR) was 0.17(95%CI:0.04-0.72).Until the endpoint of our study,88.3%of patients presented with clinical and laboratory adverse events probably or definitely were attributed to antiretroviral treatment.The common adverse effects were nausea and vomiting(37.2%),poor appetite(35.6%)),fatigue(30.9%),skin rash(24.5%),peripheral neurotoxicity (21.8%),anemia(53.2%),hepatoxity(17.0%),and so on.87.2%of adverse effects were moderate,however there were 12.8%patients changed the initial regimens owing to adverse effects.According to the results of the generalized estimating equation,after adjusting the other factors,the incidence of nausea and vomiting,poor appetite and shin rash decreased over time;patients with clinical symptoms at baseline had higher incidence of poor appetite,shin rash and headache;female patients had higher incidence of nausea and vomiting;patients infected through commercial blood had higher incidence of fatigue;Patients with a CD4 cell count＞=50cells/μl at baseline had higher incidence of fatigue and headache.[Conclusions]ART is effective for HIV-infected adult people in Wenxi county despite initiation of treatment during the advanced stage of disease;Adverse effects are common with ART in HIV-infected people,including some drug-specific adverse effects,and thus regular monitoring is required according to the special time regularity of adverse effects.2.Study of the life status and quality of life among people living with HIV/AIDS in a rural area of Shanxi province[Objective]The study aims to understand the life status and quality of life(QOL) among people living with HIV/AIDS in a rural area of Shanxi province,so as to provide scientific evidence for promulgate the "Four Free and One Care" policy and better the quality of life and for future researches on QOL.[Methods]A cross-sectional study was conducted.All patients were HIV-positive and aged above 18 years.MOS-HIV scale,CES-D scale and ADL scale were used to ascertain quality of life,depression,activity of daily living and so on among PLWHAs.Data entry was using Epidata software and data analyses were performed using SPSS 11.0 software.[Results]A total of 195 PLWHAs were interviewed.The mean age was 45.6 years (25-64 years).Among them,119(61.0%) were male and 76(39.0%) were female. 152(77.9%) were married,and the majority of them(82.6%) were infected with HIV through commercial blood/plasma donation.Their main economic source were farm (62.6%) and working outside(33.3%).about 58.5%had the monthly incoming under 400 yuan.47.2%lived at the poor house.After they knew their status,71.8%received life aids,58.9%reduced school and miscellaneous expense of Compulsory education.Among those who were married,132 reported sex during the past six months and 87.1%consistently used condoms.Among those where were not married.4 reported sex during the past six months and 3(75%) of them consistently used condoms.For the last sexual act,condom use was reported by 92.3%of those who were married and 75%of those who were not married.146(74.9%) were on treatment.among them,90.4%used first line regiments, 94.5%did not forget taking medicine,the taking time and Medicine to diet request, 67.1%were Directly Observed Treatment by their family members and 97.9% examined their health regularly.163(83.6%) had different clinical symptoms and the depression rate were 66.4%.At dimensions involving Mental Health Summary Scores,General Health Perceptions,Energy,Quality of Life,Mental Health and Health Transition,men have got higher scores than women;At dimensions including Physical Functioning,Energy and Physical Health Summary Scores,younger groups have higher scores;At dimensions containing General Health Perceptions,Quality of Life,Mental Health, Physical Functioning and Role Functioning,higher incoming groups have higher scores;at all dimensions,patients with depression had lower scores than those without depression.[Conclusions]the majority of HIV/AIDS patients had poor economic status and suffered severe stigma;The proportion of condom use was high among PLWHAs, however there were still 7.7%-25.0%with risky sexual behaviors;the adherence rate were high but they still presented more clinical and depression symptoms;The MOS-HIV scale was suitable among PLWHAs in a rural area;Paying more attentions to Feminine,older,low incoming PLWHAs and patients with depression,will be helpful in enhancing quality of life on the population.

The Clinical and Experimental Research of Shuang Hong Cream Cure Rhagadia of Hand and Foot

Sun, 27 Mar 2011 13:31:22 +0000

1.ObjectiveRhagadia of Hand and Foot is a common dermatosis,easy occur in exposed part such as the skin of hand and foot.The damage is crack of different size and depth.The occurrence of this disease is related to the epidermal incrassation,dryness,external stimulate and local activities.Chinese medicine practitioners believe that,due to the exogenous wind and cold pathogens,this disease is caused by the disorders of Qi,Blood running sluggish, Dry,brittle and frequent friction of skin,or the meridian surface lack of nutrients.For light patients,this disease only involve the epidermis and affect skin appearance;For hard patients,this disease can involve the dermis or subcutaneous tissue.It cause bleeding and paining that affect patient\' s partial functional activities and bring the pain that affect the patient\'s normal work and daily life.With the development of society,people\'s social activities are becoming more frequently.Rhagadia of skin have brought more embarrassments and inconveniences to the people\'s social activities.Now the main therapy for skin diseases is using medicine in local part,including urea ointment,salicylic acid ointment,etc.These therapies shall moisten the skin and alleviate the symptoms.However,it is not a deal healing for improving and promote the bleed,the pain and the other symptoms which caused by skin lesions.The main medicament ingredient of Shuang Uong Cream isDaemonoropsdracoB1 andRhodiolaroseaL.It\' sdermatological drug made by our hospital,After years of clinical observation,it trulyhasagoodeffect.In order to further know the curative effect of Shuang Uong Cream,and provide theoretical evidence for this medicine clinic extend,We decide to research this subject.2.method2.1Clinical observationAll cases,amount to 80,are Outpatients and Inpatient hospital sick be accepted treatment in WUHAN institute of traditional Chinese medicine dermatology department during November,2007 and December,2008.According to the order of examination,all case are divided into two teams:therapy team and control team.Two teams of case are compared at size of age,sex,course of disease,etc. The difference does not have significance meaning,have comparativity.Treatment course last for 2 weeks.During this,therapy team are given external Shuang Uong Cream and control team are given externalurea ointment.All test complied with the ruleof random and double blind.Curative effect,subjective symptom and skin rash are analyzed statistically according to the scone standard after cure.All the cases,according with the standards,shall be selected into the experiment.Before and after treatment,analysis and compare symptoms and signs of patient\' s treatment.Counting files are tested withχ~2 test and measure files with t test.2.2Experiment Adaptive feeding rats aweek after,use of 10%chloral hydrate intraperitoneal anesthesia 0.3ml/100g,10%sodium sulfide go back hair,At the backof the right and left sides of the cut 2 cm x 2cm-sized full-thickness skin,Caused the wound model,sub-cage feeding.Will be a successful model rats were randomly divided into four groups:blank control group,matrix groups,treatment group and control group,nine in each group.Modeling the first two days beginning medication,every day, for 20 days.Treatment group given Shuang hong cream,control group given urea ointment.The rats in the post-traumatic 7,14,21 days,cut wound tissue, made of FN immunohistochemical specimens were observed.High-power microscope,the organization has brown staining were positive,5 per high power field of the number of positive cells.Analyze.the datas by SPSS 13.0,and measure files use t test.3.Result3.1 Clinical observationThe therapy group convalescing rate is 55%,the control group convalescing rate is 25%.The therapy group convalescing rate is higher than the control group,has very obviously difference after statistics analysis(p<0.01).3.2 ExperimentThe result of immunohistochemistry:Compare the number of FN positive expresion between the two group,after statistics analysis, the difference is obviously(p<0.01),the number of FN in the therapy group is more than the one in the control group.4.COonclusion To cureRhagadia of Hand and Foot,the Shuang HongCream is better than traditional 10%urea ointment.It will get a great curative effect,and it is Non-toxic side effects,easy to use,worth promoting in clinic.Shuang hong cream can speed up the process of FN to promote wound healing purposes.

DNA Methylation in the Disturbance of Spermatogenesis Caused by Exposure to Chemical in Neonatal SD Rats

Sun, 27 Mar 2011 13:31:20 +0000

During germ cell development,the building of DNA methylation patterns plays an important role in the maintaince of male reproduction.The change of DNMT activity and expression may induce abnormal DNA methylation and further interfere with normal spermatogenesis.It has been noticed for a long time that many germ cells in testis were apoptosis during the fetal and early postnatal ages.Cell cycles and apoptosis were regulated by gene.However,change of the DNA methylation level in gene promoter region could influence the expression of these genes and might induce apoptosis.Although DNA methylation and apoptosis could regulate the spermatogenesis,the molecular events governing these facts have not been clearly characterized.5-Aza-2 -deoxycytidine(5-Aza-CdR) is a cytidine analogue which can incorporate into replicating DNA,and indirectly caused genomic hypomethylation It was also reported that 5-Aza-CdR can cause reproductive dysfunction in laboratory animals.Cd and PCB153,persistent bioaccumulative pollutants in environment,can be absorbed into the bodies through food chain.Cd and PCB have been demonstrated a variety of adverse effects including reproductive development,and they also have been reported changing DNA metyltransferase in vitro.Gonocytes resume into mitosis at early neonatal stage,and many of them differentiate into spermatogonia,while others degenerate at this time.The maintance and de novo methylation are important for spermatogenesis.Therefore neonatal exposure to 5-Aza-Cd,Cd and PCB153 models were used to investigate global DNA methylation,DNMT activity and expression of Dnmts and P53 promoter methylation,which helps to explore the role of DNA methylation in chemical induced spermatogenesis dysfunction.Neonatal male rats were randomly divided into 10 groups,24 rats per group and received oral administration of 5-Aza-CdR(0.025,0.25mg/kg),PCB153(0.025,0.25, 2.5mg/kg) and Cd(1.0,2.0,4.0mg/kg) from PND3 for five days.12 rats were killed 24h after the last administration.The remains were fed until 12 weeks.Testes were taken for histological study,immunohistochemistry detection and DNA methylation assay. After exposure for 5d,5-Aza-CdR significantly reduced body weight of rats, which compared with control group,while in adult rats of 12-week age,only 0.25mg/kg dose group lost weight.But there was no significant difference between different dose groups as far as the organ coefficient and anogenital distance were concerned.Although there was no significant abnormal in adult histology,the exposure to 5-Aza-CdR did repress the spermatogenesis and reduced the rate of daily sperm production.The event that 5-Aza-CdR exposure decreased global DNA methylation may happen via repressing the expression of Dnmt3a and lowering DNMT activity.5-Aza-CdR also decreased P53 promoter region methylation level while increased P53 expression therefore it could induce apoptosis.5-Aza-CdR exposure significantly increased apoptosis compare with control group further confirm the thesis.Exposure to Cd resulted in losing weight of rats at PND8 and then it presented no significant change till12-week.No obvious abnormal events happened in histology of PND8 testis.While there was obvious abnormal in 12-week testis,Histologic study showed that there was less muration sperm in tubules and the arrangement of cell was disorder.And sperm heads count from testis homogenates showed significantly decrease in Cd exposure groups compared with control.Cd exposure decreased global DNA methylation may via de novo methylation represse the expression of Dnmt3a, Dnmt3b and therefore lowered DNMT activity.Cd exposure significantly increased apoptosis in testis compared with control group.But it was different with 5-Aza-CdR,because P53 independent may explain why Cd induced apoptosis.As far as PCB153 was concerned,there was no obvious change in body weight, histology of testis both at PND8 and 12 week.And there was no significant change in DNA methylation and apoptosis and related gene expression.But this study did find the daily sperm production decrease at adulthood.This suggested there may be other pathway such as endocrine that can affect spermatogenesis.This study indicated that DNA methylation could be affected by chemical during neonatal exposure via multiple pathways.The abnormal pattern of DNA methylation could cause reproduction dysfunction and disturb the spermatogenesis.Further studies are warranted to increase our understanding of the effect ofDNA methylation in relation to the underlying mechanisms of chemical induced disturbance of spermatogenesis.

Outcome Evaluation of Qinba Health Project

Sun, 27 Mar 2011 13:31:18 +0000

ObjectivesThe overall objective is to evaluate the impact of Qinba health project on maternal health services utilization in there areas. The specific objectives are to find out the factors which influence the maternal healthcare utilization in these areas according to the Behavioral Model of Andersen Health Service Utilization, to compare the utilization of maternal care between the intervention group and control group, then to give some advices to improve the utilization of the maternal care in these areas.Materials and Methods(1) Source of materials: The data in this research was collected from the post intervention household health survey for Health projectⅧin 2006 by MOH. The survey was carried out in three provinces, including 4 counties in Ningxia, 5 counties in Shaanxi and 5 counties in Sichuan respectively, with a total of 14 counties. There were 4 intervention townships and 2 control townships which were randomly selected from each county, and then there were 2 villages randomly selected from each township, and next there 40 households randomly selected from each village. The women who had live birth from 2002 to 2006 were included in this study, finally this study sampled a total of 1653 cases, 1145 cases of the intervention group and 508 cases of the control group.(2) Content of the study: This study included part of content of the household survey, including the demographics and socio-economic status of the maternity and her family, the accessibility and utilization of maternal health services and other related information.(3) Methods: To describe these influencing factors according to behavior model of Andersen health services utilization and compare the utilization of maternal health care services between the intervention group and control group and use the multi-level model to test the effect of intervention and other influencing factors on the utilization of maternal health care. Results(1) The women\'s houses were far away from the hospitals, and the women needed a lot of time to get there. The proportions of women\'s house to the clinic within 1 km distance were 49.04% and 52.36%, the proportions of women\'s house to the clinic within 10 mininutes\' distance were 62.88% and 68.90%. The average distances between the women\'s house and the township hospitals were 9.4 km and 6.6 km, the average time needed to get the township hospitals are 36.6 mininutes and 26.3 mininues. The average distances between the women\'s house and the county hospitals were 48.4 km and 36.3 km, the average time needed to get the county hospitals were 115.7 mininutes and 80.1 mininues. Generally speaking, the women\'s houses in the intervention group were farther than the control groups, and they also needed more time to reach the maternal health care services than the control group. The annual average household income was quite low in these areas, the annual average household income is 1450.0 Yuan and 1707.5 Yuan; so it was hard for them to afford the maternal care services; and the situations in the intervention group was even worse than the control group. So the accessibility of maternal health care in the intervention group was lower than the control group.(2) The proportion of prenatal visit and the proportion of early prenatal visit were gradually increasing in these areas from 2002 to 2006. The proportion of prenatal visit in the intervention group increased from 87.06% to 97.02%, and in the control group, it increased from 83.75% to 93.10%. The proportion of the first prental visit within 13 weeks of gestation in the intervention group increased from 35.37% to 53.88%, and in the control group, it increased from 42.42% to 48.57%. And the extent of the change in the intervention was greater than in the control group. The average times of prenatal visit in the intervention group increased from 2.8 to 3.6, and in the control group, it increased from 2.4 to 3.4. The difference between the intervention group and control group were statistically significant after the adjustment of other influencing factors.(3) The proportion of hospital delivery increased with the time in these areas from 47.06% to 68.44% in the intervention group, and increased from 45.00% to 73.91% in control group After adjusting confounding factors, the probability of hospital delivery in the intervention group was a little more than in the control group, the OR is 1.17, though the difference is not statistically significant.(4). The proportion of postnatal visit at least more than one time increased from 66.27% to 76.83% in intervention group and increased from 61.25% to 71.30% in control group. The average times of postnatal visit increased from 1.5 to 1.9 in intervention group and increased from 1.3 to 1.4 in conrol group. The times of postnatal visit in the intervention group were more than in the control group after the adjustment of other influencing factors, and the difference was statistically significant.(5) The accessibilities and utilization of maternal care services were quite different among different provinces and counties, and the utilization in the same county were quite similar, the ICCs for average prenatal visits, proportion of hospital delivery and average postnatal visits were 0.2355, 0.0821 and 0.1879 respectively, and the similarity were statistically significant.(6) According to the index of Composite prenatal care utilization, the proportion of insufficient in the intervention group and control group were 65.76% and 65.94% respectively, but from 2002 to 2006, this percentage decreased from 77.65% in 2002 to 57.14% in 2006 in the intervention group, and from 75% in 2002 to 60% in 2006 in the control group. According to the index of Composite prenatal care utilization, the probability of the hospital delivery for the appropriate or adequate is 2.3 times and 1.9 times of the insufficient, the average times of post-natal visits was 0.39 times and 0.59 times higher than the insufficient, and all the differences were statistically significant.Conclusions and Suggestions(1) There were more utilizations in the intervention group than the control group even though the accessibility in the intervention group were worse in the control group, so the intervention had improved the utilization of the maternal health care services by improving health care systems and economic accessibility of the poor maternal family.(2) The social and economical development in study areas was slower than the other rural areas. There is still big room for improvement in the area of maternal care provision and utilization. We should invest more to change women\'s belief in maternal care utilization and improve the access to maternal care in these areas.

A Cost-effectiveness Analysis of Peginterferon and Nucleoside Analogues for HBeAg-Positive Chronic Hepatits B

Sun, 27 Mar 2011 13:31:15 +0000

BACKGROUND:Hepatitis B virus infection is an important public health issue.A variety of pharmaceuticals are currently approved for the treatment of hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB),but their relative economic value is unclear in China.The goal of this analysis was to compare the cost effectiveness of adefovir(ADV),entecavir(EYV),lamivudine(LMV),pegylated interferon alfa-2a and telbivudine(LDY).METHODS:We conducted a cost-utility analysis from a Chinese payer perspective over a lifetime time horizon using a Markov model,in a hypothetical population with HBeAg-positive CHB and a mean age of 30 years.We evaluated 1 year of treatment duration of pegylated interferon alfa-2a and 4 years of treatment duration of adefovir,entecavir,lamivudine,and 2 years treatment of telbivudine,with the use of salvage therapy for drug resistance.Disease progression probabilities,costs and quality-of-life data were derived from the published literature.Sensitivity analyses were conducted to evaluate uncertainty in the results.All costs were valued in RMB, year 2009 values.Costs and QALYs were discounted at 5%per anum.RESULTS:Initiation with peginterferon alpha-2a(PEG-IFNα-2a)(9.84 QALYs) and telbivudine(9.64 QALYs) provided the largest treatment benefits overall, followed by adefovir(9.49 QALYs).Initiation with PEG-IFNα-2a for 48 weeks resulted in higher discounted quality-adjusted life years(9.84 QALYs) and total healthcare costs RMB 163,044.With an increase of 0.20 QALYs compared with the next highest QALYs(telbivudine),giving an incremental cost-effectiveness ratio of RMB 7,455.CONCLUSIONS:Treatment with peginterferon alfa-2a for a defined duration of 48 weeks,although its retail price is more expensive than lamivudine,adefovir, entecavir,and telbivudine,it provides improvement in health outcomes,with the greatest clinical and economic value below the current China cost-effectiveness threshold,followed by telbivudine.

Study on Quality of Life among Community Breast Cancer Survivors and Community-based Intervention Strategy in Shanghai

Sun, 27 Mar 2011 13:15:37 +0000

BackgroundBreast cancer is the most prevalent cancer among women in the world today. There are approximately 1.3 million new diagnosed breast cancer patients every year and approximately 0.46 million women die of breast cancer,which has made breast cancer the leading mortality cause of cancer among women.Today,quality of life (QOL)is considered as an important outcome indicator of cancer rehabilitation.More and more researches have shown that the effect of clinical treatment would be increased if the QOL assessment is applied appropriately.QOL can also be the prognostic factor of cancer recovery.However,among existing QOL researches of breast cancer,most of them are new diagnosed cases on hospital base.Actually QOL of breast cancer may differ according to the stage,other physical situation and recurrence situation of the survivors.Researchers can choose proper study population according to different research objectives.Along with the technologic advancement of the diagnosis and treatment of breast cancer,there are more and more community breast cancer survivors.QOL of community breast cancer survivors should be a great concern of researchers.Besides,the quantitative methods are often applied in the QOL research of breast cancer,while the qualitative researches are limited.The questionnaire,the most frequent method of QOL research always gives us the results in number way.We would also like to know how the breast cancer survivors describe their experience from diagnosis,operation,adjuvant therapy to rehabilitation.Objective The study is to assess the QOL of community breast cancer survivors comprehensively and to discuss possible related factors of QOL through both quantitative and qualitative methods;to describe the survivors\'need for community-based rehabilitation intervention from perspectives of both survivors and health care providers;to explore feasible community-based rehabilitation intervention model and to provide recommendations for the community-based breast cancer intervention. Methods1.The quantitative study of QOL among community breast cancer survivors:390 breast cancer survivors were asked to fulfill the questionnaires when the community doctors visit her at home.2.The qualitative study of QOL among community breast cancer survivors:In-depth interviews were conducted among 9 community breast cancer survivors.3.The intervention strategy of community-based rehabilitation for breast cancer: in-depth interviews were conducted among 9 community breast cancer survivors and 3 community health service providers.And literature anaylsis was also used.Results1.The current situation of QOL among community breast cancer survivors:the overall score of QOL based on FACT-B questionnaire is 102.21±16.62,accounting for 70.98%of the full score.The scores differ among different domains of QOL. Multiple regression analysis showed that economic situation,the severity of disease, social support situation and current treatment all have an independent,significant association with overall QOL and differential domains of QOL.Social support can improve the QOL of community breast cancer survivors;the related symptoms and side effects of treatment will bring negative influence to different domains of QOL; Self-adjustment is the internal motivation of breast cancer to improve their QOL.2.The need of breast cancer survivors for community-based intervention:the problem of current community cancer registration and visit system are:lack of user-friendly visit way;lack of efficient information platform;insufficient capacity of community health service providers for community-based breast cancer intervention; and shortage of funds for community-based breast cancer intervention.Community breast cancer survivors are expecting to return to the society;to get user-friendly rehabilitation and categorized health services;to obtain more economic support,to take part in more accessible,effective and various group activities;and to psychological care instruction. 3.The new model of community-based intervention should take comprehensive social support and classified rehabilitation instruction into consideration to increase knowledge of breast cancer survivors,to enhance their confidence and to improve the QOL.Conclusion and suggestionThe overall QOL of Shanghai community breast cancer survivors is quite good, but QOL of different domains have differences.As a comprehensive assessment indicator,QOL can reflect the survivorship of community breast cancer survivors. The new model of community-based intervention will address the inevitable requirement of modern medical development as well as benefit the most breast cancer survivors.The comprehensive social support and classified rehabilitation instruction are two main strategies.

Construction and Identification the Therapeutic Chimeric Antibodies with Broad-spectrum Neutralization to Highly Pathogenic Avian Influenza H5N1 Virus

Sun, 27 Mar 2011 13:15:35 +0000

The highly pathogenic avian influenza (HPAI) is a severe infectious disease caused by H5N1 virus. As the virus spread worldwide in recent years, the poultry industry has suffered large economic loss and the virus has broken the restriction of interspecies transmission to infect human. The current problem of H5N1 is the high mortality and the lacking of effective drugs. The pandemic virus strains with different antigenicity prevalence at the same period due to the persistent mutation. It is necessary to develop drugs of wide range to H5N1 to meet the need of long-term strategic reserve.Neutralizing antibodies is an effective way to control viral diseases. However, the neutralizing antibody to H5N1 has limit spectrum to a few strains because of the highly frequent mutation of virus genes, especially the hemagglutinin (HA) genes, which restricts the application of neutralizing antibodies.We has constructed a H5-specific antibody panel with hemagglutinin inhibition(HI) to 41 strains of virus belonging to 10 clades and some of them had broad-spectrum protection to mouse in vivo. In the paper, four strains of mouse antibodies designated as 4D1, 10F7, 8H5 and 13D4 with high activity in neutralizing assay are humanized to reduce the human anti-mouse antibody (HAMA) in therapy, which is fundamental to the clinical application. It is also expected to construct a platform for therapeutic antibody humanization including gene modification, construction of expressing vector, stable cell strain of highly expression, scale-up culture with serum-free and antibody purification.In our research, the variable genes were obtained from murine hybridoma cells by RT-PCR and combined with the constant region of human IgG1. The genes were then inserted into pcDNA3.1 to transiently expressing chimeric antibody. The activity was tested by HI, ELISA and Immunofluorescence. To accumulate the appropriate amount of antibody for further research, we used Flp-In expression system which could stably express antibody with high efficiency by FRT targeted integration. After the screening procedure, we obtained several stable cell lines and the production of antibody increased 3-4 times.To fulfill the purification and scale-up production, we developed the CHO culture with serum-free system, including nutrient content in medium, roller bottle culture. We found that the addition of BSA could increase the production significantly, especially 3% BSA addition which could increase by 8 times but influenced the purification. The expression strategy could be optimized considering the purity and yield of antibody. The affinity chromatography by protein A was a specific and effective way for antibody purification. We explored eluting buffer with different pH and obtained the appropriate one with final purity of 95% and yield of 90%.To confirm the humanization and expression procedure had no influence on the activity of variable genes, the purified antibody from serum-free culture was tested by HI and neutralizing assay to several strains of H5N1 virus to test the broad-spectrum neutralization. The protection assay in mouse was performed to the 13D4cAb with broad-spectrum neutralization. It showed that the 13D4cAb had 100% therapeutic activity to 4 clades of virus.In summary, we successfully constructed four strains of chimeric antibodies to HPAI H5N1. They maintained the broad-spectrum neutralizing activity as murine antibodies and reduced the HAMA in therapy, which laid a foundation to antibody therapy for H5N1 infection.

Preparation and Characterization of Chitosan Derivates as Anti-tumor Drug Delivery System

Sun, 27 Mar 2011 13:15:33 +0000

Sustained/Controlled release drug delivery system (SCDDS), one of the fast-developing technologies over the past two decades in international medical community, by changing the dosage form or drug delivery methods, can overcome the shortcomings of traditional anti-cancer drugs. Although research on China\'s anti-cancer drugs and development of controlled-release drug delivery system have improved a lot, there is still a big gap when compared with developed countries abroad, therefore, it is of great significance to acquire independent intellectual property rights of sustained/controlled release drug delivery system for anti-cancer drugs. Chitosan and its derivatives are very promising candidates as ustained/controlled release carrier-materials for a variety of drugs, due to low toxicity and biological activity, which can have a dual effect when combined with drug treatment. It is reported that chitosan and its derivatives can be used as antitumor targeting-drugs-carrier. That drugs coupled with chitosan and its derivatives or chitosan and its derivatives as raw materials are mainly methods for the preparation of anti-tumor drug particulate formulations.The main work includes: (1) Establishment of tumor-targeted drug carriers by modified chitosan, and use X-ray diffraction, scanning electron microscopy, transmission electron microscopy, infrared spectroscopy and so on to analyze modified nanoparticles, research and analysis the influence of preparation methods and experimental conditions on the physical and chemical properties of chitosan; (2) Hydroxycamptothecin as a model drug to study modified carriers, such as water solubility, particle size, preparation methods, drug release characteristics of drug delivery systems, etc.; (3) MTT experiment to study carriers and drug loaded particles on the destruction of liver cancer cells; (4) Construction of S180 sarcoma model in mice to study the inhibitory to sarcoma solid tumor of drug-loaded system; (5) Construction of H22 liver cancer model in mice, using in-vivo-imaging system to study the targeting behavior of the drug system in vivo. The main findings are as follows; (1) Successful synthesis of water-soluble N-succinyl-chitosan and a new type of water-insoluble N-succinyl-chitosan under microwave irradiation; (2) Use of dialysis and membrane emulsification method to prepare a variety of HCPT-loaded N-succinyl-chitosan nanospheres, which can achieve different levels of sustained-release effect; (3) Water-soluble N-succinyl -chitosan can be accumulated in the lungs; (4) Water-insoluble N-succinyl-chitosan has a better inhibitory effect on S180 sarcoma solid tumor and destruction effect on H22 hepatoma cells, showed targeting behavior to liver tumor tissue in vivo experiments.

Amelioration of Multiple Low Doses of Streptozotocin Induced Diabetes by Inhibition of Aldose Reductase

Sun, 27 Mar 2011 13:15:31 +0000

TypeⅠdiabetes mellitus is an islet-specific autoimmune disease resulting from the specific destruction of insulin-producing pancreaticβ-cells,culminating in a state of hypoinsulinemia and hyperglycemia.Inhibition of aldose reductase have been shown to modulate a number of immune functions,including suppression of nuclear factor-κB mediated transcription in lipopolysaccharide treated cells of macrophage, and further reduction of the production of interleukin -1 and tumor necrosis factor-α. The aim of this study was to determine the effect of inhibition of aldose reductase on type 1 diabetes mellitus.Administration of multiple low doses of streptozotocin(40 mg/kg intraperitoneally for 5 consecutive days) to mice resulted in phenotypes similar to that of typeⅠdiabetes mellitus.Together with the injection of streptozotocin,mice were co-treated with inhibitor of aldose reductase(50 mg/kg/day) or not for 10 days. Blood glucose level was assayed periodically.Serum insulin level was determined at the end of experiment using insulin Enzyme-linked immunosorbent assay kit.Western blot was performed with serum samples to determinded the antibodies level in serum. Aldehyde fuchsin staining and hematoxylin and eosin staining were used for histopathological examination of pancreas.20 days after the initiation of streptozotocin treatment,mice receiving both streptozotocin and inhibitor of aldose reductase treatments had significant lower blood glucose levels than the streptozotocin treated but inhibitor of aldose reductase untreated mice.At the end of experiment,the serum insulin level for inhibitor of aldose reductase and streptozotocin co-treated mice was almost 2 folds as much as that of the streptozotocin treated but inhibitor of aldose reductase untreated group.The total antibodies in serum for inhibitor of aldose reductase and streptozotocin co-treated mice were greatly reduced than that of the streptozotocin treated but inhibitor of aldose reductase untreated group.Pancreatic hematoxylin and eosin staining and aldehyde fuchsin staining showed that multiple low doses of streptozotocin treatment caused significant pancreatitis,islets atrophy,and significant reduction of the number of pancreaticβ-cells.These histological lesions,however,was significantly alleviated by inhibitor of aldose reductase treatment.We further investigated the effect of inhibition of aldose reductase on the immune response of mouse peritoneal macrophage.Mouse peritoneal macrophage was isolation from wild type mice and aldose reductase knockout type mice. Lipopolysaccharide was added to stimulate mouse peritoneal macrophage to increase its immune response.Neutral red test and reverse transcription-polymerase chain reaction for tumor necrosis factor-αwere done to value the immune ability of macrophage.Both of inhibitor of aldose reductase treatment and knockout of aldose reductase can decrease the lipopolysaccharide stimulated increase of tumor necrosis factor-αmRNA level and also the phagocytosis of macrophage.Our results indicate that inhibition of aldose reductase may ameliorate multiple low doses of streptozotocin induced diabetes through suppressing immune response.

Studies on Bioactive Constituents of the Gorgonian Muriceides Collaris

Sun, 27 Mar 2011 13:15:29 +0000

A gorgonian,also known as sea whip or sea fan,is a sessile animal of the Subclass Alcyonaria,Class Anthozoa,Phylum Coelenterata found throughout the oceans of the world,especially in the tropics and subtropics from inter-tidal to as deep as 4,000 meters.There are 45 species from seven families of gorgonians in China, mainly located in Hainan,Guangdong,Zhejiang and Taiwan coast.Studies in the past half century showed that a variety of novel and bioactive chemical constituents were gained from the gorgonian.The gorgonian Muriceides collaris were collected from the coast of Weizhou Island,Guangxi province,China in April,2006.To the best of our knowledge,this is the first report of the chemical constituents and biological activities from the genus Muriceides.In our ongoing program to search for natural products with unique structural features and pronounced biological activities,this paper systematically investigated on the chemical constituents and biological activities of the gorgonian Muriceides collaris.Isolation and structure determination:Monitoring by bioassay-guided isolation,the MeOH extract of the gorgonian Muriceides collaris was chromatographed on silica gel,TLC and Sephadex LH-20,and semi-preparative HPLC,respectively,then thirty-two compounds were isolated and purified from the gorgonian Muriceides collaris.Twenty-seven compounds were identified on the basis of spectral methods(1D-NMR,2D-NMR,MS,IR,etc.) and their physicochemical properties.Among them,there were seven new compounds(1-7) and four new natural products(8-11).These compounds were structurally elucidated as fourteen guaiane sesquiterpenoids,namely,2-tropolonylguaiazulene(1),collarisin A(2), collarisin B(3),collarisin C(4),2-formyl-8,9-dihydro-linderazulene(5), 2-formyllinderazulene(6),guaiazulene-2,9-dione(9),2-formylguaiazulene(10), 2-methoxylguaiazulene(11),guaiazulene(12),4-forrnyl-7-isopropyl-10-methyl -az ulene(13),sesquiterpene ketolactone(14),guaianediol(15),guaia-l(5),6-diene (16);and two indene sesquiterpenoids,namely,6-formyl-5-isopropyl-3 -hydroxymethyl-7-methyl-1H-indene(7),5-isopropyl-3,7-dimethyl-1H-indene-1-one (8);and one cholesterol(17);and one batyl alcohol(18);and three benzoyl derivatives,namely,benzoic acid(19),diisobutyl phthalate(20),1, 2-benzenedicarboxylic acid bis(2-ethylhexyl) ester(21);and two miazines,namely, uracil(22),thymine(23);and four nucleosides,namely,2\'-deoxyuridine(24), 2\'-deoxythymidine(25),thymidine(26),2\'-deoxyadenosine(27).Biological activities:Sixteen sesquiterpenoids(1-16) were evaluated for their cytotoxicities against P388 and BEL-7402 cancer cell lines by the MTT method.The results indicated that compounds 9-10,12 showed cytotoxicities in certain extent. Among them,compound 9 had moderate cytotoxicities against P388 with the inhibition rate 57.46%at concentration of 50 ug/mL and weak cytotoxicities against BEL-7402 with the inhibition rate 15.42%at the same concentration.Compound 10 exhibited strong cytotoxicities against P388 and BEL-7402 with the inhibition rate 95.67%and 82.52%at concentration of 50μg/mL.Compound 12 showed weak cytotoxicities against P388 and BEL-7402 with the inhibition rate 21.94%and 9.45% at concentration of 50μg/mL.In addition,compound 9 exhibited potent antibacterial activity against vibrio anguillarum,vibrio harveyi and vibrio alginolgticus; compounds 10,16 had significant antibacterial activity against vibrio anguillarum.Moreover,possible biosynthetic pathways of sesquiterpenoids were postulated according to their structures and literatures in the paper.Based on the results of this research,we could do further more.

