Magazine abstract

The Effect of Plasma Triglyceride on Platelet Aggregation

Thu, 11 Mar 2010 16:59:55 +0000

Objective:to explore the effect of different plasma triglyceride on the platelet aggregation.Methods:100 patients with hyper-triglyceridemia(TG≥1.7mmol/L) were collected and 68 patients with normal-triglyceridemia(TG＜1.7mmol/L) were collected.The platelet aggregation induced by adenosine diphosphate(ADP) on a 2-channel light aggregometer was measured;Comparing respectively:(1) the PAGT of the patients who had never taken Aspirin in two groups(45,30);(2) the PAGT of the patients who have always taken Aspirin in two groups(55,38);(3)in hyper-triglyceridemia group,the PAGT of the patients with different level of the plasma triglyceride in subunit.Result:The PAGT of the hyper and normal triglyceridemia groups who had never taken As-pirin(45,30) were(75.77±7.15)%,(58.48±8.31)%,respectively,(P＜0.05);The PAGT of the hyper and normal triglyceridemia groups who have always taken Aspirin(55,38)were(56.15±13.65 )%,(48.64±12.75)%,respectively,(P＜0.05);In the hype-triglyceridemia group,the PAGT of the patients in subunit who have always taken Aspirin were:from 1.7 to 2.26mmol/L,(54.04±14.07) %;from 2.26to2.8 mmol/L,(65.86±7.02)%;greater than 2.8mmol/L,(65.86±7.02)%,then,the first and the third subuint were statistically significant(P＜0.05).Couclution:(1)The level of the plasma triglyceride might have an effect on platelet aggre-gation.(2)The inhibit function of the drug,eg.Aspirin on platelet aggregation might be weakened by plasma hyper-triglyceride.

Experimental Study on Expression of a New CD44 Variant in Multidrug Resistance Breast Cancer Cells and the Role of the CD44 Variant to Human Breast Cancer Cells

