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 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.

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 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 evaluate MRI in judging the therapeutic effect of argon-helium cryoablation therapy for prostate cancer. Methods The clinical data and imaging materials of 16 patients with prostate cancer , who had received ultrasound-guided argon-helium knife cryogenic treatment at authors ’ hospital during the period from March 2012 to Oct. 2014, were retrospectively and comprehensively analyzed. The preoperative and postoperative laboratory test results were compared with MRI findings, and the ablation effect was assessed, focusing on the surgical residue, metastasis, etc. Results One months after the surgery, MRI demonstrated that satisfactory ablation extent was obtained in all patients, the ablated tumor tissue was characterized by long-T2 signal and no obvious recurrence could be found on DWI and DCE-MRI. A slight decrease of PSA level was observed. Six months after the surgery, MRI revealed that the prostate size was significantly reduced, PSA level was markedly decreased and no obvious evidence of recurrence was observed. No severe postoperative complications, such as urethral necrosis, urethral stricture or urethro-rectal fistula, occurred. During the 6-month following-up time, one patient died from other causes. Conclusion MRI has excellent clinical application value in estimating the ablation extent and in judging postoperative recurrence or metastasis of prostate cancer after argon-helium cryoablation treatment.

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 investigates the efficacy and tolerability of second line treatment with S-1 plus thalidomide in patients with advanced pancreatic cancer.Methods Sixty patients hospitalized in Department of Oncology of Cangzhou Central Hospital from July 2010 to October 2012 were included in this study.All the patients were diagnosed as having pancreatic carcinoma.The patients were randomly divided into two groups,one group was treated with S-1 alone,and the other group was treated with S-1 plus thalidomide.Then the efficacy and toxicity of two groups was evaluated.Results The disease control rates were 36.7％ and 46.7％ in the S-1 alone group and the S-1 plus thalidomide group,and the difference between the two groups was not statistically significant (P =0.31).The progression free survival (PFS) was 2.9 months and 3.3 months,and the difference between the two groups was statistically significant (P ＜ 0.05),the Karnofsky score,pain,sleep and weight improvement rate was 63.3％,46.7％,66.7％ and 53.3％ in combination group,which were significantly better than those in control group (30.0％,13.3％,30.0％ and 20.0％),and the difference between the two groups was statistically significant (P ＜ 0.05).The major adverse events were nausea,vomiting,fatigue and drowsiness,mainly of grades Ⅰ ～ Ⅱ.Conclusions S-1 plus thalidomide as second line treatment of pancreatic cancer can prolong the PSF of patients with advanced pancreatic cancer with excellent safety,and patients' quality of life is also improved.

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