Study on the Isolation, Identification, Bioactivity and Secondary Metabolites of Mangrove Actinomycetes

Sun, 27 Mar 2011 13:15:28 +0000

Nature products are important source of Medicine.Actinomycetes have a distinguished advantage in the field of antibiotics:about 75%of antibiotics are produced by actinomycetes,from the secondary metabolites of endophytic actinomycetes,a lot of compounds with new structures and great bioactive diversities could be found.Mangrove plants as important new resources for potential pharmaceutical because of their ecosystems that straddled the land and the sea,from freshwater to seawater,often with distinct microorganism species,have gradually attracted more and more attention of natural product chemists.From the leaves and roots of two mangrove plants(Kandelia candel and Aegiceras corniculatum)which collected from Fugong,Jimei,Haicang in Fujian Province,55 mangrove actinomycetes(endophytic or epiphytic)were isolated.In the investigation of antimicrobial activities,26 strains(about 47.3%of the total isolates)which showed antimicrobial activities against one or more sensitive microbes(Bacillus subtilis,Bacillus pumilus,Escherichia coli, Staphlococcus aureus,Candida albicans,Aspergillus niger)were screened out by using the agar diffusion assay.Using the method of MTT(Methyl Thiazolyltetrazolium),the isolates were screening for antitumor strains in vitro. The results showed that 5 strains(9.1%of the total isolates)could inhibit the growth of Hela and HepG2 tumor cell lines.The identification of the isolates by 16S rDNA assay indicated that only one strain belonged to Nocardiopsis while all the others belonged to streptomyces. The diversity of AHBA synthase gene was investigated using a pair of primers, AHBA R1-AHBA F.The KS domain of each isolate was amplified.6 strains could be amplified and amplification products displayed 700bp DNA bands, indicated DNA of these strains might have AHBA synthase gene.The strain Nocardiopsis sp.(No.3)and another bioactive strain streptomyces sp.(No.18)were selected to study their secondary metabolites. 13 compounds were isolated from strain No.3,including 3 new compounds(3C, 3Y-10)which were pyranones and 10 known compounds(3A,3Y-5,3Y-7, 3Y-14,3L-A,3L-M,3L-G,3Y-18,3Y-19,3Y-20).Within the 10 known compounds,3A,3Y-5,3Y-7,3Y-14 were derivatives of tryptophan,3L-A was glycerol 2-aminobenzoate,3L-M was hydroxybenzamide,3L-G was glycerol acetate,3Y-18 was acetyltyramine,3Y-19 and 3Y-20 were Cyclic dipeptides. From strain No.18,4 known compounds(18-C,18-F,18-E,18-G)were isolated,which were Methyl p-hydroxyphenylacetate,Dimethylα-ketoglutarate,2,6-dideoxypyranoside,2,6-dideoxypyranose,respectively.In the later investigation of the antimicrobial and antitumor bioactivities,the new compound 3C showed no antimicrobial bioactivity at 50μg/disk but weak antitumor bioactivity on Hela cell,the inhibition ratios were 2.73%and 7.39% when the concentrations of 3C were 10ug/mL and 20ug/mL.Our study indicated that mangrove plants included numbers of potent actinomycetes with antitumor and antimicrobial bioactivity,and that they were reliable resources for novel metabolites with antitumor and antimicrobial bioactivity at the same time.

The Effects of Leptin, Insulin and IGF-2 on the Expression of PKB and P~(70)S6K Protein in the Trophoblastic Cells and the Signaling Pathway

Sun, 27 Mar 2011 13:15:24 +0000

ObjectiveSince the birth weight is closely related to adult diseases,Present investigations on the birth weight indicated that the birth weight was regulated by nutrition、the structure and function of placenta,growth factors and hormones,among growth factors and hormones the most representative factors were leptin,insulin and insulin-like growth factor -2(IGF-2),these three factors have certain regulative functions in the birth weight where is the mechanism for the kind of regulatory functions? PI3K/PKB/mTOR signaling pathway can regulate the growth and differentiation of the trophoblastic cells by regulating the phosphorylation of p~(70)S6K.This investigation discussed whether the effects of leptin,insulin and IGF-2 on the birth weight have correlations to the PI3K/PKB/mTOR pathway by studying the expression of PKB and p~(70)S6K protein in the trophoblastic cells and the regulatory functions of specific inhibitors in the PI3K/PKB/mTOR signaling pathway,namely LY294002 and rapamycin.Material and MethodsThe villi were collected from the normal pregnant women had been pregnant for 6 to 8 weeks were digested with 0.25%trypsin at 4℃for about 40 mins.25 IU/ml DNase I was added for 10-15 mins after the centrifugation.Then it was sieved by a strainer mesh of 100 screen mesh and subjected to a density gradient sedimentation with 2.5% and 1.25%BSA for 45 mins.The trophoblastic cells in the underlayer were collected after the centrifugation and they were inoculated in a culture flask that had been overlaid typeⅠcollagen after another centrifugation.The culture flask was incubated in an incubator containing 5%CO_2 at 37℃and the relative humidity was 99%.The culture solution was changed once for every two or three days according to the conditions of the cells.The trophoblastic cells during the first trimester in good conditions were selected and DMEM nutrient solution containing 10%fetal bovine serum was changed after the cells were starved for 24 hours.The cells were subjected to treatments with 10nmol/L leptin;6nmol/L insulin and 25nmol/L IGF-2 for 24 hours, respectively.20μmol/L LY294002 and 100nmol/L rapamycin were added to the other group for 24 hours simultaneously.The cells(5-6×10~6) were collected after 24 hours for use.According to the procedures for Kaiji kit of total protein extraction.The protein concentration was determined by Coomassie brilliant blue method.Western blot was used to detect the expression of PKB and p~(70)S6K in the trophoblastic cells after dealt with leptin,insulin and IGF-2;the effects of leptin,insulin and IGF-2 on the expression of PKB and p~(70)S6K were re-examined after the addition of rapamycin and LY294002. Gene Tools from Syngene gel image analysis system was used for the systematic result analysis,the data were represented by(?)±s,and SPSS13.0 statistical software was used for ANOVA,and the difference was statistically significant(P＜0.05). Results1.Compared with control group,the expression of PKB and p~(70)S6K in test group were significantly raised(dealt with leptin,insulin and IGF-2)(P＜0.05).2.The expression of PKB and p~(70)S6K in the trophoblastic cells after added LY294002 or rapamycin were significantly decreased in test group(dealt with leptin, insulin and IGF-2)(P＜0.05).Conclusion1.Leptin,insulin and IGF-2 can promote the expression of PKB and p~(70)S6K protein in the trophoblastic cells and the promoting effects were significantly inhibited after the specific inhibitor in the PI3K/PKB/mTOR signaling pathway,namely rapamycin and LY294002,were added.2.The effects of leptin,insulin and IGF-2 on the birth weight may be realized partly by the PI3K/PKB/mTOR signaling pathway.

The Application and Clinical Value Analyss of Intraoperative Positioning by Fiberoptic Ductoscopy in Patients with Nipple Discharge

Sun, 27 Mar 2011 11:36:27 +0000

ObjectivesNipple discharge is one of the three most common symptoms of breast diseases, the rate of occurrence of which is between 3%-8%.Since both benign and malignant breast diseases may with the symptom of nipple discharge,the clinical diagnosis determine further treatments.Nowadays,the wide clinical uses of fiberoptic ductoscopy(FDS) make it possible for us to observe the pathological changes of large and middle ducts and the opening part of small ducts directly by an amplificatory view,which provide particular references to the doctors with qualities, numbers and locations of the diseases pre-operation.After all,in clinical practices, the pathological finding rate pro-operation was often much lower than the clinical finding rate by FDS pre-operation.At the same time,considering the important function of breasts\' location,how to decrease the harm of operation to meet the needs of better appearance and minimally invasive as well as how to upgrade the pathological finding rate became an important clinical lesson.In this investigation, we discussed the practical value and advantages of orientation using FDS during operation by comparing with the traditional operation.MethodsWe divided 60 outpatients cases with nipple discharge into observation group and controlling group randomly.In the former group,we chose minimally invasive operation by locating the neoplasm with FDS.On the other hand,the patients of the controlling group received the traditional operations.Then,the effects including operating time,incision length,specimen size and pathological finding rate were evaluated.ResultsWe found that the average incision length of observation group was 2.67±0.24cm while that of the controlling group was 3.47±0.26cm.Average time for operation was 48.00±7.14min of observation group while 33.00±7.60min of controlling group.Average weight of specimen was 7.73±6.02g of observation group while 43.40±5.89g of controlling group.Pathological diagnosis of observation group included 24 cases of intraductal papilloma,4 cases of ductal epithelial papillary hyperplasia,1 case of mammary duct expansion and 1 case of mastopathy,the pathological finding rate is 93.3%.By contrast,pathological diagnosis of controlling group included 18 cases of intraductal papilloma,2 cases of ductal epithelial papillary hyperplasia,2 case of mammary duct expansion and 8 case of mastopathy,the pathological finding rate is 66.7%.ConclusionsComparing with traditional operations,minimally invasive operations under the assistance of FDS had the advantages of small incision,less damage of glands,and higher rate of pathologic finding,which had great clinical significances in treating breast-duct diseases.

Studies on the Constituents of Brucea Javanica

Sun, 27 Mar 2011 11:36:25 +0000

Brucea is a genus with ca. 6 species mainly found in tropical eastern hemisphere areas. Brucea mollis and B. javanica, the only two species in China, are employed as traditional herbal medicines because of their antitumor and antimalaria activities. The green herbal medicinal plant B. mollis. is distributed in southwestern China (Yunnan and Guangzhou Provinces). The herbal medicinal plant B. javanica has more biological activities. This plant, called‘Yadanzi’, distributed in south of China (mainly Guangxi and Guangdong Provinces), shows antitumor, antimalaria, and antigastric ulcer activities. Recently, much attention has been paid to the Brucea genus and its chemical constituents because of the multifaceted activities. Extensive studies of the genus Brucea have led to the identification of many compounds, such as nigakilactones, alkaloids, triterpenoids, and flavonoids.In this dissertation, we investigated the chemical constituents from the aerial parts of B. javanica. Forty compounds were isolated from the aerial parts of B. javanica by silica gel, Sephadex LH-20 and preparative HPLC and thirty-three compounds were identified as follows by spectral analysis: BJ-1:β-sitosterol BJ-2:Daucosterol BJ-3:Stigmasterol BJ-4:cis-Oleic acid BJ-5:Palmitic acid BJ-6:Crotonic acid BJ-7:Vanillic acid BJ-8:(?)-threo-guaiacylglycerol-8-O-4\'-(coniferyl alcohol) ether BJ-9:(20R)-O-(3)-α-L-arabinopyranosyl-pregn-5-ene-3β,20-diol BJ-10:α-D-Glucopyranoside, (3b, 20R)-3-hydroxypregn-5-en-20-yl BJ-11:Luteolin BJ-12:Apigenin BJ-13:Quercetin BJ-14:1, 3-Propanediol, 2-[4-(3-hydroxy-1-propenyl)-2-methoxyphe- noxy] BJ-15:Bis(2-ethylhexyl)phthalate ?BJ-16:Bruceajavanin C BJ-17:Cucumegastigmanes I BJ-18:1-Propanone, 2, 3-dihydroxy-1-(4-hydroxyphenyl) BJ-19:Dis(2-methylpropyl) ester BJ-20:3, 4′-Dihydroxypropiophenone BJ-21:Dysosmarol BJ-22:Adenosine BJ-23:Hirsutrin BJ-24:Hyperin BJ-25:Bruceoside A BJ-26:Bruceoside B BJ-27:Bruceine D BJ-28:Bruceine E BJ-29:Yadanzioside A BJ-30:Yadanzioside G BJ-31:Citroside A BJ-32:1H-Isoindole-1-acetic acid BJ-33:OxanosineAmong them, BJ-16 was a new compound, BJ-10 and 14 were new natural compounds, and BJ-8, 11, 12, 15, 17, 18, 19, 20, 21, 22, 31, 32, 33 were isolated from this plant for the first time.The nine compounds from this plant were evaluated for inhibitory effect on NO production in LPS-stimulated RAW264.7 cells. The results showed that the compounds BJ-16, BJ-26, BJ-27, BJ-28 and BJ-29 displayed significant inhibitory activities on NO production in LPS-stimulated RAW264.7 cells with aminoguanidine as control.

PEG Induced Cloud Point Extraction

Sun, 27 Mar 2011 11:36:23 +0000

When an aqueous micelle solution of a nonionic surfactant is at a temperature above its cloud point or in the presence of certain additives, phase separation occurs to form a surfactant-dilute phase and a surfactant-rich phase or coacervate phase. Such a system is called a cloud point system (CPS). The cloud point system has also been utilized as a separated method, which is known as cloud point extraction (CPE). Generally, in order to implement extraction/reaction at a relatively lower temperature, the nonionic surfactant with low hydrophile-lipophile balance (HLB) value is used. However, it’s difficult to strip organic compounds from nonionic surfactant with low HLB, which makes it difficult for the product recovery and nonionic surfactant reutilization.A novel PEG induced cloud point system (PEG-CPS) with high molecule PEG and nonionic surfactant with relatively higher HLB has been developed. The experimental results suggest that in PEG-CPS, a CPE can be carried out with a relatively higher HLB (TX-100) value nonionic surfactant at about room temperature. High boiling point 1-naphthol has been used as a model organic solute, and the recovery in nonionic surfactant rich phase of PEG-CPS is 90%. A volume ratio of surfactant rich phase to organic solvent iso-butyl alcohol on 5:3 has been proposed to extract surfactant and solute into Om phase of WinsorⅡ, and the recovery of surfactant and solute are both about 96%. After removal of organic solvent and resolving the residual organic solute and surfactant with water, the solution has been used to WinsorⅠextraction with volume ratio of solution to ethyl ether on 5:12 at 6℃, and the recovery of surfactant and solute are both above 79%.PEG-CPS has been developed as a novel medium for whole cell microbial transformation, and the downstream process has also been researched. The experimental results suggest that in PEG-CPS with 15% hydrophilic TX-100 and 6.4% PEG20000, the yield of product is about 6.38g/L. The microemulsion extraction has been used as the separation technique, i.e., WinsorⅠwith volume ratio of supernatant to ethyl ether on 5:12 at 6℃has been conducted to strip product/substrate from nonionic surfactant/PEG; WinsorⅡwith volume ratio of water phase of WinsorⅡto iso-butyl alcohol on 5:3 at 25℃has been conducted to separate nonionic surfactant and PEG.

A Novel Hydrogel System Based on Chitosan: Preparation and Application in Pharmaceutics

Sun, 27 Mar 2011 11:36:21 +0000

Chitosan, a polysaccharide derived from naturally abundant chitin, gets much attention currently for biomedical application. Hydrogels based on chitosan are attractive systems for drug delivery and tissue engineering, which combine biodegradability and biocompatibility, especially those with the ability to form in situ gel.The present work focuses on a novel gelation system composed of chitosan and NaHCO3, which shows thermal-induced sol-gel transition. The factors that may influence the process of gelation were investigated, such as the NaHCO3 concentration, the temperature and the storage condition. And the gelation mechanism was also discussed and proposed. The gelation time was determined by the vial tilting method. Rheological measurements were carried out to evaluate the storage and loss modulus of both sol and gel. The pH-meter was applied to evaluate the pH value of both sol and gel.The chitosan/NaHCO3 system can exhibit sol-gel transition at the concentration of NaHCO3 ranged from 0.08mol·L-1 to 0.12mol·L-1. Increasing the concentration of NaHCO3 or the environment temperature would speed sol-gel transition of such system, while sol-gel transition would be delayed if the mixture was sealed with wax or sealed under the CO2 atmosphere. The pH value of the gel was higher than that of the corresponding sol, and increased along with the concentration of NaHCO3. It is proposed that the pH value of the system was a key factor to trigger sol-gel transition, which may affect the ionization equilibrium between -NH2 and -NH3+ group in chitosan molecule. As the pH value became higher, more -NH3+ turned to -NH2, then polymer chains entangled to form a three-dimensional network, as a result of the formation of chitosan hydrogel finally.The structure of the gel was investigated by SEM, and its erosion behavior was studied by membrane-free method. The freezing-dried hydrogels showed porous structure. The concentration of NaHCO3 had great influence on the inner morphology of hydrogels, which affected the erosion of the gel.The in situ gelation of this chitosan system was tested in vivo by dorsal subcutaneous injection in rat. Results showed that the gel could form in situ during 3h after injection and maintain morphology in 30 days. The formed chitosan hydrogel was studied as drug carrier, using dipyridamole as the hydrophobic drug and 5-Fluorouracil as the hydrophilic one. In vitro release behavior of the drug-loaded hydrogel coincided with the erosion trend of the corresponding hydrogel, which was influenced by the formulation of gelation system. The amount of drug loaded in the gel also had an impact on the drug release rate, that increasing drug loading would lead to a decrease of the drug release rate.

The Research of Detection Method for Early Lung Cancer Based on the Magnetic and Piezoelectricity Biological Sensor

Sun, 27 Mar 2011 11:36:19 +0000

The morbidity and mortality rate of lung cancer is the highest among all cancers, but there is no an effective detection method for the early stage on of lung cancer. Therefore it is significant to seek a detection method for early lung cancer cells to increase the five-year survival rate of patients.Quartz crystal microbalance (QCM) is a micro-instrument based on the characteristic that the oscillating frequency of piezoelectric quartz crystal is very sensitive to the load on the surface, it can sensitively detect the little change in the micro reactions and can translate it to the oscillation frequency. QCM provides an easy and quick manner for getting the real-time information for the micro reaction system, so it has become a key issue in biosensor area.In this paper , a nano-electronic detection device is based on the principle that the vibration frequency of piezoelectric quartz crystal is sensitive to the load on its electrodes surface , also combine with the control ability of FPGA (Field Programmable Gate Array ),single-chip and improving the load response and biochemical reactions that nano-magnetic beads have. The system is designed and developed via the development tools Borland builder 6.0, and it can realize the measurement of frequency of piezoelectric quartz crystal sensor. It consists of the following modules: micro-reactor, signal function circuits, signal processing and control module, the signal acquisition analysis system. The system provides a friendly window to realize the real-time sampling, data acquisition and data displaying via four channels. It can also real-time monitor, process and store the real-time oscillation frequencies of the sensor array. So the system has the ability of qualitative and semiquantitative analysis of cancer cells.According to the measurement results of the distilled water ,NaCl and lung cancer CPA-Y2 solution , we get the following conclusions: the measurement precision of the system is down to 1Hz; the repeated utilization rate of a single piezoelectric sensor can be as high as 7-10 times; The system has a good linear relation between the frequency changes of QCM and the concentration of the NaCl ,CPA-Y2 solution concentration ,as well as it can accurately reflect the quality of biological material. This system will have important value of reference for early diagnosis of lung cancer.Furthermore, we will try to improve the detection efficiency of the system via increasing the Detection channels and the detecting container to improve detecting capacity.

Studies on the Active Constituents of Ainsliaea Fulvioides

Sun, 27 Mar 2011 11:36:18 +0000

The genus Ainsliaea belongs to the Compositae family and consists of around 70 species. Among them, 44 species are distributed in the south regions of China, such as Sichuan, Guizhou and Yunan provinces. Most of them have been used as a folk medicine for the treatment of rhumatism, traumatic injuries, edema, stomachache.So far, several sesquiterpenes, triterpenes and flavonoids, have been isolated from this genus. Ainsliaea fulvioides is a medicinal herb growing mainly in south of China and no phytochemical studies on this plant have been reported. In this dissertation, we investigate the chemical constituents from A. fulvioides systematically, anti-inflammatory and anti-tumor activities. Thirty-two compounds were isolated from A. fulvioides by silica gel, Sephadex LH-20, preparative HPLC and preparative TLC and twenty-eight compounds were identified as follows by spectral analysis (1H-NMR, 13C-NMR, 2D NMR, EI-MS,ESI-MS,HRESI-MS, and IR) and the relative stereochemistry of two new compounds were determined by the X-ray diffraction: TE-1: daucosterol TE-2: 4-methylbenzoic acid TE-3: 1-heptadecanol TE-4: 1-dodecanol TE-5: n-dodecane TE-6:β-sitosterol TE-7: octadecanoic acid *TE-8: ainsliadimer B TE-9: gochnatiolide B TE-10: ainsliadimer A TE-11: umbelliferone *TE-12: ainsliatrimer A *TE-13: ainsliatrimer B TE-14: aescutin TE-15: gochnatiolide A TE-16: 4-hydroxy ethylbenzoate *TE-17: ainsliatone A TE-18: 1, 2-benzenediol TE-19: spinacetin TE-20: 4-hydroxylbenzaldehyde TE-21: 3,4-dihydroxy-benzaldehyde TE-22: luteolin TE-23: ayanin TE-24: apigenin TE-25: ameliaroside TE-26: piceol TE-27: quercetin-3-O-β-D-glucoside TE-28: syringinAmong them, TE-8, TE-12, TE-13, and TE-17 were new compounds. Up to now, TE-12 and TE-13 were the first two representatives of naturally occurring guaianolide-type sesquiterpene lactone trimers. TE-17 was a new nor-sesquiterpne lactone. Others were isolated from this plant for the first time. A proposed biogenesis for these trimers was illustrated by using natural Diels–Alder reaction in this thesis.Six sesquiterpenes from this plant were in vitro evaluated for their anti-tumor activities against four human cancer cell lines, A549, LOVO, CEM and MDA-MB-435 (MDA) using MTT assay with DOX (doxorubicin) as positive control. These compounds showed potent cytotoxicites against four human cancer cell lines, and TE-15 showed significant cytotoxicites with IC50 values of 3.24,0.17,0.01 and 0.13μg/ml respectively. Three sesquiterpenes from this plant were tested for the inhibitory activity against the production of NO in RAW264.7 macrophages stimulated by lipopolysaccharide (LPS) and against NF-κB activities in pNF-κB-luc-293 cells induced by LPS. The results showed that TE-12 had the 50% inhibitory rate against NF-κB activities at 1μg/ml.

Studies on the Active Contituents of Viburnum Propinquum and Viburnum Erosum

Sun, 27 Mar 2011 11:36:16 +0000

The genus Viburnum (Caprifoliaceae) includes ca. 200 species, widely distributed in temperate zone in northern and hemisphere subtropical regions. About 74 species were widely distributed in China, especially in Southwest China. Several species are employed as folk medicines to treat different diseases, such as cough, diarrhea, rheumatoid, arthritis and tumefaction. Extensive studies of the genus Viburnum have led to the identification of many compounds, such as diterpenoids, triterpenoids, iridoids, monoterpenes, sesquiterpenes, flavonoids, and lignans, including a number of compounds with good antioxidant and anti-tumor activity.Viburnum propinquum Hemsl. is a kind of evergreen shrubs, distributed in Shanxi, Gansu, Zhejiang, Jiangxi, Fujian, Sichuan, Taiwan, Guizhou, Yunnan provinces of China. It has been used in Chinese folk medicines to treat Fractures, bone-setters injury, trauma hemorrhage. Viburnum erosum Thunb. is a kind of deciduous shrub, distributed in east China, Shanxi, Guangdong, Guangxi and Southwest of China. It has been used in Chinese folk medicines to treat stomatitis, eczema.In this dissertation, we investigate the chemical constituents from the branches and leaves of Viburnum propinquum Hemsl. Besides, some of compounds from the title plant have been evaluated their antioxidant activity. Fifteen compounds were isolated by Silica gel chromatography, Sephadex LH-20 chromatography, and all of the compounds were identified by spectral analyses as follow: VP-1: 3, 4, 2′, 4′-tetrahydroxy-trans-chalcone, VP -2: 3, 4, 2′, 4′-tetrahydroxy-trans-chalcone-2′-O-β-D-glucoside, VP -3: (7-αH, 8′-αH)-4, 4′, 8α, 9-tetrahydroxy-3, 3′-dimethoxy-7, 9′-epoxylignan, VP -4: Taraxerol, VP -5:β-sitosterol, VP-6: Stigmasterol, VP-7: 3β, 28-dihydroxy-12-ursene, VP-8: Ursolic Acid, VP-9: Daucosterol, VP-10: 4, 2′, 4′-trihydroxy-dihydrochalcone, VP-11: 4, 2′, 4′-trihydroxy-dihydrochalcone-2′-O-β-D-glucoside, VP-12: Quercetin, VP-13: Dihydroquercetin, VP-14: 7, 3′, 4′-trihydroxy-flavanone, VP-15: Eriodictyol. Among them, VP-2, VP-3 were new compounds, and VP-1, VP-4, VP-10, VP-11, VP-13,VP-14, VP-15 were isolated from Viburnum genus for the first time. We also investigate the chemical constituents from the branches and leaves of Viburnum erosum Thunb. Eight compounds were isolated by silica gel chromatography, Sephadex LH-20 chromatography and identified by spectral analyses as follow: VE-1:β-sitosterol, VE-2: Ursolic Acid, VE-3: 5, 3\', 5′-trihydroxy-7-methoxy-dihydro-flavone, VE-4: Daucosterol, VE-5: (-)- lyoniresinol 3α-O-β-D-glucopyranoside, VE-6: (+)- lyoniresinol 3α-O-β-D-glucopyranoside, VE-7: Adoxoside, VE-8: Loganin. Some compounds from Viburnum propinquum Hemsl were evaluated for their antioxidant activity. Compounds VP-1, VP-2, VP-12, VP-13, VP-14 exhibited antioxidant activities in 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical, and lipid peroxidation assays.

The Key Technology Research on Mammograms Computer-aided Diagnosis

Sun, 27 Mar 2011 11:26:54 +0000

Mammography is especially valuable for early detection of breast cancer. This thesis summarizes the research on computer-aided diagnosis of breast masses, including breast boundary segmentation, image enhancement, and fast detection of breast mass regions.To separate breast region from background and decrease the impact of the background on our algorithm, we use double threshold and morphological method based on watershed transformation to extract breast border. Furthermore, we make use of different image enhancement methods to augment mammogram contrast. The enhancement can make the spicules detection more precise and make the abnormality regions more accessible to physician.Based on breast border extracting and image enhancement, we detect the possible centers of mass regions from the breast region. The characteristic of breast mass is that its gray level decreases from the center to outside, especially, the malignant mass has the spicules feature. We employed the multi-scale generic neighborhood operator to calculate the image gradient. Our algorithm is evaluated by comparing the detected mass centers with the mass centers from golden standard. The results demonstrate that within 2 false positive per image, our algorithm can detect more than 90% of masses. We develop the algorithm in our lab, employing the image database of DDSM. It will be better if it can be applied in clinical practice.

Study of Simulated Phosphene Characteristics Based on Tactile Perception

Sun, 27 Mar 2011 11:26:53 +0000

As the rapid development of visual prosthesis, the study about the evaluation of the effect after prosthesis implantation becomes an active area of artificial vision research in these years. Phosphenes which are generated by the stimulation of implanted visual prosthesis are elements of the image which the patient will see after implantation, so the study of the phosphene’s characteristics becomes more and more important. The first question is that how we could help the blind to describe the phosphene’s characteristics precisely, so dependable data could be provided to the researcher. The focus of this paper is that a study on an evaluation system of phosphene’s characteristics based on tactile perception which is a kind of important function for the blind, taking simulated phophene as the research object.This paper includes two parts: the first part is the study of simulated phosphene positioning based on tactile perception. In this part, a tactile board based on rectangular coordinate is used as the tactile guidance. The whole positioning system and a series of psychophysical experiments are designed. The results of the accuracy, dispersion and response time are analyzed to evaluate the performance of the system. It is concluded that the simulated phosphene positioning system with tactile board is accurate and reproducible in evaluating simulated phosphene. Then a new tactile guidance based on polar coordinate which conform the human physiological requirement better is used in the following research. All of the tactile guidance system and designation of the psychophysical experiments is improved. The second part is a study of the evaluation of simulated phosphene’s based on tactile perception,. At the first stage of the study, the effect of different tactile boards, distributions of simulated phosphene and distances on the evaluation of the phosphene size is studied. The conclusion is that the method of the evaluation of simulated phosphene size based on tactile perception is available. The error of size evaluation of simulated phosphene decreases in the near distance using tactile board with round holes.

Association, Expression Studies and Meta-Analysis of Genetic Variations Implicated in Schizophrenia

Sun, 27 Mar 2011 11:26:51 +0000

Schizophrenia (MIM 181500) is one of the most severe psychiatric disorders, with a worldwide incidence of 1%. Data from twin, family, and adoption studies provide strong evidence that schizophrenia is predominantly a genetic disorder with high heritability. However, the genes evoking schizophrenia remain unclear.Convergent evidence has suggested that oxidative stress and free radicals may be involved in the pathophysiology of schizophrenia. Glutamate-cysteine ligase, modifier subunit (GCLM) provides a significant protective mechanism against oxidative stress. The purposes of the current study were to determine the role of genetic variations and expression levels of GCLM in susceptibility to schizophrenia in the Chinese population. We conducted a case-control association study on GCLM involving 797 Chinese subjects and three tag-SNPs using directly sequencing. An expression study using quantitative real-time polymerase chain reaction (RT-PCR) was also carried out in a subgroup of 180 samples using peripheral blood leukocytes. No significant difference was detected in the association study for any of the three SNPs or haplotypes. The expression of GCLM was significantly lower in the clozapine-treated schizophrenia patients compared to the control group (P=0.0018) and higher in the chlorpromazine-treated subgroup compared to the control group (P=0.0259). The expression levels varied greatly with respect to drug-treatment, suggesting that GCLM may be one of the drug targets, further reminding me that variants in the gene locus may be responsible for a certain therapy. The association analysis did not support the theory that GCLM was related to schizophrenia in the Chinese Han population. However, we could not exclude the role of oxidative stress and GSH-related gene in the pathology and etiology of schizophrenia, and antipsychotics-treatment should be taken into consideration in the further investigation of such genes.2’,3’-Cyclic nucleotide 3’-phosphodiesterase (CNP), one of the most promising candidate genes for schizophrenia, plays a key part in the oligodendrocyte function and in myelination. Our study aims to investigate the relationship between CNP and schizophrenia in the Chinese population and the effect of other factors on the expression level of CNP in schizophrenia. Five CNP SNPs were investigated in a Chinese Han schizophrenia case-control sample (n=180). The results were included in a meta-analysis. RT-PCR was conducted to examine CNP expression levels in peripheral blood lymphocytes. Factors including gender, genotype, sub-diagnosis and antipsychotics-treatment were found not to contribute to the expression regulation of the CNP gene in schizophrenia. Our meta-analysis yields no positive evidence. The results suggest that the CNP gene may not be involved in the etiology and pathology of schizophrenia in the Chinese population.

Visualization of Blood Vessels by Evans Blue and Its Application in the Study of Spinal Cord Injury

Sun, 27 Mar 2011 11:26:49 +0000

Spinal cord injury (SCI) often leads to lifelong disability of the patient, resulting in suffering of the patient and burden of the family and society. At present there is still no effective therapy. Therefore it is difficult and important to study the pathology and therapy of SCI. The pathological process includes the primary injury and the secondary injury. The primary injury causes mechanical trauma of cells and tissue, and also diversified noxious agents which are cytotoxic damage adjacent spinal cord segments and lead to the secondary injury. The necrotic wave spreads in the direction of rostra and cauda, and leads to a greater injury several times more than the original injury. Therefore the mechanism, pathological changes and therapy of the secondary injury are of great importance. Ischemia is the important pathological mechanism for the reduction of spinal cord blood will exacerbate SCI. To study ischemia in the secondary spinal cord injury, the angiography that displays the blood vessels and associates with the immunohistochemical staining is the essential study method.At present, methods used to display blood vessels mainly contain ink perfusion, immunohistochemical method, tannic acid-ferric chloride staining, enzyme histochemical method and so on. However, they all have disadvantages, for example, unable to do immunohistochemical staining, troublesome or fluorescence quenching and so on. We use Evans Blue to dispaly blood vessels. Evans Blue is a fluorescent dye, which is blue at the bright field and red under the excitation of green light with fluorescent microscope. It is used to study the breakdown of the blood brain barrier. Evans Blue can bind to plasma albumin forming Evans Blue-albumin complex, and then combine with the receptor for albumin on the membrane of vascular endothelial cell. Therefore Evans Blue can be perfused to display blood vessels theoretically.There are two sections in the experiment. First we established the method. The experiment animals are C57BL/6 mouse and SD(Sprague-Dawley)rat. 12 C57BL/6 mice are divided into 2 groups: control group, uninjury perfusion group. After injection of EB through the femoral vein (the control group are given normal saline instead), the mice are perfused with normal saline and paraformaldehyde, sliced. Then one slice is dehydrated and transparentized with 100% ethanol and fixed with neutral balata, another slice is done HE staining, and the third slice is done immunostaining for GFAP. 15 SD rats are divided into 2 groups: Post-fixation group and Prefixation group. The method for post-fixation group likes C57BL/6 mouse, including control group, 5% Evans Blue group and 8% Evans Blue group. Prefixation group is divided to two groups: 5% gelatin-Evans Blue group and gelatin-Potassium chromium sulfate-Evans Blue-albumin group. After perfusion with normal saline and paraformaldehyde, the rats are respectively injected solutions to display blood vessels. After gelatin becomes hard, the tissue is taken out, sliced, dehydrated and transparentized with 100% ethanol, fixed with neutral balata or done immunohistochemical study. The results can be summarized as follows: When C57BL/6 mice are given 0.5ml 5% Evans Blue, the blood vessels in the spinal cord are clearly displayed. For Post-fixation group of SD rats, 5ml 8% Evans Blue is enough to show the blood vessels clearly. For prefixation group, when given 5% gelatin-Evans Blue, the blood vessels are fully displayed. Immunohistochemical study can be done in gelatin-Potassium chromium sulfate-Evans Blue-albumin group.In the second section, we use the method in spinal cord injury. The experiment animal are SD rats. Two weeks after injury, they are operated respectively by post-fixation and prefixation method. The results indicate the configuration of blood vessels between injury area and normal area, also the breakdown of BBB can be observed. Immunohistochemical study can be done on neighboring slide. In prefixation group, when given 5% gelatin-Evans Blue, the blood vessels are clear and there is little Evans Blue diffusion next to the center of injury. When given GCEA solution, not only the blood vessels are displayed, but also immunohistochemical study can be done. However, the effect of blood vessels is not so good as 5% gelatin-Evans Blue group.In conclusion, we study method to visualize blood vessels with Evans Blue systemically and the results reveal that Evans Blue is a sensitive fluorescent dye to display blood vessels in spinal cord. Different fixation method and additive can bring different effects. Some method can do immunohistochemical study and this is very important for spinal cord research. You can use one method or combined method as you need in research.