Thu, 11 Mar 2010 08:39:38 +0000

Objective:To clone CD44 gene from multidrug resistant human mammary carcinoma cells(MCF-7/Adr)and construct its eukaryotic expression vector which lays the groundwork for the experiments with multidrug resistance,invasion and proliferation. Secondly,to analyze the relationship between CD44v expression and invasion of breast cancer cell line MCF-7,and to explore possible molecular mechanisms of MMP-2 and MMP-9 regulated by CD44 vatiant.Finally,to study the relationship between CD44v expression and the proliferation of MCF-7/CD44v cells.Methods:①The expression of CD44 gene and protein in the MCF-7 and MCF-7/Adr cells was detected by RT-PCR and flow cytometry.②The full length cDNA encoding CD44 was obtained by RT-PCR from the total RNA isolated from the MCF-7/Adr cells.In order to get CD44 gene contained the site of the two restriction enzymes EcoRⅠand KpnⅠ,the primer contained the site of the two restriction enzymes was used for polymerase chain reaction(PCR).Treated the PCR product with 1%Agarose DNA gel electrophoresis,then reclaimed and purified the CD44 gene according to the directions.③After purification,the cDNA fragment was cloned into the pMD19-T simple vector.The recombinant plasmid pMD19-T-CD44v was transformed into E.coli DH5αfor amplification,and then digested with two restriction enzyme EcoRⅠand KpnⅠafter being extracted with Plasmid Mini Preparation Kit.The nucleotides sequencing of the CD44 gene was confirmed by DNA sequencing.④The eukaryotic expression vector pcDNA3.1 and the recombinant plasmid pMD19-T-CD44v were digested with EcoRⅠand KpnⅠ,respectively,then connected by T4 DNA ligase and amplificated in DH5α.The eukaryotic expression vector pcDNA3.1-CD44v was identified by the two restriction enzyme and nucleotides sequencing.⑤The recombinant vector pcDNA3.1-CD44v was transfected into MCF-7 cells by Lipofectamine,and the expression changes of CD44 gene and protein were detected by Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) and flow cytometry at 48 hour later,respectively.⑥After incubation with HA for 24 hour,all the cells in each group were collected for the detection of flow cytometry and RT-PCR.The conditioned medium were collected,clarified by centrifugation,and subjected to gelatin zymography.⑦The number of the cells through the artificial matrix membrane in every group was counted to compare the change of the invasion capability among the expermental groups.⑧Molecular mechanism of MMP-2 and MMP-9 expression regulated by CD44 vatiant was detected by western blotting.⑨.The number of the cells was counted by microscope to detect diverse proliferation among the experiment groups.Results:①.CD44 gene variant expressed in MCF-7/Adr cells,but not in MCF-7 cells.②The CD44 gene contained the site of the two restriction enzymes EcoRⅠand KpnⅠwas introduced into pMD19-T simple vector.After the identification and sequencing,the reconstructed plasmid was confirmed containing the sequence of CD44 gene variant which contains 1 to 4 exons,16 to 17 exons,and 1 to 205 basees of 18 exons.The new gene sequence was sent to NCBI for publication,registered number is FJ216964.③The new gene sequence was successfully cloned into recombinant vector pcDNA3.1 and identified by the two restriction enzyme as well as sequencing,the sequences of recombinant plasmid pcDNA3.1-CD44v were the same as that of FJ216964.④After transfered recombinant vector pcDNA3.1-CD44v to MCF-7 48 hour, RT-PCR and flow cytometry detected the mRNA and protein level of CD44 gene variant were obviously up-regulated.The expression ratio of CD44 protein in MCF-7/CD44v cells is 70.0±2.5%.Cells after transfection were screened by the conditioned medium G418 to obtain positive cell clones.Stable G418-resistant clones were obtained 2 week later and the expanded cells were then used for subsequent studies.⑤Overexpression of CD44v by transfecting tumor cells with CD44v cDNA increases MMP-2,MMP-9 expression as well as invasion capability of MCF-7 cells(P<0.05);⑥Our studies also indicate the activation of MMP-2 and MMP-9 secretion could be blocked by CD44 blocking anti-body.Pretreated MCF-7/CD44v cells with the neutralizing antibody against CD44 could strongly block the invasion of MCF-7/CD44v cells.⑦Finally,it is important to note that CD44v could inhibit the proliferation of MCF-7/CD44v cells.Conclusion:①The expression vector pcDNA3.1-CD44v was constructed successfully and could be expressed in human mammary carcinoma cells.A new CD44 gene vatiant was found in MCF-7/Adr cells(GeneBank NO.FJ216964);②Our study may start a new approach to study the function of CD44 gene vatiant and provide a method of gene therapy.③CD44 vatiant could increase invasion capability of MCF-7 cells though influence expression of MMP-2 and MMP-9.④HA could integrate With CD44 vatiant and regulate expression of MMP-2 and MMP-9,which increased invasion capability of MCF-7 cells through Ras/MAPK signaling pathways.④Over expression of CD44 gene vatiant could inhibit proliferation of MCF-7/CD44v cells.

Dynamic Changes and Effects of IL-31 in Lung Tissues of Experimental Mice with Pulmonary Fibrosis

Tue, 09 Mar 2010 17:00:54 +0000

Objective To investigate the IL-31 protein expressions in lung tissues of experimental mice with pulmonary fibrosis(PF) induced by bleomycin in various periods and to study the effects of IL-31 in pulmonary fibrosis.Methods Forty eight male mice were randomly divided into two groups:control group(n = 24) and experimental group(n = 24),and poured respectively by 0.9%NaCl saline and bleomycin solutions(5 mg/kg).6 mice of each group were sacrificed on the 7th,14th,21th,28th day and the lung tissues were harvested respectively.The alkaline hydrolysis method was used to test the content of hydroxyproline. Real-time PCR examined the expression of IL-31 mRNA.Western Blotting method was used to analyse the protein expression of IL-31.Results1.The content of hydroxyproline in lung tissues in experimental group were higher than those in control group on the 14th,21th,28th day(P＜0.05).2.The protein and mRNA expression of IL-31 in lung tissues in experimental group were higher than those in control group(P＜0.05).3.The expression of IL-31 mRNA in lung tissues in experimental group were higher than control group from the 7th day,peaked on the 21th day,and lower on the 28th day.4.The protein expression of IL-31 in lung tissues in experimental group were higher than control group from the 7th day,peaked on the 21th day,and lower on the 28th day.Conclusion The expression of IL-31 were high at different stages of pulmonary fibrosis.It played an important role especially at the middle and late stages in pulmonary fibrosis.

