Objective To investigate the expression of GABA_A receptor in the process of activation of hippocampal neurons and explore the epileptogenesis. Methods The activated hippocampal neuron products (experiment group) and the serum-free-medium (control group) were injected into rat's lateral ventricle, respectively, then we observed the behaviors, electroencephalographic (EEG) and immunochemical changes of GABA_A receptor in rats. Results 10~30 min after administration of pentetrazole (PTZ)-activated hippocampal neuronal products, Ⅱ～Ⅲ class epileptic behaviors were found in the experiment group. EEG showed many short distance moderate amplitude sharp waves, sharp and slow wave complex. The behaviors of rats and EEG results were normal in the control group. GABA_A receptor positive cells extensively spread over cerebral cortex, hippocampal CA_1 and dentate gyrus in the control group, and the expression of GABA_A receptor significantly decreased in the experiment group.Conclusion Activated hippocampal neuronal products could induce epilepsy. One of its mechanisms might be related to the decrease of GABA_A content.

5000 copy/ml were both significantly higher than those in the controls.The levels of HBV-DNA positive in cultured supernatants of PBMC from suffering grave hepatitis B were higher than those in the sera by qualitative PCR of the same period.They were similar between the positive rate of quantitative PCR of HBV-DNA in sera and FQ-PCR in cultured supernatants of PBMC from hepatitis B types.They were not obviously related with the levels of ALT and TBiL.Conclusions The IL-12 and ? IFN secretion increased when the HBV-DNA of PBMC from carriers of HBeAg were at high-reproducing state.The HBV-DNA positive rate in cultured supernatants of PBMC of grave hepatitis detected by FQ-PCR was higher than that in sera by qualitative PCR.

Objective To analyze the consistency of transcutaneous perianal ultrasonography (TPUS) and pelvic magnetic resonance imaging (MRI) in diagnosing perianal lesions of Crohn's disease (CD), and to evaluate the value of transcutaneous perianal ultrasonography in detecting perianal lesions of CD. Methods A cohort of 102 patients diagnosed as Corhn's disease were enrolled from August 2008 to August 2010. Perianal abscess and fistula of these CD patients was diagnosed by ultrasonography and MRI system. Statistics was performed with SPSS 11.5 software for X2 test. The consistency was analyzed with Kappa test. Results The mean onset time of perianal lesions in CD was -0.443 year (95％CI:-1.659～0.773 year) before typical symptoms showed up. There was no significant difference in detecting perianal lesions of CD between transcutaneous perianal ultrasonography and pelvic magnetic resonance imaging (P ＝ 0.706, Kappa ＝ 0.541). If pelvic magnetic resonance imaging was considered as the golden standard in detecting perianal lesions of CD,the sensitivity (Sen), specificity (Spe), Youden's index, positive predictive value (PPV) and negative predictive value (NPV) of TPUS were 72.73％, 82.61％, 0.55, 66.67％ and 86.36％ respectively.Furthermore, there was no significant difference between transcutaneous perianal ultrasonography and pelvic magnetic resonance imaging in detecting perianal abscess ( P ＝ 0.706, Kappa ＝ 0.496) and fistula (P＝0.655, Kappa＝0.546) of CD. Conclusions Perianal lesions occur in the entire course of CD. There was favorable consistency between transcutaneous perianal ultrasonography and pelvic magnetic resonance imaging in detecting perianal abscess and fistula of CD. Transcutaneous perianal ultrasonography can be used as an additional method in detecting and evaluting perianal lesions of CD.

Objective To explore the value of application of choledochofiberscopy in the diagnosis and treatment of the extrahepatic bile duct disease, and the effect on reducing the incidence of the postoperative residual stone in biliary ducts. Methods According to the case history and ultrasonography,if the common bile duct(CBD) diseases suspected,the CBD was explored by intraoperative choledochofiberscope(IOCF). During the procedure,a biliary passage mirror inducer apparatus and biliary tract probe which were manufactured by ourselves were used. Results During LC,IOCF was performed on 385 cases of the 10 396 LC cases,and possitive findings were dicovered in 102 cases(26.49%). Among those positive patients, 67 cases belonged to stricture of the lower biliary tract; 5 cases were Mirizzi syndrome; 2 cases were carcinoma of the periampulla; 1 case was primarily carcinoma of the bile duct; 1case was ascarisis of the biliary system. Conclusions IOCF is a good inspect technique with high success rate and clear image of bile duct, it can discover the common duct diseases which are difficult to be diagnosed through the routine examination.At the same time, it can provide the locative and qualitive diagnosis, determine reasonable methods of operation,and effectively provent postoperative complications.

