Objective To explore the protective effect and probable mechanism of JNK inhibitor SP600125 on hippocampal neurons in rats with status epilepsy following lithium?pilocarpine. Methods 48 Wistar rats,in accordance with the random number table,were divided into control,status epilepticus ( SE) and JNK in?hibitor SP600125 group ( SP ) . HE staining and fluorescent TUNEL method were used to observe pathological changes and neuronal apoptosis in the hippocampal area of rats in each group. Western blot was applied to detect the phosphorylation expression of JNK and its downstream effector molecule c?JUN in hippocampal tissues of rats in each group. Results Compared with control group,neuronal loss and apoptosis in CA3 area of hippocampus in SE group were significant (percentage of TUNEL positive cells (26.34±3.04)%, P<0.05). The mortality of rats was significantly decreased and neuronal loss and apoptosis were obviously reduced in SP group than in SE group ( mor?tality in SP and SE group :6.25%,37.5% respectively, P<0.05). Meanwhile,the expression levels of phospho?JNK and phospho?c?JUN were significantly increased in hippocampus of rats in SE group ( The relative OD values respectively 0.447±0.025,0.552±0.035, P<0.05 compared with Control group). After treated with SP600125 in SP group,the phosphorylation levels of JNK and c?JUN were obviously decreased ( The relative OD values respec?tively 0.211±0.016,0.237±0.028, P<0.05 compared with SE group). Conclusion JNK inhibitor SP600125 may play an important protective effect on neurons in the rat hippocampus after status epilepticus through inhibition of JNK and c?JUN phosphorylation.

BACKGROUND:Gene transfection of cells includes virus and non-virus vector.As virus vector has some issues,such as safety and immunological rejection,the present study explored lipofectamine and electroporation transfection methods.OBJECTIVE:To establish genetic engineering cells using human brain-derived neurotrophic factor(hBDNF)gene transfected bone marrow mesenchymal stem cells(BMMSCs)by lipofectamine or electroporation,and explore its characteristics and expression in vitro.METHODS:Lipofectamine method:The BMMSCs were obtained from the tibias and femurs of the guinea pigs.The third passage BMMSCs were cultured with plasmid-lipofectamine mixture for 6 hours,followed by fetal bovine medium for 48 hours.Immunohistochemistry was performed for transient expression.G418 was added after 48 hours.Electroporation method:BMMSCs were trypsinized and resuspended with serum-free medium.Cell suspension was added into electrotransformation pool,and plasmid was added.The electrotransformation pool was moved between electrodes.After transfection for 48 hours,gene transient expression was detected.G418 was added after 48 hours.Brain-derived neurotrophic factor expression was detected by immunohistochemistry and RT-PCR.RESULTS AND CONCLUSION:Immunohistochemistry showed that BDNF transient expression was 5.80% by lipofectamine and 24.29% by electroporation.Cells almost died at 14 days following lipofectamine transfection.Stable expression cell lines of BDNF engineered BMMSC were successfully established by electroporation,with 90% expressive rate by immunohistochemistry and expression of BDNF mRNA by RT-PCR.Genetic engineering cells using BDNF transected BMMSC were established by electroporation whereas failed by lipofectamine,and the expressed BDNF was confirmed by immunohistochemistry and RT-PCR in vitro.

Objective To investigate the function of bone-marrow mesenehymal stem eells(BMSCs) differ-entiating into neuron-like cells in vitro after transfected by human brain-derived neurotrophie factor (BDNF) gene. Methods Human BDNF genes were cloned and recombinant pcDNA3. 1(-)-BDNF plasmids were construc-ted. BMSCs from five guinea pigs were isolated and cultured while their morphologies were observed by microscope. The surface antigen was detected by flowcytometry. BDNF genes were transfected into BMSCs with electroporation, and the transfected BMSCs were induced by ratinoie acid(RA)after bolted by Geneticin-418 (G418), then the dif-ferentiated BMSCs were identified by immunocytochemistry. Results The culture ceils demonstroted the typical mor-phology and surface antigen of BMSCs. The transfected cells expressed neuron- specific enolase(NSE), Nestin and glial fi-brillary acid protein(GFAP) also secreted BDNF. Conclusion BMSCs transfected by BDNF genes can differentiate into neu-ron- like cells in vitro, electroporation can enhame the transfection efficiency, RA can promote cell induction.

Hepatocellular carcinoma(HCC)has high incidence rate,recurrence rate after surgecal treat-ment,as well as fatality rate in China .HCC is not sensitive to radiotherapy or chemotherapy and no effective treat-ment is available for HCC patients at advanced stage .Sorafenib is the first effective molecularly targeted drug for the treatment of HCC,which represents a milestone in the treatment of HCC .However,it also shows drug resist-ance during clinical application .Therefore,it is important to investigate the mechanism of drug resistance and its molecular markers for HCC .In this review ,we summarize the current status of the studies in these fields .

Objective To observe the effects of Panax notoginseng saponins and aspirin on platelet aggregation and analyzed resistance probability.Methods One hundred and twenty patients with cerebral infarction were di-vided randomly into Panax notoginseng saponins group and the aspirin group (each group 30 cases),respectively treated with adenosine diphosphatc (ADP),arachidonic acid (AA) to induced and detect the largest platelet aggre-gation (MAR).All patients received platelet aggregation test in hospital on the first and 14th day.Results In Pa-nax notoginseng saponins group and the aspirin group platelet aggregation induced by gents in the same role did not show statistical significance.There were resistances,semi-resistance phenomenon.Conclusions Panax notogin-seng saponins on platelet aggregation and the impact of aspirin was not significant; it may be used as anti-platelet aggregation drug,but there is also the resistance phenomenon.By combining Panax notoginseng saponins other an-ti-platelet drug anti-platelet aggregation of the long-term clinical.It might solve the resistance problem through large-scale clinical trials.