The Influence of Hypoxic Exercise on Upstreams of mTOR in Skeletal Muscle

Tue, 09 Mar 2010 08:39:39 +0000

Aims:To study the effect and mechanism of hypoxic exercise on PKB/mTOR signal pathway in skeletal muscle,it tested the effect of hypocic exercise on Upstreams of PKB/mTOR signal pathway.Materials and methods:Adult male Sprague-Dawley rats(12 weeks old)were randomly divided into four groups:normoxia sedentary group(NS),normoxia Exercise group(NE),hypoxic sedentary group(HS),hypoxic exercise group (HE).The hypoxia circumstance was normal pressure hypoxia,oxygen concentration is 13.6%,which equivalented 3500m above sea level.HS and HE group constantly lived in hypoxia.Tested total protein、3-kinase(PI3K) and phosphorylation of protein kinase B of extensor digitorum longus(EDL) muscle on the 3rd day、7th day、14th day and 28th day.Results:(1) After 3 days,hypoxia inhibited total protein of PI3K significantly(p<0.05);After 7 days,exercise significantly promoted total protein of PI3K(P<0.01) while hypoxia inhibited it evidently(P<0.01) After 14 days,exercise significantly promoted total protein of PI3K(P<0.01)and phosphorylation of PKB(P<0.01) significantly, in the meanwhile,hypoxia inhibited both of them evidently;and the situation was the same as 14th day\' s after 28 days.(2) After 3 days,there was no difference between all groups about the total protein content.After 7 days,total protein content of NE group was higher than HS(P<0.05)and HE group(P<0.05).And after 14 days,HE group(P<0.01)and NE(P<0.05) group was higher than HS group.After 28 days,the total protein content of all the four groups had no significantly differences(P<0.05).Conclusions:Exercise promotes total protein of PI3K and phosphorylation of PKB significantly while hypoxia inhibits them evidently.Hypoxic exercise reduces the inhibition of hypoxia two both total protein of PI3K and phosphorylation of PKB,which means hypoxia represses muscle protein synthesis.

A New Assay Method-The Application of Plasma 1,5-Anhydroglucitol in Diabetes Mellitus

Mon, 08 Mar 2010 16:59:16 +0000

Objective:To evaluate a new marker of 1,5-anhydroglucitol(1, 5-AG) for short-term glycemic monitoring with liquid chromatography-tandem mass spectrometry(LC-MS/MS) method.Method:1,5-AG concentrations from 243 diabetic patients (NIDDM),267 healthy subjects and 69 aged non-diabetic patients were determined by LC-MS/MS method.The correlation of 1,5-AG to other glycemic markers including hemoglobin Alc(HbAlc,n=327), fructosamine(FA,n=63),fasting blood glucose(FBG,n=174),and postprandial glucose(PPG,n=138) was evaluated.Results:Significant reduction of 1,5-AG level was observed in diabetic patients versus in control group(p＜0.001).Significant between-sex difference was observed in the control group(p＜0.001). The normal reference intervals were set as 14.13-43.81μg/ml for male and 7.77-30.15μg/ml for female subjects.There was significantly negative correlation of 1,5-AG to HbAlc,FA,FBG,and PPG with correlation coefficient(r) values of-0.578(n=327,p＜0.001),-0.539 (n = 63,p＜0.001),-0.409(n = 174,p＜0.001),and -0.503(n = 138,p＜0.001),respectively.Good correlation was observed among plasma, serum,and finger-prick blood samples(r＞0.980,p＜0.001).The LC-MS/MS method showed good linearity with r＞0.999 at 1-50μg/ml of 1,5-AG in plasma.The intra-,inter-assay precision(%) were less than 6.37%,8.75%,respectively,and accuracy ranged 96.2-103.1%.Conclusion:1,5-AG can be applied as a maker of short-term glycemic control,combined with existing ones,for full glycemic monitoring in diabetic patients.The LC-MS/MS method is simple, specific and cost-effective for 1,5-AG determination in clinical application.

