AIM: To observe the enhanced green fluorescent protein (EGFP) gene expression and cytotoxicity in human adipose derived adult stem cells (hADSCs) by pEGFP-N1, Ad5-EGFP and rAAV-2/1-EGFP, and investigate the suitable gene-transferred vehicle. METHODS: The experiment was conducted at Chinese National Human Genome Center and the Third Hospital of Peking University from January to July 2006. ①After the patients and their relative were informed consent, the subcutaneous adipose tissue was obtained from the patients undergoing routine total hip joint replacement in Department of Orthopaedics, Third Hospital of Peking University. pEGFP-N1 was provided by Clotech Company, Ad5-EGFP and rAAV-2/1-EGFP by Vector Gene Technology Company. ②hADSCs were cultured in vitro after isolated from the adipose tissue after dissected and digested with type I collagenase. ③hADSCs of passage 3 were infected with pEGFP-N1, Ad5-EGFP and rAAV-2/1-EGFP and the EGFP expression and the cell toxicity were observed. ④Twenty-four hours after being transfected, 5?104 cells were reseeded in a 24-well plate and the solution was changed three times every week. The growth curves of each group were drawn. Normal non-transfected cells served as control. The influence of different transfection ways on the growth of hADSCs was observed. RESULTS: ①Comparison of transfected efficiency with different ways: pEGFP-N1 transfection showed a higher cytotoxicity and lower efficiency of 10.5%; Ad5-EGFP could efficiently transfect hADSCs (multiplicity of infection=5?102, 82.5%); when MOI was 0.05); however, the growth capability of hADSCs was decreased significantly in the pEGFP-N1 transfection group compared with the control, and the differences were significant at day 3-10 after transfection (P

Objective To investigate the related risk factors referred to hematoma enlargement in acute stage of spontaneous cerebral hemorrhage.Methods 104 stroke patients were divided into the observation group(hematoma enlarged group) and control group (without hematoma enlarged group).The course of disease,mean arterial pressure,the bleeding amount of the first time,hematoma shape and mortality were compared between the two groups,and the risk factors of hematoma enlargement of cerebral hemorrhage patients were analyzed.Results (1) The course of disease,mean arterial pressure,hematoma shape and the bleeding amount were significantly different between the two groups(t =9.5431,11.4499,6.1847,10.7233,all P ＜ 0.05).(2) The multivariate logistic regression analysis showed that factors of influence on hematoma enlargement were mean arterial blood pressure,the course of disease and hematoma shape.(3)The mortality of the observation group was 36.53 ％,which was significantly higher than that of the control group (19.23 ％) (χ2 =3.8731,P ＜ 0.05).Conclusion High level of mean arterial blood pressure,short time of interval to onset,irregular shape of hematoma and frequent vomiting are main risk factors of hematoma enlargement in patients with intracerebral hemorrhage at acute stage.

Objective:To investigate the chemical constituents in the roots of Angelica dahurica var.formosana.Methods: The dried rhizomes of Angelica dahurica var.formosana were chopped and extracted with 80% EtOH for 3 times under reflux.The EtOH extract was isolated by chromatography on silica gel using a gradient solvent system(PE:EtOAc & CHCl_3MeOH) and Sephadex LH-20.The structures of the contents were elucidated by nuclear magnetic resonance and MS spectral analysis.Results: Seventeen compounds were identified,namely,isoimperatorin(1),imperatorin(2),bergapten(3),alloimperatorin(4),xanthotoxol(5),cnidilin(6),byakangelicin(7),bergaptol(8),byakangelicol(9),pabulenol(10),oxypeucedanin hydrate(11),desmodimine(12),palmitic acid(13),stiamasterol(14),?-sitosterol(15),?-daucosterin(16),and sucrose(17).Conclusion: Compounds 8 and 12 have been isolated from Angelica dahurica var.formosana for the first time.Compound 12 has been firstly isolated from Umbelifera plants.All the courmarin compounds belong to linear 6,7-furanocourmarins.

