Objective To investigate the protective effect of Qiweibaizhu Powder against HRV infection of intestinal mucosal epithelial cells of suckling mice.Methods The 5-day-old NIH mice were made HRV diarrhea model by oral infection,and randomly divided into six groups:normal group(NG),model group(MG),Qiweibaizhu Powder group(BG),ribavirin group(ZG),suckling mice in each group were instilled corresponding drugs 3 times/d(NG and MG with normal saline) through the mouth.5 d after treatment,suckling mice were killed,small intestine stool was taken to test HRV,and pathological changes in small intestinal mucosa were observed by HE staining.Results Intestinal faeces HRV clearance of BG was significantly better than MG(P

Objective To explore the surgical treatment of bile duct necrosis.Methods Clinical data of 94 cases of bile duct necrosis treated in this hospital from May1990 to December 2008 were retrospectively analyzed.Results There were no death or severe complications such as biliary fistula and massive hemorrhage in these patients.Conclusion Bile duct necrosis should be treated with a proper surgical approach based on its features.

OBJECTIVE:To establish a method for the analysis of chemical compositions in Jinqiancao granule. METHODS:HPLC-ESI-Q-TOF-MS was adopted. The chromatographic conditions:the column was Inertsil ODS-4 with mobile phase of 0.1%formic acid- methanol (gradient elution) at a flow rate of 0.8 ml/min,column temperature was 30 ℃,and the injection volume was 15 μl. MS conditions:ion source was ESI(negative ion mode),endplate offset voltage was -500 V,capillary electrophore-sis was 3500 V,carrier gas was helium gas,atomization and drying gas was high purity nitrogen gas at a flow rate of 6 L/min and a pressure of 1.0 bar,drying air temperature was 180℃. Scanning range was 50-1500 m/z. ChemBioDraw Ultra13.0 and Bruk-er data analysis software 4.0 were used to analyze the chemical composition molecular formula. RESULTS:A total of 27 kinds of chemical components were identified in Jinqiancao granule,involving sucrose,uridine,gallic acid,new chlorogenic acid,(-)-epi-gallocatechin,protocatechuic acid,chlorogenic acid,schaftoside,caffeic acid,schaftoside,vanillc acid,ferulic acid,hyperin,ros-marinic acid,rutin,isoquercitrin,astragalin,quercetin-3-rhamnoside,myricetin,4-hydroxybenzoic acid,quercetin,naringenin, luteolin,kaempferol,isorhmnetin,apigenin and emodin. CONCLUSIONS:Caffeic acid,rosmarinic acid,vanillic acid,(-)-epigal-locatechin,uridine and emodin are firstly found and reported in the chemical compositions in Jinqiancao granule.

Objective To assess the causes and methods of surgical treatment of hepatolithiasis reoperation Methods The clinical data of 81 cases of hepatolith reoperation were analyzed retrospectively. Results The main causes of hepatolith reoperation include biliary stricture,biliary tract variation,cholangiocarcinoma, etc. The chief reoperation patterns were hepatic lobectomy, Roux-en-Y hepaticojejunostomy, and lobectomy combined with Roux-en-Y hepaticojejunostomy.A follow-up of 2 months to 12 months showed excellent (outcome) of 93.8% of cases. According to postoperative cholangiograph,the retained stone rate was 6.2%. Conclusions When treating cholelithiasis ,we should follow the following principles: remove stones, relieve biliary stricture, correct biliary variation, resect abnormal liver, and establish adeguate biliary drainage.

OBJECTIVE:To systematically review the difference of phlebitis induced by Alprostadil injection with different ad-ministration routes,and to provide evidence-based reference for clinical rational use. METHODS:Retrieved from PubMed,EM-Base,Cochrane Library,CBM,CJFD,VIP and Wanfang database,RCTs about phlebitis induced by Alprostadil injection with dif-ferent administration routes were collected. Meta-analysis was conducted by Rev Man 5.2 statistical software after literature screen-ing,data extraction and quality evaluation according to Cochrane System Evaluator's Manual 5.1.0. RESULTS:A total of 20 RCTs were included,involving 2562 patients. The results of Meta-analysis showed that the incidence of phlebitis induced by intravenous injection was significantly higher than that induced by intravenous dripping [OR=4.11,95%CI(1.59,10.67),P=0.004] and intrave-nous pump [OR=3.50,95%CI(1.50,8.16),P=0.004]. The incidence of phlebitis induced by general apparatus infusion was signifi-cantly higher than that induced by fine filtering infusion [OR=0.03,95%CI(0.01,0.08),P<0.001],with statistical significance. CONCLUSIONS:The incidence of phlebitis induced by low-concentration of Alprostadil injection or fine filtering infusion is low-er,and that of intravenous injection is higher.