Objective To summarize the clinical data of cardiac transplantation in 28 patients of our hospital,and study the effectiveness of operation and the methods of immunosuppressive regime.Methods From January 2000 to May 2005,in our center,28 patients were subjected to orthotopic cardiac transplantation.In early stage(before 2003) ciclosporin A and mycophenolate mofetil were used for immunosuppression.In late stage(after 2003) Daclizumab and mycophenolate mofetil were used for immunosuppression.Results Orthotopic cardiac transplantations were done successfully in 28 patients.There was no haemorrhage,twist and right heart insufficiency postoperatively.In the patients before 2003,the complications of hepatic-renal inadequacy to varying degrees occurred in 6 cases and infection in 3 cases respectively.In the patients after 2003,there were no complications of hepatic-renal inadequacy and infection.Conclusions Cardiac transplantion is an effective way for end stage cardiomyopathy.Immunosuppressive effect of Daclizumab and mycophenolate mofetil is satisfactory.

Objective To study the cytotoxicity of the membrane material of heart assist pump. Methods The membrane materials tested incladed polyurethane (group A) and medical polychloroethylene (group B),and a blank control group with none of the material. L-929 cells (the mouse fibroblasts) were cultured on either material in groups A and B,and 6 cell cultures were without any material. After 24,48 and 72 hours,the growth of cell in groups A,B and the control was observed,and the number was counted. Educts of two materials used in groups A and B was respectively added to culture medium,then the OD was measured every 24h and proliferation rate was determined by MTT test. Results L-929 cells grew very well in groups A and B with less death. The cell counts showed no difference between 2 groups at each time. The OD of group A was always higher than that of group B after 24,48 and 72h ( P

Objective To investigate the function of bone-marrow mesenchymal stem cells(BMSCs) differentiating into neuron-like cells in vitro after transfected by human brain-derived neurotrophic factor (BDNF) gene.Methods Human BDNF genes were cloned and recombinant pcDNA3.1(-)-BDNF plasmids were constructed. BMSCs from five guinea pigs were isolated and cultured while their morphologies were observed by microscope. The surface antigen was detected by flowcytometry. BDNF genes were transfected into BMSCs with electroporation,and the transfected BMSCs were induced by ratinoic acid(RA)after bolted by Geneticin-418(G418),then the differentiated BMSCs were identified by immunocytochemistry. Results The culture cells demonstroted the typical morphology and surface antigen of BMSCs. The transfected cells expressed neuron-specific enolase(NSE),Nestin and glial fibrillary acid protein(GFAP) also secreted BDNF.Conclusion BMSCs transfected by BDNF genes can differentiate into neuron-like cells in vitro,electroporation can enhame the transfection efficiency,RA can promote cell induction.

Objective To evaluate the electrophysiological change in patients with acute tetrodotoxin(TTX) poisoning.Methods The electromyogram, motor nerve conduction velocity(MCV), sensory nerve conduction velocity(SCV),F wave,H reflex and somatosensory evoked potentials(SEP) were randomly detected in 58 patients with acute TTX poisoning.Results 22 patients with TTX(37.9%) detected by the electromyogram showed mainly polyphase irregular waves;MCV and SCV were weakened; SCV was even more remarkable,the latency of distant MCV action potentials was prolonged obviously, the abnormality rate of nerve conduction velocity was much higher than that of fibrillation,the detectable rats of positive wave was high,the low abnormality rate of F wave and H reflex suggested that the ill TTX poisoning involved the nerve roots;the abnormality rate of SEP was 56.9%.Conclusion TTX poisoning can company with the damage of central nerves, the measure of electroneurophysiology can be used to observe the extent,course and range of nerve system damage in patients with acute TTX poisoning, and it is one of the early detection means of this disease.

Objective To observe the clinical effects of endoscopic great saphenous vein harvesting(ESVH) in coronary artery bypass grafting. Methods Endoscopic great saphenous vein harvesting was performed in 89 patients undergoing coronary artery bypass grafting(CABG) from April 2004 to May 2005.An incision 2 cm in length was made on the knee joint and the great saphenous vein was harvested using the VasoView-5 system.Clinical data of the 89 patients(Endoscopic Group) were compared with another 38 patients undergoing conventional great saphenous vein harvesting for CABG from April 2003 to March 2005(Conventional Group) in respect of complications of the leg, the recovery time to walking,and the patency rate at 6 postoperative months.Results The number of harvested vein grafts in the Endoscopic Group was 2~3(mean,2.6).Complications of the leg were significantly less in the Endoscopic Group(6/89) than in the Conventional Group(8/38)(?~2=4.197,P=0.040).The occurrence of pain and numbness of the leg was 7/89 in the Endoscopic Group and 36/38 in the Conventional Group(?~2=89.740,P=0.000).The occurrence of swelling of the leg was 9/89 in the Endoscopic Group and 30/38 in the Conventional Group(?~2=59.299,P=0.000).The recovery time to walking was significantly shorter in the Endoscopic Group(2.3?0.9 d) than in the Conventional Group(3.4?1.6 d)(t=-4.952,P=0.000).There was no significant difference in the patency rate at 6 postoperative months between the Endoscopic Group(96.0%,48/50) and the Conventional Group(95.3%,19/20)(?~2=0.000,P=1.000).Conclusions Use of endoscopic vein harvesting in coronary artery bypass grafting decreases the incidence of postoperative leg-wound infections,post-operative pain,and length of hospital stay.