Objective To assess the clinical efficiency of percutaneous coronary intervention (PCI) on cor-onary artery disease patients with complicated lesions (CAD-CL). Methods A total of 252 patients of CAD-CL treated with PCI from August,2002 to September,2009 were enrolled in the study. Clinical data on prognosis e. g, success rate, complication occurrence and major adverse cardiac events were observed. Results Two hundred and forty-four patients with CAD-CL(including chronic obstruction, bifurcation lesion, long lesion and restenosis) were implanted stent successfully(96.83%), with no residual stenosis or residual stenosis less than 20%. There were col-lateral obstructions in 32 patients and artery secret compartment in 48 patients, and restenosis in 15 patients. One patient of acute anterior myocardial infarction complicated by cardingenic shock was in low blood pressure status after two anterior descending artery stent placement, and died after temporary pacemaker implantation and other positive rescue measures. One patient occurred subacute stent thrombosis on the third day, and was cured after occlusive vas-cular recanalization through emergency PCI. Conclusions Percutaneous coronary intervention for coronary artery disease patients with comphcated lesion is safe and effective.

Objective:To synthesize disubstituted pyridine and I-substituted propynyl carbamates and study their antimicrobial activities, searching for more potent and less toxic antimicrobial agents.Methods: The title compounds were synthesized through the process of electrophilic and nucleophilic substitution. Antimicrobial test in vitro was determined with 13 kinds of common mildews and bacteria(Aspergullus niger, aspergillus flavus, Aspergillus versicolor,Trichoderma viride, Paecilomium varioti Bainier, Chaetomium globsum, Penicillium citrinum, Cladochytrium clodospoium, Escherichia colo, Staphylococcus aureus, Pseudomonas fluorescens Bacillus fluorescens, and Bacillus megatherium). The chemical structures of all compounds were determined by IR, 1 HNMR and elementary analysis.Results: Seven disubstituted pyridine and I-substituted propynyl carbamates obtained were firstly reported. All compounds showed antimicrobial activity, especially compound 3e, who had more potent activity compared with the that of 3-iodo-2-propynyl-butyl- carbamate (IPBC). MIC of the compound 3e was between 2?10 -6 g/ml to 30?10 -6 g/ml . Conclusion: Compound 3e has the best antimicrobial activity and should be futher studied.

Objective To explore the effects of Shihusan on the intracellular free calcium of human retina cells and the protection from the damage of glutamate. To investigate its function and mechanism in retinal degeneration diseases, and to offer the basis on which to treat the similar disease clinically. Design Experimental study. Participants Retinal cells from normal human eye. Methods Cultivation of human retinal neural cells was perfonned. The changes of fluorescent density of intracellular free calcium of cultured cells labeled with Fluo3/AM before and after adding Shihusan and verapamil were detected with laser scanning confocal microscopy. In addition, the effect of Shihusan on calcium overload provoked by glutamic acid was observed. Main Outcome Measures The level of intracellular free calcium of retinal cells. Results The retinal cells adhered, spread and grew well in vitro. The fluorescence intensity increased 88% in glutamic acid group, decreased 14.8% and 57.3% in Shihusan group and verapamil group respectively. Shihusan couldn't decrease the intracellular Ca2+ that was increased by glutamic acid, but could inhibit the increasing tendency induced by glutamic acid. The difference was significant compared to control group (P=0.000). Conclusions Shihusan might decrease the level of intracellular free calcium in cultured human retinal cells, and resist the damage of glutamate. Its mechanism was related possibly to resisting calcium overload and inhibiting apoptosis.

BACKGROUND: Osteogenic ability of bone morphogenetic protein-2 has been wel documented in many experiments, but a series of factors are involved in osteogenesis induction that is a complex network adjustment process. OBJECTIVE: To quantitatively determine the level of insulin-like growth factor I during the lumbar spinal fusion of rabbits induced by recombinant human bone morphogenetic protein-2. METHODS: Sixty adult male New Zealand white rabbits were randomly divided into three groups: bone autograft, bone al ograft or composite bone (bone al ograft with 75 μg recombinant human bone morphogenetic protein-2) was implanted into the L5-6 intertransverse process of rabbits, respectively. At days 7, 14, 21, 28, 35 after implantation, formed cal us was taken to detect the expression of insulin-like growth factor I using real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: In the three groups, the expression of insulin-like growth factor I gradual y increased with implantation time, peaked at 28 days and then decreased. At 7 days after implantation, the expression of insulin-like growth factor I was higher in the autograft group than the composite and al ograft groups (P < 0.05); at 14 days, the expression of insulin-like growth factor I was higher in the autograft and composite groups than the al ograft group (P < 0.05); at 21, 28 and 35 days, the expression of insulin-like growth factor I was higher in the composite group than the autograft and al ograft groups (P < 0.05). These findings indicate that recombinant human bone morphogenetic protein-2 can improve the expression of insulin-like growth factor I effectively during the lumbar spinal fusion.

