Cyanide is a hypertoxic compound widely used in industry and militqry forces in the world. Hemorrhagic shock, the most severe syndrome, in the battle injuries can result in a higher fatality rate. Combind injuries of chemical intoxication and wound have been reported frequently in accidents or disrasters. However, no detailed research on the combind effects of sodium cyanide intoxication and hemorrahgic shock and their treatment was seen up to now.  Object: Physiological、biochemical and molecular biological techniques would be used to determine the interaction between hemorrhagic shock and cyanide intoxication. Further studies were carried out to clarify the influences of the combind injuries on cardia function and oxdative stress.  Methods: Combind injury animal models were established in rats and rabbits. The function of respiration and circulation were recorded with electrophysiological system. Myocardium enzymogram and blood gas were analyzed biochemically. Spectrophotometry was employed to detect SOD、MDA and ROS in serum and NO and iNOS in cardiac muscle. Western blot were also used to further the study of iNOS.  Results: The LD50 value of sodium cyanide decreased obviously in the animals with combind injuries. There was an extention in Q-T stage .Cardiac muscle zymogram and blood gas were changed. The value of PH、PaO2 and HCO3- decreased manifestly. The activity of SOD decreased, however, iNOS activity and the amount of MDA、ROS and NO increased. There were a significant difference among intoxication group、hemorrhagic shock group and combine injury group.there was a amelioration after the use of 4-DMAP.  Conclusion: Hemorrhage and hemorrhagic shock elevated the toxicity of sodium cyanide and improve its injury to cardiac function. Disorder in the course of cardiac muscle repolarization and obvious increase in the amount of cardiac muscle enzymes can cause

  Intestinal epithelial cells (IECs) are the major component of the intestinal mucomembranous barrier, and play important roles in the body owing to its many kinds of functions such as absorption, immunity, and secretion, and so on. IECs are sensitive to ionizing radiation (IR), and they are one of the main targets in case of whole-body irradiation. Intestinal wound induced by large does of IR can form, thus make the intestine lose its barrier function, impose intestinal infection of endogenous bacteria, and cause mult-organ dysfunction syndrome. It is the main reason of high death rate of intestinal-type radiation disease. Ionizing irradiation can give rise to a series of biochemical events inside of the cell. Free radicals that are derived from intercellular water interact with DNA and proteins, thus either inducing or reducing gene expression, as well as proteins. These proteins are associated with many important cellular process including DNA repair, apoptosis, cell cycle control, and oxidative stress response.Some studies have showed intestinal epithelial can reconstruct to some extent after IR (non-lethal). It is possible to study the protein related to injury and reconstruction of small intestinal epithelium irradiated byγ-Ray. With the accumulating evidence in the literature that new protein are found to be implicated in radiation response, the injury by radiation will be significantly reduce.  Our topic is supported by NSFC,the point is to research the mechanism underlying the radiation response of small intestine epithelial cells.we have used the comparative proteome approach to isolate the protein, ERp29, which expression was regulated by ionizing radiation in mouse .The same results are found in IEC-6 cells. Through the literature retrieval, there is no report about the function of ERp29 by ionizing radiation.Combined the known function of ERp29, we suspect that ERp29 is a gene associated with injury and reconstruction in ionizing irradiation.Therefore, it is necessary to study the function of ERp29 in injury and reconstruction of intestinal epithelium cell irradiated byγ-Ray.  RNAi is the process of using specific sequences of dsRNA to knock down the

  Objective:The arsenic trioxide- polylactic-co-glycolic acid nanoparticles (As_2O_3 -PLGA -NP)were prepared by optimal technological procedure,its distribution and pharmacokinetics characteristics were investigated with animal experiments.  Methods: 1. As_2O_3-PLGA-NP were prepared by w/o/w double-emulsion evaporation technique.Various factors related with the entrapment ratio and nanoparticle diameter were studied.The uniform experimental design U_(15) (15~5) was chosen to optimize the preparation technology of nanoparticles.The particle diameter,surface morphology,release property in vitro and stability of the optimized nanoparticles colloid were evaluated by electron microscopy,release test in vitro and stability test.  2.100 mice,weight 20-25g,were divided randomly into 20 groups,and 5 mice each group,which were injected As_2O_3-PLGA-NP or As_2O_3 at the dose of 2mg/kg in candal vein.At different times after injection ,the lung,liver,kidney,spleen and heart were rapidly removed, arsenic concerntration in tissues respectively were measured by hydride generation atomic fluorometry spectrophotometry(HG-AFS). Pharmacokinetic characteristics of As_2O_3-PLGA-NP and As_2O_3 in liver of mice were analyzed by PKS computer programs.  3.6 rabbits,weight 2.3-2.5kg,with cross-over design were injected As_2O_3-PLGA-NP or As_2O_3 at the dose of 1mg/kg.The arsenic concerntration in plasma were measured by HG-AFS at different times after injection.Pharmacokinetic characteristics of As_2O_3-PLGA- NP and As_2O_3 in plasma of rabbits were analyzed by PKS computer programs. Results:1. As_2O_3-PLGA-NP were prepared by w/o/w double-emulsion evaporation technique with emulsion by ultrasonic,then evaporating at room temperature for 3 hours.The factors that influence the nanoparticles entrapment ratio and diameter was investigated.The analysis showed (1)drug/polymer;(2) polymer concentration;(3) ultrasonic intensity;(4)PVA concentration,which were the main factors that influence the entrapment ratio of As_2O_3-PLGA-NP.Under the optimal manufacture conditions, As_2O_3 was entrapped 53.19±1.82%,the drug loading was 0.64±0.02%,the mean diameter of the obtained