An Empirical Study on Customer Satisfaction Assessment of Pediatric Inpatient

Mon, 08 Mar 2010 08:40:01 +0000

customer satisfaction has already become a vital topic to many scholars all over the world. in the service area,survival and development pressure force more and more enterprises to put customer satisfaction as their competition strategies.This thesis was started with the overview the study of Service Quality,Customer Satisfaction and Customer Loyalty,which will be used in the model building.Then the survey on the pass research was showed,the Customer Satisfaction Assessment Model was established based on Pediatric,and the hypothesis mutual effect and relationship of each variable were also proposed.The thesis also explained the how the modeling variables comes.And the questionnaire design and its content were described based on the paper.Then the text makes a synopsis elucidation on the data analysis method of this study.212valid questionnaires were collected within two weeks.After collecting the questionnaires,programming was used to transfer the original data into the SPSS analysis data.The data was tested so that it can meet the needs of validity and reliability.Exploratory Factor Analysis and Confirmatory Factor Analysis(CFA) were used to test the Service Quality Modeling,and Amos software was used to verify the mutual effect between Service Quality,Secondly Factors,Customer Satisfaction and Customer Loyalty.The study shows that the model performs well on explaining that how the service variables contribute to the Customer Loyalty.Meanwhile,the limitation of this thesis and follow-up research were discussed in the end of this paper.

Research on Function of Small RNA in Shigella

Mon, 08 Mar 2010 04:29:23 +0000

Shigella spp., commonly called Bacillus dysenteriae, are gram-negative, facultative anaerobes, non-spore forming pathogens. The pathogenesis of S. flexneri is based on its ability to invade and replicate within the colonic epithelium, which results in severe inflammation and epithelial destruction. Shigella include 4 groups and 37 serotypes.In our country the predominant group of Shigella spp is S. flexneri (86%) and the predominant serotype is S. flexneri 2a (80%). Upon infection, humans develop severe abdominal cramps, fever, and frequent passage of bloody stools. According to statistics of WHO, Shigellosis caused 1.5million deaths and over 164 million cases each year with the majority of cases occurring in the children of developing countries. While in China, there were about 20 million cases each year, and S. flexneri 2a was responsible for 50-70% of those cases. The traditional treatment to dysentery is the use of antibiotics. But more than 95% strains have the resistance to various antibiotics. The best treatment is the use of vaccine, but few are known about pathogenesis of Shigella and the immune response of host till now. Thus it is important to research on S. flexneri 2a for our sanitary and epidemic prevention.Prokaryotic small RNAs (sRNAs) that are not ribosomal transfer or messenger RNAs were initially identified in the intergenic regions of genome. SRNA is a kind of newly discovered 50-400 nt small RNAs that do not encode proteins. In recent years, many computational strategies have identified hundreds of sRNA candidates in prokaryote and these sRNA have many kinds of biological functions.They conduce bacteria to regulate physiological systems and accommodate the change of surroundings .Thus research on the function of sRNA is very important. Small RNA maybe represent a new style of Gene Expression Regulation.Now people have realized that sRNAs have participated many gene regulations of organisms in bacteria, such as translational activation, translational suppression, housekeeping sRNAs, quorum sensing, protein degradation ,ion dynamic equilibrium, glycometabolism and expression regulations of growth-dependend outer membrane proteins, the sRNAs are considered the regulators "over" the genomes. Although now more sRNAs have been found in bacteria rapidly, most of their