Objective To explore the molecular regulation mechanism of asporin in the matrix synthesis and secretion of the intervertebral disc,and to clarify its role in degenerative lesions of intervertebral disc.Methods There were 8 cases of intervertebral disc tissue in patients with severe intervertebral disc herniation (including typical clinical symptoms,signs and Pfirrmann's grade Ⅲ).There were 6 male and 2 female with an average age of (20.25 ± 3.37) years old (ranged from 11 to 28 years).After primary culture and redifferentiation in alginate beads,cells were reseeded and treated with different concentrations of TGF-β1 for 6,12,18 and 24 h.Total RNA extracted from the cells and was subjected to real-time PCR analysis to examine the expression of asporin.After silencing the expression of endogenous asporin by siRNA,the cells stimulated 24 h with TGF-β1.Total RNA extracted from the cells and was subjected to real-time PCR analysis to examine the expression of asporin,collagen Ⅱ and proteoglycans.After treatment of specific p38 inhibitor or ERK inhibitor for 12 h,cells were stimulated with TGF-β1 for 24h.Protein extracted from the cells by protein extraction kit to examine the level of asporin.Results In the primary intervertebral disc cell experiment,TGF-β1 stimulation induced asporin transcription significantly in a dose and time dependent manner.After 24 h stimulation,a significant difference between different concentration groups (5,10 and 15 ng/ml) was observed,2.754±0.24,3.651 ±0.319 and 4.583±0.38,respectively (F=24.782,P=0.001).Knockdown of endogenous asporin led to the upregulated expression of aggrecan and collagen]Ⅱ (aggrecan:t=7.387,P=0.002,collagen Ⅱ:t=4.443,P=0.0113).Specific p38 inhibitor was used to block p38 phosphorylation,and TGF-β1 on asporin induction was significantly inhibited.Conclusion Our results have verified a functional feedback loop between TGF-β1 and asporin in human intervertebral annulus cells indicating that TGF-β1 can increase asporin expression,whereas asporin inhibits TGF-β 1 signaling pathway by negative feedback,thereby inhibiting TGF-β1 mediated synthesis of extracellular matrix,and TGF-31 can increase asporin expression by p38 in human intervertebral disc cells.

Objective To investigate the gene expression of PIWIL2 in the bladder urothelial carcinoma (BTCC) and siRNA interact on PIWIL2 gene expression in human bladder cancer cell line BIU-87.Methods Semi-quantitative reverse transcription polymerase chain reaction (qRT-PCR) was applied to detect the PIWIL2 expressions in tissues of BTCC (46 cases),cystitis glandularis(21 cases),adjacent non-cancerous tissues (17 cases) and normal bladder tissues (7 cases). 3 specific siRNA targeted PIWIL2 gene were synthesized after designed and transferred. After siRNA was transferred into BIU-87 cells, MTI and TUNEL methods were applied to detect the proliferation inhibitory rate (IR) and apoptosis index (AI) in BIU-87 cells,qRT-PCR and Western blot were used to examine effects of siRNA on the expressions of the PIWIL2 gene and protein,respectively.Results The expression rate of PIWIL2 mRNA in BTCC tissues was 76.08 ％(35/46) and significantly higher than those in the cystitis glandularis tissues (42.86 ％,9/21),adjacent non-cancerous tissues (41.17 ％,7/17) and normal tissues (7.14 ％,1/14) (P =0.008,P =0.010,P =0.000).The IR [(37.52±8.84) ％,(64.36±9.64)％] and (62.94±8.43) ％] and AI [(26.18±5.42) ％,(38.75±6.19) ％ and (40.02±5.64) ％] of BIU-87 cells in the siRNA 1～3 groups were respectively significantly higher than those [(1.97±0.02) ％ and (3.35±0.47) ％] in the control group(P=0.000),and expressions of PIWIL2 mRNA and protein in the siRNA groups were both lower than those in the control group. Moreover, the effects of siRNA 2 group and siRNA 3 group on inhibiting PIWIL2 expression, IR and AI of BIU-87 cells were stronger than siRNA 1 group. Conclusion The over-expression of PIWIL2 suggested that it played an important role in the mechanism of development and malignant progression of BTCC. The siRNA of transcription can significantly inhibit its expression, induce cell apoptosis and inhibit the growth of BIU-87 cells which might provide the experimental evidence for the gene targeting therapy of bladder tumor.