Analysis to DNA Methylation Status of the Promoter of Human Telomerase Reverse Transcriptase (hTERT) in Gastric Carcinogenesis

Sun, 27 Mar 2011 11:26:47 +0000

ObjectiveWe analyze the specific methylation status of hTERT promoter in gastric carcinogenesis and analyze the relationship among the methylation status of hTERT promoter,the expression of hTERT and telomerase activity.We hope this study may provide experimental data and academic information for the early diagnosis of gastric cancer.MethodsGenomic DNAs were extracted from normal gastric mucosa samples,precancerous lesion samples and gastric cancer samples.Genomic DNAs were modified using sodium bisulfite.After the modification the unmethylated cytosine in the CpG sites of hTERT promoter were changed to uracil.The modified gemomic DNAs were amplified by PCR with primers that did not contain CpG sites.After PCR amplification,each PCR product was sequenced.Via matching the sequencing results and the original sequence,the staus of each samples were obtained.PCR were carried to identify the expression of hTERT.TRAP was carried to test the activity of telomerase among the three groups.Results1.The promoter region of hTERT in gastric cancer was more methylated than in the precancerous lesions and normal gastric mucosa..The differences have statistic significance.(P＜0.05)2.hTERT was absent in normal gastric mucosa and its positive rate was higher in gastric cancer than that in precancerous lesions.The differences have statistic significance.(P＜0.05)3.The activity of telomerase was not detected in normal gastric mucosa and its positive rate was higher in gastric cancer than that in precancerous lesions.The differences have statistic significance.(P＜0.05)ConclusionThe promoter region of hTERT in gastric cancer was more methylated than in the precancerous lesions and normal gastric mucosa.This may suggest that the degree of methylation status of hTERT promoter was increased gradually during gastric carcinogenesis and it may be a potential biological maker in early diagnosis of gastriccancer.During the gastric carcinogenesis the expression of hTERT and the telomerase activity was increased.This may suggest that the methylation of hTERT may have influence on the expression of hTERT and the activity of telomerase.

Effect of Different Doses of Daidzein on Serum Lipids and Liver LDLR Expression in Ovariectomized Rats and Its Relation with ERs

Sun, 27 Mar 2011 11:26:45 +0000

Background.Phytoestrogen is widely used for the health of perimenopausal women.There are more and more studies about it.Isoflavone,as one of phytoestrogens,is reported of effects on serum lipids.And Daidzein,as an important ingredient of isoflavone,its effects on serum lipids and liver LDLR expression and the role of estrogen receptors in this process need to be researched.PartⅠEffects of Different Doses of Daidzein on Serum Lipids and Liver ERs and LDLR mRNA Expression in Ovariectomized RatsObjectiveTo investigate the effects of different doses of daidzein on serum lipids and liver ERs、LDLR mRNA expression in ovariectomized rats.Materials and methodSixty-six four-month-old female Sprague-Dawley(SD) rats were randomly allotted to one of the following groups:ovariectomized group,sham-ovariectomized group,E_2 group,low-dose daidzein group,medium-dose group and high-dose daidzein group. The OVX operation is believed success when there are no Keratinocytes in vaginal smear in the next five days from the 7~(th) day after operation.Then Sham group and control OVX were administrated with distilled water,whereas other groups received valerate estradiol(800μg/kg·d) and different doses of daidzein(10,50,200 mg/kg·d) by gavage respectively for three months.These parameters were measured after the sacrifice of rats.Serum lipids including TG,LDL-c and HDL-c were measured.The expressions of mRNA of ERα,ERβand LDLR in the liver were measured by realtime-RT-PCR.Results1、Serum LDL-c:LDL-c in OVX group is higher than Sham group,compared to OVX group,the LDL-c in E group,M-Dai group,and H-Dai group are lower,the LDL-c in daidzein are decreased with the increase in the dose of daidzein,and thers are significant difference between the groups.2、Serum TG:the TG in E_2 group is higher compared with Sham group and OVX group,while the TG in H-Dai group is lower than Sham group and OVX group, and the TG in H-Dai group is significantly lower than M-Dai group and L-Dai group.3、Serum HDL-c:the HDL-c in OVX group are lower than Sham group and E_2 group, there is no significant difference between Sham group and E_2 group,TG in daidzein groups are lower than Sham group,although there are no significant difference when compared with OVX group.4、The levels of ERαand ERβmRNA:ERαmRNA in OVX group is decreased compared with Sham group and E_2 group,the level is increased in M-Dai group and H-Dai group than OVX group,the level in daidzein groups are increased with the increase of daidzein doses,and there are significant difference between the daidzein groups.5、The level of LDLR mRNA:LDLR mRNA in OVX group is decreased compared with Sham group and E_2 group,the level is increased in M-Dai group and H-Dai group than OVX group,the level in daidzein groups are increased with the increase of daidzein doses,and there are significant difference between the daidzein groups.Conclusions1、Daidzein exerts some befinical effects on serum lipids in ovariectomized rats,and the effect of daidzen on LDL-c is dose dependent,it is more powerful with the dose increased.2、ERβmRNA expresion is not detected in all the six groups.Daidzein increases ERαand LDLR mRNA expression in the liver of ovariectomized rats3、Daidzein\'s effect on serum LDL-c is in accordance with its effects on liver ERαand LDLR mRNA expression,that is,the effects are more significant when the dose increased,so we speculate that the LDL-c-lowering effect of daidzein may be realized by increasing liver LDLR expression through the ERαpathway. PartⅡEffect of Daidzein on LDLR mRNA Expression in HepG2 cell and its relation with ERsObjictiveTo investigate effects of daidzein on LDLR expression in HepG2 cells and the role of ERs in the regulation of LDLR expression.Materials and methodsIn order to determine the concentition of daidzein,we analysed the proliferation of HepG2 cells by MTT,then calculated IC50.We treat HepG2 cells with different concentitions within(IC5010~(-9) M、10~(-7) M and10~(-5) M),and measure LDLR protein expression by western-blot.Then we block ERs with ICI182780 and block ERαwith MPP,and again measure LDLR protein expression by western-blot.Results1、Compared with the control group,ERαand LDLR protein expression were higher in10~(-7)M daidzein group and 10~(-5)M daidzein group than the control group and 10~(-9) M daidzein group.ERαand LDLR protein expression in 10~(-5)M daidzein group were higher than10~(-7) M daidzein group.2、Compared with 10~(-5)M daidzein group,LDLR protein expression decreased after ERs or ERαwere blocked,but there were no difference between the two blocking groups.Conclusions1、Dadizein increases LDLR protein expression in HepG2 cells,and it is concentition dependent.2、Daidzein increases LDLR protein expression through ERα. Summary1、Daidzein exerts some befinicial effects on serum lipids in ovariectomized rats.2、Daidzein increases LDLR protein expression through ERα.

Analysis of Genetic Polymorphism in Duffy-binding Protein among Chinese Plasmodium Vivax Isolates

Sun, 27 Mar 2011 11:26:43 +0000

OBJECTIVETo investigate the genetic polymorphism of PvDBP RⅡin chinese yunnan province.METHODSStudy sites of Yunnan province is a malaria year-round transmission of P.vivax and P.falciparum.P.vivax genomic DNA was extracted from dried filter paper blood spots which obtained by using QIAamp DNA mini kit.The full-length open reading frame of the PvDBP RⅡfrom P.vivax genomic DNA was amplified with a polymerase chain reaction.The PCR products were directly sequenced.Computer-based algorithms were used to analyze neucleotide and amino acid sequences and the sequence alignments were done with BioEdit software.Intrapopulation and interpopulation diversity indexes were analyzed by DnaSP version 4.5 and MEGA 4.1 software.RESULTS1.Nineteen samples of PvDBP RⅡfrom P vivax genomic DNA was amplified successfully with the full-length 600bp(853-1452bp),which coded 200 amino acid(codon285-484).2.Compare to DBP Sal-Ⅰsequence,there were 13 variant sites in isolates from Yunnan province,with the frequency of 2.17%(13/600),accordingly,12 variant amino acids were found,with the frequency of 6.00%(12/200).Among all the variant sites found,one was synonymous,i.e.A1134G.There were 9 kinds of haplotype found,also 9 amio acid types,A1151G-G1169A-T1270A(D384G-R390H-L424I)was the most predominant one(26.3%).No insertion or deletion was found in both levels of base pair and amino acid.3.Compare with foreign isolates,the variant amino acids sites of DBP found in isolates from Chinese Yunnan were among those foreign ones,no geographic specificy was found.4.Data of nucleotide diversity showed thatπ=0.0091(SE:0.0028);dn/ds=6.168＞1(P＜0.05);Tajima\'s D:1.21360,P＞0.10.Fu and Li\'s D~* test statistic:1.48091 (P＜0.05),Fu and Li\'s F~* test statistic:1.62640(P＜0.05).DISCUSSIONCompare the genetic polymorphism of PvDBP RⅡamong China,South America, Africa and region of Middle East,variant sites of Chinese isolates were relatively limited,suggesting relative conserve founction of PvDBP antigen in China.From our results,we found 3 new sites in Yunnan isolates compare to Zhejiang and Hubei ones,although the low frequency,there was some suggestion on the studies of genetic polymorphism of PvDBP RⅡamong our country.Also,we found 2 sites occurred frequently in association,i.e.N417K and W437R,which implied that vaccine developed targeting these sites might be available to malaria endemic regions in our country.Following the statistic tests departure from neutuality showed evidence that the observed polymorphism might be trending natural selection,dn/ds predicting that positive selection may be occurring in regionⅡof PvDBP favouring the fixtion of amino acid replacement in isolates from Cinese Yunnan.Being the limitation of samples and analysis method,this is only a glimpse at the overall population in our country.CONCLUSION1.Compare the results of South America,Africa and region of Middle East,variant sites of Chinese isolates were relatively limited,there were some similar with isolates in South Korea located in the same continent of Asia.In China,there were some difference in variant sites between Yunnan and Zhejiang,Hubei province. 2.Our analysis showed evidence that the observed polymorphism might be trending natural selection,dn/ds predicting that positive selection may be occurring in regionⅡof PvDBP favouring the fixtion of amino acid replacement in isolates from Cinese Yunnan.

Clinical Study on the Relationship between Expression Patterns of HBcAg in Hepatocytes and Response to Interferon-alpha in HBeAg-postive Chronic Hepatitis B

Sun, 27 Mar 2011 08:01:25 +0000

Objective:(1) To observe the relationship between expression patterns of HBcAg in liver tissue of Chronic Hepatitis B with the inflammation activity in liver tissue and efficacy of interferon-alpha by clinical cases.(2) To investigate the best timing of antiviral therapy and to improve the response rate of IFN-a therapy.Methods:216 cases of patients with HBeAg-postive chronic hepatitis B from Hepatopathy Department in Xiamen Traditional Chinese Medicine Hospital affiliated to Fujian College of Traditional Chinese Medicine during February,2006 to September,2007 were conducted liver biopsy,determined expression patterns of HBcAg in liver tissue by immunohistochemistry staining method,classified into four groups:nuclear localiazation of HBcAg group,cytoplasmic localiazation of HBcAg group,nuclear localiazation of HBcAg dominant group and cytoplasmic localiazation of HBcAg dominant group.All the patients were treated with 5MU IFN-a subcutaneously thrice a week for 6 months.To observe and detect between pre and post treatment:(1) serum alanine transaminase(ALT), serum aspartate aminotransferase(AST);(2) serum HBVM(Electrocherniluminescence kit);(3)serum HBVDNA(fluorescence quantitative PCR);(4) predict the outcome of treatment.Statistical methods:x~2 test,logistic regression analysis,and so on.Results:(1)The 216 cases of patients accorded with the inclusive criteria were classified into nuclear localiazation of HBcAg group(35),cytoplasmic localiazation of HBcAg group(58),nuclear localiazation of HBcAg dominant group(54) and cytoplasmic localiazation of HBcAg dominant group(69).There was no statistical difference among four groups in sexes,age,disease course and serum ALT、serum AST、serum HBVDNA with t-test(P＞0.05).(2) The cytoplasmic localiazation of HBcAg group and cytoplasmic localiazation of HBcAg dominant group based mainly on moderate and severe inflammation in liver tissue,showed obvious piecemeal necrosis and lymphocytic infiltration.While the nuclear Iocaliazation of HBcAg group and nuclear localiazation of HBcAg dominant group based mainly on mild inflammation.(3) The serum ALT、AST of cytoplasmic localiazation of HBcAg group were close to upper normal limit and had obvious statistical difference(P＜0.01)after IFN-a therapy,cytoplasmic localiazation of HBcAg dominant group were close to upper normal limit too after IFN-a therapy and have statistical difference(P＜0.05) compare with untreament,while nuclear localiazation of HBcAg group and nuclear localiazation of HBcAg dominant group had no statistical difference compare with untreament(P＞0.05).(4)The seroconversion rate of HBeAg/HBeAb of cytoplasmic localiazation of HBcAg group was highest(70.7%)after IFN-a therapy;and cytoplasmic localiazation of HBcAg dominant group took the second place(58%);the seroconversion rate of the other two groups were low;The seroconversion rate have obvious statistical difference between the cytoplasmic localiazation of HBcAg group and the nuclear localiazation of HBcAg group,the same as the cytoplasmic HBcAg localiazation of dominant group and the nuclear localiazation of HBcAg dominant group(P＜0.01).(5)49 cases of patients\'HBVDNA quantitative was less than 1000 copies/ml in cytoplasmic localiazation of HBcAg group after treatment,and 54 cases in cytoplasmic localiazation of HBcAg dominant group whose HBVDNA quantitative was less than 1000 copies/ml.There were still 28 cases of patients\'HBVDNA quantitative more than 1000 copies/ml in nuclear localiazation of HBcAg group and 44 cases in nuclear localiazation of HBcAg dominant group.The HBVDNA negative rate had obvious statistical difference between the cytoplasmic localiazation of HBcAg group and the nuclear localiazation of HBcAg group,the same as the cytoplasmic localiazation of HBcAg dominant group compare with the nuclear localiazation of HBcAg dominant group(P＜0.01).(6)Most patients of cytoplasmic localiazation of HBcAg group achieved complete response and only ten of them had no response.In cytoplasmic localiazation of HBcAg dominant group,40 cases of patients achieved complete response,15 cases achieved partial response and 14 cases had non-response.Most cases of patients in nuclear localiazation of HBcAg group and nuclear localiazation of HBcAg dominant group had non-response,were 19 and 22 cases respectively.(7) the higher whose inflammation grade in liver tissue was before therapy,the higher proportion of complete response can gain.(8) The expression of cytoplasmic localisation of HBcAg in hepatocytes was independent influencing factor of IFN-a efficacy by logistic regression analysis. Conclusion:1.The inflammation activity in the liver tissues of cytoplasmic localisation of HBcAg group and cytoplasmic localisation of HBcAg dominant group was higher than that was expressed in nuclear localiazation of HBcAg group and nuclear localisation of HBcAg dominant group.2.The antiviral effect of IFN-a of cytoplasmic localisation of HBcAg group and cytoplasmic localiazation of HBcAg dominant group was obviously better than which of nuclear localisation of HBcAg group and nuclear localisation of HBcAg dominant group.(3) the higher whose inflammation grade in liver tissue was before therapy,the higher antiviral effect can gain.(4) The mechanism of obvious antiviral effect in cytoplasmic localisation of HBcAg group maybe related to the immune clearance phase and immune activation.And,the expression of cytoplasmic localisation of HBcAg in hepatocytes bennifit the reponse of IFN-a.

Distribution of Mesenchymal Stem Cells Transfected with Hepatocyte Growth Factor in the Rats Kidney with Unilateral Ureteral Obstruction

Sun, 27 Mar 2011 08:01:23 +0000

Obstructive nephropathy(ON) is a common disorder in urinary surgery,which major pathologic feature is renal interstitial fibrosis.In children,obstructive nephropathy is the primary cause for end stage renal disease(ESRD) and represents 16.1%of all pediatric kidney transplantation.Bone marrow-derived mesenchymal stem cells(MSCs) are characterized with self proliferation and multi-lineage differentiation.MSCs can differentiate to histocyte originated from mesoderm,endoderm and neuroectoderm,include osteoblast, cardiocyte,nerve cell and renal cell.It has been shown that systemic delivery of MSCs could protect mice with acute renal failure from renal function impairment and severe tubular injury.Hepatocyte growth factor(HGF) as a determiner facilitate adult stem cells differentiate to renal cell,and posses the ability to prevent apoptosis of renal tubular cell.In this sdudy,bone marrow-derived MSCs of SD rats were separated and cultured in vitro,and then transfected with Ad-HGF.Rats model with unilateral ureteral obstruction(UUO) were established and Ad-HGF transfected MSCs were injected into the rats with UUO.Finally,we observe whether MSCs transfected with HGF can immigrate and localize in the rat\'s kidney with UUO,which may provide a theoretical clue for gene and cell therapy for obstructive nephropathy.Materials and methodsBone marrow derived-MSCs of the male SD rats were separated and cultured in vitro,and the phenotypic identification of CD34 and CD90 were detected by flow cytometry.MSCs were transfected with Ad-GFP in different MOI in order to select an optimal MOI.Transfection efficiency was detected by flow cytometry.The expression of HGF in MSCs transfected with optimal MOI was measured with ELISA assay.Sixteen female SD rats with UUO were randomly divided into two groups: transplantation group and control group.Ad-HGF transfected MSCs or saline were injected into the rats by vena caudalis.Kidney tissue of the rats were collected at the end of the 7th day and 14th day after operation.The distribution of Y chromosome in the kidney was determined by in situ hybridization.Results1.After 7-9 days,primary MSCs can be passaged.After generation,cell proliferation rate increase,after 4-5 days,MSCs can be passaged again.MSCs surface antigen by flow cytometry show CD34(—),CD90 99%.2.Transfection efficiency of Ad-GFP to MSCs is 73.29%when MOI equaled 400.3.The expression of HGF in MSCs reached the highest level after being transfected with Ad-HGF for 48h.4.Y chromosome positive cells were found only in the obstructed kidney of the transplantation group.The positive cells mainly distributed in the tubular cells.Conclusions1.We can isolate and purify MSCs by means of attachment culture method.2.MSCs were ideal genetic carrier for HGF gene therapy.3.MSCs transfected with HGF can immigrate to the rat\'s kidney with UUO,and mainly distributed in the region of renal tubular epithelial cells.

Experimental Study of Cyclosporine A-containing Cubosomes on Corneal Rejection in the Rat Penetrating Keratoplasty Model

Sun, 27 Mar 2011 08:01:21 +0000

Objectives:1.To observe irritant reaction of different cyclosporine A eye drops on rabbit eyes.2.To establish a rat model of penetrating keratoplasty;To observe the immunosuppressive effect of cyclosporine A eye drops on immunological rejection after corneal transplantation in rats,and evaluate the effect of cyclosporine A-containing cubosomes on corneal allograft rejection.Methods:1.Draize Eye Irritation Test:Pre-selected healthy adult New Zealand white rabbits,animals in each group are 4.Drugs test were dropped in the right conjunctival sac of each animal,while the left conjunctival sac of each eye were dropped control drugs.Drugs were administrated 50μl per eye,every 15min for 2h.At 1h,3h,6h,24h,48h,72h after last administration,evaluate irritant reaction of rabbit conjunctiva,cornea,iris with slit-lamp and score.2.Experimental study of different cyclosporine A eye drops on corneal rejection in the rat penetrating keratoplasty model:Penetrating keratoplasty was performed with Lewis rats as recipients and Wistar rats as donors.55 Lewis rats as recipients were randomly divided into five groups:control group,cubosomes without drug treated group,l%CsA-containing cubosomes treated group,0.5%CsA-containing cubosomes treated group and 1%CsA-containing olive oil treated group.The drugs were administrated 20μl each time and 5 times per day after operation for ten days.The corneal grafts were inspected with slit-lamp and scored every day.At the 10th day postoperatively,the rats of each group were killed respectively.The corneal grafts were examined with HE staining.Results:1.The groups of nanoparticles and cubosomes did not have irritant reaction, only the individual had short-term conjunctival hyperemia and restored after about 1h. The group of olive oil had a little more irritant reaction.There was slight conjunctival hyperemia in about 6h,and iridal hyperemia in one eye.These phenomenons disappeared after 24h.At the same time they blinked frequently.There was more obvious conjunctival hyperemia in two prescription of microemulsion-gel group,and iridal hyperemia in partial eyes.But corneal opacity wasn\'t observed.At the same time they blinked more frequently.2.Compared to control group and cubosomes without drug treated group,two prescription of CsA-containing cubosomes could extend the survival time of corneal grafts and reduce rejection,and the differences had Statistical significance. Pathological examination revealed that lymphocytic infiltration and neovascularization was less.Conclusion:1.Microemulsion gel and olive oil had mild irritation,while there is almost no irritant in nanoparticles and cubosomes group,which is in line with the requirements of ophthalmic preparations.2.0.5%and 1%CsA-containing cubosomes can reduce the rate of rejection of the rat penetrating keratoplasty model,and extend the survival time of corneal grafts.

Epidemiological Survey of Cataract among Adults Aged 40 of Above in Gongshan County of Yunnan Province

Sun, 27 Mar 2011 08:01:19 +0000

Objective To estimate the prevalence of cataract,surgical coverage rate and bilateral cataract-blindness burden among adults aged 40 or above in Gongshan county of Yunnan provice,China,for evaluating the cataract status and blinding situation,and to expore the risk facors for cataract by collecting related information.Methods Cluster sampling was used in randomly selected individuals at 26 villages in Gongshan county.Individuals on roster was 3070 and aged 40 or elder was 1236,and 1117 received questionnaire and ocular clinical examination.Cataract was graded clinically using the Lens Opacity Classification System(LOCS)Ⅲsystem. The available data were analyzed by univariate and multivariate Logistic regression model.Results1.The prevalence of cataract was 23.81%among adults aged 40 or above.The prevalence of cataract was increasing by aging.The prevalence of cataract in individuals aged 70 or above and illiterates was significantly higher statistically.2.When the bilateral pinhole vision＜3/60 was defined as the blindness criterion, the bilateral cataract-blindness burden was 1.25%,and cataract blindness surgical coverage rate was 50%.3.When the bilateral presenting vision＜6/60 was defined as the blindness criterion, the bilateral cataract-blindness burden was 25%,and cataract blindness surgical coverage rate was 12.89%.The cataract surgical coverage rate were much lower and the bilateral cataract-blindness burden much higher in woman,illiterates and persons aged 70 or above.4.Univariate factor analysis showed:age,education,BMI,income,ultraviolet ray, hypertension were associated with any cataract,cortical cataract,nuclear cataract and postsubcapsular(PSC) cataract.Occupation was associated with nuclear cataract and PSC cataract.Education and BMI associated any cataract,cortical cataract and PSC cataract.The Lisu nationality and the Nu nationality were assotiated with any cataract and cortical cataract.Sex,smoking,drinking are not associated with all types of cataract.Multivariate Logistic regression showed:age was independent risk factors for all types of cataract,occupation was independent risk factor for nuclear cataract and PSC cataract,hypertension was independent risk factor for nuclear cataract, income is independent risk factor for PSC cataract.Conclusion The bilateral cataract-blindness burden was higher.Cataract blindness is a serious public health problem between in aged individuals and illiteracy.Many identified risk factors for cataract in the population have potential for being modified through public health intervention.

The Expression of HIF-1α and VEGF in the Retinas of Diabetic Rats

Sun, 27 Mar 2011 08:01:17 +0000

Diabetic retinopathy is the most common complication of diabetes resulting in blindness.The study about pathogenesis has become the hot topic.The abnormal increased angiogenesis promoters is thought as the major cause of diabetic retinopathy,vascular endothelial growth factor(VEGF) is an important factor involved.In the recent years,it has been found that hypoxia-inducible factor-1(HIF-1) was a key point in the modulating vascular endothelial growth factor signal transduction pathway under hypoxic condition.However,whether these factors also have expression in the early diabetic retinopathy is ready for further study.ObjectivesThe purpose of this study was to detect the spatiotemporal expression of HIF-1αand VEGF in retinas of streptozocin(STZ) induced diabetic rats.To approach the mechanism of nonproliferative diabetic microangiopathy was also concerned.Methods130 SD rats were randomly divided into two groups:control group(C) and diabetes group(DM).70 rats were received vena caudal injection of STZ as diabetes group,and the rest were taken as controls.The rats were euthanized and eyes were enucleated for paraffin section with HE staining,examining the lens and retina espectively at 2,4,6,8,10,12 weeks after STZ injection.After peeling off retina of rats with careful manipulation under the microscope,RT-PCR and immunofluorescence were taken to detect the expression of HIF-1αand VEGF in retina respectively at 2,4,6,8,10,12 weeks after STZ injection.ResultsSTZ induced diabetic rats had lasting and stable hyperglycaemia,displaying with the obvious signs of "polydipsia,polyphagia,hyperdiuresis and loss of weight". There was significant difference of weight and plasma glucose of rats between the two groups at different time points(P＜0.05).The epithelial cells of lens are in regular arrangement in control groups.There was no sign of morphologic alteration at 4 weeks after STZ injection and mild signs of cataract at 8 weeks after STZ injection.All lens of diabetes eyes were detected to cataract alteration at 12 weeks after STZ injection.There was no significant sign of morphologic alteration of retina in control groups. The retinal structure of diabetic rats is similar as control eyes at 2 to 6 weeks after STZ injection by HE staining.There was hypocellularity of the nuclear layer in diabetes group at 8 weeks after STZ injection and there were signs of thinness, dissolution and cells loss in the retinas of diabetic rats at 12 weeks after STZ injection.HIF-1αmRNA were expressed in normal retina and began to increase at 2 weeks after STZ injection(P＜0.05).The longer duration of the disease,the more expression of HIF-1αmRNA.The expression at 10 weeks after STZ injection began to decrease(P＜0.05).The expression of HIF-1αat 12 weeks after STZ injection was lower than at 10 weeks after STZ injection(P＜0.05).The expression of HIF-1αprotein in retina was found in control group.It mainly localized in the ganglion cell layer,outer plexiform layer.HIF-1αprotein began to increase at 2 weeks after STZ injection.It mainly localized in the ganglion cell layer,inner plexiform layer,inner nuclear layer,outer plexiform layer.The longer duration of the disease,the more expression of HIF-1αprotein(P＜0.05).The expression of HIF-1αprotein reached peak at 10 weeks and decreased after 12 weeks after STZ injection(P＜0.05).VEGF mRNA were also expressed in retinas of normal rats,the expression began to increase at 2 weeks after STZ injection(P＜0.05).The longer duration of the disease,the more expression of VEGF.The expression at 12 weeks after STZ injection is 4 fold compared with control animals(P＜0.05).The expression of VEGF protein in retinas was found in control group.It mainly localized in the ganglion cell layer,inner plexiform layer,inner nuclear layer,outer plexiform layer,outer nulear layer.VEGF protein began to increase at 2 weeks after STZ injection.The longer duration of the disease,the more expression of VEGF protein(P＜0.05).Conclusions(1) STZ induced diabetic rat is a reliable animal model which can be used to imitate human nonproliferation diabetic retinopathy.(2) The expression of HIF-1αand VEGF can be detected on retina of early diabetic rats. (3) The protein expression trend of HIF-1αand VEGF is similar as the expression of mRNA.It was showed that hypoxia played an important role in DR.(4) HIF-1αmay regulated the expression of VEGF by combinating with hypoxia response element of VEGF on the retina of early diabetic rats.Further study is needed.

Relation between HPV16/18 and E-cadherin, β-catenin in Cervical Intraepithelial Neoplasia

Sun, 27 Mar 2011 08:01:15 +0000

IntroductionCervical cancer(CC) is one of the most frequently founded malignant tumors in women world wide.Current evidences implicated high risk humn papillomavirus(HPV) especially HPV16/18 infection as a causative part of the natural history of cervical carcinogenesis.HPV can be found in 99.7%of cervical cancer patients.It\'s critical for precaution of cervical cancer to detect HPV.However only a part of the patients who contracted HPV may advance to cervical cancer.E-cadherin is mediated by a class of cell-cell adhesion,with the organizational structure to maintain the integrity and polarity of the calcium-dependent transmembrane protein.E-cad function is not only required the existence of calcium ion,but also the needs of their Ligand catenin (catenin,cat) can be combined into a complex role,β-cat and E-cad direct binding,α-cat-mediated E-cad /β-cat and actin cytoskeleton-linked protein.Therefore,the maintenance of the polarity of epithelial cells and play an important role in differentiation,E-cadherin function may result in decrease or loss of cells-the role of intercellular adhesion reduced,resulting in tumor occurrence and development.In cervical intraepithelial lesions and cervical cancer in expression was low in the high expression of the normal control group,so if the factors on the etiology of cervical cancer and HPV biomarkers factor E-cadherin,β-catenin co-testing,to high-risk groups for cervical cancer screening,may have important significance.In our study,we detect HPV16/18 by the hybridization in situ and E-cadherin,β-catenin by immuohistochemical analysis,to research the value of them in cutting down the mortality of CC,and to supply rationale to exploitation of convenient samples. Materials and Methods1.Materials(1) specimen600 cervical liquid-based cells remained samples were obtained from the Department of gynecology in the First Affiliated Hospital of China Medical University, during Nov 20067to Nov 2008.All of them were treated as CC or CIN,Among them 13 samples as control gropes.All of them possess corresponding histological samples as controls.The patients with CC were 40,with CINI were 21,with CINⅡ\Ⅲwere 19. The average age is 48.4(range,26～77 years).The standard of cytology grouping based on histodiagnosis.All patients had not the history of the operation on cervix,pregnancy and radiation therapy in the cavity of pelvic.(2) Main agents,equipment and sourceKit of HPV16/18 in situ hybridization was purchased from MAIXIN biological technology company in Fuzhou,antibody of E-cadherin,β-catenin protein was friendly presented by Jin woo Kim professor in Korea.Sub-type HPV-DNA detection kit purchased from Cape company.S-P Kit and DAB Kit were obtained from MAIXIN biological technology company in Fuzhou.2.Methods(1) ImmunohistochemistryDetection of E-cadherin,β-catenin by Immuohistochemical analysis S-P methods. Result determination:The overexpressed staining was in the membrane.The number of staining cells were graded as follows:10-24%(+);25-49%(++);50-74%(+++);≥75% (++++).(2) In situ hybridizationDetection of HPV16/18 by Hybridization in situ.The masculine staining was in the nucleus.The overexpressed staining was in the cytoplasm.The number of staining cells were graded as follows:10-24%(+);25-49%(++);50-74%(+++);≥75%(++++). (3) HPV-DNA hybridization diversion type detectionDetection of HPV-DNA expression in the various subtypes.Results in a corresponding subtype of the template on a solid dot the region is the emergence of positive expression of the subtypes(4)Detection of HPV16/18 by western-blot.Stamples were incubated on ice for 30 min in an appropriate RIPA buffer(Solarbio) with 1%PMSF(Solarbio).The stamples was centrifuged at 15000g for 45 min at 4℃, and the supematant was fractionated by electrophoresis on a 15%PAGE and transferred to a nitrocellulose membrane.After blocking with 5%skimmed milk,the membrane was incubated with the appropriate primary antibody overnight at 4℃,and with labeled secondary antibodies(1:2000,Chemicon,USA) for 2h at 37℃.Proteins were visualized by DAB(Perking Applygen Co.,Ltd.) according to the manufacturer\'s protocol.(5) Statistical analysisThe results of the various groups usingχ2 test and Fisher\'s exact probability test,P <0.05 statistical significance.Results1.Expressions of E-cadherin,β-cateninExpressions of E-cadherin,β-catenin in cervical exfoliated cells and cervical tissue by Immuohistochemical analysis S-P methods.Membrane E-cadherin,in normal cervical exfoliated cells and high expression in cervical tissue,cervical lesions in each group there are different levels of expression:E-cadherin,in normal,CIN,invasive carcinoma of the expression rates were 84.6%(11/13),72.5%(29/40),27.5%(11/40),β-catenin in normal,CIN,invasive carcinoma of the expression rates were 76.9% (10/13),67.5%(27/40),32.5%(13/40).As the extent of cervical intraepithelial lesions showed a downward trend to increase2.Expression of HPV16/18 by Hybridization in situ. HPV16/18 was overexpressed in cervical cancer82.5%(33/40) and CIN 50.0%(20/40) compared with it in normal15.4%(2/13),P<0.05.The expression of HPV16/18 in cervical cancer and precancerous lesion correlated well with cervical neoplasia(CIN).It\'s consistent both in cervical exfoliated cell and in cervical tissue.3.HPV-DNA hybridization diversion type detection to detect the expression of HPV subtypesHPV-DNA test results of the subtypes in the corresponding subtype of the template region appear solid dots,which is positive for the subtype.HPV-DNA of various subtypes in normal cervical exfoliated cells are not expressed or weakly expressed in cervical lesions in each group there are different levels of expression:600 patients were positive for HPV detection were 245 cases,the positive rate was 40.83%(245/600),including the composite infection,specific infections shown in Table 1.Table 2.HPV21 subtypes,in addition to HPV43,HPV44 There are 19 kinds of subtypes have been detected,the highest rate of infection is HPV16(87/245),other common type for the HPV58(36/245),HPV6(33/2-45), HPV53(28/245),HPV18(20/245),HPV31(19/245),HPV52(19/245),cp8304(16/245) and HPV11(15/245).HPV infection in women can be seen in our province is the main subtype of HPV 16,HPV58 and HPV6.4.Expression of HPV 16/18 by westernHPV16/18 was overexpressed in cervical cancer85%(34/40) and CIN62.5%(25/40) compared with it in normal23.1%(3/13),P<0.05.The expression of HPV16/18 in cervical cancer and precancerous lesion correlated well with cervical neoplasia(CIN). It\'s consistent both in cervical exfoliated cell and in cervical tissue.Conclusions1 In CIN and CC,The positive rates of E-cadherin,β-catenin decrease with the aggravation of cervix affection. 2 In CIN and CC,The positive rates of HPV16/18 increase with the aggravation of cervix affection.3 In CIN and CC,the expression rate of E-cadherin,β-catenin and HPV16/18 showed a negative correlation,It will be more accurate for screening CIN and CC to detect them together.4 Testing HPV16/18 by western has high sensitivity,especially in CIN.It is suitable for questioned case by this way.It helps testing HPV16/185 The possible role of HPV16/18 in the remained samples of cervical liquid-based cells is coincident with that in cervical tissues.There is great feasibility in screening cervical affection by utilization of the remained samples of cervical liquid-based cells.6 HPV-DNA hybridization diversion typing detected 600 eases of women in our province of the positive rate of HPV infection to 40.83%(245/600),infection is the main subtype of HPV16,HPV58 and HPV6.