  Objective: Sepsis is the most frequent cause of the death in serious patients. Although great attentions have been paid in treatment. It still keeps a high mortality about 50%-60%. CpG DNA is recognized as a key molecule during the pathophysiology of sepsis. CpG DNA can bind to TLR9(toll-like receptor 9) of monocytes and macrophages, then induces cytokines such as TNF-αand IL-6 to release, which at last lead to sepsis or even multiple organ dysfuction syndrome and death. Therefore, preventing CpG DNA from binding to its receptor is considered as the most promising strategy.  Clinical experiences and studies had revealed that a lot of traditional Chinese herbs possess the anti-inflammatory effect. However, there are too complicated chemical components in the herbs, and the material bases of the anti-inflammatory effects are unclear, which limits the uses of the herbs in clinic. Therfore, targeting on CpG DNA, we screened the effective components extracted from the herbs, and investigate their pharmacologic effect to treat sepsis.  Methods: (1) CpG DNA, the target, was immobilized on the biotin cuvette of biosensor. The biosensor technology was applied to screen anti-inflammatory traditional Chinese herbs targeting on CpG DNA. (2) The activites of the fractions were isolated by silica gel chromatography and high performance liquid chromatography(HPLC)from rhubarb. After the activites of the fractions were evaluated, the most active fraction was confirmed. (3) In vitro, the abilities of Fraction D binding to CpG DNA and Lipid A were measured, and its inhibition on TNF-αrelease from RAW264.7 cells induced by CpG DNA and LPS were observed, too. (4) In vivo, the protection of Fraction D on mice challenged with heat-inactived E coli was observed.  Results: (1) The platform to screen the anti-inflamatory effect of traditional Chinese herbs was established. Among the seventy-eight traditional Chinese herbs, fourteen herbs possessed the higher binding ablity for CpG DNA, and nine herbs possessed larger content of effective components. (2) The active fraction D(Fraction D)from rhubarb was

  Background and Objective:  Atherosclerotic cardio-cerbral vascular diseases are still the most severe hazard to people’s health and life. Effective drugs improving the stability of atherosclerotic plaque were deficiency. Accumulating evidence supports a central role for inflammation in preclinical atherosclerosis invading mononuclear cells release enzymes (eg, matrix metalloproteinases [MMPs]) that degrade collagen and elastin, thereby allowing cells to invade by disrupting matrix layers that otherwise stabilize developing plaque. Clot forming and inflammatory pathways then work in tandem to accelerate local macrophage and T-cell activation, which contributes to plaque erosion or rupture. But until now, the prospective studies about the relation between inflammatory factors and the stability of atherosclerotic plaque were unclear. So, inflammatory AS model was established to observe the change of the stability of atherosclerotic plaque between inflammatory AS model and AS model, and the effects of total saponins of Panax Notoginseng on the stability of atheosclerotic plaque were studied as well.  Methods:  Rabbit atherosclerotic models were set up by hypercholestermia diet for twelve weeks. Then the AS rabbits were divided into six groups randomly (n=8): control group; PNS groups (administrated ig with PNS 15, 45, 120mg/kg, sid, respectively); Aspirin group (Aspirin 12 mg/kg, ig, sid ); Losartan group(losartan 25mg/kg, ig, sid). All the rabbits were fed with common forage from 13th week to the end point of the experiment. Another 32 rabbits were injected intraperitoneally with zymosan A(10mg/kg, Om.bid) from 13th week to the end point of the experiment and devided into four groups eg, inflammatory group, PNS group (45mg/kg), Aspirin group and Losartan group. Feeding with common forge from the first week to the end point of the experiment, the rabbits in the normal group were injected intraperitoneally with normal saline(4ml/kg) and administrated intragastricly with normal saline(5ml/kg) from 13th to the end point of the experiment week. TNF-αand IL-6 level in plaques and serum were determined with immunohistochemical and ELISA method,