Objective To observe the effects of ( - )-epigallocatechin-3-gallate (EGCG) on human prostate cancer xenografted tumor growth and connexin43 expression in nude mice,and explore the mechanism of the EGCG on prevention for prostate cancer.Methods The methyl thiazolyl tetrazolium and annexin-V/PI double-labeled flow cytometry methods were used to observe the growth inhibiting rate (IR)and apoptosis rate (AR) of human prostate cancer cell line PC-3 which was treated by EGCG at different concentration (10,20 and 40 mg/L,respectively).The scrape-loading fluorescence dye transfer method was applied to assess the gap junction intercellular communication (GJIC) through fluorescence microscope.PC-3 cells were subcutaneously transplanted to establish tumor-bearing nude mice model.A total of 32 mice were randomly divided into four groups,both control group and three treatment groups were treated with different doses of EGCG ( 10,20 and 40 mg/kg,respectively).After two weeks,the mice prostate tumor tissues were taken out.The tumor wet weight was measured and tumor growth inhibiting rate was calculated.The tumor microvascular density (MVD) and apoptosis index (AI) were detected by the immunohistochemical techniques and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling techniques,respectively.Semi-quantitative reverse transcription polymerase chain reaction was used to examine the expression level of the Cx43 mRNA.Results EGCG at concentration ( 10 and 20 mg/L) could significantly inhibit the proliferation[(22.33 ±4.62)％,(38.67 ±5.67)％ vs (3.47 ±0.31 )％,P ＜0.01],induce the apoptosis [(7.84 ± 1.37 ) ％,( 24.53 ± 2.28 ) ％ vs ( 2.17 ± 0.70 ) ％,P ＜ 0.01] and enhance the GJIC of PC-3 cells.EGCG of different doses could inhibit prostate cancer xenografted tumor growth,induce tumor cells apoptosis and inhibit angiogenesis.EGCG ( 20 and 40 mg/kg) could effectively up-regulate Cx43 mRNA expression in xenografted tumor (0.58 ± 0.08,0.80 ± 0.07 vs 0.42 ± 0.04,P ＜ 0.0 ).The effects had significant correlation with the dose-dependent of EGCG ( P ＜ 0.05 ).Conclusions EGCG could up-regulate the Cx43 expression and enhance the gap junction intercellular communication mediated by Cx43 in the prostate tumor,which provide the experimental evidence for the mechanism of its effectively inhibiting the prostate cancer growth.

E2 is an envelope glycoprotein of Classical swine fever virus (CSFV) and contains sequential neutralizing epitopes to induce virus-neutralizing antibodies and mount protective immunity in the natural host. In this study, four antigen domains (ABCD) of the E2 gene was cloned from CSFV Shimen strain into the retroviral vector pBABE puro and expressed in eukaryotic cell (PK15) by an retroviral gene expression system, and the activity of recombinant E2 protein to induce immune responses was evaluated in rabbits. The results indicated that recombinant E2 protein can be recognized by fluorescence antibodies of CSFV and CSFV positive serum (Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China) using Western blot, indirect immunofluorescence antibody test (IFAT) and ELISA, Furthermore, anti-CSFV specific antibodies and lymphocyte proliferation were elicited and increased by recombinant protein after vaccination. In the challenge test, all of rabbits vaccinated with recombinant protein and Chinese vaccine strain (C-strain) were fully protected from a rabbit spleen virus challenge. These results indicated that a retroviral-based epitope-vaccine carrying the major antigen domains of E2 is able to induce high level of epitope-specific antibodies and exhibits similar protective capability with that induced by the C-strain, and encourages further work towards the development of a vaccine against CSFV infection.

Objective To study the proliferation,collagen production and related gene expression in keloids and normal skin fibroblast.Methods Isolated primary cells of keloid fibroblasts (KFb,n=12) and normal human dermal fibroblasts (NFb,n=12) were identified,the cell viability and proliferating potential and the cell cycle were detected,and the difference on the collagen synthesis between KFb and NFb were compared.The expression of cell cycle-associated genes such as p21,p16,and p27 was dectected by real-time fluorescent quantitative PCR.Results The phase contrast optical microscopy imaging showed that both KFb isolated from keloid tissues and NFb from normal skin tissues possessed classic and similar fibroblast morphology.But there was a significant difference between cell proliferation,Hyp [(2.30±0.10) μg/ml vs.(1.66±0.13) μg/ml,P＜0.05] and collagen levels [(17.19±0.75) μg/ml vs.(12.37±0.94) μg/ml,P＜0.05].Compared with NFb,KFb exhibited more percentage of G2/M phase cells [(5.90±0.62)％ vs.(16.94 ％±1.93)％,P＜0.05]and less percentage of G0/G1 phase cells [(90.24 ±2.27)％ vs.(75.65±1.92)％,P＜0.05].Cell cycle related genes p16,p21 and p27 were low expressed.Collagen type Ⅰ was highly expressed at mRNA levels in KFb than that in NFb [0.84±0.11,1.32±0.2,1.69±0.12,4.33±0.27 in KFb vs.1.43±0.13,2.56±0.26,2.89±0.37,1.40±0.12 in NFb,P＜0.05].Conclusions There are cell dysfunction and abnormal cellular dynamics in keloid fibroblasts.The formation of keloid likely involves aberrant interactions of some genes that affected its development at different extents.