functions are still unknown. Many researchers analyzed that bacteria maybe regulate their physiological system by sRNAs to accustomed to survive in rapidly changing environment. Therefore understanding these sRNAs may open a new research area, because they may represent a new level of the regulation of gene expression.Functional analysis indicate sRNA is a uncared-for aspect in bacteria expression regulation.Rencent research on the sRNA’s functions supply the blank of ions dynamic equilibrium, glycometabolism, growth-dependend outer membrane protein expression regulation.Study on the function of sRNA can promote the study of physiology, phyletic evolution, pathogenic mechanism in S. flexneri 2a. The research on expression of sRNA also can provide a new approach for vaccine of shigella. Studies show sRNA in bacterias play an important role in bacteria metabolism, environment , quorum -sensing and bacteria virulence expression.First, according to the characters of the detected E.coli sRNAs, we use bioinformatics methods :sequence alignment between genome , secondary structure of RNA and un-dependency terminator analysis to predict sRNA in pCP301.We also use Northern blot to validate the predicted sRNA.We got 6 sRNA in pCP 301 by machine learing-based prediction method and choose 5 sRNA in S. flexneri 2a genome from internet and literatures(http://www.sanger.ac.uk).In order to clarify the function of sRNA in shigella, the deletion mutant S. flexneri 2a 301ΔsRNA::Kan were constructed with modifiedλ-Red recombination system. The growth curve of the sRNA mutants and the wild-type presented non-significant difference in LB medium and ferrum limitated medium.Compared with wild strain, the metabolism and growth presented non-significant difference.Furthermore, we evaluated the effect of sRNA to the virulence of S. flexneri. In this study ,we use cell culturing (in vitro) and animal tests (in vivo) to evaluate the function of sRNA. HeLa cell line was used for invasion test of sRNA mutants.Sereny test with guinea pig and lung invasion test with Balb/c mouse were used, respectively, to verify the virulent of wild type and mutants. It had been demonstrated that the three sRNA mutants retained the ability to invade mammalian cells in sereny test and tissue model, on the other hand, ln3 RNA mutant evoking time and inflammation intensity were less efficient than those of wild-type but invasiveness of C0719 sRNA and IS061 sRNA mutants were more efficient than that of wild-type.The comparative proteomics includes the proteomic research of different cultivation, the proteomic research of different strains and the proteomic research between mutants and wild-type. In our study, 2D-gel electrophoresis was used to identify the differential expression of the mutant and the wild-type strain at various growth phases. Moreover, different protein spots between sRNA mutants and wild- type were excised from gels, in-gel digested,and then indentified by using atrix- assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/ TOF) in combination with Mascot search program. It had been found that through the comparative proteomics of eight mutants in stationary phase of 37℃and sf301ΔIS061 in exponential phase of 37℃,we found 31 differential expressed protein spots.We analyzed the functions of these proteins and found the deletion of these sRNAs didn\'t make differential expression between the mutants and the wild-type strain.But we also found a differential expressed protein OspC3 that may influence the virulence of Shigella and another differential expressed protein OmpA,whose expression may be regulate by sRNAs.In conclusion,we have predicted 6 sRNAs in pCP301 and use Northern blot to validated them;we have got 8 sRNA mutants and analyzed the pathogenesis of sRNA mutants by using cell culturing (in vitro) and animal tests;we also combine with com- parative proteomic to analyze the function of sRNA’s expression regulation. The sRNA mutants in this research form the basis and provide valuable information for future sRNA function analysis.