The Expression of DMBT1 Gene in Prostate Cancer and Its Clinical Significance

Sun, 27 Mar 2011 08:01:12 +0000

Part One Screen the treated and untreated prostate cancer cell lines by Genome MicroarrayObjectives:To identify some genes that may be regulated by methylation during the prostate tumorigenesis.Methods:After treating prostate cell lines LNCaP and PC-3 with DNA methyltransferase inhibitor--5-aza-2\'-deoxycytidine(5-aza-dc),we used the Whole Human Genome Oligo Microarray to screen the treated cells and the controlled groups to identify some genes which restored expression after anti-methylation.Results:The expression of DMBT1 gene(Deleted in Malignant Brain Tumor)was observed to be significantly up-regulated in treated PC-3 cell lines than controlled cell lines, however no similar expression change was observed in both treated and controlled LNCaP cell lines.Conclusions:Anti-methylation with DNA methyltransferase inhibitor(5-aza-dc)is able to up-regulate the expression of DMBT1 in PC-3 cell lines.This implies that the expression of DMBT1 gene may be related to DNA methylation.Further tissue evaluation was necessary to confirm the expression of DMBT1 gene in prostate cancer.PartⅡThe expression of DMBT1 in prostate cancer cell lines and tissues and its clinical significanceObjectives:To observe the expression of DMBT1 gene in both prostate cancer cell lines and tissues,then to evaluate the potential correlation between DMBT1 expressions and clinical and pathologic features such as age,PSA,clinical stage,tumor grade and bone metastasis.Methods:The expressions of DMBT1 in both mRNA and protein levels were detected by real-time quantitative PCR and western blot in prostate cancer cell lines(LNCaP & PC-3) and prostatic tissues from 16 benign lesions and 39 carcinomas.The correlation between gene expression level and clinical/pathologic features was analyzed using Spearman Correlation method by the software SPSS 13.0.Results:The mRNA expression of DMBT1 in both PC-3 and LNCaP cell lines were down-regulated compared to normal tissues and the fold changes in LNCaP cell lines was significantly obvious.Similar to the results of genome microarray,anti-methylation with 5-aza-dc may significantly restore the expression level of DMBT1 in PC-3 cell lines than LNCaP cell lines.Compared to the normal prostatic tissues,both mRNA and protein expressions of DMBT1 gene were significantly down-regulated in whole prostate cancer tissue samples.The statistical results showed that the expression levels of DMBT1 were correlated to clinical stage and osseous metastasis,but not correlated to patient age,PSA level and tumor grade at diagnosis.Conclusions:Real-time quantitative PCR is recommended to detect the mRNA expression compared to traditional RT-PCR.DMBT1 is down-regulated in prostate cancer tissues and cell lines,but the mechanism is unknown.The DMBT1 gene is confirmed to be a potential biomarker in identification of hormonal independent subgroup from hormonal dependent prostatic carcinomas,as well as the prognostic value with its correlation to clinical stage and tumor grade.It is necessary to elucidate the regulation mechanism of DMBT1 and its contribution in cell physiology and tumorigenesis.

Construction of Ankle and Foot Three-Dimensional Numerical Model and Analysis of Finite Element of Talus in Ectocondyle Unstable Situation

Sun, 27 Mar 2011 07:56:29 +0000

Purpose:To Create three-dimensional numerical model of ankle-foot joint(Finite element model).Based on this to analysis the rule of redistribution of talus\'s Von Mises stress when lateral ankle is in unstable situations,and to study the critical range of risk in talus.Methods:One normal healthy volunteers was performed ankle scan using GE light speed 16.The scanning range included whole ankle and distal tibiofibula.Using MATLAB and ANSYS software,to establish normal human ankle-foot three-dimensional numerical model.The applicability of the model was tested by applying load and boundary condition,and the data was compared with the other relative articles and clinical treatment study,to verify its validity.Based on the Finite element model of the normal people,adapt different restriction and loading regime,to construct different conditions of chronic extramalleolus unstablitied(53 in the unstable condition in the combination of the internal rotation ankle-foot and the varus).Result:1.Foot ankle orthogonal anisotropic three-dimensional numerical model was set(total node:96748.unit sum:488482).2.The major talus Von Mises stress mainly located in under normal walking condition on articulatio talonavicularis,neck of talus,evagination and processus aborolis of the talus,articulatio subtalaris and posterior articular facet of the tibial astragaloid joint,the maximum Von Mises stress is mainly on the evagination and processus aborolis of the talus.3.Comparing to the normal situation,the internal and rearward Von Mises stress decrease.The outside and the head and neck part was bigger.The bigger stress centers on the articulatio talonavicularis,neck of talus,evagination and processus aborolis of the talus.The range of the bigger stress become narrow,which leads to increases of the local stress. 4.Compared to the varus,the talus Von Mises stress is mainly affected by internal rotation.5.The talus fracture\'s risk value is within 60 mm internal rotation,the stress on the extramalleolus and the neck of talus will be over 80 MPa,(which was close to or above the bone eortex\'s yield limit(83 MPa)),when internal rotation larger than 60 mm and easily caused bone destruction. Conclusion:1.Establishment of a normal orthotropic finite element model of ankle-foot,with a high degree of geometric and physical similarity.2.Definition of the normal stress distribution of ankle-foot.3.When Chronic lateral ankle is instabe, the posterior talus has less stress than normal,head and neck and Lateral of talus has more stress and increased local stress,which was easy to cause Micro-fractures in intra-auricular and injury of surrounding cartilage.4.Internal rotation has a greater impact on the talus than Varus.Internal rotation should be mainly corrected when correcting clinically chronic lateral ankle instability,when internal rotation was more than 60 mm,lateral malleolus and talar neck was most common location to fracture.5. The model had provided a biomechanical basis for clinical treatment of chronic ankle instability.

The siRNA Targeting BAK Inhibit Apoptosis of DC That Induced by Glioma and the Mechanism

Sun, 27 Mar 2011 07:56:26 +0000

Object Dendritic Cells co-culture with glioma,To study the apoptosis of Dendritic Cells that induced by glioma.To study the vector expressing siRNA targeting BAK inhibit the apotosis of Dendritic Cells and to speculate the possible pathway of the apoptosis.Methods The vector that expressing siRNA targeting BAK was transfected into DC by the lipofectamine2000 and the cells stably expressing the siRNA were selected by G418.To choose the high performance vecteor that interference BAK.Then the DC that stably expressing the siRNA were co-culture with glioma and observe the apoptosis of DC.Investing the relate gene for example caspase3 caspase8 and the cytochrome c in endochylema,to speculate the possible pathway of the apoptopsis.Resuits When the DC2.4 co-culture with glioma lead to the apoptosis.When the vector were transfected into DC2.4,the expression of BAK were degraded(western blot:A:0.83,B:0.14,C:0.75,D:0.89,un-transfecte:0.92; RT-PCR.- A:0.42,B:0.08,C:0.39,D:0.46,un-transfecte:0.49 P＜0.05).When the DC2.4 that stably expressing siRNA target BAk co-culture with glioma,the expression of caspase 8 was not change(a:0.78 b:0.75 c:0.77,P＞0.05) to tmexpression siRNA one,but the expression of caspase 3 and the cytochrome C (caspase3:a:0.67 b:0.70 c:0.2;Cytc:a:0.70 b:0.71 c:0.17 P＜0.05)in endochylema was less than the unexpression siRNA one.Conclusion The glioma can induce the apoptosis of DC2.4.The pathway of apoptosis may be the mitochondrial pathway; The vector that expresse siRNA targeting BAk can degrade the apoptosis of DC2.4.

The Relationships between Brain-derived Neurotrophic Factor, Cognition and Clinical Status in Patients with Schizophrenia

Sun, 27 Mar 2011 07:56:24 +0000

Objective:Our study attempted to examine the serum brain-derived neurophic factor(BDNF) concentrations and cognitive functions in schizophrenic patients and their changes after treatment.It also explored the relationships between serum BDNF, cognition and the clinical status in patients with schizophrenia.Methods:Totally 111 patients with schizophrenia who were successively admitted into the psychiatric hospital were enrolled into the study.The serum BDNF concentrations were assessed at baseline in these patients and were compared with healthy controls(control group 1).Eligible patients entered into the 4-week treatment study and the serum BDNF,the Wisconsin Card Sorting Test(WCST),the Personal and Social Performance Scale(PSP),the Positive And Negative Syndrome Scale (PANSS) and the Clinical Global Impression(CGI) were assessed before and after treatment.For the WCST comparison,another normal sample(control group 2) was enrolled and WCST was assessed.Antipsychotic medications and their dosage were chose by treating pshysicians.A well-trained researcher blind to the treatments evaluated the rating scales and cognitive measures.The serum BDN concentrations were measured by enzyme linked immunosorbent assay(ELISA).All the data were analyzed with t test,rank sum test, correlation analysis and analysis of variance(ANOVA).The significance level was set at 0.05.Results:1.The serum BDNF levels was significantly lower in patients with schizophrenia compared to normal controls(20.67±5.78ng/ml vs.23.72±5.61ng/ml,t=4.310, p＜0.01).2.The serum BNDF levels were significantly increased in schizophrenic patients after 4-week treatment(22.51±6.05,t=-2.027,p=0.046),close to normal level.3.The patients had significantly poor performance in the numbers of categories, correct responses,perseverative errors and non-perseverative errors of WCST compared with healthy controls(p＜0.01).4.The mean PSP score in patients with schizophrenia was 46.05 at baseline, indicating a remarkable impairments in social cognition,and was significantly increased to 71.95 after treatment(t=12.733,p＜0.01).5.Correlation analysis revealed no association between the increase levels of BDNF and improvement in cognition and symptoms of schizophrenia patients.Conclusion:1.The serum BDNF is downregulated in schizophrenia patients compared to healthy controls and is significantly increased after 4-week antipsychotic treatment.The results support the view that BDNF might have an important role in schizophrenia.2.Schizophrenic patients demonstrate impairments in neural and social cognition, which could be improved with short-term treatment of antipsychotics.3.The parallel improvement of cognitive functions to the BDNF increase after treatment suggest that antipsychotic medication may upgrlate BDNF functioning.

Morphologic Changes in Neurons and Expressing Alterations in β-catenin and GSK-3β in Brain Regions of Rat Autism Model

Sun, 27 Mar 2011 07:56:22 +0000

Autism is one of the most serious of the pervasive developmental disorders and is characterized by deficits in social relatedness,communication,and cognitive flexibility.Onset of the unusual behavior typically occurs in infancy and is fully present by the child\'s 3 years.In 2 to 3-year-old,a significant developmental characteristic to the majority of autistic patients is the greater head circumference than the normal children with same age,while these wloud be followed by the less head circumference in adolescent than the normal.The incidence is approximately 4 in 1000 live births with a 4:1 predominance of males to females.The cause of autism is still unclear,and may be genetic or environmental.Nongenetic causes are speculated to involve prenatal interruptions in the normal signaling pathway of brain development.Genetic causes may arise from mutations in genes along this developmental pathway.Wnts are a large class of signaling molecules implicated in a variety of processes throughout development such as stem cell self renewal,cell-fate determination,axon guidance,and synaptogenesis.There are over 16 Wnt proteins in vertebrates,which all have a conserved cysteine residue that is palmitoylated before the proteins are secreted.In the canonical pathway,Wnt binds to the Frizzled coreceptor,which signals downstream to Disheveled(Dvl).Activated Dvl then inhibits a trio of proteins, GSK3β,Axin and APC.Inhibition leads to dephosphorylation ofβ-catenin,which triggers its activation and allows its translocation to the nucleus,where it binds to TCF/LEF transcription factors.It has been suggested that abnormalities of this signaling transduction may be present in schizophrenia,unipolar or bipolar depression and autism.It also has been shown in animal experiments that mice completely deficient of Dvl-1 display abnormalities in social behavior and sensorimotor gating characteristic to several psychiatric disorders including autism.Sinceβ-catenin represents the key protein that transduces activation of the Wnt pathway into the nucleus while GSK-3βis the central negative regulator of cytoplasmicβ-catenin levels,the present study was designed to examine these two signaling proteins,β-catenin and GSK-3β,in an animal model of autism as following:Ⅰ:An animal model of autism was obtained in offspring of the Wistar rat by a single intraperitoneal injection of sodium valproate to the rat at the pregnancy day of 12.5.The present study examined the expression of two signaling proteins,β-catenin and GSK-3β,which take the key position in Wnt pathway,in prefrontal cortex, hippocampus and cerebellum of the autism model of rat with different ages by means of the Western blot technique.The results showed that,the level of expression ofβ-catenin was higher in animal model tissue than in control with the exception of the hippocampus and cerebellum at the age of postnatal(P) 7,and there was a trend to a greater difference of expression in these regions during the course of brain development.GSK-3βprotein levels tended to be less in the animal models when compared to controls.The only exception to this was in cerebellum in which no difference was observed between models and controls.Consistent withβ-catenin,the difference was greater with the development of the rats.These results indicated that the Wnt signaling pathway was over activated in the autism models,and this activation might be the one of the causes of abnormal development of neurons in the brain of autism.Therefore,the present results suggested that Wnt signaling pathway plays an important role in the etiology of the autism.Ⅱ:For evidence from the morphology to our results,the present study selectively observed the GSK-3βimmunoreactive neurons of cerebellum by immunohistochemical staining.The same animal model of autism was obtained by VPA treated pregnant Wistar rats.We examined the expression of GSK-3βin the cerebellum at the age of P7,P14,P21,P28 and P90.The results showed that the immunoreactivity of the GSK-3βwas contained in the body of the cerebellar Purkinje cells.At P7,the GSK-3βimmunoreactive neurons exhibited similar distribution patterns and morphology in both the model and contral animals.Howere,differences with statistics significance in number of neurons between the model and contral rats were boserved in P14,P21,P28 and P90,and the patterns of distribution of the GSK-3βimmunoreactive neurons was observed in a diffusing mannar in animal models compared to controls.These results indicated that the GSK-3βexpression in the cerebellum significantly reduced,which is consistent with the western blot results in partⅠof the present study.Accordingly,the reduction of the GSK-3βis bound to cause the reduction of theβ-catenin degradation in the Wnt signaling pathway,and therefore leading to Wnt signaling enhanced.Additionally,Golgi staining was carried out to show the morphology and number of the pyramidal neurons in prefrontal cortex at the age of 90d.The results showed that the density of pyramidal neurons increased in the animal model.The top dendrites of the neurons in the model animals were shortened,and the dendrites were excessively branched in disorders.This morphology of the pyramidal neurons might increase the volume of the cortex,and cause abnormal neuronal networks both in local neurons and the long-distance between brain regions, and further cause the imbalance of excitory and inhibitory neuronal activity.To sum up,the present study suggested that the Wnt signaling pathway in the brain of autistic patients might be over-activated,and this,in turn,might cause the number and morphological changes of the neurons,for example,the excessive proliferation of the neurons,and then lead to the increased volume of the brain.Further more,the over-growed neurons might affect the role of the environment to the brain development,which lead to incorrect neural connections,and impede the formation of normal local and long-distance neuronal networks between brain regions,and eventually the autistic symptom,impairment in social behavior,emotion and language, occur.

Study on the Optimizational Model of Management for Hypertension in Community

Sun, 27 Mar 2011 07:56:19 +0000

Objective:A large number of domestic and foreign experiences have shown that hypertension management in community was the effective way.In recent years,some Community Health Service Center through the way such as screening the health records of residents,making a sample survey in high-risk groups to determine the intervention and management of objects for hypertension.And then following-up, intervention and evaluation for the hypertensive patients were carried out by the team of general practitioners and got some preliminary results.How to continue and effectively promote the work of community-based hypertension management, however,has become difficult.In the present study,we explored the effective way of hypertensive management in community,in order to improve the effective interventions and the treatment compliance for the hypertensive patients,and also explore intervention strategies on high-risk groups and general population,more effectively to control the occurrence and development of the hypertension,as well as increase the control rate of blood pressure and improve the patients\' life quality.Methods:964 community residents aged 18-year-old and over were inquired by stratified and cluster sampling.They were divided into the intervention group and the control group randomly.Measuring blood pressure,clinical diagnosis,establishing card,following-up,evaluation were carried out in the intervention group through two ways of clinic and community(clinicians\' workstation and team of general practitioners),and information software was applied together to complete prevention, cure and management for hypertension.The control group still carried on the original model.The data was analysed by x~2 test.Results:At present,the prevalence rate of hypertension of≥18-year-old residents in Changfeng Community was about 22.8%,significantly higher than the level of national and Shanghai.After intervention,the rate of awareness,control rate of blood pressure,rate of standardized treatment were increased obviously in the intervention group.And also the patient compliance and satisfaction were improved obviously than that in the control group.Conclusion:The prevalence of hypertension in community is still high,the optimizational model of management for hypertension is obviously effective,this model is practicable and deserves to be spread.

The Effect of Autonomic Nervous System Adjustment between Traditional and New Health Qigong Yijinjing and sEMG Research

Sun, 27 Mar 2011 07:56:17 +0000

Objective:To explore the traditional "health qigong yijinjing" and new "health qigong yijinjing" on the autonomic nervous system adjustment effect and the difference between them.Through the comparison of traditional and new "health qigong yijinjing" twelve movement of the surface myoelectric signals of muscle group related,then find the difference of muscle strength among the process of twelve movement.As the theoretical basis to further explore "health qigong yijinjing" and improve the fitness of physiological functions.So,To inherit and carry forward the Chinese traditional health qigong and provide a theoretical basis and reference.Method:46 students of Wuhan Institute of Physical Education were tested,including 33 males and 13 females.The test was carried out after 2 months traditional "health qigong yijinjing" practice and new "health qigong yijinjing",respectively.HRV signal respectively recorded before and after exercise\'s sit stage,and during the exercise.10 students from Wuhan Institute of Physical Education Department of Wushu whom had yijinjing exercise foundation were tested.Using surface electromyography to record each muscle group electronic signal on the surface of representative passive or active muscles of twelve movement,such as the forearm,the lateral muscles,during practice the traditional "health qigong yijinjing" and new "health qigong yijinjing",respectively.Take the largest amplitude,RMS amplitude and IEMG values,SPSS13.0 package used for statistical analysis of data.Result:1.In the course of HRV detection during practicing new "health qigong yijinjing", R-Rmean value from 771.83±98.21 in the stage sit-in before practice to 641.60±73.92 in practice;TF value from 114.67±99.02 to 300.46±156.41;CV from 9.41±2.13 to 11.94±2.62;LF/HF value from 2.20±1.60 to 8.63±5.24;NLF from 61.69±16.06 to 86.77±6.37;NHF from 38.30±16.06 to 13.23±6.37;Stage sit-in after practice,RR mean was 789.31±111.33,TF was 110.01±91.70,CV was 6.93±2.78,LF / HF was 2.47±1.74,NLF was 64.91±16.05,NHF was 35.09±16.05;RMSSD in the whole process at a relatively high level of stability(the stage sit-in before practice:32.29±13.84,in practice: 34.14±10.32,Stage sit-in after practice:33.71±16.70).And the differences of HRV parameters between traditional "health qigong yijinjing" and new "health qigong yijinjing" were not significant.2.The RMS(0.36±0.1,0.28±0.23V) of forearm,lateral muscle groups during practicing traditional "health qigong yijinjing" "Weituo pestle" potential was significantly higher than the new(0.12±0.02,0.15±0.11 V);There were subjects in new"Claw of wings",the first half of force is not enough force only after the half. Conclusion:1.Practicing "health qigong yijinjing" can improve the vagus nerve to enhance the tension in the heart of the vagus nerve protective effect,and the adjustment of cardiac autonomic nervous system of new and traditional "health qigong yijinjing" are equally effective.However,new "health qigong yijinjing" was more easily to learn,so,it should be popularized and Long-term practiced.2.The muscle strength,muscle strength size and the degree of uniformity of strength of new\'s twelve movement are more reasonable than the traditional.And during practice, not only need every potential movement still need to master correct posture strength.EMG analysis can help practitioners to practice a more accurate grasp of the essentials.3.As the HRV and sEMG analysis for testing and evaluation technology are simple, non-invasive and quantitative,it can reflect the overall body and local health status and functional level.By the use of these technologies to research the effects of traditional fitness exercises on human body physiology,for the role of fitness exercises to provide a physiological basis.

Ghrelin Inhibits Apoptosis Induced by Oxidative Stress in Adult Rat Cardiomyocytes, Throught the PI3K-Akt Signaling Pathway

Sun, 27 Mar 2011 07:56:11 +0000

Ghrelin inhibits apoptosis induced by high glucose and in adult rat cardiomyocytes,through the PI3K-Akt signaling pathwayObjective The aim of the present study was to observe the impact of ghrelin on apoptosis induced by high glucose and sodium palmitate and its mechanism of anti-apoptosis.Methods isolate and culture cardiomyocytes from hearts of adult rats.Use high glucose and sodium palmitate to stimulate oxidative stress of cardiomyocytes. Apoptosis was detected by annexin-v-FITC/PI binding assay and Caspase 3 Activity assay.Reactive oxygen species were tested by DCFH-DA fluorescent probe. Phospho-Akt,phospho-Erk,NFκB were identified by ELISA assays.Expression of anti-apoptosis genes wwere detecteded by realtimePCR and western blot.Results①ghrelin inhibited apoptosis induced by high glucose and sodium palmitate in cardiomyocytes(p＜0.05),but it could not reduce the oxidative stress in cardiomyocytes.②anti-apoptotic effect of ghrelin could be blocked by wortmannin(p＜0.05),but PD98059 had no influence on it.③activation of extracellular signal-regulated kinase-1/2 and Akt serine kinases by ghrelin,peaked at 10min and 20min respectively.Ghrelin stimulated transposition of NFκB to nucleus and peaked at 30min.④transposition of NFκB to nucleus stimulated by Ghrelin was inhibited by wortaminin.⑤The expression of c-iap was 17.9 times compared with control group after ghrelin stimulating for 8h.And the expressions of bcl-2、bcl-xl and c-fos were elevated 12.2,4.4 and 4.3 times respectively after ghrelin stimulating for 20h when compared with contrls.⑥after stimulating for 24h with 1μghrelin,the protein levels of IAP-1 and Bcl-2 in cardiomycytes increased 3.67- and 2.59-fold that of control group.Conclusion These findings provided the evidence that ghrelin could inhibit apoptosis induced by oxidative stress in cardiomyocytes,but ghrelin could not reduce oxidiative stress in cells.And ghrelin produced anti-apoptotic effects through activation of PI3K/Akt pathway which promoted phosphorylation of BAD and activation of NFκB to increase expression of anti-apoptotic genes. Decreased expression of p85αin myocardium of spontaneous diabetes ratsObjective The aim was to study the molecular and cellular mechanism of cardiomyopath in long-term diabetes.Method OLETF rats and LETO rats were used as experimental and control group. Gene chip,real-time PCR,western blotting analysis were applied to show the different expressions of P85αbetween OLETF and LETO rats.Results compared with LETO rats,the concentrations of average plasma insulin in OLETF rats was lower(0.27±0.13 vs.2.39±0.34 ng/ml,p＜0.000),the triglyceride and cholesterol level were higher(1.73±0.34 vs.0.48±0.08 mmol/L,p＜0.000; 4.41±0.21 vs.2.25±0.75 mmol/L,p＜0.001),and the velocity of left ventricular diastole of OLETF rats decreased by 30%(p＜0.05).The result of microarray assays indicated an 8-fold fall-off in the expression of p85αmRNA in myocardium of OLEFT rats.The real-time PCR assays and immunoblot analysis further supported the point with the gene transcription and protein expression levels of p85αdecreased by 81.5%and 43.2%respectively(p＜0.05 and p＜0.001).Conclusion these results indicated that reduction of gene transcription and protein expression of p85αin myocardium of OLETF rats was related to impaired construction functions.

Pharmacogenomics Study of UGT1A6 Gene in Different Chinese Ethnic Groups

Sun, 27 Mar 2011 06:02:44 +0000

The human UDP-glucuronosyltransferase enzymes (UGTs) superfamily constitutes important drug metabolizing enzymes in the human. It has been estimated to be involved in the metabolism of many endogenous substrates and extragenous substrates, as well as in the metabolism of about 30% of clinically prescribed drugs. The members of the UGT superfamily demonstrate greater inter-individual and interethnic variations in humans. Genetic polymorphism is the common cause of inter-individual and interethnic differences in the metabolism of the same substrate. This polymorphism influences both drug response and disease treatment (such as schizophrenia, cardiovascular disease, hypertension and cancer etc). An understanding of the genetic variation will improve the ability to predict a patient’s drug responses, adjust the therapeutic regimen and decrease the adverse drug reactions.With the availability of complete human genomic sequence, more and more people has considered that the variations in genomic DNA sequence, i.e. gene polymorphisms are the most important determinants for disease susceptibility, clinical phenotype diversity and different response to pharmacotherapies and environmental factors. To date, many UGT1A6 alleles (haplotypes) have been publicized and mainly come from Caucasians and African-Americans as well as Japanese and Korean. There are little known about Chinese population and different Chinese racial groups.We carried out the first systematic screening of polymorphisms of the gene in an SNP analysis involving 1074 Chinese subjects from three ethnic groups, namely Han, Dong and She, using direct sequencing. A total of 16 SNPs were detected including 9 known and 7 novel SNPs. The novel SNPs were 73G>A (V25I), 89T>G (L30R), 222A>C, 657C>A, 773A>T (D258V), 1040A>G (N347S) and 1467C>T. In addition, we detected, for the first time in the Chinese population, SNPs 105C>T, 308C>A (S103X), 627G>T, IVS2+15T>C and 1088C>T (P363L). Linkage disequilibrium and haplotype analysis was also made. Strong linkage disequilibrium was observed among 19T>G, 315A>G, 541A>G and 552A>C. There were seven haplotypes whose frequencies were more than 0.01 in one or more of the three ethnic groups. The analysis provided a comprehensive picture of the kinds and frequencies of SNPs of UGT1A6 in the population, including 541A>G and 552A>C which are associated with low enzyme activity.These studies may have an important significance for the understanding of genotype and phenotype relationships of UGT1A6. The further analysis of UGT1A6 SNPs will reveal the differences in individuals and ethnic groups. The applications of such information are particularly relevant for pharmacogenomics, in which knowledge of SNPs in UGT1A6 genes may lead to individualized drug dosing, decreased the adverse drug response and improved therapeutics. Moreover, an understanding of the genetic variation that exists in various populations will aid in tailoring health care to different populations, especially Chinese minority ethnic groups.

The Changes of Cerebral CT Perfusion for the Patients with Cerebral Patent Artery Stenosis before and after Stent Placement

Sun, 27 Mar 2011 06:02:42 +0000

Purpose:The purpose of this study was to assess the change of CT Cerebral Perfusion\' s parameter and the prognosis of clinical symptom in symptomatic patients with cerebral patent artery stenosis before and after stent placement.Materials and Methods:Ten patients with unilateral cerebral patent artery stenosis(≥65%) underwent brain CT perfusion and the National Institutes of Health Stroke Scale(NIHSS) assessement 48 hours before and after the stent placement.Cerebral blood volume(CBV)cerebral blood flow(CBF) and time to peak(TTP) values were measured.Results:the change of CT Cerebral Perfusion\'s parameter and the prognosis of clinical symptom were observed in the middle cerebral artery areas supplied by the stenotic artery after stent placement:A significant decreased rTTP was observed in 9 patients,1 patient unchanged(p=0.01＜0.05).rCBV and rCBF increased in 5 patients,rCBV and rCBF decreased in other 4 patients,Decreased rCBV and increased rCBF were observed in 1 patient.Decreased NIHSS were observed in 7 patient(P=0.004＜0.05).The Pearson correlation between the CTP and the prognosis of clinical symptom is r=-.062,p=0.864＞0.05.Conclusion:Perfusion CT is a useful technique for the assessment of the cerebral perfusion in symptomatic patients with cerebral patent artery stenosis before and after stent placement.Pearson correlation between the CTP and the prognosis of clinical symptom is uncertain..

Treatment of Chronic Actinic Dermatitis with 0.1% Tacrolimus Ointment

Sun, 27 Mar 2011 06:02:40 +0000

PartⅠClinical response of chronic actinic dermatitis to 0.1%tacrolimus ointmentObjectiveTo evaluate the efficacy of 0.1%tacrolimus ointment in the treatment of chronic actinic dermatitis.MethodsWe have recruited 40 patients with chronic actinic dermatitis.They were all male patients,the mean age being 57.8±11.1,and the mean course of disease being 85.2±126.1 months.0.1%tacrolimus was applied topically to the lesion in the sun-exposed areas twice a day.Follow up was conducted every two weeks for one month.Those who couldn\'t tolerate the therapy dropped out of the study.At the baseline and each subsequent visit,symptom and sign scores were recorded,and the total effective rate was calculated.At the same time,dermatology quality of life index (DLQI) and VAS for pruritus symptoms were assessed at the significance level of 0.05.Results1 patient who got aggravated after the treatment was rated as ineffective.39 patients who completed the treatment improved at different degrees.The total effective rate was 62.5%.At the same time,DLQI and VAS were improved significantly(P＜0.05).Conclusions0.1%tacrolimus ointment is effective in the treatment of chronic actinic dermatitis. PartⅡThe mechanism of topical tacrolimus in the treatment of chronic actinic dermatitisObjectiveTo investigate the mechanism of chronic actinic dermatitis treated with 0.1% tacrolimus ointment.Methods15 selected patients who gave the informed consent received biopsy before and after the topical therapy.Immunohistochemical investigation was carried out to detect the change o Langerhans cell(surface marker CD1a) and inflammatory dendritic cell(surface marker CD11b),as well as the expression of chemokine receptor7 (CCR7)in the lesions,at the significance level of 0.05.ResultsImmunohistochemistry showed that the expression of CD1a,CD11b,CCR7 in the skin lesions were significantly down-regulated(P＜0.05).ConclusionsLangerhans cells and inflammatory dendritic cells play a role in the pathogenesis of CAD.0.1%tacrolimus ointment can inhibit Langerhans cells as well as inflammatory dendritic cells,and down-regulate the expression of chemokine receptor7 in CAD leisions.

The Correlation of Follicle Stimulating Hormone Receptor Genetic Polymorphism to Ovarian Serous Cancer and Its Prognosis

Sun, 27 Mar 2011 06:02:37 +0000

Ovarian cancer is one of the common gynecologic malignant tumors.Because of lack of early diagnosis method and effective treatment to this disease,ovarian cancer becomes a main disease that threatens women\'s health and life.While etiology of ovarian cancer is still unclear,some studies suggested follicle stimulating hormone (FSH),an important female sex hormone,not only stimulates the development of follicles in reproductive period,but also has a close relationship with ovarian cancer. FSH promotes the growth of tumor cells by the specific binding to the follicle stimulating hormone receptor(FSHR).There are many single nucleotide polymorphisms(SNPs) in FSHR gene.The Ala307Thr and Ser680Asn are the most common SNPs in the polymorphisms due to the linkage disequilibrium of exon 10.As a result,amino acid alteration related to the corresponding SNPs might affect the post-translational modifications of the FSHR protein,and hence the function of the receptor including FSH efficacy.Different genotypes of FSHR were related to different basal level of serum FSH.And the FSHR polymorphism might play a role in ovarian carcinogensis.Therefore,it\'s necessary to investigate the FSHR polymorphism.And the results might support the diagnosis and treatment of ovarian cancer at the genetic level in the future.To date,few studies concern the relationship between SNPs in FSHR gene and ovarian cancer.Neither do they achieve consistent conclusions.To further investigate their relationship,this study mainly consists of two parts.(1) Using the "polymerase chain reaction-restriction fragment length polymorphism"(PCR-RFLP) method to investigate how the SNPs in FSHR exon 10(Ala307Thr and Ser680Asn) impact the susceptibility of ovarian serous cancer.(2) Collecting and reviewing the ovarian serous cancer patients\' medical information to estimate the relevance of the SNPs in exon 10(Ala307Thr and Ser680Asn) and the clinical prognosis of ovarian serous cancer. Part 1 SNPs of FSHR and the Susceptibility of Ovarian Serous CancerObjective:This study investigated the relationship between the SNPs in FSHR exon 10(Ala307Thr and Ser680Asn) and the susceptibility of ovarian cancer.Methods:This is a case-control study.In the analysis,ovarian tissues were collected from 35 patients with ovarian serous cancer and 22 people with normal ovaries as control.With the PCR-RFLP method,genotyping analyses were performed on Ala307Thr and Ser680Asn in FSHR exon 10.The results were analyzed by statistical test such as exact test,chi-square test,and non-conditional Logistic regression with two categories.Results:(1) The distribution of three genotypes of FSHR Ala307Thr(Thr/Thr, Ala/Thr and Ala/Ala) was statistically different between the patients with ovarian serous cancer and those with normal ovaries(P=0.004).The percentage of Ala carriers in case group was 97.1%,while it was 68.2%in control group.And the rate of Ala carry was significantly different in two groups after age-adjusted(P=0.022, OR=13.637,95%CI=1.466～125.850).(2) The distribution of three genotypes of FSHR Ser680Asn(Asn/Asn,Ser/Asn and Ser/Ser) was statistically different between ovarian serous cancer patients and normal ovaries group(P=0.004).The percentage of Ser carriers in case group was 68.6%,while it was 31.8%in control group.And the rate of Ser carry was significantly different in two groups after age adjustion(P=0.008, OR=4.964,95%CI=1.529～16.109).(3) The ratio of haplotype of Ala307-Ser680 was statistically different in two groups(P=0.031,OR=5.217,95%CI=1.040～26.165).Conclusion:Ala307Thr and Ser680Asn in FSHR exon 10 correlate with the susceptibility of ovarian serous cancer.307Ala carriers and 680Ser carriers were associated with significantly increased the risk of development of ovarian serous cancer.Moreover,the haplotype of 307Ala-680Ser was also associated with higher cancer risk. Part 2 Relevance of SNPs of FSHR and Prognosis of Ovarian Serous CancerObjective:This study investigated the relevance of the SNPs in FSHR exon 10 (Ala307Thr and Ser680Asn) and prognosis of ovarian serous cancer.Methods:The medical histories of patients with ovarian serous cancer and their follow-up information were reviewed so as to examine how the SNPs in FSHR exon 10(Ala307Thr and Ser680Asn) affect the prognosis of ovarian serous cancer patients. Statistical tests,including exact test,Kaplan-Meier test and Log-Rank test,were conducted to analyze how the SNPs at the two sites(Ala307Thr and Ser680Asn) affect recurrence,survival time,grades,staging,lymph node metastasis,and serum CA125 level in the initial diagnosis.Results:(1) The genotype distribution of Ala307Thr and Ser680Asn in FSHR gene exon 10 was of no significant difference in recurrent and non-recurrent group.And the ratio of 307Thr carriers and 680Ser carriers were also no statistically different in these two groups.(2) At the end of follow-up(Dec.31,2008),the mid-survival of the 35 patients with ovarian serous cancer was 42 months(8～79 months),and the 5 year survival rate was 40.0%.The genotype distribution of Ala307Thr and Ser680Asn in FSHR exon 10 was of no significant difference between life time over 5 years and less than 5 years.And 307Thr carriers and 680Ser carriers were not associated with the life time of ovarian serous cancer.(3) There was no statistical difference in different age of onset and staging of the genotype distribution of Ala307Thr and Ser680Asn in FSHR exon 10.And they had no association with serum CA125 level in initial diagnosis and lymph nodes metastasis.However,the distribution of Ala307Thr was statistically different in grade 3 and grade 1～2 group(P=0.02).The distribution of Ser680Asn had a differentiation trend along different pathologic grades(P=0.078).Conclusion:Ala307Thr and Ser680Asn in FSHR exon 10 had no significant correlation with prognosis of ovarian serous cancer patients.