Objective To investigate the expression of CD40-CD154 co-stimulatory pathway in peripheral circulation and intestinal mucosa in patients with inflammatory bowel disease (IBD),the difference between the expression of CD40-CD154 in patients with IBD and that in healthy controls,and the correlation between CD40-CD154 levels and disease activity. Methods A total of 62 patients with Crohn's disease (CD),64 patients with ulcerative colitis (UC) and 56 healthy controls were enrolled. Enzyme-Linked Immuno Sorbent Assay (ELISA),SYBR-green real time PCR and immunohistochemical assay were respectively applied to evaluate expression of CD40-CD154 in plasma,mononuclear cells of peripheral blood and intestinal mucosa. Results Levels of CD40 (P=0. 000) and CD154 (P=0. 001) in plasma,mononuclear cells of peripheral blood and intestinal mucosa were significantly higher in patients with CD and UC than in healthy controls. However,no correlation between disease activity and CD40-CD154 expression in peripheral circulation or intestinal mucosa was detected (P ＞ 0. 05 ). Conclusion CD40-CD154 pathway activation is found in plasma,peripheral blood mononuclear cells and intestinal mucosa in patients with IBD,but is not correlated with disease activity.

Objective To explore the function of infliximab in inducing remission in Crohn's disease and the effect of the inducing remission were followed up. Methods Ten patients with Crohn's disease received a infliximab, 5-aminosalicylic acid (5-ASA) and Azathioprine (AZA) therapy for inducing and maintenance remission. Crohn' s disease activity index (CDAI), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), alanine aminotransferase (ALT), apartate aminotransferase, (AST), total bilirubin (TBil), conjugated bilirubin, (CB), creatinine (Scr) were evaluated at week 0, 10, 22 and 50. Simple endoscopic score for Crohn's disease (SES-CD) were evaluated at week 0, 10 and 50. Adverse reactions were also evaluated. Results At week 10, all patients achieved remission. The indicators of CDAI, CRP, ESR and SES-CD were significantly declined than those at week 0 (P＜0.01). The follow-up was terminated in one patient due to the relapse at week 30. At week 50, the indicators of CDAI, CRP, ESR and SES-CD in six patients a little bit increased compared with those at week 10, but no statistic significant (P=0. 2001、0. 0600、0. 1328、0. 4230 respectively), but significantly declined compared with those at week 0 (P =0.0005、0.0087、0.0054、0. 0163 respectively). No severe adverse reaction was observed in all patients.Conclusions Infliximab showed an exact efficacy in inducing remission in Crohn's disease. And 5-ASA and AZA were effective for maintenance remission in part of the patients after infliximab induced remission.

Objective To understand the Helicobacter pylori ( Hp) infection eradication rate of standard tri-ple therapy in Guangdong Meizhou and the drug resistance situation for metronidazole ,clarithromycin ,amoxicillin and levofloxacin ,in order to look for the treatment countermeasures in Hp eradication failure .Methods 297 cases of Hp positive patients because of gastrointestinal symptoms to our hospital examined from April 2011 and March 2013,were randomly assigned into three standard triple therapy groups:A ( OCA ) group and B ( OCM ) group and C ( OCL ) group.The Hp eradication rate was analyzed .Patients with primary treatment failure were selected as group D (OBAL),proceed to (PPl+B+A+L)7 d therapy,the Hp eradication rate was analyzed .230 Hp strains were isola-ted and cultured from 297 cases received the first eradication therapy and 87 cases received again eradication therapy . The minimum inhibitory concentration (MIC) of metronidazole,clarithromycin,amoxicillin and levofloxacin were tested by E-test,in order to determine the resistance of these four antibiotics in clinical isolated Hp strains .Results With intention-to-treat(ITT) analysis,the Hp eradication rates of group A (OCA),group B(OCM) and group C(OCL) were 72.0%(72/100),63.0%(63/100) and 72.2%(70/97),respectively.With per-protocol(PP) analysis,the Hp eradication rates of group A (OCA),group B(OCM) and group C(OCL) were 72.7%(72/99),64.3%(63/98),73.7%(70/95),respectively.The eradication rate among three standard triple therapy groups had no obvi-ous difference (ITT:P=0.278,PP:P=0.288,P>0.05).With ITT analysis,the Hp eradication rate in the quadrup-le therapy group D(OBAL) was 92.0%(80/87).With per-protocol(PP) analysis,the Hp eradication rate in the quadruple therapy group D(OBAL) was 97.6%(80/82),which was higher than that of the three standard triple ther-apy groups(ITT:P=0.000,PP:P=0.000).In 230 clinical isolated Hp strains,the resistant rates of levofloxacin,amoxicillin,clarithromycin and metronidazole were 6.08%(14/230),6.52%(15/230),25.65%(59/230), 70.87%(163/230),respectively.Of those 37 strains were mixed resistance,the mixed resistant rate was 16.09%(37/230).The resistant rate of metronidazole was higher than levofloxacin , amoxicillin and clarithromycin ( P =0.000,P<0.01),the resistant rate of clarithromycin was higher than levofloxacin and amoxicillin (P=0.000),no statistically significant difference between amoxicillin and levofloxacin (P=0.848).Conclusion The Hp resistance is similar to the national average in Guangdong Meizhou ,the eradication rate of standard triple therapy is lower than 80%,contain bismuth agent of quadruple therapy is good rescue therapy .