Objective To investigate the curative effect of sodium nitroprusside in the treatment of EV71-related pneumoedema/pneumorrhagia and hypotension in this study. Methods This was a retrospective study of a total 10 children with EV71-related pneumoedema/pneumorrhagia treated with sodium nitroprusside using case-control methods. The clinical manifestations and outcomes of the 10 children (present cohort) were compared with those of 8 children (past cohort) who had EV71-related pneumoedema/pneumorrhagia without sodium nitroprusside. Results Among these 10 patients,all were at the appearance of pneumoedema/pneumorrhagia, cardiac arrhythmia and hypotension. Two patients showed severe cardiac arrhythmia, two patients showed cardiac arrest for many times. All 10 patients were treated with mechanical ventilation and other conventional treatments. At the first time of the manifest of hypotension,sodium nitroprusside were put into practice in all 10 patients. Ten patients were treated with intravenously sodium nitroprusside in the stage of hypotension by maxi30 min ～ 1 h,the patients showed an improvement in vultus,pulse and peripheral circulation and decrease of heart rate and elevation of blood pressure after 30 min ～2 h,but at least 2～6 h later,the blood pressure tended to stabilize at normal standard. Conclusion Hypotension is the intensive stage in EV71-related hand,foot and mouth disease ,and the shock syndrome caused by acute left ventricular disorder is related to sympathetic nerve activity. Sodium nitroprusside can effectively reduce the cardiac afterload,and correct shock and improve the prognosis.

Objective:To analyze the curative effects of laparoscopic radical prostatectomy for the prostate cancer and the its effect on the level serum hormones levels.Methods:86 patients with prostate cancer were selected and divided into the control group and the observation group with 43 cases in each group according to the draw method.The control group was treated by open radical prostatectomy,while the observation group was treated by laparoscopic radical prostatectomy.The operation indicators,serum follicle stimulating hormone (FSH),luteinizing hormone (LH),dihydrotestosterone (DHT),free testosterone (FT),total testosterone (T),prostate specific antigen (T-PSA,F-PSA) levels,CD3+,CD4+,CD8+,CD4+/CD8+ before and after the surgery as well as the occurrence of postoperative complications were compared between two groups.Results:The operation time of observation group was significantly longer than that of the control group,and the blood loss,bowel function recovery time,hospital stay,pain scores of observation group was significantly lower or shorter than those of the control group (P＜0.05).No significant difference was found in the serum FSH,LH,DHT,FT,T,T-PSA,F-PSA levels before and after the surgery between two group(P＞0.05).The CD3+,CD4+,CD4+/CD8+ of observation group was significantly higher than those of the control group (P＜0.05),the CD8+ of observation group was lower than that of the control group(P＜ 0.05).The incidence rate of postoperative complication in the observation group was significantly lower than that of the control group (P＜0.05).Conclusion:Laparoscopic radical prostatectomy could play a similar effect with the open surgery on the tumor control,it could improve the level of serum testosterone,immune function with high safety.

Objective:To discuss the effect ofChinese herbal medicine ofremoving toxic and blood stasis and nourishing yin on lymphocyte apoptosis in spleen ofMRL/lpr mouse and expression ofCyt-C and Bcl-2.Methods:50 2-month-old male MRL/lpr mouse and 10 2-month-old male C57BL/6 mouse were randomly divided into six groups:high, middle and low dosage groups, pre-treatment group, model group and control group.After 30days treatment, the lymphocyte apoptosis in spleen oflupus mouse and expression ofCyt-C mRNA and Bcl-2 mRNA were detected.Results:The percentages oflymphocyte apoptosis in model group were(57.18?1.72)% lower than that in normal group(78.99?3.76)%(P