  Staphylococcus aureus (S. aureus), one of the most frequently isolated bacterial pathogens in hospital infections, is also a common pathogen producing biofilm. Data obtained recently from intensive-care units in Germany indicated that S. aureus is the second most frequent cause of catheter-related septicemia (15.4%). S. aureus producing the biofilm could be high resistant to antimicrobial agents and avoid the immune killing and clearance. Moreover biofilm could act as“niduses”of acute infection if the mobilized host defenses can not eliminate the planktonic cells, which can be released at any time during the infection,leading to underlying chronic infections and even more severe consequences. Therefore, to know how biofilm comes into being in S. aureus and other pathogens has presently become a hot topic.  Despite the great efforts have made, the mechanism of biofilm forming by S. aureus had not still been known much about. The studies so far in biofilm forming showed that the attachment of the bacterial cells to the polymer surface was the first step,which may occur very rapidly. And thereafter multilayered cell clusters formed by the growth dependent accumulation of bacteria were surrounded by a slimy matrix induced by polysaccharide intercellular adhesion(PIA)whose synthesization was catalyzed by products encoded by ica operon. This process was regulated by multiple factors and in multiple levels. Among them sigB, theσfactor code gene was thought to be the key factor since it had been demonstrated could regulate directly not only the expression of ica but also control the expression of sarA and/or agr. However recent studies found that the S. aureus whose sigB was deleted by homologous recombination is still able to form biofilm. Since all the experimental bacteriia in the literature were limited to study on some specified strains, the strains isolated from clinic during July 2004 to December 2004 in our hospital had been used as the target in this study. The resistance to antibiotics and the presence of biofilm of these strains were identified and the genes related to the forming of biofilm were amplified by PCR. Then the homologous recombination plasmid of sigB gene was constructed from Staphylococcus aureus.

  Background  In recent years, the number of patients suffering from allergic asthma has increased , and allergic diseases including asthma have become a social problem affecting medical costs and quality of life. Allergic asthma is a complex disorder characterized by airway inflammation,bronchial hyperresponsiveness and reversible airway obstruction. Elevated numbers of activated Th2 cells, mast cells and eosinophils in the bronchial mucosa cause certain features of asthma, including increased serum IgE levels in allergic asthma. The available data suggest that there are many potential susceptible genes for allergic asthma, including genes for cytokines, receptors, transcription factors,immune recognition.  Asthma may be based on an inflammatory mechanism involving T-helper type 2 cytokines, such as IL-4 and IL-13, through their receptors.Upon release of IL-4 or IL-13, binding of the cytokine to it receptor at the cell surface induces receptor dimerization and activation of the Janus tyrosine kinase 3 and the transcription factor STAT6 . Tyrosine phosphorylation of signal transducer and activator of transcription 6 mediates homodimerization, triggering movement to the nucleus, and leading to its specific binding to promoter sequences in the DNA . STAT6 DNA-binding sites have been identified in the promoter regions of several IL-4-inducible genes. This can result intransactivation of the target genes, which have central roles in IgE synthesis and the development of bronchial hyperresponsiveness.  The signal transducer and activator of transcription 4 on chromosome 2q32.2–q32.3 is known to be essential for mediating responses to interleukin 12 in lymphocytes and regulating the differentiation of T helper cells.IFN-γbelongs to a cytokine class that affects the immune function, in fact it plays a major role in the induction of the immune-mediated inflammatory response.  Objective  To study the 2964G/A variant of the Signal transducer and activator of transcription 6