Primary Research of Activation of the Exogenous Blood Coagulation in Patients with Chronic Urticaria

Sun, 27 Mar 2011 06:02:34 +0000

Background:Chronic urticaria(CU) is a common skin disease,whose etiology is not very clear yet.As the data we already know,25%-50%are autoimmune urticaria,which is caused by the autoantibodies against the alpha subunit of the high-affinity IgE receptor on mast cell and autoantibodies against IgE,but other 30%-40%are thought to be idiopathic for no identified causes.Auto serum skin test(ASST) can be carried out to determine these autoantibodies,whose sensitivity and specificity are just about 80%.What is interesting is that in patients with CU caused by physical factors been excluded the positive rate of ASST is about 50%,while the positive rate of APST is to 95%.There were also cases reporting injecting heparin subcutaneously or using warfarin to treat patients with CU got good effect.More studies showed heparin could inhibit reaction of skin to auto serum and inhibit histamine-releasing of basophilic granulocyte in vivo.So some scholars did research of CU and blood coagulation,the results revealed concentrations of plasma D-dimer,FⅦa and F_(1+2) were higher in patients with CU than in normal controls,but concentration of plasma FⅫa had no difference between patients and controls.There was a positive correlation between plasma FⅦa content and F_(1+2) content.Biopsies of skin lesion showed expression of TF was higher in cells with severe inflammation.These founds indicated abnormal activation of exogenous coagulation pathway in patients with CU.Recently,the hot research points finds many factors of blood coagulation have complicated relations with inflammation.On this foundation,we did primary research to determine if patients with CU have activation of exogenous blood coagulation pathway and discuss the roles coagulation factors playing in pathogenesis of CU. Objective:To know common sick status and DQLI of patients with CU.To evaluate the role of blood coagulation in the pathogenesis of chronic urticaria by investigating the PT,APTT,concentration of plasma D-dimer, concentration of plasma activated factorⅦin patients with chronic urticaria and evaluate the relationship between concentration of plasma D-dimer and concentration of FⅦa,at the same time,the correlation between concentration of FⅦa and the disease activity and correlation between concentration of TAT and the disease activity are to be confirmed.By doing this,we hope to discover how coagulation parameters act in CU and find possible therapy clues of incurable CU.Methods:Studied 30 patients with CU and 30 normal controls and investigated common sick status and DQLI of patients,whose disease activity was also estimated.Both in the 30 patients and 30 healthy subjects,PT and APTT were measured by coagulation method,plasma D-dimer content was determined by turbidimetric immunoassay,plasma FⅦa content was investigated by ELISA. Four correlations,which are plasma FⅦa and TAT content,TAT and D-dimer content,concentration of FⅦa and disease activity and concentration of TAT and disease activity,were studied.Results:Common sick status,DQLI is showed in chart 1 and chart 2,while picture 1 shows disease activities.The results of PT,APTT,plasma FⅦa,TAT and D-dimer content are in chart(5-9).Compared with the normal controls, there was no significant difference of PT and APTT in patients with chronic urticaria(P＞0.05,separately).But the plasma D-dimer content was higher in patients than controls(P＜0.05) and plasma FⅦa content was significant higher in patients than that in controls(P＜0.001).There was a positive correlation between concentration of FⅦa and TAT(P＜0.001),the similar correlation also existed between concentration of TAT and D-dimer(P＜0.001),and the level of FⅦa,but not the level of TAT(P＞0.05) was significantly correlated with the disease activity(P＜0.05).Conclusion:From analysis of this study,CU does much negative influence to patients\' life quality.And elevated concentrations of plasma FⅦa,D-dimer and TAT and the positive correlations between FⅦa and TAT content,TAT and D-dimer content prove abnormal activation of exogenous blood coagulation. While the fact that concentration of plasma FⅦa and disease activity has positive correlation,which doesn\'t exist between plasma TAT content and disease activity,suggests that the activation of blood coagulation may play a part in the pathogenesis of chronic urticaria.Coagulation factors,especially FⅦa,may involve in the pathogenesis of chronic urticaria.

Studies on Effective Components and Antimicrobial Effect of Hottuynia Cordata Thunb

Sun, 27 Mar 2011 06:02:32 +0000

Houttuynia cordata Thunb is a genus of perennial herbs.It has a great potential for development and is also one of the valuable wildlife resources for its nutrition,health care and medical functions and has been formally identified by the National Ministry of Health as both food and medicine.In domestic reports,much controversy and extensive clinical application of Houttuynia cordata are announced.This research is mostly concentrated on its volatile constituents, and a considerable part on other ingredients,in addition,clinical pharmacology,separation and extraction of the chemical composition, reproduction,cultivation and other aspects of Herba houttuyniae are also studied.All the facts show that many researchers have great interests in the study of Houttuynia cordata and it is a attraction or a difficulty to make an investigation on Houttuynia cordata for a long time between past and present in a hot spot.This paper is aimed at comparing different components including flavinoids and volatile constituents in Houttuynia cordata to explore similarities and differences of chemical compositions in Houttuynia cordata in different areas of Guizhou.A domestic study which is conducted for the first time on restraining effect of vegetative pathogen of volatile oil in Houttuynia cordata showed volatile components in Houttuynia cordata have good restraining effects on Rhizoctonia solani, Gibberella saubinetii,Fusarium graminearum Schwabe,Phytophthora capsiei Leonian,Magnaporthe grisea which are all fungus.This study provided theoretical and scientific basis for further development of Houttuynia cordata for its high value in rationale.This paper is divided into the following five parts:The first part gives us an overview of this article which introduces with circumstances origin,distribution,biological characteristics, chemical composition,pharmacological effects,medical,food value,health care functions and development prospects of the Houttuynia,together with purpose and significance of this paper.The second part aims at researching on total content of flavonoids in Houttuynia cordata and different extraction methods are tried in this experiment,the methods are boiled with water,extracted with alcohol, soshi extraction and ultrasonic extraction,respectively.Taking total flavonoids as exploring index,influences of different methods to extracted content of total flavonoids are compared.The best extraction condition of ultrasonic extraction method is investigated,together with extraction technic of total flavonoids and stability of this technic.In this part,contents of flavonoids compounds in different places on Houttuynia cordata are also studied.The results showed that the highest concentrationt of flavonoids was obtained from leavies,secondly stems, and the lowest concentrationt of of rhizomes from same place.But concentrationt of flavonoids have diversity from different places.Application of Shimadzu HPLC is introduced in the third part to determine contents of seven flavonoids,and they are; Kaempferol-3-O-α-L-rhamnopy,hyperoside,quercitrin,rutin,quercetin, kaempferol and isorhamnetin.Using 50 percent ethanol as extraction agent,acetonitrile and 0.1%phosphoric acid solution for gradient elution, separation is conducted in Dumas C18 column(4.6 x 250 mm,5μm), selecting PDA-wide wavelength ultraviolet detector to scan,good sensitivities of seven flavonoids at wavelength 320 nm are obtained,, and methodological study results has proved that this determination is reliable for its good sensitivity,high accuracy,exclusive and stability, what is more,determination results can objectively reflect contents of seven kinds of flavonoids,therefore,conclusions in this paper play an important role on exploitation of Houttuynia cordata as a kind of medicine.The results showed that the concentrationt,of kaempferol-3-O-β-D-rhamnopy,hyperoside,quercitrin,rutin,quercetin, kaempferol and isorhamnetin have remarkably diversity. Constituents and their similarities of volatile oil in Houttuynia cordata produced in Guizhou are analysed with the help of Shimadzu GC-MS modeled QP2010 in the fourth part.The results revealed that high similarities appear on fingerprint images of 10 batches of samples, generated common modes and 12 common eigenpeaks in fingerprint images have the ability to estimate qualitative standards of volatile oil in Herba houttuyniae.Resemblances of 10 batches of Houttuynia cordata samples all lie upon 90 percent,and no outlier ones are found,so the samples accord with requirements from State Administration and Superintendence for Drugs.Bacterial inhibited activity of volatile oil in Houttuynia cordata is investigated in the fifth part with the aid of five fungus,they are vegetative pathogens named Rhizoctonia solani、Gibberella saubinetii(mont.)Sacc、Fusarium graminearum Schwabe、Phytophthora capsiei Leonian、Magnaporthe grisea.Results of this part indicate that volatile oil in Herba houttuyniae shows different bacterial inhibited concentrations(IC50) on different kinds of pathogen.The smallest concentration belongs to Rhizoctonia solani,with the value 81.24μg/mLwhile the highest belongs to Magnaporthe grisea with the value 320.21μg/mL,and bacterial inhibited concentrations for other three pathogens are 209.86,312.75,245.09μg/mL,respectively.

Study on HIV Knowledge, Attitude and Behavior among Migrant Fertile Women in Minhang District

Sun, 27 Mar 2011 06:02:28 +0000

1.The HIV KAP intervention study in the migrant fertile women in the Transient Population Family-planning Service CenterObjective:To improve their HIV knowledge,Voluntary Counseling and Testing (VCT) knowledge and willingness,to change their behaviors and to reduce the HIV infection risk by teaching the migrant fertile women HIV knowledge and interventing their behaviors,in the Transient Population Family-planning Service Center.Methods:An epidemiology intervention study was implemented in the Transient Population Family-planning Service Center of Minhang District.The convenient sampling method was adopted to carry out the questionnaires survey(the baseline and the final).After the 1-year intervention,the intervention effectiveness was evaluated by comparing the participants\' HIV knowledge,attitude,behaviors,VCT knowledge and willingness before and after the intervention.Results:During the questionnaires survey(the baseline and the final),four hundred participants was inquired respectively.Baseline survey indicated that,most migrant fertile women lived with their husbands and didn\'t change their dwelling place frequently.Only 9%of the participants often used condoms last month.Only 1%thought it was possible for them to be infected HIV.Free VCT was known by 38.8%,and 49.7%participants knew the VCT service,but they didn\'t know where to get the service.61.3%of the participants accepted the VCT service after they were informed that VCT was free.Logistic regression analysis show:the higher their education degree was,the more they knew HIV knowledge.The high HIV prevention awareness of condoms and choosing condoms as contraception method promoted the participants to use condoms.The participants who had more HIV knowledge and knew VCT were more likely to accept VCT service.After the intervention,the participants kept more HIV knowledge than pre-intervention.The awareness of thinking to be infected by HIV(4.8%) increased obviously compared to pre-intervention(P＜0.05,).The rate of using condoms went up to 12.8%at the final survey.64.8%of the participants acknowledge the VCT,69.1%could gave the answer of the testing location,and VCT willingness(82.8%) went up obviously,and 2 participants accepted free VCT service. Conclusions:The intervention in the Transient Population Family-planning Service Center increased the migrant fertile women\'s related HIV knowledge, condoms using,VCT knowledge and willingness.Carrying out HIV intervention in the Transient Population Family-planning Service Center is an effective way to protect migrant fertile women from being infected by HIV.2.The HIV KAP observational study in the migrant fertile women in the physical examination clinic of the Community Medical Service CenterObjective:To know the migrant fertile women\'s HIV KAP,VCT knowledge and willingness and the change without direct intervention,in the Community Medical Service Center.Methods:This study was an observed study.The study site was the physical examination clinic of the Community Medical Service Center of Xinzhuang of Minhang District.The objects of study were 15-49 year-old migrant fertile women who attended exams in the physical examination clinic.The convenient sampling method was adopted to in the twice cross-sectional survey.The HIV KAP,VCT knowledge and willingness of the participants in the baseline and final survey without direct intervention were evaluated.Results:631 and 656 participants were visited in the baseline and final survey respectively.The first cross-sectional survey indicated that,the migrant fertile women who came to the physical examination clinic less lived with their husbands or boyfriends and often change their dwelling place,35.1%unmarried participants have ever had sexual activities,VCT knowledge rate was low(46.9%),62.g%accepted the VCT service after they were informed that VCT was free.Logistic regression analysis show that the participants who had more HIV knowledge,and who were more aware of being infected HIV and knew VCT more were more likely to accept VCT service.The second survey indicate that,without direct intervention,the HIV knowledge, VCT knowledge(50.5%) and willingness(66.9%) of the migrant fertile women who came to the physical examination clinic increased a little,which was not statistical significant).The rate of condoms used by unmarried participants in the latest sexual activity(52.0%) was higher compared to 1 year ago(44.4%),but it was not a big distance.The rate of condom used in the latest sexual action in married participants (25.5%) was similar to 1 year ago(24.8%).Conclusions:The sexual activities rate among the unmarried migrant fertile women was high,but the rate of condoms using was comparatively low.Both the free VCT knowledge and willingness among the migrant fertile women were low.Without direct intervention,the HIV KAP and VCT knowledge and willingness among the migrant fertile women increased slightly during last year.

Grading System for Diagnosis and Treatment of Intracranial Nongerminomatous Malignant Germ Cell Tumors

Sun, 27 Mar 2011 05:01:34 +0000

Object:To discuss the clinical feature,treatment,and prognosis of intracranial nongerminomatous malignant germ cell tumors(NGMGCTs).Methods:The records of 39 patients who were treated in Shanghai Huashan hospital between 1995 and 2007 were reviewed retrospectively.According to the classification of Matsutani in 1997,they were grouped into intermediate prognosis group and poor prognosis group based on the histology of the tumor.Clinical manifestation,diagnosis, treatment and outcome were analysis in each group.Results:In these 39 cases,there were 15 mix germ cell tumors,15 immature teratomas, 7 embryonal carcinomas and 2 yolk sac tumors.All patients were treated surgically.The tumor was totally removed in 29 cases,sub-totally in 5,and partially in 3.Biopsy was performed in the other 2 cases.34 patients(87.2%) were followed up.The common 5-year survival rate was 36.8%.The 5-year actuarial overall survival rate for patients in the intermediate prognosis and poor prognosis groups were 42.6%and 0%. Chemotherapy combined radiotherapy has significant relationship with the prognosis of intermediated prognosis group(P=0.039).The 5-year survival rate of immature teratoma patients who received gamma knife surgery after surgery was 100%.It had significant difference(P=0.0049) compared to the 5-year survival rate of patients who did not receive gamma knife surgery.Conclusions:NGMGCTs can be divided into intermediate group and poor group base on the prognosis(P=0.0003).Embryonal carcinoma can be classified to the intermediate prognosis group because of its similar prognosis with immature teratoma and mixed tumors mainly composed of germinoma or teratoma.Surgery is the first choice for NGMGCTs because treatment should be base on tumor histology.For patients in the intermediate prognosis group,combined treatment,including surgical resection, radiotherapy,chemotherapy and gamma knife surgery was effective.

Study on the Measles Epidemic Change and the Current Immunity Situation of Population in Jiading District

Sun, 27 Mar 2011 05:01:33 +0000

Part 1 The incidence situation of the measles in Jiading Districtfrom 1953 to 2007[Objectives]The goal of the study is to learn about the epidemic characteristics of the measles in Jiading District,providing the basic information for studying the measles immunity situation,and prevention and control of meales.[Methods]The measles epidemic data in Jiading District from 1953 was collected and analyzed,and attention was paid to the descriptive analysis for the epidemic reporting data and the monitoring data of the measles from 1992 to 2007.[Results]There are 50594 measles cases in Jiading District from 1953 to 2007. The incidence rate from 1953 to 1965 remained at a relatively high level,the average incidence is 884.84 per 100000,and the incidence rate in 1959 achieves a peak above 3000 per 100000.The incidence rate started reducing rapidly from 1966.and the average incidence rate from 1966 to 1981 was 114.08 per 100000.But the incidence rate varied with a large scale,and the incidence rate reached a peak in 1980,achieving 606.30 per 100000.The incidence rate reduced to a low relatively level rapidly afterwards,and the average incidence rate from 1982 to 1991 was 0.62 per 100000. The total of 503 Measles cases was diagnosed from 1992 to 2007.in which 104(20.7%) were local population with the incidence rate of 1.31 per 100000,and 399(79.3%) was non-native population with the incidence rate of 9.59 per 100000. The epidemic situation presented a rising trend with the sporadic onsets predominating in that period,and the incidence rate of non-native population was higher than that of local population.The positive correlation between the measles cases and the non-native population quantity was absorbed.The local cases mainly concentrated in adults above 20 years old and 0-8 month infants,accounting for 83.65%,and the non-native cases mainly concentrated in 8-month-old to 7-year-old children and adults above 20 years old,accounting for 72.43%.In the local population, the incidence rate of the 0- year- old group infants was the highest,achieving 60.70 per 100000,and the 20- year-old group adults rank second.[Conclusions]The measles epidemic situation was well under control after the usage of measles vaccine in 1965.The epidemic situation presented a re-rise in 1992, the onset age of measles trended to be \'\'bidirectional shifting",low-age children and adults above 20 years old is much easier to be infected.The incidence rate of non-native population is the emphasis and difficulty of measles control and prevention in Jiading District.Part 2 The measles immunization coverage situation of the children in Jiading District[Objectives]The goals of the study is to learn about the measles immunization coverage situation of the local and non-native children in Jiading District,and to analyze the correlation between measles incidence and the immunization coverage.[Methods]To investigate the measles immunization coverage situation,the vaccination data was collected from 2003.applying the PPS sampling(the capacity proportion probability sampling,PPS) to collect 210 local children and 210 non-native children at the age of 8 months to 4 years.[Results]The measles immunization coverage rate of the local children is higher than that of the non-native children in 2003.The MV1 immunization coverage rate of 8-month-old to 4-year-old children is 98.33%in 2007.and the MV2 immunization coverage rate is 94.41%.The immunization coverage rate and the qualified rate of the local children are higher than that of the non-native children in the same age group.The average vaccination month age of the local children is 8.44 months,which of the non-native children is 9.23 months.The immunization qualified rate of MV1.MV2 of the non-native children is 92.38%and 82.91%respectively,and that of MV1.MV2 of the local children is 81.43%and 59.59%respectively.The difference of the qualified rate of MV1 between the local children and the non-native children has statistical significance.The difference of immunization constituent ratio of MV and MMR also has statistical significance.The main reasons of non-vaccination are to vaccinate ahead of time(71.43%) and unknown to the vaccination place(28.57%).[Conclusions]The immunization coverage situation presents imbalanced between the local children and the non-native children.The situation of measles vaccination in local children is better than that in the non-native children.The immunity barrier still needs to be reinforced.Health education on the Planned Immunity and vaccination in time should be strengthened.To eliminate the immunity blank,leak-check and revaccinate should be done well.Part 3 The measles immunity level of population in Jiading District[Objectives]The goal of the study is to learn about the serum antibody level of population in Jiading District,and to observe the correlations between the serum antibody level and the incidence,so as to develop the immune prevention and control aiming the high risk population.[Methods]General population:Ma Lu Town where non-native population centralizes is selected for the study site,and cluster sampling is applied for selecting local population sample and non-native population sample,considering they are in the same class and the same workshop.The sample of 8- month infants group are selected from the same EPI clinics,according to the visiting sequence.Pregnant woman:Healthy pregnant women were selected from MCH hospital in Jiading District from October 10,2007.according to the hospitalizing time order. Enzyme-linked Immunosorbent Assay(ELISA) is applied in the study.[Results]General population:There are total of 375 cases,in which 188 cases are local population,and 187 cases are non-native population.The measles antibody positive rate of population in Jiading District is 89.60%.and the difference of the positive rate between the local population and the non-native population has no statistics significance.The measles antibody positive rate of 8- month infants is the lowest in all age-groups,and that of 20-40 year-old adults is lower.The proportion of those with vaccination records in the cases of measles antibody negative is 61.54%, and it is 33.33%who have vaccination records more than one time.The positive rate is only 11.11 percent in 8-month-old infants when the time is no more than 10 days from the first-dose vaccination coverage.The difference of the positive rate between the vaccination times has statistics significance,and the positive rate rises with the vaccination times incerases.The positive antibody geometric average titer(GMT) of the local population is lower than the non-native population,and the difference has statistics significance.The difference of the antibody titer level has statistics significance among age-groups.The antibody titer level of 3-year old is the highest at 1489.29,and there is a downward trend with age,but increase suddenly in 30 to 40-year-old group.Pregnant woman:From 400 healthy pregnant women,we obtained 395 serum samples,in which 197 women are local population and 198 women are non-native population.The investigation showed that the measles antibody positive rate is 92.15%,and the rate of the antibody negative to low protection level(1:200) is 28.86%.The measles antibody positive rate of local population is higher than that of the non-native population.The antibody geometric average titer(GMT) of the local pregnant woman of antibody positive is lower than that of non-native women.[Conclusions]8-month infants and 20 year-old adults are the high risk population of the measles infection,especially that the 8-month infants without vaccination as well as before the immune response after vaccination are very susceptible to measles,so the protection for them should be strengthened.The immune failure occurs sometimes,and the vaccination quality should be improved. The measles antibody level of the pregnant woman should be improved,so as to reduce the incidence rate of infants before vaccination.Part 4 conclusions and policy suggestions[Conclusions]The epidemic situation presented a re-rise in 1992.The onset age of measles trended to be "bidirectional shifting",and children of old age group and adults above 20 years old are much easier to be infected.The immunization coverage situation presents imbalanced between the local children and the non-native children, and the measles antibody level of the pregnant woman is ineffective to prevent the incidence of infant.The immunity barriers still need to be reinforced.The non-native population is the emphasis and difficulty of measles control and prevention in Jiading District.[Policies and suggestions]Attention should be paid to the measles control and elimination.Management of the monitoring and reports of measles should be strengthened,and epidemic situation of measles should be handled in time.The study on the measles vaccination for women of child-bearing age before marriage still needs to be developed further.We should strengthen health education on measles,strengthen standard construction of EPI clinics,develop standard vaccination,and enforce quality control.To improve the implementation effect of the immunity strategy,the management of the non-native population should be strengthened.

The Exploratory Development of CT Perfusion in the Course of Surgical Treatment for Ischemic Cerebrovascular Disease

Sun, 27 Mar 2011 05:01:29 +0000

Objective To investigate the applicative value of CT perfusion in the course of surgical treatment for ischemic cerebrovascular disease,and how to improve accuratissime and reliability of CT perfusion.Methods CT perfusion imaging was performed in three teams patients preoperatively and postoperatively,who accepted bypass surgery or stent implant for ischemic cerebrovascular disease,and the machine was siemens semsation 64 multislice computed tomographyic.In the course of operating CT perfusion,the method of drawing the region of interest and the evaluative method for parameters of CT perfusion imaging were used,that was all summarized in this research.The final result was analysised by statistics process.Results The CBF and CBV in the operative side of most patients in three teams were descent and TTP were prolong before treatment,this condition was consistent with indicated of bypass operation and stent implant.Postoperative rCBF of all patients were improved obviously,this proved the effect that bypass operation and stent implant improved the cerebral blood flow in ischemic region were very significant;postoperative rCBV of moyamoya team and stent team were all higher than preoperative rCBV,but the diversity of rCBV in non-moyamoya team were not obvious;Postoperative rTTP of moyamoya team was shorter than preoperative rTTP,the diversity of rTTP of non-moyamoya team and stent team were not obvious.Conclusion The result of CT perfusion was important indicated of surgical treatment for ischemic cerebrovascular disease;CT perfusion was very value in evaluating therapeutic effect of surgical treatment for ischemic cerebrovascular disease; drawing the region of interest exactly was important for ensuring the accuratissime of CT perfusion;calculating the relative magnitude was the principal method for evaluating parameters of CT perfusion.

The Inspiration of Taoist Health Thought to the Development of Residents Physical and Mental Health

Sun, 27 Mar 2011 05:01:27 +0000

Taoist thought is a kind of health related to people\'s livelihood and life philosophy. It integrates the Chinese idea of cultural resources used for health, and civil secular philosophy put Unicom Living together to form their own unique cultural identity, has profound meaning of life and the spirit of inclusive values. Chinese traditional sports in its formation and development of the process learned a lot of Taoist health idea of intelligence. Today, the Taoist thought of China\'s still have a significant impact on the theory and practice of modern sports development.Urban Community Sports of our country are an important component of fitness activity is carried out major development model, urban community sports development aimed at increasing the level of physical and mental health of all the people, to improve the urban culture and entertainment life of urban residents to meet the growing demand for PE, as well as set up a solid feeling of community, and promote harmonious society plays the role of a positive impact. At present, the research about urban community sports lack for the conducts form the traditional Taoism thought angle, so the article trying from the Taoism thought unique angle and using many methods such as material analysis, historical analysis, and system analysis and so on, study the influence of the Taoism thought to the urban community sports practice and theory.

A Quantitative Assessment of the Effects of Normal Sun Protection Education in the Photosensitive Patients

Sun, 27 Mar 2011 05:01:26 +0000

Objective:To evaluate the influence of normal sun protection education on the sun exposure habits of the photosensitive patients and to assess the preventive and therapeutic effects of this education for the chronic actinic dermatitis(CAD) and polymorphous light eruption(PLE).Methods:The patients with CAD or PLE were assigned to either control group or intervention group by the random digits table.Usual suggestions about sun protection methods were provided to all the subjects at their visits and all the subjects could voluntarily charge of free broad-spectrum sunscreen(SPF30+,PA+++) in the whole study year.The intervention group was given two normal sun protection educations in addition.The first education was held at the beginning of spring by lecture about the knowledge of ultraviolet radiation,the introduction of photosensitive diseases (including CAD and PLE),the normal sun protection methods and the knowledge of sunscreen.The education manual was given out to the intervention group at the same time.The other reinforced sun protection lecture was held at the beginning of summer All the subjects completed an interview at enrollment and at 12 months after enrollment.Both interviews included a questionnaire about sun exposure habits and a questionnaire about severity and therapy of CAD/PLE in the past year.The intervention group completed another interview about sun exposure habits at 1 month after the first normal sun protection education.SPSS 11.5 was used for statistical analysis.Results 1:①At 1 month after the first sun protection education,the light exposure habits(self-control of sun exposure time,shadow seeking behavior,avoidance of man-made ultraviolet source),the physical sun protection behaviors(the usage of sunglasses,the cognition and usage of proper clothing) and the usage of sunscreen (the frequency,the location and the dosage) of the intervention group improved significantly from haseline(p＜0.01).②At 6 months after reinforced education,the cognition and usage of proper clothing and the shadow seeking behavior further improved and had statistical significance compared with those behavior at 1 months after normal education(p＜0.05).③At 1 year after enrollment,the light exposure habits,the physical sun protection behavior and the proper usage of sunscreen of the intervention group improved significantly compared with that of the control group (p＜0.01).Results 2:①The severity degree of CAD(the number of the patients with persistent eruption for whole year,the number of the patients with generalized lesions, cumulative exacerbation time) was significantly different between pre- and post-intervention(p＜0.05).And in the study year,the severity degree of CAD in the intervention group decreased statistically compared with that in the control group (p＜0.05).②The need of drug treatment and the health care cost of the patients with CAD in the intervention group were significantly different between pre- and post-intervention(p＜0.05).And in the study year,the need of drug treatment and the health care cost of the patients with CAD in the intervention group decreased statistically compared with that in the control group(p＜0.05).Results 3:①The severity degree of PLE(the number of months with eruption in the preceding year,the times lesions provoked,the seasons with eruption,the facial incidence,the duration of sun exposure required to provoke the eruption) was significantly different between pre- and post-intervention(p＜0.05).And in the study year,the severity degree of PLE in the intervention group decreased statistically compared with that in the control group(p＜0.05).②The need of drug treatment and the health care cost of the patients with PLE in the intervention group were significantly different between pre- and post-intervention(p＜0.01).And in the study year,the need of drug treatment and the health care cost of the patients with PLE in the intervention group decreased statistically compared with that in the control group (p＜0.05).Conclusion:Normal sun protection education including two lectures and education manual was a more effective way of improving sun protection habits of the photosensitive patients.With improvement of sun protection behaviors in the study year,the severity of CAD/PLE decreased,the need of drug treatment and health care cost were also reduced.This study suggested that more attention to sun protection education should be taken by the dermatologists in the treatment of photosensitive diseases.

Design of Portable Instrument for Analyzing Pulse Wave Based on ARM9

Sun, 27 Mar 2011 05:01:24 +0000

Cardiovascular disease is the primary illness to human health,so finding one means to reflect our normal cardiovascular parameters is very important.Because the pulse wave can imply lots of information about the human cardiovascular system.By detecting and analyzing the pulse wave signal,we can accurately obtain the user\'s cardiovascular parameters.Based on this information,we can objectively assess the state of the user\'s cardiovascular function,which is of great significance in forecasting and treating cardiovascular diseases or related diseases.By analyzing the current situation of clinical instruments,we designed a portable apparatus based on ARM9 for analyzing pulse wave in this paper.The users can easily and accurately obtain their cardiovascular parameters from this instrument.The main research works are as follows:1.Studying the working principle of the pulse wave,such as the medical theory, the measurement and analysis method and the current situation,and setting up the whole system including the hardware development platform and the operating system.2.Developing the embedded system based on Linux,including building the host development environment,transplanting the Bootloader,configuring and compiling the kernel as well as the production of the root file system.3.Programming the graphical user interface based on MiniGUI.Firstly,we built the development environment of MiniGUI.Then we designed the graphical user interface and transplanted the program to the target system after debugging.4.Completing the design of the module for blood pressure measurement. According to oscillometric method,the framework of the module was designed. C8051F330 single chip is the core of the whole module.Then the hardware and software design was completed on this system.5.Designing the system applications,including the serial communication programs of pulse wave acquisition module and blood pressure measurement module with ARM9,the program for analyzing the pulse wave signal and so on.6.Finishing the debugging of the whole system,including the debugging and calibration of the blood pressure measurement system,the debugging of the two serial communication programs,the debugging of the graphical user interface and so on. Finally,we achieved the desired functions in this instrument.

Study of Screening in Vitro for Nanofiber Scaffold Materials in Tissue Engineered Nerve

Sun, 27 Mar 2011 05:01:22 +0000

Part 1:Culture in vitro and identification of schwann cell line.Objective:comparing with normal schwann cells,we observed the cell morphous and regularity,and detected the purity and genetype of schwann cell line cultured in vitro, to prove the possibility of application of screening the scaffold materials for peripheral nerve.Methods:Culture the schwann cell line RSC96 conventionally,and observe the cell morphous and regularity;Evaluate the purity by immunofluorence staining of S-100; Detect the expression of gene S-100、GDNF、BDNF、NGF、CNTF、PMP22 of schwann cell line through RT-PCR.Result:Schwann cell line RSC96 expressed the typical cell morphous of schwann cell,with the fusiform shape and spindly bipolar.The growth regularity was similar with the normal schwann cells while the course was abbreviated.All cells were positive by immunofluorence staining,which demonstrate 100%purity,and the distribution of S-100 consistented with the regularity of normal schwann cells. RT-PCR proved the expresion ofgene S-100、GDNF、BDNF、NGF、CNTF、PMP22 of schwann cell line,which were the significant gene for normal schwann cells.Conclusion:Schwann cell line RSC96 has the similar cell morphous,growth regularity and genetype with normal schwann cells,with the 100%purity,high speed of proliferation and lower degeneration.So schwann cell line RSC96 is more suitable to make use of the screening scaffold materials in vitro compared with others. Part 2:Identification in vitro of nanofiber scaffold materials made of PLLA and co-polymer with PCL.Objective:to identify and compare different kinds of composition of PLLA, P(LLA-CL)(75:25) and P(LLA-CL)(50:50) and different preparing technology for nanofiber scaffold materials for peripheral nerve,through schwann cell line,and finally choose the best one for the animal test in vivo.Methods:Observe the microstructure of three kind of nanofiber scaffold materials by scanning electron microscopy,compared with microstrcucture of extracellular matrix. Evaluate the biocompatibility of them through proliferation test,attachment test and observing of cell morphous.At the same time find the regularity between the orientation of nanofiber and alignment of cells.Analyse the expression of gene S-100、GDNF、BDNF、NGF、CNTF、PMP22 of schwann cell line which was cocultured with nanofiber scaffold materials using Real Time PCR,compared with the gene expression of schwann cell line which was not cocultured.Result:The cocultured schwann cell line attached and proliferate well,which indicate the favorable biocompatibility of three kind of scaffold materials,in which P(LLA-CL)(75:25) was the best one.The result was relative to the fact that the microstructure of P(LLA-CL)(75:25) is more similar to ECM\'s,which can be identified by SEM.At the same time we found that the cells can align along the orientation of nanofiber.And also,three kind of scaffold materials gived no impact on gene expression of S-100、GDNF、BDNF、NGF、CNTF、PMP22 of schwann cells.Conclusion:Compared with PLLA and P(LLA-CL)(50:50),P(LLA-CL)(75:25) has more favourable biocompatibility,with mircostructure more similar with ECM,and also has no obvious impact on gene expression.Especially,the orientation of nanofibers can lead cells to align regularly.These results demonstrate that the oriented nanofiber scaffold materials made of P(LLA-CL)(75:25) can be used in animal test.