The Effects of Ruan Mai Ling on Myocardium、Vascular Remodeling in Spontaneously Hypertensive Rats and on the Migration of Vascular Smooth Muscle Cells

Sun, 07 Mar 2010 12:49:18 +0000

Objective1. To observe the effects of Ruan Mai Ling(RML),one of Chinese traditional herbal composition,on the remodeling of myocardium and vessels in spontaneously hypertensive rats(SHR).2. To evaluate the expression of Cavα1C,SERCA-2a,IP3R-1 and RyR-3 in heart and thoracic aorta under the treatment of RML.3. To investigate the effects of blood serum containing decoction of RML on the migration of vascular smooth muscle cells(VSMCs) induced by platelet derived growth factor-BB(PDGF-BB).MethodsPartⅠThirty-six twelve-week-old male SHRs were randomized into 4 groups:high- dose(RH:3ml·kg-1·d-11) and low-dose RML treated group(RL:1ml·kg-1·d-1),untreated control(SHR),losartan treated positive control(LOS:10mg·kg-1·d-1),normotensive WKY control.Systolic blood pressure was examined when administered for 12 weeks.Wet weight of left ventricle to body weight ratio(LVM/BW)was assessed on sacrifice.Myocardial collagen fibers in the sections were observed under light microscope.Both the wall-to-lumen area ratios(W/L)and the thickness of the wall to the radius of the lumen(TW/Rl)of thoracic aorta and mesenteric arteries(3th branch)were calculated. PartⅡThe mRNA levels for Cavα1C,SERCA-2a,IP3R-1 and RyR-3 in heart and thoracic aorta were detected by RT-PCR.The amount and distribution of IP3R-1 and RyR-3 protein were determined by immunohistochemistry.PartⅢCultured VSMCs derived from the aorta of 8-week-old rat were used.Blood serum containing decoction of RML and normal blood serum from rats were prepared.Modified Boyden chamber technique was employed for VSMCs migration accessment.ResultsPartⅠ1. After 12 weeks treatment with RML,systolic blood pressure in both groups was markedly lower than that in SHR Group(175.0±4.1,182.5±4.6 vs 208.9±9.3 mmHg,P<0.01).2. Collagen volume fraction(CVF)in the epicardium was significantly reduced by the treatment with RH group(0.03±0.01 vs 0.05±0.02,P<0.01),while no marked difference in RL group was found;CVF in the endocardium was markedly decreased by the treatment with both RML groups(0.03±0.01,0.02±0.01 vs 0.05±0.01,P<0.01); After 12 weeks treatment with RML,CVF in the perivascular in both groups was significantly decreased than that in SHR group(0.04±0.01,RL:0.04±0.02 vs 0.08±0.02,P<0.01).3. In thoracic aorta,the W/L and the Tw/Rl in SHR group were markedly higher than that in RH group(W/L:0.16±0.01 vs 0.20±0.02,P<0.01;Tw/Rl:0.09±0.01 vs 0.11±0.01,P<0.05),while no marked difference in RL group was found.4. In mesenteric arteries(3th branch),the W/L in SHR group was markedly higher than that in RML group(0.29±0.03,0.32±0.10 vs 0.42±0.09,P<0.05);The Tw/Rl in SHR group was markedly higher than that in RH group(0.18±0.07 vs 0.25±0.06, P<0.05),while no marked difference in RL group was found. PartⅡ1. After treated with RML,The mRNA of Cavα1C in ventricle were reduced significantly in comparison with SHR group(0.24±0.02,0.24±0.05 vs 0.35±0.06, P<0.05),while no marked difference in thoracic aorta was found.2. Compared with SHR group,the mRNA of SERCA-2a in both ventricle and thoracic aorta was not influenced by the treated with RML.3. After treated with both RML groups,The mRNA of IP3R-1 in ventricle was not influenced in comparison with SHR group;But the mRNA of IP3R-1 in thoracic aorta was decreased by both RML groups(1.12±0.08,RL:1.07±0.11 vs 1.36±0.12,P<0.05).4. Compared with SHR group,the mRNA of RyR-3 in thoracic aorta was not influenced by the treated with RML.5. In left ventricle,the amount of IP3R-1 protein in both RML groups was reduced significantly in comparison with SHR group(0.014±0.003,0.014±0.006 vs 0.027±0.006,P<0.05),while no marked difference in thoracic aorta was found.6. In thoracic aorta,the amount of RyR-3 protein in RL group was down- regulated significantly than that in SHR group(0.025±0.007 vs 0.195±0.050,P<0.05), while no marked difference in RH group was found.PartⅢ1. The migration of VSMCs was promoted by PDGF-BB(10 ng/ml)compared with control(59.20±13.26 vs 6.96±1.32,P<0.01).2. Compared with 5% normal blood serum,blood serum containing 5% RML could restrain the migration of VSMCs,with inhibition rate at 36.5%(93.20±14.68 vs 147.40±30.17,P<0.01);Blood serum containing 10% RML could restrain the migration also,with inhibition rate at 52.4%(59.90±9.05 vs 125.80±35.94,P<0.01).Conclusions1. RML may reduced the arterial blood pressure in SHR.2. RML may inhibit cardiac collagen fibers accumulating and decreased the cardiovascular remodeling in SHR. 3. The expressions of calcium regulatory protein such as Cavα1C and IP3R-1 were decreased in myocardium.4. The expressions of calcium regulatory protein such as Cavα1C,SERCA-2a, IP3R-1 and IP3R-1 were down-regulated in thoracic aorta.5. RML may decreased the expressions of calcium channels such as Cavα1C and IP3R-1 in myocardium.6. RML may reduced the expressions of calcium channels such as IP3R-1 and RyR-3 in thoracic aorta.7. PDGF-BB(10 ng/ml)may induce the migration of VSMCs.8. Blood serum containing RML(5%～10%)could restrain the migration of VSMCs.