  Of numerous forms of DNA damage which can block cell growth, O6-Alkylguanine is the major product formed in DNA by the reaction of alkylating agents. But the DNA repair protein O6-methylguanine–DNA methyltransferase (MGMT) existing in cells can revert the DNA damage induced by alkylating agents is the most important reason for the alkylating agents resistance especially for nitrosoureas and for the decreasing killing efficiency in tumor cells after treatment of alkylating agents, it’s the probably molecular mechanism of nitrosoureas resistance for many tumors.  To increase tumor’s sensitivity for alkylating agents, Kreklau etcetera tried to inhibit the MGMT activity in tumor cells by inhibitor but simultaneously the MGMT activity in peripheral blood monocytes also be inhibited and the cumulation of such kind of immuno-reacted inhibition can lead to marrow toxicity. Yoshinaga etcetera reported that tumor’s resistance for alkylating agents can be reverted by inhibiting expression of MGMT gene through antisence or ribozyme strategy but also has two questions: one is the low efficiency and the other is the blindness in antisense sequence selecting. In recent years, RNA interference comes to be the most efficient manner for gene silence, siRNA has a special double string complementary structure and especially siRNA eukaryotic expression vector can express in cells during a long period of time and more stable than antisense RNA, so that the siRNA has an inimitable, possibly more perfectible effect for regulating gene expression when compared with antisense RNA or ribozyme strategy.  In present study, we synthesized 3 single DNA string coding hairpin siRNA and cloned them into downstream of H1.2 promotor of pRNATin-H1.2/Neo vector to construct a recombinant vector with target gene fragment in it, then transfected the vector into HeLa S3 cell by lipid Lipofectimine 2000 mediating.  Assessed the inhibiting effect of MGMT by semi-quantitative RT-PCR and real time quantitative RT-PCT methods, assessed by MTT the variation of HeLa S3 cell sensitivity to

  Since 1950’s, organic pollution has become more and more serious following the process of global industrialization and the abuse of chemicals. Most of the organics were part of persistent organic pollutants (POPs), which were harmful to human health and ecological system. It reminds us the situation is very serious that main rivers in China have been polluted by organics on different levels. The Three Gorges Project is a great work to take advantage of the water resources. But it will certainly affect the ecological environment when it was built up. Jialing River locates in the reservoir region and is the main water supply of Chongqing City. If we could know the features and toxicity of pollutants in the river clearly, we will offer scientific evidences in evaluating the safety of the water and protecting people in Chongqing and cities along the down-stream of the Yangtze River.  With the organic pollution becoming worse in recent years, both governments and the public have paid great attention to it. Many labs all around the world have begun their researches to detect the toxicity of organic pollutants. Some researchers cared about toxicity of single compound. Others interested in approaching the harm of mixtures extracted from river water. Many zoic, cellular and molecular techniques were used to verify the features of different organic extract. However, few researchers have tried to do some work on systematic observation of animals in chronic or subchronic toxicity test. But only chronic taking with suitable doses can imitate the actual mode of people drinking water and provide believable data to direct our measures in environmental protecting.  In our study, organic extract from Jialing River was collected by method of solid phase extraction. A subchronic toxicity test for rats was adopted to investigate the main toxic target organs. At 2L, 12L, 72L/(kg.bw) doses of pouring into stomachs every two days, pathological changes, hematological parameters, analysis of urine, serum biochemical parameters and indexs related to free radical damages were observed. Based on these results, we expanded our research to more details. The genetic, reproductive, neural and immune

  Sulfur mustard, the widely used vesicant by foreign armed forces, has been demonstrated to be transformed into a reactive sulfonium electrophile which, in turn, can be attached to various biomolecules such as proteins, nucleotieds and enzymes. Consequent to these changes, a systemic toxicity is always the most important out put of mustard after absorption and distribution from its local contaminating area of the body. Among the sensitive organs and tissues to mustard, the small intestine has been regarded as the most vulnerable one because of its importancy in maintaining the balance of the internal environment. Previous reports described that intestinal mucous membrane barrier could be damaged after burn, which was the primary factor of infection development in intenstine and leading to death. In the modern war, the opportunity of multi-weapon attack will be high and the combined injuries of SM and burn will bring more difficulties to cure. However, there wasn’t any report about the damage on intestine induced by this kind of injury.  Objective: Using the small intestine of rats as a main target, we investigate the early injury of small intestine in rats administrated with burn, SM or SM plus burn, and hope to understand the mechanisms of toxic action of SM or SM plus burn on the intestine. Searching for pharmacological protection of NAC, a scavenger of oxygen free radical, on intestinal mucous membrane damaged by burn, SM or SM plus burn and offering more theory about the diagnosis, prevention and cure measure.  Method:⑴Establishing a in vivo model of rats damaged by burn, SM or SM plus burn. The LD50 value of sulfur mustard was determined by modified Karber’s method. (2)Using HE stain, transmission electron microscope, immunohistochemistry, spectrophotometry, RT-PCR, and changing of morphology, ultrastructure and permeability of the small intestine, apoptosis on the intestinal epithelium, oxidative/deoxidative state in rats post-injury treated or un-treated with NAC were observed.  Results: (1) The LD50 values of SM under the conditions of single intoxication and