A Preliminary Study on Quality of Life and Detection of Endothelin-1 in the Plasma and Skin Tissue Fluid on Patients with Vitiligo

Sun, 27 Mar 2011 04:31:44 +0000

Objective:1.To evaluate the effect of vitiligo on quality of life in Chinese patients.2.To study the role of endothelin-1(ET-1) in the pathogenesis and prognosis of vitiligo,level of ET-1 in the plasma and skin tissue fluid of patients with vitiligo was detected.Materials and Methods:1.We collected eligible data from 1025 patients with vitiligo by a uniform questionnaire about clinical and epidermiology study,of which 786 patients were evaluated by the measurement of quality of life.And we collected samples of plasma (n=96) and lesion tissue fluid(n=25) from vitiligo patients.30 healthy people and 25 normal skin tissue fluid were set as controls.2.We evaluated the effect of vitiligo on quality of life in Chinese patients by DLQI.3.We measured the levels of ET-1 in the plasma and the tissue fluid from vitiliginous lesion and the normal skin by the method of radio immunity.4.Statistical analysis was performed by one-way ANOVA and LSD-test.Results:1.The overall mean DLQI score for vitiligo was 7.32±5.68,which meant vitiligo had moderate effect on patient\'s life.The individual proportions of five grades were:2.60%,33.08%,30.66%,20.99%,2.67%.2.The disease duration(Spearman=0.1548),lesion area(Spearman=0.1973) and treatment time(Spearman=0.1021) were independently related to the DLQI. Gender was significantly associated with the DLQI score,women reached a higher overall score(P=0.0035).Examined across different marriage status groups,the total DLQI score for vitiligo changed a lot(P=0.003,married>single).During the disease type groups,universal type got the highest DLQI score,while localized one was on the contrary.We observed itching feeling,white hair and cost effect on DLQI score.3.The level of ET-1 in the plasma of patients with vitiligo was significantly higher than the normal controls(P=0.0000).We found no interaction of disease type (universal and non-universal,P=0.0707;segmental and non-segmental,P=0.1908) and disease stage(P=0.4046) on the level of ET-1 in the plasma.And the level of ET-1 in the skin tissue fluid did not differ significantly between vitiliginous lesion and the normal skin(P=0.5880).Conclusion:1.Vitiligo had moderate effect on patient\'s life.The disease duration,lesion area and treatment time were independently related to the DLQI.A significant association between gender or marriage status and DLQI score.We observed disease type,itching feeling,white hair and cost had great effect on DLQI score.2.The level of ET-1 in the plasma of patients with vitiligo was significantly higher than the normal controls.And the level of ET-1 in the skin tissue fluid did not differ significantly between vitiliginous lesion and the normal skin.

Effects of Massage on Basic Fibroblast Growth Factor during the Injury and Repair of Skeletal Muscle

Sun, 27 Mar 2011 04:31:42 +0000

Purpose:Basic fibroblast growth factor (bFGF) is a growth factor which is generally distrbuted in skeletal muscle,and have biological effect on skeletal muscle growth and devel-opment,and play a critical role on physiological and pathological process of skeletal mus-cle.Present study indicates that bFGF modulates skeletal myoblast functons and plays a key role in myoblast proliferation and inhibiting its differentiation.Skeletal muscle injury is the frequency disease in sports.There are many treatment methods on skeletal muscle injury , massage is a safe economic and effective treatment method.Present study indicates that mas-sage can increase skeletal satellite cells during the skeletal muscle injury and regeneration , and promote skeletal muscle recovery .The present study investigated the time course changes in bFGF protein expression and the effects of massage on bFGF in skeletal muscle injury and reparative process,discussed the mechanism of action on massage healing skeletal muscle injury.Method:Twenty one adult male Sprague-Dawley rats are divided into seven groups at random: control group(n=3);acute traction injury group (n=12),which was further divided into 1h、2d、5d、7d postinjury groups;massage group (n=6) was divided into 3d and 5d postmassage groups. Rats’gastronemius acute traction injury model are established by self-made facility. Massage treatment started on second day postinjury. The bFGF expression change was meas-ured by immunohistochemistry and image analysis system.Results:(1)In rats’gastronemius,bFGF protein level increased in acute traction injury group at 1h and 2d postinjury ,reached the peak on 7th day after injury.(2)In rats’gastronemius,bFGF expression level lower afer 3-day-massage than spontaneous recovery group on synchroniza- tion ,also there was a big difference between 5-day-massage group and spontaneous recovery group on synchronization.(3) By macroscopic observation , massage reduced the adhesions not only between skin and muscle but also between gastroenemius and soleus muscle,and reduced the accrementition of connective tissue. By light microscope observation, massage reduced the inflammatory reaction and cellular necrosis and cicatricle repair after muscle injuried, enhance -mented muscle cell regeneration and muscle tissue repairation,shortened the repair phase.Conclusion:The time course changes in bFGF protein level was showed after acute traction injury . Massage had an effect on bFGF expression and had a significant effect on injuried skeletal muscle.

The Study on Service and Quality Control in the Filed of Hospital Pharmacy Based on the Value Chain Management

Sun, 27 Mar 2011 04:31:40 +0000

The theory of Value Chain is the efficient tool to research on the competitive advantages since Michael Porter published it in 1980s.The theory of the Value Chain is adopted in all the fields and takes the overwhelmed advantages over the competitors.The Value Chain starts with the customer and ends with them,therefore, it\'s strategically important for the enterprises management.The further the reforming of Health System develops,the more the medical market opens,and the severer competition in the market is.As we all know,the quality control is the core of hospital management.As a part of the medical services, the pharmaceutics is the main field for the hospital to provide the pharmaceutical services and plays an important role to make sure the quality and safety of the leechdom and to improve the efficiency of the medical.For some reasons,the quality control of the hospital pharmacy can\'t meet the requirement of the pharmaceutical services.The paper analyses the current situation of hospital pharmacy and the problems and challenge to face to in the reforming of Health system.It also analyses the inner and vertical value chains of the hospital pharmacy by the theory of Value Chain and discusses services and management of hospital pharmacy based on the Value Chain Management.By the discussion,we can find the following:The pharmaceutical service consists with the theory of Value Chain.The Pharmaceutical service should be attached the importance to the value creation with the core of patients.It is very important for improvement of the value of pharmaceutical service to change role and service orientation of Licensed Pharmacist,strengthen the serving thought and skills of pharmacist,improve enlarge services.The changes of pharmaceutical management require us to make the pharmaceutical knowledge and expertise catching up with the time development. The pharmacists\' professioncy should be made full use and the pharmacists should be encouraged to attend the whole chain of the hospital medicaling so as to make the "Five Rights" come true,that is,Right Patient,Right Medicine,Right Dosage,Right Way and Right Time,in every fields related to the medical using in the hospital.,so that the value of pharmaceutical services could be improved.The paper will contribute to the guidance and reference of quality control of hospital pharmacy.

Review of Diagnosis and Therapy for Pemphigus and Pemphigoid with Measurement of Serum Cytokines in Pemphigus Patients

Sun, 27 Mar 2011 04:31:36 +0000

OBJECTIVEPemphigus is relatively common among all the autoimmune bullous diseases and it\'s pathogenesis is quite complex.Nowadays,with the help of rapid development of Immunology and Molecular biology we could understand it further on the level of cytokines and cell immunity.However,the symptoms of pemphigus and pemphigoid are sometimes similar with other diseases,which makes it difficult to be diagnosed.Once onset,the severity of the disease could be quite different,which demands different criterion for treatment,ln this study,we aim to estimate ways of making diagnosis and purpose to have a general idea of the properly dosage of corticosteroid.1.A retrospective study of clinical diagnosis and therapy in 282 cases with pemphigus and 366 cases with pemphigoidMaterials and Methods:To analyze the diagnosis and therapy of pemphigus and pemphigoid.Results:The mean first-onset-age of pemphigus was smaller than pemphigoid(p＜0.001).In 175 patients of pemphigus confirmed by direct immunoflorescence(DIF),IgG deposition were 100%positive,while C_3 92.0%.In 223 patients of pemphigoid,the proportion of C_3 positive was 99.1%.①To patients of pemphigus,coincidence proportion of clinical diagnosis,pathology and DIF was 68.8%,while the proportion of the later two was 80.7%.②To patients of pemphigoid,consistent rate of clinical diagnosis,pathology and DIF was 62.8%,while the proportion of the later two was 78.1%.The patients were mainly treated by corticosteroids with a dosage of 0.5～1.5 mg·kg~(-1)·d~(-1)(prednisone).Conclusion:For cases of pemphigus and pemphigoid which can\'t be confirmed by clinical features and pathology,DIF is an important diagnostic method.Doctors in community hospital with limited funds could mainly test IgG and C_3 when the diagosis of pemphigus was doubtable.Similarly,they could mainly test C_3 if they consider pemphigoid as first diagnosis. Corticosteroid is the chief way for controlling the two diseases,while intra venous immunoglobulin(IVIG) could be added when necessary.2.Measurement of serum interleukin-4,6,10,17 and interferon-gamma levels in patients with pemphigusMaterials and Methods:Serum levels of interleukin-4,6,10,17 and interferon-gamma were determined by double antibody sandwich enzymelinked immunosorbent assay in 33 patients with pemphigus and in 34 normal controls.Results:Serum IL-6 and IL-17 levels were higher in patients with pemphigus than those of controls(p＜0.05).Serum levels of IFN-γand IL-10 were lower in patients with pemphigus than those of controls(p＜0.05).Conclusion:IL-6,IL-10,IFN-γand IL-17 might be involved in the pathogenesis of pemphigus.IL-17 might be a key factor among cytokines above.

The Application of Intercostal Nerve Transfer in Obstetric Brachial Plexus Palsy

Sun, 27 Mar 2011 04:31:34 +0000

Objective To study the therapeutic effect of intercostal nerve transfer for restoration of elbow flextion in infants with obstetric brachial plexus palsy.And to assessment the pulmonary function of the children after transfer of the intercostal nerve.Methods Between December 2001 and July 2008,a total of fifty-seven patients with obstetric brachial plexus palsy(OBPP) underwent intercostal nerve(ICN) transfer.The mean patient age at surgery was 6.8 months and mean follow-up period was 2.0 years.33 cases of them were randomly selected to take an examination of pulmonary function(operation group),the mean age was 33.1 months.A control group with 31 patients related to OBPP took the pulmonary function examination at mean 32.9 months.All of the control group have never underwent the nerve transfer of intercostal nerve or phrenic nerve.The function of elbow flexion was evaluated in all cases,which include muscle power and appearance time of elbow flexion.Muscle power was assessed using Kawabata improved British Research Council Muscle Movement Scale grades of M0 to M4.Grade M0 signifies no contraction and M1 a trace of contraction.Grade M2 is used to indicate active flexion more than 90°with gravity eliminated and Grade M3 active flexion more than 90°against gravity.Grade M4 was recorded when there was active movement against gravity and resistance.Appearance time of elbow flexion was indicated by the first time when we observed the elbow flexion(＞M1) after operation. Pulmonary functional parameters including vital capacity max(VCMAX), forced expiratory volume in one second(FEV1),forced expiratory flow 25-75(FEF25-75),airway resistance Rrs5(R5),reactance Xrs5(X5),tidal volume(VT/kg),time taken to reach peak expiratory flow to total expiratory time(TPTEF/TE),volume taken to reach peak expiratory flow to total expiratory volume(VPEF/VE).Those who could collaborate with the examination,checked VCMAX、FEV1、FEF25-27 when they were elder than 4.5 years and the others checked R5 and X5.TPTEF/TE and VPEF/VE were examed in the patients who could not collaborate with the test.We observed daily clinical syndrome in operation group.Results There were 24 patients followed up more than 2.0 years.Fourteen patients were used 4 ICNs,four of them gained biceps muscle power to the level of Grade M4,six patients exhibited Grade M3 and three Grade M2.Mean follow-up period of the 14 patients was 32.3 years.There were 9 patients using 3 ICNs,followed up by 45.6 months.Eight of them gained biceps muscle power to the level of Grade M4 and one Grade M3.We performed 4 ICNs transfer to musculocutaneous nerve(MCN) in four patients and three of them reached M4.There were two patients gained M4 in 10 children who underwent 4 ICNs transfer to the anterior division of the upper trunk(AUT).We observed 54/57 patients gained active elbow flexion at average 7.8 months after surgery.Twelve of them were used 3 ICNs,whose elbow flexion were found out at mean 7.0 months.We transferred 3 ICNs to MCN in seven patients at average 6.6 months old.Mean 7.0 months later,active elbow flexion were observed.Four ICNs were used in 40 patients,their elbow flexion were discovred 7.4 months after surgery.We transferred 4ICNs to MCN in 24 infants at average 10.5 months old.After mean 6.3 months,we observed active elbow flexion.The other 16 patients whose 4 ICNs were transferred to AUT flexed their elbow at average 9.1 months after surgery.In operation group 4 of 33 were examed VCMAX、FEV1、FEF25-27 and 3 of all were checked R5 and X5.We checked TPTEF/TE and VPEF/VE in other 26 patients.In control group 2 of 31 were examed VCMAX、FEV1、FEF25-27 and 2 of all were checked R5 and X5.We checked TPTEF/TE and VPEF/VE in other 27 patients.There were 17 patients in operation group and 17 patients in control group,whose pulmonary fuction were abnormal.The difference are of no statistical significance(X~2test,X~2=0.07&0.13,p＞0.05).All the patients in operation group appeared to have no difficulties in daily living.Conclusions Transfer of intercostal nerve for restoration of elbow flextion in infants with obstetric brachial plexus palsy presented excellent results and did no harm to the pulmonary function of the patients. We should approach the anastomotic stoma to biceps brachii to improve the effect.Using 3 ICNs or 4 ICNs made the result no difference,but if the age of the patient was old,we recommended using more ICNs to gain the better effect.

Glutamine: The Effect of Gastrointestinal Protection in Newborn Rats

Sun, 27 Mar 2011 04:31:27 +0000

Objectives:Recent literatures have focused on the role of glutamine in the area of nutrition and metabolism.We carried on this Gln-related research with the expectation to better nourishment support to critically-ill children.To observe the changes of the intestinal tract of the maternal breast-fed newborn rats,we deprived glutamine(Gln) artificially by depressing glutamine synthetase(methionine sulfoximine,MS) in mother and/ or baby rats.Also,newborn rats model with endotoxemia were used to observe the effect of Gln supplementation to intestinal function.Methods:First part of the experimentNewborn rats were divided into 5 groups.All rats were breast-fed.Group 1,the control group,consisted of rats and maternal rat without MS injection into the abdominal cavity.Group 2 consisted of rats with maternal use of MS through intraperitoneal injection from the 3rd day of given birth for 6 days.Group 3 consisted of rats with maternal use of MS from the first day given birth for 6 days.Group 4 consisted rats who were injected MS from the 3rd day on,maternal rat used MS from the 3rd day of given birth,both for 6 days.Group 5 consisted of rats with MS from the 3rd day of life until day 9,without maternal use of MS.All newborn rats were killed.Small intestine was harvested for morphological studies.Changes of the small intestine villi were observed by light microscope and electron microscope.Membrane fluidity of intestinal epithelial cell was examined, too.Second part of the experimentNewborn rats were divided into 4 groups.Group 4(Gln completely-deprived model) from the first part experiment as experiment set and the normally fed rats as control set.Gln completely-deprived model:rats who were injected MS from the 3rd day on,maternal rat used MS from the 3rd day of given birth.normally fed rats:rats and maternal rat without MS injection into the abdominal cavityGroup 1 consisted Gln completely-deprived rats whom used Gln through intraperitoneal injection from the 3rd day for 6 days.Group 2 consisted Gln completely-deprived rats without Gln supplementation for 6 days.Group 3 consisted normally fed rats whom used Gln through intraperitoneal injection from the 3rd day for 6 days.Group 4 consisted of rats and maternal rat without use ofmethionine sulfoximine All newborn rats were uesd LPS through intraperitoneal injection in the 7th day of experiment.After 6 hours the rats were killed.Small intestine was harvested for morphological studies.Changes of the small intestine villi were observed by light microscope and electron microscope,and the membrane fluidity of intestinal epithelial cell was examined.Results:In the first experiment,the morphology of small intestine was significantly different in each group.In the control group small intestine villi and microvilli were slender、developed、arranged closely and intestinal structure was clear under optical and electric microscopics,the membrane fluidity of intestinal epithelial cell was best in all groups.By the contrast,the small intestine villi and microvilli in group 4 were badly destroyed and arranged,the membrane fluidity of intestinal epithelial cell was worst among all groups.In the second part of the experiment,the morphology of small intestine of rats with endotoxemia was significantly different in each group.In group 2 intestinal structure was not integral,small intestine villi and microvilli were the shortest and badly destroyed under optical and electric microscopics,the membrane fluidity of intestinal epithelial cell was worst in all groups.However,due to the additional Gln supplementation,the destroyed small intestine villi and microvilli of group1 were alleviatived compared to group 2,so does the membrane fluidity of intestinal epithelial cell.The morphology of small intestine was not significant changed between group 3 and group 4,but both of them were better than the former two groups.Conclusions:(1) The experiment showed that Gln deprivation can really change the gastrointestinal structure and function of newborn rats.(2) The 4th group will be the most suitable animal model for Gln deficiency model in breast-fed baby rats.(3) The study preliminary certified that Gln supplement could protect the gastrointestinal structure and function in newborn rats during endotoxemia.

Clinicopathologic Initial Study and Investigation of Mechanism of Perineural Invasion in Adenoid Cystic Carcinoma

Sun, 27 Mar 2011 04:31:22 +0000

Obective1.To analyze the clinicopathologic features of adenoid cystic carcinoma(ACC), specially its morphologic features,and try to find out the prognostic factors which were related to perineural invasion.2.To investigate the mechanism of perineural invasion,and provide the theoretical reference for improving the therapeutic efficacy and living quality.Methods1.The subjects of this retrospective study were 42 patients suffered from ACC between 1983 and 2007 in the Stomatology Hospital of Tianjin Medical University. The clinic pathologic factors were collected including age,sex,primary tumor site, size,histological features,perineural invasion and TNM stage.The statistic analysis was performed between those parameters and perineural invasion.2.By immunohistochemical stain,to investigate the expressions of SMA、GFAP、GDNF、Ret and Ki-67 in ACC,and the statistic analysis was performed between those antibodies and perineural invasion.Results1.Among 42-case ACC,25 female and 17 male.Age is from 22 to 74.Average age is 47.29.Besides,18 cases in major salivary glands,23 cases in minor gland,and 1 case in other location.Plate gland is the most organ.Pain is the frequent symptom,presented in 54.76%cases.Perineural invasion can be found in 30 cases,and the rate is 71.43%,including 2 cases with vascular metastasis,1 case with lymphatic metastasis.In clinic diagnosis,9 cases were diagnosed as ACC,11 cases malignant pleomorphic adenoma or malignant salivary tumor,6 cases pleomorphic adenoma,2 cases gingival carcinoma,others without diagnosis.There are 5 cases inⅠstage,8 cases inⅡstage,13 cases inⅢstage,16 cases inⅣstage.There were no statistic differences between PNI and age,sex,primary tumor site. But there were statistic differences betweenⅠ+Ⅱstage andⅢ+Ⅳstage(P＜0.05), also between cribriform type and tubular type(P＜0.05),between cribriform type and solid type(P＜0.05),but no statistic difference between tubular type and solid type (P＞0.05).The Schwann\'s cell marker GFAP and the myoepithelial cell marker SMA proteins were co expressed in kytoplasm of the same onco-myoepithelial cells.Both GDNF and Ret were expressed in kytoplasm of the carcinoma cell of ACC. GDNF was expressed in neural fibro.But Ret was not expressed in neural fibro.In carcinoma cell of ACC,there were significant correlations between GFAP、GDNF and PNI、histological features、TNM stage(P＜0.05).There was no significant correlation between Ret and histological features、TNM stage(P＞0.05),but between Ret and PNI(P＜0.05).There was no significant correlation between Ki-67 and PNI (P＞0.05),but between Ki-67 and histological features、TNM stage(P＜0.05). Besides,There were significant correlation between GDNF and GFAP(P＜0.05), between GDNF and Ret(P＜0.05).But there was no significant correlation between GDNF and Ki-67(P＞0.05).Conclusions1.PNI is the biological behavior of ACC,whose incidence rate is high.There were significant correlations between PNI and TNM stage、histological features.The symptom of nerve involvement is usual in ACC.With the classic symptom,it is easy to diagnose.GFAP is expected to become an useful index to evaluate the PNI of ACC.2.PNI is a complex process.The Schwann cell differentiation occurs in onco-myoepithelial cells of ACC,and it may be the pathological histological base of PNI in ACC.GDNF combining with its receptor Ret improves the PNI of ACC.The sigal way with GDNF and Ret plays an important role in mechanism of perineural invasion in ACC.3.GDNF and Ret are expected to become useful indexes to evaluate the PNI of ACC,so that it is early to know the PNI.And it is possible to inhabit the sigal way to improve its treatment effectiveness.4.Ki-67 can be used to evaluate the severity of ACC.The relation between Ki-67 and PNI need a deep study.

Glucocorticoid Amplifies IL-2-induced Expand the Ratio of CD4+CD25+to CD4+CD25-and Inhibit the Allogeneic Reaction

Sun, 27 Mar 2011 04:12:06 +0000

Glucocorticoid amplifies IL-2-induced expand the ratio of CD4+CD25+to CD4+CD25-and inhibit the allogeneic reactionObjective:1.To determinate the effect of the short-term administration of DXM and IL-2 in vivo on the mixed lymphocyte reaction.2.To establish the method to expand the ratio of CD4+CD25+to CD4+CD25- by DXM and IL-2 in vitro.Methods:1.Seven days after administration of DXM(5mg/kg/day,3days) and IL-2 (400,000IU/mouse/day,3days) to C57BL/6N mice,mixed lymphocyte reaction was taken by fresh spleen cells from C57BL/6N mice and untreated BALB/c mice.2.In vitro,fresh spleen cells from untreated C57BL/6N mice were cultured with IL-2,and added DXM 4 hours later.The cells were made Flow cytometry analysis 20 hours after coculture.Results:1.In vivo,short-term simultaneous administration of DXM and IL-2 markedly expanded functional suppressive CD4+CD25+regulatory T cells in murine spleen and inhibited mixed lymphocyte reaction obviously.2.In vitro,the combination of DXM and IL-2 can selectively enhance the proportion of CD4+CD25 regulatory T cells,the ratio of CD4+CD25+to CD4+CD25-was also changed.Conclusions:The administration of DXM and IL-2 inhibit mixed lymphocyte reaction.And this because the combined use of DXM and IL-2 expand the ration of CD4+CD25+to CD4+CD25-.

Feasibility Study of Closed Reduction in Young Children Older Than 18 Months with Developmental Dislocation of the Hip

Sun, 27 Mar 2011 04:12:05 +0000

Background:Developmental dislocation of the hip(DDH) is one of the most common lower limb abnormalities among young children,earlier diagnose and treatment,especially conservative one,can bring into almost satisfactory morphous development and near-normal function of the hip joint in the future. To date,wide identification on the treatment has been accepted universally, which is conservative treatment(or closed reduction) under the age 18 months and surgery(or open reduction) above the age 36 months.However,great conflicts lie in the treatment between the age 18 months and 36 months.Objective:A retrospective review of hip joint development after closed reduction in young children below and above 18 months old with DDH by ways of radiological parameters has been done to observe whether the development between these two groups is alike or not,accordingly,The purpose is to see whether it is feasible to apply closed reduction to young children above 18 months old with DDH or not.Methods:Seventy eight patients(97 hips),with 70 girls(86 hips) and 8 boys (11 hips),59 unilateral hips and 38 bilateral hips.were observed after closed reduction by X-ray parameters.All patients received combined adductor tenotomy,closed reduction,modified frog-position(or frog-position) plaster fixation and abduction device fixation in Children\'s Hospital of Fudan University during January,2003 and December,2007,when they were less than 36 months old.The patients were classified into two groups according to the age at reduction,below and above 18 months respectively.X-ray parameters of hip were measured through anteroposterior pelvis rediograph in 6 months,12 months,24 months and 36 months after closed reduction,including acetabular index(Al),medial joint space(MJS),ossific nucleus(ON) of femoral head, center-head distance discrepancy(CHDD),shenton\'s line,together with avascular necrosis(AVN) and residual acetabular dysplasia(RAD).The average duration of follow-up was 25.2 months(6months to 60 months).Results:There is no statistical differences of changes on radiological parameters of AI.MJS.CHDD and ON after closed reduction between these two groups(p＞0.05,a=0.05),and the shenton\'s line becomes unstable,from breaking before reduction,intact 6 months after reduction to breaking again when children begin to walk,while the rate of AVN is slightly lower in the group above 18 months(p=0.04,a=0.05) and RAD is higher than that of the group below 18 months(p=0.01,a=0.05).Conclusion:Closed reduction is still available in DDH above the age 18 months with lower rate of AVN while higher of RD.

Quantitative Sensory Test and Its Application in the Diabetic Polyneuropahty

Sun, 27 Mar 2011 04:12:03 +0000

Objects:To have a cognition of Quantitative Sensory Test(QST) and its application in the Diabetic Polyneuropathy(DPN).Methods:We recruited 45 non-insulin dependent diabetes mellitus(DM) patients in Hua Shan Hospital and 25 age、sex、height matched healthy people from November, 2008 to February,2009.We conducted clinical evaluation,including:DM duration、the control condition of fasting blood glucose(FBG)、Neuropathy Symptom Score(NSS),Physical Examination:Neuropathy Disability Socre(NDS) of Low limbs and Nerve Conduction Velocity(NCV) test;Cold sensation and heat pain threshold tested by limit method and quantitative vibration threshold tested by level method using Neuro Sensory Analyzer Model TSA-Ⅱmachine in thenar eminence、proximal anterior thigh and dorsum of foot to evaluate the DM patients\' nerve function.Results:1.All the 45 DM patients\' cold sensation threshold in proximal anterior thigh and dorsum of foot were lower than control group,heat pain threshold in these two places were higher than control group and quantitative vibration threshold in index finger and first toe were higher than control group.All of the above had the statistically significance.Furthermore,the difference of both cold sensation and heat pain threshold in the distal limbs were most obvious.But there were no difference in the ability of detecting small fiber nerve lesion themselves between cold sensation and heat pain threshold.2.According to the result of NCV and socres of NSS and NDS,there were 42 out of 45 DM patients had diagnosed as length-dependent diabetic polyneuropathy(LDDP), the incidence was up to 93%,but the abnormality rate of NCV was only 27%;Pain was the first of these 42 LDDP patients\'chief complaints(69%) and the most kind of pain was twinge(45%).The next chief complaint was numbness(62%);Of all the body parts,incidence of distal lower limbs sensation lesion was the high,incidence of ankle reflex disappearance was highest.Besides,the incidence of foot swelling and cutaneous change were also not low.3.In 42 LDDP patients,the quantitative vibration threshold in dorsum foot correlated with DM duration,the heat pain and quantitative vibration threshold in dorsum foot both correlated with NDS socre.There was no correlation between all the parameters of QST and NSS score.4.In 42 LDDP patients,the thermal threshold(including cold sensation and heat pain) and quantitative vibration threshold of Index finger correlated with the amplitude of both peroneal and sural nerve.The quantitative vibration threshold of first toe correlated with the amplitude of sural nerve.There was no correlation between all the parameters of QST and distal latency of peroneal nerve。5.Thirty out of 42 LDDP patients had normal NCV results.Compared these 30 patients with age、sex and height matched health people,their cold sensation thresholds in proximal anterior thigh and dorsum of foot were lower than control group,the heat pain threshold in these two places were higher than control group, both had statistically significance.Furthermore,the difference in dorsum of foot was the most obvious.The quantitative vibration threshold in index finger and first toe were higher than control group statistically.6.According to the results of NCV,we divided 42 LDDP patients into normal NCV and abnormal NCV groups.Compared these two groups,we found the scores of NSS and NDS,heat pain threshold in dorsum of foot and quantitative vibration threshold in index finger and fist toe in abnormal NCV group were higher than normal NCV group, all had statistically significance.7.According to the condition of fast blood glucose(FBG),we divided 42 LDDP patients into good FBG control and bad FBG control groups.There was no statistically difference in all the parameters between these two groups.8.In the 42 LDDP patients,compared the patients with metabolic syndrome (MS)with the others without metabolic syndrome,the age was older in metabolic syndrome group.This result was also found in the groups of with and without hypertension group or with and without high body mass index(BMI) group.But there was no statistically difference in thermal and quantitative vibration threshold of all the three places between with and without MS groups,with and without hypertension groups,with and without higer body mass index groups and with and without hyperlipema groups.Conclusion:1.QST was a steady instrument to apply in the examination of peripheral neuropathy including DPN.It can predict the possible incidence of DPN,detect early phase DPN and reflect the severity of DPN.Its abnormality detection rate was high and its evaluation of DPN was thorough.It can serve as the supplementary of NCV,because it detected small fiber and large fiber lesion simultaneously.Cold sensation and heat pain threshold had the same detection ability of small fiber lesion. The changes of threshold in dorsum of foot were the highest in three places,this indicating the length dependent characteristic of LDDP honestly.The methods in this study was good enough to apply in the diagnosis of DPN.2.DPN featured small fiber and large fiber lesion simultaneously and length dependent.There was no obvious difference between the lesion of Aδfiber and C fiber.Small fiber lesion existed before the abnormality detected by NCV.The factors including control condition of FBG and metabolic factors didn\'t influence the degree of DPN nerve lesion.

The Study of Quality Standard of Mongolian Medicine Aquilegia Viridiflora Pall

Sun, 27 Mar 2011 04:12:01 +0000

Objective:To provide experimental evidence for the estalishment of quality standards of the Traditional Mongolian medicine \'Loudoucai\'.Methods:1.The textual research of herbalism and pharmacognostical study on \'Loudoucai\'.2.The microscopical identification,physichemical identification and thin layer chromatographic(TLC) methods were used for the qualitative analysis.3.The content of luteolin and caffeic acid in Aquilegia Viridiflora pall.were measured by HPLC method and the content of amino acid was measured by amino acid analyser.4. The content of moisture,total ash,acid-insoluble ash and extract were also measured. Results:The textual research of herbalism and pharmacognostical study on \'Loudoucai\' showed that the traditional Mongolian medicine \'Loudoucai\' was the dry herb of the perennial plant Aguilegia viridifloa Pall.The microscopic characteristics of the roots,stems,leaves,and powder of Aquilegia Viridiflora pall were detected by microscopical identification;The existence of alkaloids and flavonoids in the herb was detected by physichemical identification;The content of berberine,luteolin,and caffeic acid were detedcted by TLC method;N-butanol - acetic acid - water(7:1:2), benzene - ethyl acetate - isopropyl alcohol - methanol - water(6:3:1.5:1.5:0.3),ethyl acetate - butanone - methanol - water(10:6:1:1) were used as mobile phase to detect berberine under 365nm wavelength.Cyclohexane-ethyl acetate-formic acid(10:10:2), n-hexane-ethyl acetate-formic acid(10:8:2),chloroform-methanol-formic acid(7:2: 0.5) were used as mobile phase,1%ethanol solution of aluminum chloride was used as color development reagent to detect luteoli.Benzene-chloroform-glacial acetic acid (6:1:0.5),chloroform-methanol-formic acid(7:1:0.1),benzene-ethyl acetate-formic acid(3:1.5:0.3) were used as mobile phase to detected caffeic acid.The chromatographic conditions for determination of luteolin: Hypersil-C18ODS(150mm×4.6mm,5μm) column;Mobile phase:methanol-0.4% phosphoric acid(55:45);flow rate:0.6ml/min;wavelength for detection:350nm; column temperature:25℃.The results showed that the content range of luteolin was 0.006875～0.034375μg with a good linear relationship with peak area(r = 0.9992); the average recovery was 95.92%,RSD was 1.95%(n = 5).The chromatographic conditions for determination of caffeic acid:ODS(150mm×4.6mm,5μm,number: E1718531);mobile phase:methanol -0.05%phosphoric acid(17:83.);flow rate: 1ml/min;wavelength for detection:323nm;column temperature:25℃.The results showed that the content range of caffeic acid was 0.0076～0.038μg with a good linear relationship with peak area(r = 0.9995);the average recovery was 96.83%, RSD was 2.00%(n = 5).In total 17 amino acids were detedted from Aquilegia Viridiflora pall.and total content of them was 4.808%,including 7 essential amino acids in the human body(total content of them was 1.689%,35.13%of total amino acids).The content of moisture,total ash,acid-insoluble ash and extract were measured as 9.57%,9.62%,0.94%and 25.2%,respectively in the herb.Conclusions: The result of textual research of herbalism and pharmacognostical was accurate.The microscopic characteristics were clear;physichemical identification method was simple and sensitive;TLC method was specific,reproducible;HPLC method was sensitive,specific,reproducible and its detecting results was reliable.