The Role of Intraoperative Facial Nerve Monitoring in Cerebellopontine Angle Microsurgery

Sun, 07 Mar 2010 11:23:47 +0000

Objective:To explore the significance of intraoperative facial nerve monitoring in cerebellopontine angle microsurgery.Method:A total of 40 unilateral cerebellopontine angle tumor patients were operated with intraoperative facial nerve monitoring(15 patients) or without intraoperative facial nerve monitoring(25 patients) and the rate of total resection and neurotomia were compared between the two groups.One week after the operation,the facial function of the two groups was evaluated and compared.Results:In the intraoperative facial nerve monitoring group(M group),the rate of total resection was 93.33%(14/15),and the rate of neurotomia was 86.6%(13/15). While,in the no monitoring group(N group),the rate of total resection was 72% (18/25),and the rate of neurotomia was 56%(14/25).The rate of total resection, especially,the rate of neurotomia in M group is higher than that in N group markedly (P＜0.05).One week after the operation,in M group,the rate of normal or near normal facial nerve function(House-Brackmann gradeⅠorⅡ) was 100%(1/1) in the small-size tumor subgroup,55.6%(5/9) in the moderate-size tumor group and 40% (2/5) in the large-size tumor subgroup;While,in N group,the rate of normal or near normal facial nerve function was 50%(1/2) in the small-size tumor subgroup,35.7% (5/14) in the moderate-size tumor group and 22.2%(2/9) in the large-size tumor subgroup.Whatever size of the tumor,the facial nerve function is better in M group one week after the operation,the rate of total facial nerve function reservation is significantly high in M group(P＜0.005).Conclusion:utilization intraoperative facial nerve monitoring can achieve significant improvement in the rate of total resection,especially the rate of neurotomia,and achieve better facial nerve function.

HELF Cells Were Stabily Transfected of Dicer and the Possible Effect of Dicer Gene on the Function of HELF

Sun, 07 Mar 2010 11:10:52 +0000

ObjectiveEpigenetics is the study of gene expression may be subject to genetic change and the genomic DNA sequence is not generated.At present,the study including:DNA methylation,histone code and chromatin remodeling and regulation of non-coding RNA.The abnormal interaction in their system can lead to the gene abnormal expression and lead to epigenetic-related diseases including the tumor.The occurrence of tumor is due to abnormal cell proliferation,in the past,the vast majority of studies focus on proto-oncogene and anti-oncogene witch encoded protein.However,with the progress of the study we found that some tumors less or no gene mutation,as well as miRNA family strong accommodated in post-transcriptional level,so that an increasing number of researchers to shift the emphasis to the relationship between miRNA and tumor.Dicer is an enzyme essential for the processing of miRNA.When the Dicer gene expression abnormal may lead to the miRNA expression abnormal.Those make proto-oncogene expression increase or anti-oncogene expression decrease and then cause the cell or tissue abnormal proliferation,resulting in the occurrence of tumor.In order to investigate the function of Dicer gene overexpression in tumor,we designed this experiment,human embryonic lung cells(HELF) were stabily transfected of pcDNA3.1-Dircer plasmid,by detecting HELF proliferation and migration to analyze the function of Dicer gene in the tumor.MethodsHELF cells were stabily transfected of pcDNA3.1-Dicer plasmid and pcDNA3.1 vector as control by lipofectamine.1:7 generated at the time of transfection 48 hours.G418 selected then picking single clone.Initial screen Dicer gene DNA horizontal conform by PCR.The transfection effects of Dicer gene on the mRNA levels were examined by reverse transcription PCR(RT-PCR),and the transfection effects of Dicer gene on the protein levels were examined by Western-blot.Cell proliferation rate and viability were measured by MTT.Reconstituted basement membrane invasion assay was utilized to evaluate the cell invasive.Analyze the function of Dicer gene in the tumor.ResultpcDNA3.1-Dicer plasmid and pcDNA3.1 vector were transfected into HELF by lipofectamine.G418 selected then picking single clone.Dicer mRNA and protein expression were obvious upregulated in pcDNA3.1-Dicer plasmid transfected HELF cells than control vector transfected cells,which illuminates cell transfection of pcDNA3.1-Dicer successfully.MTT incorporation rate were increased significantly was apparent in pcDNA3.1-Dicer plasmid transfectedHELF cells.Transwell migration assay results showed HELF cells migration of pcDNA3.1-Dicer plasmid transfection was increased significantly compared with vector pcDNA3.1 transfected HELF cells.ConclusionStabily transfected HELF of pcDNA3.1-Dicer plasmid can lead to upregulate Dicer mRNA and protein expression obviously,which showed transfection HELF of pcDNA3.1-Dicer successfully.MTT incorporation rate were increased significantly was apparent in pcDNA3.1-Dicer plasmid transfected HELF cells and Transwell migration assay results showed HELF cells migration of pcDNA3.1-Dicer plasmid transfection were increased significantly compared with vector pcDNA3.1 transfected HELF cells.