The Analysis of the Influencing Factors and Clinical Significance of Coronary Collateral Circulation in Patients with Chronic Total Occluded (CTO) Coronary Artery Disease

Sun, 27 Mar 2011 04:11:57 +0000

Objective:The development of coronary collateral circulation(CCC) was a complicated process,and was influenced by many factors.The objective of the study was to investigate the significance of coronary collateral circulation in patients with chronic total occluded(CTO) coronary artery disease and factors influencing collateral channel development.Methods:The retrospective study was to assess the relationship between the coronary collateral circulation(CCC) and factors influencing clinical features coronary angiographic characteristics by means of univariate analysis and logistic regression analysis in specimens from patients with CCC group A(n=205) and patients with non-CCC group B(n=111) according to the Cohen and Rentrop\'s method which are used to score the CCC commonly.The clinical factors such as age,gender,cigarette use,alcohol use,blood pressure,cholesterol,duration of angina pectoris,angiographic characteristics of CTO arteries are concerned.Results:1 The result of CAG revealed that The proximal lesion promotes the development of collaterals of coronary artery.2 Univariate analysis indicated that the level of fasting plasma glucose(FPG) among the CTO patients,was lower in CCC group than non-CCC group(7.65±1.82 vs.10.65±3.86,P＜0.01);Furthermore,logistic regression analysis proved the result with a formula:Y(CCC)=11.341-2.150(FPG)+2.443(duration of the angina)(P=0.001).3 Duration of angina(＞6months) is significantly higher in group A than that in group B.Furthermore,logistic regression analysis proved the result with statistics(P =0.001).4 Electrocardiogram:Both group A and B had extensive ST segment depression,which revealed the possibility of chronic coronary occlusions with multivessel disease.Abnormal Q waves(26.3%vs 64.9%) were less frequently observed in group A than in group B.5 Univariate regression analysis revealed that there was negative correlation between other factors such as gender,age,cigarettes,alcohol,hypertension,the level of serum total cholesterol and CCC development.Conclusions::①The proximal lesions promote the development of collaterals of coronary artery.②Duration of angina(＞6 months) is an important factor for the presence of collateral vascular in chronic coronary occlusions.③There are some points that revealed the possibility of chronic coronary occlusions with multivessel disease.The points conclude permanent history of exertional angina and extensive ST segment depression formed.④The presence of proximal lesion of location and diabetes mellitus was independent predictor of collateral development.The diabetes mellitus restrains the development of collaterals.

The Analysis for the Characteristics and Risk of Coronary Myocardial Bridging and the Evaluation for the Role of hs-CRP between Myocardial Bridging and Coronary Artery Lesion

Sun, 27 Mar 2011 04:11:55 +0000

Objective:The objective of the study is to analyse the clinical characteristics as well as the coronary angiographic features of the patients with myocardial bridging(MB),evaluate the risk of MB for the coronary diseases,proceed the Logistic analysis including MB and other risk factors for coronary diseases,assess the correlation between coronary angiographic features of MB and cardiac events,and also explore the role and significance of hs-CRP for MB and cardiac events.Methods:We enrolled 474 patients and divided them into myocardial bridging(MB) group(n=330) and non-MB group(n=144) according to coronary angiographic results.We studied the clinical features and angiographic findings of MB patients by means of univariate analysis.We excluded 18 patients for incomplete datas,and divided the rest 456 patients,according to the angiographic results,into non-lesion group(n=163),one-lesion group(n=107),two-lesion group(n=82),and three-and-above lesion group(n=104).We grouped 115 patients into lesion-proximal-to-MB-in-LAD group and non-lesion-proximal-to-MB-in-LAD group for the sake of atherosclerosis degree there.Logistic analysis was made,including MB and other risk factors,to evaluate the risk of MB for its proximal lesion.Also,we divided 250 patients into myocardial infaction(MI) group and non-MI group,and assessed the risk of MB and other factors for MI.We measured mural coronary artery (MC) length,MC reference diameter(MC-RD),MC compression diameter(MC-CD), compression ratio of diameter(CR-diam),and compression ratio of area(CR-area) in 298 patients of atypical group(n=90),stable angina group(n=58),unstable angina group(n=77),acute myocardial infarction group(n=45),and others(n=28) excluding 32.We used ranksum test to make the evaluation between angiographic measurements with cardiac events.266 patients were detected high sensitivity C-reactive protein(hs-CRP) and were divided into MB group(n=210) and non-MB group(n=56). 210 patients were further subgrouped into lesion-proximal-to-MB group(n=113) and non-lesion-proximal-to-MB group(n=97).These 113 patients were divided into atypical group(n=6),stable angina group(n=14),unstable angina group(n=14),acute myocardial infarction group(n=18),and others(n=61).Results:The univariate analysis showed that the patients with MB were younger than non-MB(P＜0.001),and possessed higher diastolic pressure(P0.030), fewer diabetes(P＜0.001),especially fasting blood glucose(P＜0.001) and 2-hour postprandial blood glucose(P=0.025),higher HDL-ch(P=0.031) and Apo-A(P=0.024), lower BUN(P＜0.001),Scr(P＜0.001) and NT-proBNP(P=0.002),and higher hs-CRP(P=0.048).And about coronary angiographic features,MB group owned more patients with non-lesion(P＜0.001) and one-lesion(P＜0.001),while fewer with two-lesion(P＜0.001) and three-and-above-lesion(P＜0.001).Logistic analysis showed that the number of coronary arterial lesions is the independent risk factor for lesion in LAD,and its odds ratio(OR) is 4.046.CRP is the independent risk factor for the occurrence of myocardial infarction,and its OR is 1.035.MB has no significant influence on the occurrence of lesion proximal to MB in LAD as well as MI.The measurements of AMI group,MC-CD,CR-diam,CR-area,were higher than those of unstable angina group(P=0.009,0.006,0.007).MB group has higher hs-CRP than non-MB(P=0.048).There was no significant difference between lesion-proximal-to-MB and non-lesion-proximal-to-MB in LAD(P=0.893).We analyzed the relation between hs-CRP and the clinical cardiac events induced by lesion proximal to MB,and we found that AMI patients have much higher hs-CRP than other patients.Conclusion:The number of coronary arterial lesions is the independent risk factor for lesion in LAD.CRP is the independent risk factor for the occurrence of myocardial infarction.MB has no significant influence on the occurrence of lesion proximal to MB in LAD as well as MI.The length of MCA has no correlation with cardiac events,but its compression degree has significant difference in different events.CRP in patients with MB is higher than non-MB.But there is no significant difference if there is lesion proximal to MB in LAD.AMI patients have much higher hs-CRP than others,and there is correlation between hs-CRP and events induced by the lesion proximal to MB.

A Study on the Change of Physiological Function and Antioxidant State in Female Rowers during the Long Term Training

Sun, 27 Mar 2011 04:11:53 +0000

Objective:This research is based on the training of the female rowers in Wuhan Institute of Physical Education ,who were preparing for the Youth National Championships in 2008. During the long term training,we observed the changes in physiological function and antioxidant state ,managed to find out the relationships between the body composition and rowing performance. We compares the changes and trends between open class and light class female rowers ,attempted to find out some rules or problems by exploring the characteristics of their training and its relationship with variation of some physical ability indices during the long term training,and to provide a reference for the development of scientific in rowing training.Methods:During the whole training term, body composition indices were tested in early winter training term and a week before the competition, analyzed the rule and the relationship with rowing performance; physiological function indices including CK,BU, erythrocyte indexes and antioxidant state indices including MDA,SOD,GSH-PX were tested early in winter training term,end of two periods, the changes and trends of indices were also analyzed systematically with the training load.Results:1. During the whole training term,open class rowers’weight and fat free mass were significantly higher than light class rowers’(P<0.01), female rowers’weight and body fat% measured pre-race were lower than the early winter training , including a significant drop in body fat%( open class P<0.01, light classP<0.05).There was a significant correlation between the average fat free mass and 8 km rowing ergometer performance(open class r=-0.440, P<0.05; light class r=-0.578, P<0.05).2. The average serum CK and BU started to rise after a decline, CK measured pre- race had a decrease compared with the early winter training (light class P<0.05), and also had a significant increase after the competition (open class P<0.01, light class P<0.05).The average BU were relatively stable, and there were no significant inter-group differences between the open class and light class.3. The Erythrocyte indexes started to decline after rising,which measured in the end of second period compared with the early winter training and the end of first period separately,the indices of Hb, HCT, MCV, MCH and MCHC decreased significantly (P <0.01), and there were no significant differences between the open class and light class.4.The serum MDA level decreased after the rise, Which measured pre- race had a significant drop compared with the early winter training (open class P<0.01, light class P<0.05), and it showed a rise after the competition (open class P<0.05); Winter training did not have significance influence on the serum SOD ,but it showed a decline after the competition (light class P<0.01);the serum GSH-PX had the tendency to rise after the winter training(light class P<0.05),which decreased significantly in the end of second period(open class P<0.01, light class P<0.05). and there were no significant inter-group differences between the open class and light class. There were significantly relevance between the average MDA value and the average CK, BU values (P <0.05), the activity of antioxidant enzymes and serum CK, BU levels were negatively correlated, including a significant correlation between the serum GSH-PX activity and CK, BU level (P <0.05).Conclusion:1. FFM of female rowers has a significant correlation with their special ability, indicating an effective improvement in body composition is a valuable physiological determinant of performance.2.Aerobic endurance training is the main form of winter training ,it decreased the blood serum creatine kinase and urea levels, the activity of antioxidant enzymes were no significant changes while blood lipid peroxidation was reducing , erythrocyte indexes increased, function level and antioxidant status of rowers were good; After Increasing in exercise intensity and Volume,training and competition make a reduction in the rowers’function level and antioxidant status.3. The indexes have markedly changes during the long term training.There are individual differences of the physiological and biochemical indexes in female rowers, and a single index can not be used as the standard of the intensity increasing. 4.In this study,the changes of two indexes of the antioxidant states including MDA and GSH-PX were comparatively outstanding indicating that these two sensitive indexes could be recommended to be monitoring indexes to see the changes of the antioxidant states during training.5.System of special training will bring significant differences in body composition between open class and light class rowers ,while there are no significant differences on the physiological and biochemical indexes between groups. Exercise capacity of rowers may depend on the body\'s own capacity to adapt and adjust to the training load Independently.

The Characteristic of Variation in Muscle Oxygenation in Intermittent Motion

Sun, 27 Mar 2011 02:55:57 +0000

Objective: This study examined the relationship of muscle oxygenation determined by using near-infrared spectorscopy with the gas metabolism and investigate the effect of intermittent recovery time in muscle oxygenation in intermittent trainning.Methods: Eight well-trained male middle-long runner do warming-up 5 minutes, then performed 4 time same exercise that running at 85% maximal oxygen uptake for 4 minutes, then rest 2 minutes on a running machina.The second and third week subjects do the same exercise but rest 4 minutes and 8 minutes respectively. Two minutes before exercise untill the end of experiment, near-infrared spectroscopy and heart-lung machine were simultaneously recorded from right quadriceps femoris and face respectively.Results: In intermittent training,muscle oxygenation decrease rapidly at the start of exercise then decrease slightly, the two variation trend significantly different(P<0.05)in every exercise,muscle oxygenation increase quickly in every rest time. The rate of increase in muscle oxygenation levels at different intermittent recovery time and the rate of variation in muscle oxygenation after different intermittent recovery time were significantly different(P<.000). Respiratory quotient also decrease rapidly when muscle oxygenation decrease rapidly, then increase untill next exercise. The analyses showed that the variation trend of muscle oxygenation was positively correlated with the variation trend of repiratory quotient(r=0.724 P<0.05).Conclusion: In intermittent trainning,the rate of variation in muscle oxygenation during exercise was significant different that refect the energy metabolism of subjects. The rate of increase in muscle oxygenation levels at different intermittent recovery time and the rate of variation in muscle oxygenation after different intermittent recovery time were significantly different. The positively correlation between the the variation trend of muscle oxygenation and the variation trend of repiratory quotient suggested that the variation of muscle oxygenation reflects the variation of gas metabolism in intermittent trainning.

The Research of Feasibility on Maximal Oxygen Uptake (VO_2max) Measuring by Constant-load Exercise

Sun, 27 Mar 2011 02:55:55 +0000

Objective: Through researching the value of VO_2max which measured by incremental-load, 100% and 110%vVO_2max constant-load exercise, in order to offer the feasibility of using constant-load exercise to measure VO_2max in laboratory. To observe the relationship between VO_2max, VO_2max Pleteau Duration, the lasting time of vVO_2max (tlimvVO_2max) and the time to exhaustion (Tlim) of male students of physical education, supply more selective measuring methods for popularizing scientific exercise.Materials and Methods: Collecting 195 VO_2max protocols measured by incremental-load exercise of male students from Wuhan Institute of Physical Education ( age: 21.87±2.20y,height:173±3.50cm,weight:64.6±4.06kg ), take the velocity which reached VO_2max when measured by incremental-load exercise as 100%vVO_2max constant-load exercise. The average value of 100%vVO_2max is 16.32±1.12km·h-1. One of 15 participated in other two tests which exercised at 100% and 110%vVO_2max constant-load exercise as long as possible until they were totally exhausted. Take the value below 5% of VO_2max as the plateau value, the time of the plateau value is defined as VO_2maxPD. The time to exhaustion (Tlim) at three tests were measured by a stopwatch. We analyzed all results by 15s and 30s respectively(The expired air were averaged by every 15s or 30s). All protocols were dealt by SPSS 13.0.Results:1. The value of VO_2max measured by incremental-load exercise, 100%vVO_2max and 110%vVO_2max constant-load exercise are 3434±359ml/min, 3554±349ml/min, 3660±428ml/min, respectively. There is no great significant difference between incremental-load exercise and constant-load exercise(P > 0.05).2. There is a great significant difference of Tlim among incremental-load exercise, 100%vVO_2max and 110%vVO_2max constant-load exercise(P﹤0.01). The value are 596.87±61.48s, 214.0±77.65s, 144.6±53.12s, respectively. The Tlim of constant-load exercise is longer than incremental-load exercise.3. There is a great significant difference of tlimvVO_2max among incremental-load exercise, 100% vVO_2max and 110%vVO_2max constant-load exercis(eP﹤0.01). The value are 93.87±44.63s, 208.07±54.65s, 144.73±52.90s, respectively. And the tlimvVO_2max of constant-load exercise is much shorter.4. There is no great significant difference of VO_2maxPD among incremental-load ex- ercise, 100%vVO_2max and 110%vVO_2max constant-load exercise(P > 0.05), and the value are 71±35.62s, 67±32.50s, 56±30.78s, respectively.Conclusions: 1. There is no great difference of VO_2max between constant-load exercise and incremental-load exercise, indicate that we can take constant-load exercise as a laboratory way to measure VO_2max.2. It can reach one’s VO_2max in2-7min, at least 2min, when measured VO_2max by constant-load exercise.3. When we measured VO_2max by constant-load exercise in laboratory, it’s more fitting to take 100%vVO_2max as the intensity of constant-load exercise than 110% vVO_2max.

The Study on Metabolic Characteristics of Partial Skeletal Muscle Oxygen by Near-Infrared Spectroscopy Detection

Sun, 27 Mar 2011 02:55:53 +0000

Introduction:Research on the relative change between inflection points of respiratory quotient (RQ) and HbO2 in skeletal muscle during the process of increased intensity sports. Compare the occurrence priority and interval regularity of the two based on time, and analyze the physiological mechanism of synchronisms.Methods:12 rowing athlete candidates are chose to test to do increased intensity sports on dynamometer (CONCEPT-II). The output power of initial load is 50W, and then increased by 50W every 3 minutes until exhaustive. Monitor the relative changes of HbO2 in Quadriceps by Near-Infrared Spectroscopy (NIRS) during sports.Simultaneously determine and record the oxygen uptake of candidates in sports, including the intake of oxygen (VO2) and exhaled volume of carbon dioxide (VCO2), by the Cardiopulmonary Function Instrument MAX-II.Results:The inflection point of RQ is relatively changing toΔHbO2. That means there is a time delay in RQ. The delay in VO2 and VCO2 relative to HbO2, which is affirmed in the single-factor variance analysis test, is do exist.Conclusion:During increased intensity sports, the respiratory quotient changes regularly accompanied with the inflection point, and content of muscle oxygen (ΔHbO2) in quadriceps changes regularly with unconventional inflection point which might denote that of respiratory quotient. Inflection point of respiratory quotient has a regular delay time relative to that ofΔHbO2. The phenomenon provides powerful evidence for us to speculate inflection point of respiratory quotient by measuringΔHbO2 in Muscle-Oxygen Monitoring System. It may provide new methods to measure oxygen uptake.

Development of the Medical Security System of the Chinese Urban Residents

Sun, 27 Mar 2011 02:55:51 +0000

China\'s medical insurance system for urban residents is an important component of our country health care system.It is also an important safety net for protecting the urban residents\' health and life in our country.According to the development of medical security system in Japan and Canadian\'s representative of the National Medical Insurance System of it.as a warning,I found a good way for our country learning from the international experience.Internationally knowing at the residents of developed countries on health care system, I have outlined the history and current situation of our country medical insurance system for urban residents.China\'s medical insurance system experienced setting up,adjusting, regulating,setting up to explore this pilot project and some stages of a comprehensive.Till today,the basic medical insurance system of "individual medical saving account combined with social risk pooling fund" has established in city.However,the issue of exposing it is clear that residents are very acute contradictions in medical treatment.In order to effectively address the "difficult to see a doctor" questions for urban residents,I have Made a lot of suggestions and comments.Also the analysis of the problem and the social economic can be awareness and other reasons.All in all,to make the best use of the system,the means in this study are essential.We should not only make some short recommendations.We must be combined with the actual situation in our country to find a viable,sustainable development road construction in favor of the people\'s livelihood.

Intravenous Thrombolysis by Tissue-type Plasminogen Activator Deletion Mutant in a Rabbit Model of Retinal Artery Occlusion

Sun, 27 Mar 2011 02:55:48 +0000

Objectives:1.To observe the nature course of experimental retinal artery occlusion established by photochemical method in a rabbit model and to validate the efficiency of the RAO model induced by this method in a certain period.2.To compare the efficiency and safety of intravenous thrombolysis by two kinds of tissue-type plasminogen activator deletion mutants in a rabbit model of retinal artery occlusion established by photochemical method.Methods:1.The nature course of the experimental retinal artery occlusion induced by photochemical method in a rabbit model10 pigmented rabbits were used in this study.The RAO was created in a random eye by a laser which central wavelength was 532nm following intravenous injection of rose bengal(5%solution,50mg/kg) through marginal ear vein.A laser was applied to a certain artery which has a distance from the accompanying vein at the optic disc margin.One hour later,the RAO was confirmed by fluorescein angiography(FFA).The animals were follow-up in the 1st、3rd、5th and 7th day by direct ophthalmoscope、fundus photography and FFA.All experimental eyes were enucleated under anaesthesia for histology examination.2.Intravenous thrombolysis by two kinds of tissue-type plasminogen activator deletion mutants in a rabbit model of retinal artery occlusion induced by the photochemical method.30 pigmented rabbits were used and randomly divided into 3 groups. experimental r-PA(0.9MU/kg)、retavase(0.9MU/kg) and injectable water (10ml) was injected respectively through the marginal ear vein 1.5 hour later after the RAO model was created.4 hours and one day later,the RAO was observed by ophthalmoscope、fundus photography and FFA respectively while the blood was collected for PT、APTT、TT、Fbg、D-Dimer before drug administration and at lst、2nd、3rd、4th hour after that.all the experimental rabbits were examined by CT to exclude the intracranial hemorrhage.One day later after thrombolysis,All experimental eyes were enucleated under anaesthesia for histology examination.Results:1.The nature course of the experimental retinal artery occlusion induced by photochemical method in a rabbit model.At 1st and 3rd day the obstructed arteries didn\'t reopen in all experimental eyes.8 and 2 obstructed arteries reopened at 5th、7th day respectively.Histological examination showed that the vessel wall was integrated under the light microscope.2.Intravenous thrombolysis by two kinds of tissue-type plasminogen activator deletion mutants in a rabbit model of retinal artery occlusion induced by the photochemical method.reperfusion rate of experimental r-PA group、retavase group and control group was 70%、60%、0%respectively after drugs administration at 4th hour as same as at 1st day.Reperfusion rate of experimental r-PA group was not significantly different from retavase group\'s.TT、PT of retavase group increased significantly respectively from experimental r-PA group\'s.Fbg of retavase group decreased significantly respectively from experimental r-PA group\'s.There was no vitreous or retinal hemorrhage and no intracranial hemorrhage in all experimental rabbits. Histological examination showed that RAO was caused by intra-arterial thrombus and the vessel wall was integrated under the light microscope.Conclusion:There was no recanalization approved by FFA after 3 days in the rabbit model of retinal artery occlusion induced by photochemical method.It suggests that the RAO model can be used for thrombolysis study in 3 days. Two kinds of r-PA shows good thrombolytic effect and less tendency to hemorrhage complication,The thrombolytic effect has no significant difference between them.Compare to the retavase group,the experimental r-PA group shows less side-effect on the coagulation and fibrinolytic system.

The Effect of Shock Wave on Rabbit Ovary

Sun, 27 Mar 2011 02:55:45 +0000

Background:Urinary lithiasis is a common disease.We can choose ESWL、ureteroscopic lithotripy or classical open surgery to deal with distal ureteral stone.There is no controversy over male patients using ESWL,but the female patients,whose ovary would be exposed to shock wave energy during treatment of distal ureteral stone during ESWL,should be cautious.Till now,there is no comprehensive evidence-based meta analysis about the injury of adjacent organs during ESWL.Objectives:to study the adverse effect of shock wave on rabbit ovary at different distance from the focus.Method:25 female rabbits(2250-3000g) were divided into 5 groups.After anesthetized,put titanium clip direct or put titanium clip around the ovary in distance of 3 or 5 cm surgically.Put the animal on lithotriopter,used X-ray to confirm the titanium clip be in F2,and started shock waves attacking.The first group,ovaries were attacked directly on focus at 16KV,2000 shock number.The second and third group,ovaries were attacked at the same condition above,but the distance from the focus were 3cm and 5cm respectively.In the last two groups,ovaries were putted at the distance of 3cm from the focus, among them the fourth group using 18 kilovolts and 2000 shock number, the fifth group using 16 kilovolts and 3000 shock number.Took out the attacked ovary,observed it with naked eye and light microscopy, observed the pathological changes and analyzed the data.Progesterone and estradiol leveas were tested before and after the ovaries were attacked by shock waves in seven rabbits.Result:On gross examination,there was no remarkable pathologic changes in ovaries at the first three groups.Ovary hyperaemia and sub-capsular hematoma were found in the fourth and fifth group.All the ovaries attacked by shock waves were found discrete low-count parenchymal hemorrhage microscopically,but no structure damage was found.In the first group,the average number of red blood cells per high power field was 64-73.In the second and third group,the average number of red blood cells per high power field was 12-38 and 1-11 respectively.There was significant difference among three groups.In the fourth group the voltage was added to 18 KV,the average number of red blood cells per high power field was 56-128.In the fifth group,the shock number was added to 3000,the average number of red blood cells per high power field was 68-186.There was significant difference between the second group and the last two groups.Progesterone and estradiol levels were tested before and after the ovaries were attacked, there was no significant difference in two groups.Conclusion:The pathologic finding of the ovary following exposure to shock waves was only scattered low-count paranchymal micro-hemorrhage,no histological structure damage was found.The severity of the injury of the ovary on focus was higher than it near the focus,the injury of ovary at distance of 3 cm or 5 cm from the focus was much more slight successively.Ovary endocrine function before and after ESW was not changed.

Functional Magnetic Resonace Imaging in Patients with Optic Neuritis

Sun, 27 Mar 2011 02:55:27 +0000

Objective To establish the role of cortical neuroplasticity changes in examinations of patients with optic neuritis by using functional magnetic resonance imaging(fMRI).We compare the the results of the fMRI imaging,the central 15 degree visual field examination and pattern-visual evoked potential(P-VEP) to investigate the diagnosis and other informations of optic neuritis by fMRI.Subjects and Methods Subjects:Seventeen patients with optic neuritis seen from 2008.3 to 2009.3 in Huashan Hospital of fudan university.Methods:static visual field perimetry with Octopus101 perimeter, pattern-visual evoked potential,fMRI and visual tests were performed. Visual tests:visual acuity,pupillary light reflex and fundus of eye. P-VEP:We tested every patient\'s both eye by using RETI-port_gamma eye electrophysiology program,and the visual stimuli subtended a field of view of about 12×16 square degrees.The latency and amplitude of P100 wave was quantitatively analyzed,fMRI:Visual stimuli:The stimuli subtended a field of view of about 12 degrees,consisted of high contrast, drifting checkerboards.Two types of stimuli were used:the annular ring, either contracting or expanding,was used to map eccentricity and the rotating wedge,either clockwise or counterclockwise,was used to map the polar angle.MRI parameters:GE signa VH/i 3.0T scanner.Functional data: GRE-EPI sequence,20 slices lying perpendicular to the caicarine sulcus. Anatomical data was obtained using 3D-SPGR sequence to acquire high resolution.Functional data was presented to the inflated surface and subsequently analyzed by SPM5 software.Results Occipital cortex,inferior temporalgyrus,and postparietalgynus angularis were activated in patients of optic neuritis.The most of the activated areas were lacated around the fissure of occipital cortex.To parts of the patients,both the activated voxel number and the average signal intensity in stimulation of affected eye were lower than those in stimulation of unaffected eye.The mean degect of central 15 degree of visual field and the number of activated voxels in cortex was inverse correlation.The amplitude of P100 wave and the number of activated voxels in cortex were positive correlation.The area of activated voxels in cortex was manifest dilatation in parts of the patient,and their clinical history promote bad prognosis.Conclusion fMRI of visual activation reliable reflects the cerebral consequences of acute optic neuritis,it might provide clinically relevant information in diagnosis and appraisal the prognosis of optic neuritis.

Hearing Results on Patients with Choronic Otitis Media after Surgical Treatment

Sun, 27 Mar 2011 01:22:09 +0000

Objective To investigate the different influence of the hearing results following surgical treatment in patients of choronic otitis media.Methods 148 cases of choronic otitis media with surgical treatment all by author were followed up retrospectively 3 monthes and 1 year after surgery from March 2007 to May 2008.66 of male cases and 82 of female cases account for all the 148 cases.Otorrhea and progressing hearing decrease were the major symptom in all cases within which 43 of tinitus,26 of ear pain,11 of vertigo,1 of total unilateral deafness,50 of bilateral choronic otitis media and 15 of revision surgery were found.PTA and CT scan were perpetrated for all these cases.Hearing assessment was based on the PTA result and 0.5KHz,1KHz,2KHz were averaged for both air conduction and bone conduction.We use postoperative bone conduction and postoperative air conduction for calculating postoperative ABG.An additional high frequency bone coduction of average value of 2KHz,4KHz was estimated.All values were exhibitted as Mean±SD and statistically worked by SPSS 16.0.All surgery treatment made by author,with canal wall down tympanipalsty on 125 cases,canal wall up tympanipalsty on 17 cases,and myringoplasty on 6 cases.84 of PORP(Partial ossicular reconstruction prothesis),3 of TORP(Total ossicular reconstruction prothesis) and 11 of tragus cartilage were placed into middle ears for ossiccular reconstruction.We regularly interpositioned the cartilage chip between prothesis and tympanic membrane.Results During the follow-up we discovered 3 cases(3/148,2%) of tiny membrane perforation and 27 cases(27/148,18.2%) of tinitus.2 of the cases which received PORP implantation complained the recuurent otitis media and one resulted extrusion of prothesis(1/87,1.1%).Post-AC for 84 of PORP implanted cases was 39.6±14.9dB,improved by 13.9±13.2dB compared with the preoperative outcome;Average ABG was 20.5±8.5dB,improved by 8.8±10.7dB,47 of the 84 cases resulted ABG value less than 20dB(47/84,55.95%);High frequency bone conduction changed by 0.7±10.5dB.Statistically it was different of AC and ABG campared preoperative value with postoperative value(P＜0.001)(tab1).No Statistic difference was found in AC of 0.5KHz,1KHz,2KHz frequency(P＞0.05);Nevertheless ABG of 2KHz showed a unsatisfactory outcome than that of 0.5KHz,1KHz(P＜0.001)(tab2).Table 3 showed the ABG bins for the PORP group.Post-AC for 11 of ossicular reconstruction with autocartilage cases was 47.1±17.3dB,improved by 5.0±19.6dB;ABG was 26.2±12.8dB,changed by 3.2±16.5dB,6 of the 11 cases resulted ABG value less than 20dB(6/11, 54.5%);High frequency bone conduction changed by 3.6±10.9dB.Statistically difference was not found between preoperative value and postoperative value(P＞0.05).Post-AC for 3 of TORP cases was 42.7±16.4dB,improved by 14.4±9.5dB; ABG was 22.8±10.7dB,changed by 12.8±3.5dB,2 of the 3 cases resulted ABG value less than 20dB(2/3,66.7%);High frequency bone conduction changed by 6.7±11.3dB.Statistically difference was not found between preoperative value and postoperative value(P＞0.05).No statistic estimation was committed due to case insufficiency.Post-AC for 6 of myringoplasty cases was 21.1±4.3dB,improved by 13.6±7.2dB;ABG was 11.1±4.2dB,changed by 6.7±4.2dB,6 of the 6 cases resulted ABG value less than 20dB(6/6,100%);High frequency bone conduction changed by 3.8±9.2dB.Statistically difference was not found between preoperative value and postoperative value(P＞0.05).17 cases perpetrated with the canal wall up tympanoplasty in which the ossicular chain was reserved.Post-AC for 17 of that cases was 31.7±9.3dB,improved by 8.2±9.5dB;ABG was 15.8±6.3dB,changed by 3.8±9.4dB.High frequency bone conduction changed by 4.0.±7.9dB.No satistic significance was found in postoperatively hearing results beween noncholesteatoma otitis media and cholesteatoma otitis media(P＞0.05),and no satistic significance beween tympanosclerosis and nontympanosclerosis(P＞0.05),also between revision surgery and first surgery(P＞0.05).There was satistically significance found when comparing unilateral COM with bilateral COM(P=0.048).Conclusions It is satisfactory results for COM with surgical treatment and it is better using hydroxyapatite prothesis for ossicular chain reconstruction with cartilage interpositioned between tympanic membrane and prothesis.Closer follow-up is needed for bilateral COM.

The Primary Study of the Perilymph Drainage to Cervical Lymph Nodes in Guinea Pigs

Sun, 27 Mar 2011 01:22:07 +0000

Objective:To ascertain the connection between the perilymph and the cervical lymphatic system in guinea pig,and to explore the perilymph\'s drainage pathways to the cervical lymph nodes.Methods:23 healthy adult guinea pigs were randomly divided into 3 groups: perilymph group(group A,n=10),middle ear group with the mucosa scraped(group B,n=10) and middle ear group with the mucosa not scraped(group C,n=3).Group A and group B were randomly divided into 2 subgroups:group A1(n=7) and group A2 (n=3);group B1(n=7) and group B2(n=3).For the guinea pigs of group A,the nanocarbon(NC) suspension was injected into the perilymph in their left scala tympani through the round window membrane;for the animals of group B,the tracer was injected into their left middle ear cavity after the mucosa scraped;for the animals of group C,the tracer was injected into their left middle ear cavity without the mucosa scraped.The right side were not injected and served as a control.24 hours after the injection of the tracer,the animals of group A1,group B1 and group C were killed by intracardiac perfusion with normal saline;and 48 hour after the injection of tracer,the animals of the group A2 and group B2 were killed.The four groups of cervical lymph nodes(mandibular,parotid,superficial ventral cervical,and deep cranial cervical lymph nodes) were dissected and collected.The cranium was removed and the intracalvarium tissues and structures were carefully observed.Acoustic vesicles of both sides were collected and opened to observe whether the middle ear cavity mucosa,cochlea and vestibule were stained.The cervical lymph nodes and decalcifying inner ear were made into frozen section and then under H-E staining.The slices were observed with the microscope to find out the stained places and the stained degree.Results:The left parotid lymph nodes and bilateral deep cranial cervical lymph nodes in the animals of group A were black stained;in the animals of group B,only the left parotid lymph nodes and left deep cranial cervical lymph nodes were slightly black stained;in the animals of group C,none of the cervical lymph nodes was stained.The nanocarbon particles was seen in the perilymph of scala tympani and scala vestibuli, the periosteum of scala tympani and scala vestibuli,spiral ligament,space around spiral ganglion and spiral vein of modiolus,the membrane of sacculus proprius and alveus communis after it was injected in the perilymph of the scala tympani,and the endolymphatic sac was also obviously stained,but there was no nanocarbon particles in the endolymph either in the cochlea or in the vestibule.There was no nanocarbon particles either in the inner ears of the animals of group B or of group C.In the animals of group A,the bilateral arachnoid around the cerebellomedullary cistern and part of the cerebral pia mater of myelencephalon were stained black,and their left endolymphatic lacs were obviously black,and their bilateral basicranial external coat of vessels and epineurium of cranial nerves were also slightly black.The animals of group B and group C had no encranial structures stained.Conclusions:There are connections between the perilymph and the cervical lymph nodes in guinea pigs.There might be two drainage pathways:one is that the tracer in the perilymph diffuses to the endolymphatic sac through the connective tissue space surrounding the endolymphatic duct,and enters into the lymphatic capillaries around endolymphatic sac and was eventually drained into the ipsilateral parotid lymph node and deep cranial cervical lymph node;the other is that the perilymph could enter into the cerebral spinal fluid(CSF) through the space around spiral vein of modiolus and spiral ganglions,and then it enters into the lymphatic capillary around cervical external coat of vessels and epineurium through basicranial external coat of vessels and epineurium of cranial nerves,and finally drained to bilateral deep cranial cervical lymph nodes.

Expression of VEGF, Flt-1, Flk-1, PCNA and CD34 in Juvenile Nasopharygeal Angiofibroma

Sun, 27 Mar 2011 01:22:05 +0000

Objective To investigate the nature of juvenile nasopharygeal angiofibroma(JNA) through compared the differences of vascular endothelial growth factor(VEGF), VEGF receptor-1/ fms-like tyrosine kinase-1(VEGFR-l/Flt-1),VEGF receptor-2/ fetal liver kinase-1(VEGFR-2/Flk-l),proliferating cell nuclear antigen(PCNA) and CD34\'s expression between hemangioma and vascular malformation.Methods 28 cases of JNA,20 cases of orbital cavernous hemangioma(all of these were sorted by microscope as vascular malformation) and 10 cases of chronic rhinosinusitis were studied respectively by immunohistochemical S-P methods,to detected the expression of VEGF,Flt-1,Flk-1,PCNA and CD34,and the correlation between the expression rates of VEGF,Flt-1 and Flk-1 was also analyzed.Results The positive rates of VEGF,Flt-1,Flk-1 expression in the endothelial cells of JNA were 96.4%(27/28),100%(28/28),100%(28/28),higher than those in orbital cavernous hemangioma which were 10%(2/20),40%(8/20),45%(9/20), there were significant difference(P＜0.05).The average microvessel density(MVD) marked by CD34 in JNA was(49.3±9.1)/HPF(high power field),higher than orbital cavernous hemangioma which was(29.1±6.7)/HPF,there were significant difference (P＜0.05).The PCNA expression in the endothelial cells of JNA and orbital cavernous hemangioma were both positive,there were no significant difference (P＞0.05).The PCNA expression in stromal cells of JNA were positive(100%),the PCNA expression in stromal cells of orbital cavernous hemangioma were negative, there were extraordinary signifycant difference(P＜0.05).Conclusion The expression of VEGF,Flt-1 and Flk-1 in JNA were higher than those in orbital cavernous hemangioma,and there was a correlation between the expression of VEGF,Flt-1 and Flk-1 in JNA.This showed us the expression of VEGF, Flt-1,Flk-1 may be the important factors and play a crucial role in the development of JNA,the nature of JNA could be a hemangioma.The expression of VEGF,Flt-1 and Flk-1 could be an effective sign to differentiate hemangioma and vascular malformation.

The Utilization of the Fourth Generation Fourier Domain Anterior Segment OCT (FD-OCT) in the Measurement of Tear Meniscus and the Diagnosis of Dry Eye

Sun, 27 Mar 2011 01:22:03 +0000

Dry eye is caused by a variety of reasons,as well as tear deficiency,corneal or conjunctival epithelial damage as chronic disorders.The cause is more complex, clinically misdiagnosed easily.The reproducibility of dry eye diagnostic criteria is not satisfied and there are many factors to interfere the results.So it is very important to search for a rapid and reliable method for diagnosis of dry eye.With the development of optical coherence tomography(optical coherence tomography,OCT) and further application in anterior segment,the fourth-generation Fourier domain(Fourier domain) OCT has beyond the characteristics of the previous time-domain(Time domain) OCT with high resolution and high speed.It has led to widespread concern researchers with a high-resolution functional imaging of anterior segment of tear meniscus.However, there is no research on the effectiveness of the Fourier-domain OCT in the diagnosis of dry eye and normal tear meniscus area.Therefore,this study will probe on the diagnostic effectiveness and tear meniscus imaging of fourth-generation Fourier domain OCT.We enroll in different ages,different types of dry eye patients,as well as normal controls to explore diagnostic effectiveness of Fourier domain OCT,in order to identify the type of dry eye which has a higher diagnostic effect.At the same time,we make the normal control group into Sub-groups in accordance with age to investigate age-related variations of human tear meniscus in Chinese.The results show that the fourth generation Fourier domain OCT can clearly image and measure the cross-sectional tear meniscus.It also has a good consistency with clinical diagnosis of dry eye examination(BUT,Schirmer test).As a non-invasive,non-contact,simple screening method,the fourth generation Fourier-domain OCT has a good application prospects in the diagnosis of dry eye,especially in aqueous tear deficiency. PartⅠResearch on the tear meniscus imaging and measurement of the anterior segment Fourier domain OCTObjective To discuss the utilization of the fourth generation anterior segment Fourier domain optical coherence tomography in the imaging and measurement of tear meniscusMethods We enrolled 146 cases(146 eyes) and 160 controls(160 eyes) in this study. The tear meniscus height(TMH),tear meniscus depth(TMD) and tear meniscus area (TMA) of each participant were measured by Fourier domain OCT(RTVue-100).We also estimated the tear break up time(BUT),performed the cornea fluorescein staining and Schirmer test,and recorded score of the dry eye questionnaire.The variability of the fourth generation anterior segment Fourier domain OCT in tear meniscus imaging and measurement was discussed.Results The average values of TMH,TMD and TMA in the patient group were 232.319±17.549(μm)、151.494±12.811(μm)、0.024428±0.003712(mm2), comparing with 328.113±19.036(μm)、220.906±12.935(μm)、0.045618±0.005623(mm2) in the control group,the difference of which reached significant (TMH t=12.2646,P=0.0000,TMD t=12.0567,P=0.0000,TMA t=10.1556, P=0.0000).The individual variability of the TMH,TMD and TMA in the patient group was less than the control group.The repeatability analysis showed a small variation in measurement.The OCT images of the bound and cross section area of the tear meniscus and the clarity of the tear fluid in the patient group were obviously different from those in the control group.Conclusion The fourth generation Fourier domain anterior segment optical coherence tomography can clearly image and measure the cross section of tear meniscus,the result of which showed a good coherence with the traditional diagnostic test(BUT,Schirmer test) and directly reflect features of the tear meniscus. PartⅡAge-Related Variations of Human Tear Meniscus in ChineseObjective To investigate age-related variations of human tear meniscus in Chinese and diagnosis of dry eye using Fourier domain anterior segment OCT.Methods.We enrolled 146 cases(146 eyes) and 160 controls(160 eyes) in this study.All the subjects in these two groups were divided into four subgroups according to different ages,respectively,group A(0～19),group B(20～39),group C (40～59) and group D(over 60).Dry eye patients have 6,60,60,20 in each group, normal controls have 6,60,60,20 in each group.The tear meniscus height(TMH),tear meniscus depth(TMD) and tear meniscus area(TMA) of each participant were measured by Fourier domain OCT(RTVue-100).We also estimated the tear break up time(BUT),performed the cornea fluorescein staining and Schirmer test,and recorded score of the dry eye questionnaire.The relativity of the clinical examinations was tested,the tear meniscus of the two groups were measured and ROC curves were drawn.Results There is a significant correlation among tear meniscus values,individual syndrome and clinical diagnoses.The mean TMH、TMD、TMA for dry-eye patients were significantly lower than those for control groups.There was negative correlation between tear meniscus values and age for Chinese healthy subjects(TMH r=-0.0649,p=0.000,TMD r=-0.0299,p=0.0024,TMA r=-0.0320,p=0.0017).The accuracy of the diagnosis of dry eye by Fd-OCT was TMH 68.95%、TMD 70.59%、TMA 70.92%.It\'s different between the age sub-groups.Area under the ROC curve (AUC) showed that overall,TMH to 0.7337,TMD is 0.7477,TMA is 0.7461.AUC of all ages there are some differences between.TMH Diagnostic value of the overall community for the 259(μm),TMD for 165.6667(μm),TMA was 0.024(mm2),the diagnosis of all ages increase community values decline with age.Conclusions Fd-OCT could instantly imply the age-related variations of human tear meniscus and has good consistency with the clinical diagnose examinations.It has different diagnostic accuracy in each age sub-group. PartⅢThe utilization of the fourth generation Fourier domain anterior segment OCT(FD-OCT) in the diagnosis of dry eyeObjective To discuss the utilization of the fourth generation anterior segment Fourier domain OCT in the diagnosis of different types of dry eye patients.Methods We enrolled 146 cases(146 eyes)of dry eye patients(DE group),102 cases(102 eyes)of aqueous tear deficiency patients(ATD group),81 cases(81 eyes)of lipid tear deficiency patients(LTD group),53 cases(53 eyes)of Sj(o|¨)gren syndrome patients(SS group)and 160 controls(160 eyes) in this study.The tear meniscus height (TMH),tear meniscus depth(TMD) and tear meniscus area(TMA) of each participant were measured by Fourier domain OCT(RTVue-100).We also estimated the tear break up time(BUT),performed the cornea fluorescein staining and Schirmer test,and recorded score of the dry eye questionnaire.The relativity of the clinical examinations was tested,the tear meniscus of the two groups were measured and ROC curves were drawn.Results The average values of clinical measurements in the patient group were quite different.The results of the diagnostic effectiveness for each group showed,SS group had the highest diagnostic effectiveness,TMH 91.19%,TMD 92.51%,TMA 92.51%; LD group had the lowest,TMH 45.49%,TMD 67.06%,TMA 46.27%.DE group was almost the same with ATD group.The area under the ROC curve(AUC) of TMA showed,DE group was 0.7855,AD group was 0.7110,LD group was 0.5279,SS group was 0.9706.Conclusion The fourth generation Fourier domain anterior segment optical coherence tomography can accurately measure all types of dry eye tears meniscus,, the result of which showed a good coherence with the traditional diagnostic test(BUT, Schirmer test) and directly reflect features of the tear meniscus.As a non-traumatic and non-contact technique,the fourth generation Fourier domain anterior segment OCT has a favorable prospect in the diagnosis of dry eye,especially in aqueous tear deficiency.

The Analysis of Cornea Refractive Surgery Based on the Finite Element Method

Sun, 27 Mar 2011 01:22:01 +0000

The construction of virtual organ is one of the frontier research areas. As one of the most important and tiny organs of human, eye should be modeled and simulated factitiously. Cornea—as one of the most important part of eye affects the refraction of the eye most. Corneal refractive surgery by changing the corneal refractive power to achieve the purpose of correcting refractive errors. It is welcomed among people because the surgery is convenient and the curative effect is remarkable. The doctor and patient are always concerned about the stress distribution before and after surgery and the effect of parameters\' changes in the operation. In this paper, the finite element models were built respectively before and after refractive surgery with the finite element software ANSYS. The stress distribution of the cornea before and after surgery, the impact of intraocular pressure to the cornea was analyzed. This can predict for the different scenarios of actual surgery, and can provide practical guidance for the refractive surgery.In this thesis, the establishment of a number of corneal refractive surgery before and after the finite element model based on the simplified model of the cornea on the physical form of the cornea; and then in the establishment of the finite element model based on the current common of refractive surgery (RK, LTK, LASIK) simulation and analysis, to explore the realization of the principle of the refractive surgery, and a detailed analysis of intraocular pressure on the impact of corneal refractive surgery and the operation parameters on the corneal refractive surgery the impact of.Followings are the innovations of this thesis:1. The finite element models of cornea before and after the RK surgery were constructed, and simulated the corneal shape before and after the RK surgery. Then analyze the effects of intraocular pressure in normal circumstances and after RK surgery to the refractive surgery, and study the impact of the depth of the surgical incision to the surgery. 2. First of all, the corneal thermal analysis under the laser irradiation was carried out to examine the cornea after laser irradiation in the heat distribution and heat transfer situation based on the characteristics of the LTK surgery; And then on the basis of the establishment of post-operative LTK finite element model, comparative analysis the intraocular pressure in the same role in different optical zone radius of the impact of surgery on the LTK and the same under the different optical zone diameter of the intraocular pressure of the impact of corneal.3. The laser in situ keratomileusis (LASIK surgery) was simplified and analyzed. First the post-operative finite element model of the cornea was set up, and then a detailed analysis of the surgical cutting of the effect of thickness on the impact of surgery and intraocular pressure effects of surgery the impact of, and RK surgery and LASIK surgery for the treatment of myopia has done.

The Techniques of Expanded Studying in Normal Blink Reflex

Sun, 27 Mar 2011 01:21:59 +0000

Objective Blink Reflex is one of the brainstem reflexes that can be evoked by light,sound or touching to cornea.It is a defensive reflex which has an important effect to protect the eyeball and face.The detection of blink reflex can evaluate the functional status not only the peripheral nerves such as trigeminal nerve and facial nerve but also the central nervous system Now the technique commonly used in clinic is elicited electrically by delivering percutaneous stimulation over the supraorbital nerve at the supraorbital foramen.Responses are recorded from surface electrode positioned over the right and left orbicularis oculi.By the electrodiagnostic recording we get the early,simple R_1 and late,complex ipsilateral R_2 as well as contralateral R_2\'.To research the different techniques and wave characteristic of applying non-invasive stimulation in Blink Reflex we selected thirty healthy volunteers and stimulated their supraorbital nerve and mental nerve then respectively recorded at orbicularis oculi and levator labli superioris.Through these different methods we can establish the laboratory data so as to offer basic physiological index for clinical using.Methods We selected 30(60 sides) healthy young volunteers.Male 14,Female 16,Age 22～28years old(average 24.1±1.5 years old).According to the following criteria:①No medically contributory conditions select.②No neurologic conditions select.③facial muscular systems are within normal limits.④No surgical or traumatic history on skull or maxillary and mandible.The machine is DISA-2000M oculomusclar electromyogram made by Danmark,the recording electrode is surface disk electrode withφ1cm.Using the electrodiagnostic techniques,orbital nerve and mental nerve were stimulated and the wave were respectively recorded at orbicularis oculi and levator labli superioris,then calculated the latency and amplitude of R_1,R_2,R_2\'.All data were dealt with statistical software SPSS13.0 for windows.There was a statistical significance when P＜0.05 occurred.Results1.The 30 healthy cases(60 sides) all obtained satisfied wave when stimulating the supraorbital nerve then recording at orbicularis oculi and levator labli superioris.Three different components of blink reflex were observed.The early ipsilateral response(R_1) was single and sharpen.The late ipsilateral response(R_2) and late contralateral response(R_2\') were constituted by several waves,the latency were long and the form were complicated.The images were consitantly and repeatable.2.The waves of stimulating mental nerve then recording at orbicularis oculi and levator labli superioris were unstable.Some were complete and others R_1 were absent.For these two kinds of method the present rate of R_1 were 67%and 60% respectively.But bilateral R_2 were all stable.3.The latencies of R_1,R_2,R_2\' were Normal distribution by the four detected methods.In statistic analyse we adopted t-test and pair t-test that the sex had not significant difference between male and female,it was the same for the left and right sides (P＞0.05).So on the subsequently statistic analyse we did not consider the sides and sex.The variation between the four groups we adopted Analysis of Variance(ANOVA).The normal value of latency was expressed by(?)±s and top limit of normal value was expressed by(?)+3s.The wave amplitude varied obviously and it was not normal distribution by D-test.So we used median to describe it.Conclusion The techniques of stimulating orbital nerve then recording at orbicularis oculi and levator labli superioris are of the same value for clinical diagnosis.The techniques of stimulating mental nerve then recording at orbicularis oculi and levator labli superioris sometimes are unstable so they can be used as supplementary examination.These detected methods are non-invisive and easy that offer impersonality and quantitated criteria.They are reliable electrophysiological technique.

Assessment before Uvulopalatopharyngoplasty and Acoustic Change after Uvulopalatopharyngoplasty

Sun, 27 Mar 2011 01:21:57 +0000

PARTⅠCorrelation between subjective assessment and objective measurement of obstructive sleep apnea hypopnea syndromeObjective To investigate the correlation between subjective assessment and objective measurement of obstructive sleep apnea hypopnea Syndrome (OSAHS).To provide accurate and systematic assessment method before uvulopalatopharyngoplasty(UPPP).Method Data of 200 patients with OSAHS were collected retrospectively. Assessment of snoring was achieved by visual analogue scale(VAS) .Assessment of excessive daytime sleepiness was achieved by Epworth sleepiness scale(ESS).The quality of life was assessed by sleep apnea quality of life index(SAQLI).The association between the scores and polysomnography(PSG) parameters in OSAHS patients was examined statistically.Result①There was a significant correlation between VAS、ESS and SAQLI scores(P＜0.01);②VAS had significant correlation with apnea hypopnea index(AHI),lowest oxygen saturation(LSaO2),cumulative percentage of sleep time with SaO2 below 90%(CT90%) and the longest time of breathing disorder(BDLon)(r=0.4834、-0.4101、0.4316、0.3979,P＜0.0001),so ESS did(r=0.4215、-0.4106、0.3007、0.2899,P＜0.01).The mean of ESS or VAS in severe OSAHS group was higher than that in mild and mederate OSAHS groups,but no diference was found between the mild and moderate OSAHS groups.The mean of ESS or VAS in severe hypoxemia group was higher than that in no,mild and moderate hypoxemia groups,but no diference was found between no,mild and moderate hypexemia groups.③There was a mild correlation between several domains on the SAQLI and several PSG parameters.There was little correlation between SAQLI and the severity of OSAHS.Conclusion①Both ESS and VAS have significant correlation with the para meters of PSG,but there is no good correlation between SAQLI and PSG examination;In order to evaluate OSAHS patients before UPPP properly, both subjective assessment and objective examination must be considered.②Both ESS and VAS have little value in assessment of OSAHS.They can be a primary screening method only for suspected patients with severe OSAHS. PARTⅡEvaluation of ApneaGraph in the assessment before uvulopalatopharyngoplastyObjective ApneaGragh is a new device for simultaneous determination of obstructive sleep apnoeic events and continuous pressure measurements for localization of upper airway obstructive sites.This study is to evaluate the clinical value of ApneaGragh in the assessment before uvulopalatopharyngoplasty (UPPP).Method Thirty-nine patients diagnosed as obstructive sleep apnea hypopnea syndrome(OSAHS) underwent a PSG and an ApneaGraph study simultaneously 2 days before UPPP.ApneaGraph\'s results were correlated with both the Friedman staging system and polysomnography statistically.Result①There was a good correlation between measurements of AG and PSG for apnea hypopnea index(AHI) and lowest oxygen saturation(LSaO2) (r=0.8771、0.7946,P＜0.01);Except for hypopnea index(HI)(p=0.021) and mixed apnea index(AI)(p=0.002),our study showed no differences between PSG and AG for other parameters;The mean of AHI on back body position was higher than side body position(P＜0.05).②The%retropalatal obstruction was 79.91%,71.53%and 78.66%in stages 1,2 and 3, respectively.There was no significant difference between stages(P＞0. 05).Conclusion ApneaGraph can serve as a portable PSG to diagnose sleep disordered breathing qualitatively.In the meantime,it also can identify the level of upper airway obstruction.It seems that ApneaGraph may be used to select patients for UPPP widely. PARTⅢAssessment of acoustics after uvulopalatopharyngoplastyObjective To determine the effect of uvulopalatopharyngoplasty(UPPP) on the voice quality.Method Twenty-six adult patients diagnosed as obstructive sleep apnea hypopnea syndrome(OSAHS) underwent the UPPP.Researches on quality assessment of speech were made preoperatively(1 week) and postoperatively (about 6 months).Patients\' subjective gradings included voice handicap index,voice-related quality of life and nasality questionnaires.The method of objective assessment was acoustic analysis of vowels.The preand postoperative articulation results were analysed statistically.Result No patient suffered from disorders such as hypernasality and articulation errors.No significant differences were found before and after UPPP regarding the questionaires\' score,fundamental frequency and Intensity(P＞0.05).But several formants changed:the third formant frequencies(F3) in vowel[o]、[e]and[u]decreased statistically,while the second formant(F2) in vowel[i]and the fourth formant(F4) in vowel [o]increased statistically(P＜0.05).Conclusion The findings of the study indicate that UPPP doesn\'t have a significant impact on daily speaking.But several formants changed, which is important in management of patients with concerns about postoperative vocal quality,such as singers and professional speakers.

Long-term Outcomes of LASIK for Hyperopic Anisometropic Anblyopia in Children

Sat, 26 Mar 2011 17:31:33 +0000

Objective:To evaluate the long-term outcomes of LASIK for hyperopic anisometropic amblyopia in children;to investigate the long-term visual quality and the factors affecting long-term visual quality of hyperopic LASIK in children.Methods:40 eyes of 40 children(age 7 to 18 years),57 eyes of 57 children(8 to 17 years) and 10 eyes of 10 children(6 to 16years) with hyperopic anisometropic amblyopia were treated by LASIK,spectacles and contact lenses respectively. All the three groups were given the routine examination included UCVA, subjective manifest refraction and cycloplegic refraction,BCVA.Then all the three groups patched the dominant eyes,and the mean follow-up were 21.65±13.09m, 18.93±8.83m,21.50±13.68m respectively.In LASIK group,Humphry Atalas 995(Carl Zeiss Meditec) was used to measure corneal curvature K1/K2,Oculus Pentacam was used to measure corneal thickness,Hartmann- Shack aberrometer (WASCA,Carl Zeiss Meditec) was used to measure the overall wavefront aberrations in scotopic pupils and the data was analyzed in 5mm diameter,IOL Master(Carl Zeiss) was used to measure the length of axis.TAKAGI CGT-1000 was used to measure the contrast sensitivity and glare sensitivity in LASIK group after the correction of refractive errors 2 weeks later.Titmus Stereo Tests was used to measure the stereopsis in LASIK group and spectacles group with the correction of refractive errors 2 weeks later.Stata 10.0 was used to analyze the data of cycloplegic refraction,BCVA,high order aberration,contrast and glare sensitivity,and stereopsis.Results:The mean postoperative SE refraction,SE anisometropia and LogMAR BCVA of the treated eyes in LASIK group were 1.22±1.34(D),1.80±1.07(D) and 0.37±0.23 respectively.All the changes postoperative were statistically significant(P＜0.05). Mean postoperative SE refraction,BCVA of the treated eyes in spectacles group and contact lenses group were 5.39±1.24(D),0.40±0.32 and 5.17±1.43(D),0.30± 0.19 respectively.The mean corneal curvature K decreased 2.09±1.20D by the latest follow up.87.5%of all LASIK eyes were within±1.00 D(including±1.00 D) of the attempt SE refraction.When preoperative central corneal thickness was not more than 522um,when optic diameter was less than 6.5mm,when the ablation depth was not less than 116um,and when the postoperative corneal curvature was not less than 46.56D,the correct assessment percent of that achieved SE was beyond±1.00 D of attempt SE refraction were 81.40%,65.12%,55.81%,67.44%respectively.The percent of stereopsis less than 60 seconds of arc was 0%preoperatively and 23.5% postoperatively.Contrast sensitivity were increased statistically significant in 0.7°、1°、1.6°and glare sensitivity were increased statistically significant in 0.7°、1°、4°(P＜0.05).The differences of Zerike coefficient Z(4,0),RMSh,RMS3, RJVIS4 and RMS5 between postoperative and preoperative were statistically significant.(P＜0.05),but all those differences were not related to age,preoperative central corneal thickness,preoperative refraction,optic diameter and the ablation depth.The difference of increased LogMAR BCVA between LASIK group and spectacles group was not statistically significant(P＞0.05).But the increased LogMAR BCVA was lager in LASIK group than in RGP group(P＜0.05).No intraoperative and postoperative complication was observed.Conclusions:LASIK for hyperopic anisometropic amblyopia in Children was safe, effective,and predictable;individual design would improved the outcomes of LASIK for hyperopic anisometropic amblyopia in Children;LASIK was alternative treatment in children with moderate and severe hyperopic anisometropic amblyopia who were unable to tolerate or failed with conventional methods of treatment;the changes of high order aberrations,contrast sensitivity and glare sensitivity after hyperopic LASIK need further research.

Rescue of Photoreceptors by BDNF Gene Transfer Using in Vivo Electroporation in the RCS Rat of Retinitis Pigmentosa

Sat, 26 Mar 2011 17:31:31 +0000

PartⅠConstruction of BDNF-GFP fusion expression plasmid vector and study on its characteristicsObjective To construct a BDNF-GFP fusion expression plasmid and study its characteristics to supply basic evidences for its future\'s application in the study of gene therapy.Methods The BDNF CDNA was amplified by genomic PCR and then were inserted into pcDNA3.1/NT-GFP-TOPO plasmid and verified by sequencing.Result The construction of BDNF-GFP plasmid was verified correctly by sequencing。Conclusion The BDNF-GFP fusion expression plasmid vector was successfully constructed.Our study could provide a new method for the construction of target vector for gene therapy.PartⅡStudy of gene transfer into retinal pigment epithelium cells and photoreceptors using in vivo electroporationObjective To investigate the feasibility and the suitable electro-pulse model of transfecting DNA into retinal pigment epithelium(RPE) cells and photoreceptors(PRs) in vivo by electroporation method.Methods pEGFP-N1,the eukaryotic expressive plasmid of enhanced green fluorescent protein(EGFP) was injected into the subretinal space of Sprague—Dawley(SD) rats and serial electric pulses were applied by in vivo electroporation.Whole mounted retinas were made one week after introduction of plasmid and the fluorescence of EGFP was photographed by fluorescent microscope.The expression of EGFP mRNA and protein was detected by reverse transcription polymerase chain reaction technique(RT—PCR) and Western immunoblot analysis.Results EGFP gene was successfully transfected into RPE cells in vivo by a series of electric pulses,and the most suitable electro-pulse model is 10V/mm electric field strength with 99ms duration to each pulse,5 pulses with 0.5s interval each other.EGFP expression in PRs was not detected at present.Conclusion These findings indicate that electroporation is an effective method for gene delivery into RPE cells in vivo,but not for PRs.This new method may be an assistant tool for gene transfer into RPE cells in vivo,explore a potential treatment for retinitis pigmentosa.PartⅢRescue of photoreeeptors by BDNF gene transfer using in vivo electroporation in the RCS rat of retinitis pigmentosaObjective To investigate the feasibility of introducing brain-derived neurotrophic factor(BDNF) gene into retinal pigment epithelium(RPE) cells in vivo by electroporation and whether this method can rescue photoreceptors(PRs) of retinitis pigmentosa(RP) in Royal College Surgeons(RCS) rats.Methods The BDNF-GFP fusion eukaryotic expressing plasmid was constructed and then was subretinally or intravitreously injected into the eyes of Royal College Surgeons(RCS) rats followed by in vivo electroporation.The expression of BDNF mRNA and protein were detected by RT-PCR and Western immunoblot analysis.The number of surviving photorecptors was counted and the TdT-dUTP terminal nick-end labeling(TUNEL) method was used to detect the apoptosis of the retinal cells at different time points after introduction of BDNF plasmid.Results After the electrointroduction,the treated eyes showed a significantly higher rescue ratio and a lower number of TUNEL-positive photoreceptors than the control eyes at various time points. Conclusion These findings provide evidence that electroporation is an effective method for genes transfer into RPE cells and the rescue of PRs can be achieved by BDNF gene transfection with electroporation.

Tissue Factor Pathway Inhibitor Effects Tissue Factor in Early Stage Atherosclerosis

Sat, 26 Mar 2011 17:31:30 +0000

Objective:The objective of this study is to evaluate whether to add the low dose of tissue factor pathway inhibitor(TFPI) long time decreases tissue factor(TF) to ApoE-/- mice by plasmid VRnew in the early stage of atherosclerosis.Methods:Feed three 8-week-old female ApoE-/- mice with west diet.Transfect mice with 60ug VRnew-TFPI plasmids at their muscle of posterior limbs after 2 weeks,then take mice\'s blood at 4th,14th and 20th day after transfaction,Add natrium citricum solution as 1:9(v:v) for anticoagulation.Measure the concentration of blood human TFPI by ELISA kit.Feed 14 6-week-old female ApoE-/- mice through the construction with west diet,According to weight of mice in 8-week to divide into 2 groups.Inject 30ul plasmid(concentration lug/ul) into muscle of every posterior limb of mice then 150V/cm square pulse shock 8 times after 2 weeks. VRnew—TFPI group(7 mice) transfects VRnew-TFPI plasmid and VRnew group(7 mice) transfects VRnew empty plasmid.Repeat every two weeks,total 6 times. 20-week-old mice are sacrificed and take aorta.Make paraffin sections.Stain TF antigens by immunohistochemistry.Employ with Image-Pro~(?) Plus V6.0 to deal with the result of immunohistochemistry,read optical density and area,count average density.Analyze the result.Results:Measure the plasma concentration of human TFPI by ELISA,4~(th) day after transfect plasmid 2.274±0.594 ng / ml;14~(th) day 1.676±0.069ng/ml;20~(th) day 0.486±0.3325ng/ml.Divide 14 8-week-old mice into two groups according to weight at that time,the average weight of VRnew—TFPI group is 18.34±1.90g and the average weight of the VRnew group is 18.85±1.39s.P＞0.05.We can observe white atherosclerotic plaque in the aorta after the mice were sacrificed,then observe plaque under microscope after HE staining.Immunohistochemistry method marks TF antigen on the aorta,Image-Pro~(?) Plus V6.0 software reads optical density and area,calculate average optical density value,VRnew—TFPI group value is 0.23±0.08 and VRnew group value is 0.34±0.10,P＜0.05.Conclusions:Given long-term low-dose TFPI,tissue factor in mice\'s aorta will be decreased in the early stage of atherosclerosis.

Arsenic Trioxide Inhibited the Growth of Trasplanted Tumor of Human Gallbladder Carcinoma and Its Mechanism in Nude Mice

Sat, 26 Mar 2011 17:31:28 +0000

Background:Gallbladder carcinoma,a common carcinoma in biliary system,lacking of specific clinical manifestation,usually diagnosed at advanced stages of the diseases and fewer can be resected in operation.Chemotherapy and radiotherapy has not shown significant activity in gallbladder carcinoma,so new agent should be applied to its treatment.Arsenic trioxide,verified as a breakthrough in the management of acute promyelocytic leukemia,has been applied to the research and the therapy of a variety of solid tumors in recent years.However,the research into the effect of As_2O_3 on gallbladder carcinoma has not been widely reported.Purpose:To investigate the inhibitory effect of As_2O_3 on the growth of transplanted tumor of human gallbladder carcinoma and its mechanism in nude mice.Methods:Twenty BALB/c nude mice,with the resembling volume of subcutaneous transplanted tumor,were randomly divided into nature saline(NS) and As_2O_3 groups. 0.2mL NS or 10mg As_2O_3/kg was injected into the tail vain of every mouse in NS group or As_2O_3 group for seven days,once per day.All mice were put to death fourteen days after the perfusion was performed,then the weight and dimension were assayed,the apoptosis cells were determined by TdT-mediated dUTP Nick-end Labeling(TUNEL) and counted as well as the expression of Bcl-2 and Cyclin D1 was detected by immunohistochemical method and Western Blot.Results:The volume and weight of transplanted tumor was 1.50±0.57cm~3 and 0.992±0.401g in NS group,which reduced to 0.83±0.39cm~3 and 0.526±0.269g in As_2O_3 group.The apoptosis cells were 22.2±3.79 in NS group,which elevated to 57.9±9.75 in As_2O_3 group.The expression of Bcl-2 and Cyclin D1 in As_2O_3 group was lower than that in NS group which was detected by immunohistochemical method and Western Blot.All the difference were significant.Conclusions:As_2O_3(10mg/kg) can significantly inhibit the growth of transplanted tumor of human gallbladder carcinoma in nude mice.The mechanism has correlation to inducing apoptosis and downregulating the expression of Bcl-2 and Cyclin D1.

Characteristics of Visual Field and Retinal Nerve Fiber Layer Defect in Advanced Glaucoma

Sat, 26 Mar 2011 17:31:27 +0000

Characteristics of Visual field defect in advanced glaucomaPurpose:To study the clinical features of visual field defect in advanced glaucoma.Methods:A retrospective non-controlled study.357 patients of primary open angle glaucoma,acute primary angle-closure glaucoma and chronic primary angle-closure glaucoma were enrolled and interviewed in our hospital between 2003 to 2008. Patients with the central 5-10 degrees of visual field or the temporal peripheral field were screened from glaucomatous visual field data.Then the shape and the distribution were analyzed.In addition,we calculated the ratio of each distribution in advanced glaucomatous visual field defects.Statistical analysis of single-factor and multivariate conditional logistic regression were performed to screen the relationship between the remaining visual field and the risk factors such as age,gender,type of glaucoma and IOP rise,high myopia,diabetes,hypertension and so on.Results:33.9%of the patients were central 5-10 degrees of visual field,66.1%of the patients were temporal peripheral field.Among the patients with central 5-10 degrees of visual field,66.1%hadn\'t affected visual acuity,while 33.9%of the patients did have.The patients with temporal peripheral field always progressed to a visual acuity of 20/200 or worse.Among the latter,it could be found 9.7%in superior field,23.3% in the inferior and 66.9%across the horizontal meridian.During statistical analysis of single-factor and multivariate conditional logistic regression,the type of the IOP rise and glaucoma has a positive correlation with the remaining visual field.Conclusions:Visual field defect in advanced glaucoma has different shapes and distributions.The type of the IOP rise and glaucoma has a positive correlation with the remaining visual field. Purpose:To correlate structure of retinal nerve fiber layer with function in advanced glaucoma.Methods:Average RNFL thickness and RNFL thickness in four quadrants and in 16 sections were measured using fourier-domain optical coherence tomography(fdOCT). 16sections were ST2(Superior Temporal),SN2(Superior Nasal),IT2(Inferior Temporal),IN2(Inferior Nasal),TU2(Temporal Upper),TL2(Temporal Lower), NU2(Nasal Upper),NL2(Nasal Lower).Results:16 patients with central 5-10 degrees of visual field and 16 with temporal peripheral field were enrolled into the study.The remaining retinal nerve fiber layer measured in all patients were:average 61.30±7.67μm,superior quadrant 78.05±9.66μm, inferior quadrant 80.02±11.91μm,nasal quadrant 44.54±9.44μm,temporal quadrant 42.60±9.68μm,the differences of RNFL thickness between normal and patients were significant(P＜0.05).The average retinal nerve fiber layer measured in patients with central 5-10 degrees of visual field and temporal peripheral field were(60.99±8.20) um,(61.52±7.54)μm respectively.There was no statistically significant difference in RNFL thickness between the two groups(P＞0.05).In the eyes with central 5-10 degrees of visual field,temporal quadrant and 3 sections(TU_1,TU_2,TL_2) thickness were higher than the other group.In the eyes with temporal peripheral field,nasal quadrant and 4 sections(SN_2,NU_2,NU_1,NL_1) thickness were higher than the other group.And the differences were statistically significant(P＜0.05).Conclusions:In eyes with advanced glaucoma,very thin but existing retinal nerve fiber layer is found on fdOCT.Such values rarely dropped below 20 to 30μm,and the correlation between structure and function was positive.