AIM:To detect the activation of macrophage autophagy caused by lipopolysaccharide ( LPS) and the possible related signaling pathways .METHODS:The macrophage cell line RAW264.7 cultured in vitro was divided into 5 groups according to the culture environment , including normal culture group , starvation-activated sautophagy group , LPS group, LPS+PI3K inhibitor (hVps34) group and LPS+mTOR inhibitor (rapamycin) group.Fluorescent expression vector pcDNA3.1-GFP-LC3 constructed in previous work was transfected into the macrophages .The fluorescence microscopy was used to detect the formation of autophagosome .The mRNA expression of autophagy-associated genes Atg5, Atg7, LC3-II and Bnip3 in the macrophages was detected by qRT-PCR.The protein levels of LC3-II, p-Akt and p-mTOR were deter-mined by Western blotting , so as to evaluate the molecular pathways of autophagy in LPS-activated macrophages .RE-SULTS:The macrophages stably expressing GFP-LC3 were successfully established , which were used to observe the auto-phagy under fluorescence microscope .Compared with normal culture group , the autophagy in starvation group , LPS +hVps34 group and LPS+rapamycin group was significantly increased .The mRNA expression levels of Atg5, LC3-II and Bnip3 were significantly increased in starvation group , LPS+hVps34 group and LPS +rapamycin group , while in LPS group, those decreased slightly .The protein level of p-Akt in starvation group , LPS group and LPS+rapamycin group was significantly increased , while p-mTOR in starvation group , LPS+hVps34 group and LPS+rapamycin group significantly declined .LC3-II expression level in starvation group , LPS+hVps34 group and LPS+rapamycin group was higher than that in control group and LPS group .CONCLUSION: LPS regulates macrophage autophagy , and its possible pathway is the PI3K/Akt/mTOR pathway, but there are some other effective regulatory pathways .

To obtain the immunized mouse spleen cells, we immnnized the female BALB/C mice I.P. with schizonts or merozoites of the cultured blood forms of Plasmodium falciparum. The immunized spleen cells were fused with the SP2/0 myeloma cells, by which we obtained two strains of hybridoma secreting monoclonal antibodies against human Plasmodium falciparum. Using indirect immunofluorescence assay, we identified that the antibodies reacted only with the surface membrane antigens of the schizonts. These hybridomas were cultured continuously in vitro for over ten months and stably produced antibodies-IgG. The two hybridoma cell lines were designated as AEB2 and AGA4, and the number of their chromosomes was 98 and 100 respectively. The hybridoma cells were injected I. P. into the paraffin oil treated BALB/C mice to obtain the hybridoma ascitic fluid containing monoclonal antibodies. The ascitic fluid inhibited the growth of P. falciparum in vitro up to 89%. The inhibition test showed that antibodies were protective.

DNA copy number variation is an important pathogenic factor of human diseases and might be involved in the pathogenesis and pathological process of inflammatory bowel disease (IBD).Aims: To investigate the copy number variation of CNTNAP3 gene and its significance in Crohn''s disease (CD).Methods: A total of 101 active CD patients admitted from Jul.2009 to Dec.2010 at Renji Hospital, School of Medicine, Shanghai Jiao Tong University were enrolled.Eighty healthy subjects were served as controls.Peripheral blood or intestinal mucosa samples of CD patients were collected, and the copy number variation of CNTNAP3 gene was screened and validated by array-based comparative genomic hybridization (aCGH, n=8) and real-time PCR (n=93);expression of CNTNAP3 encoding protein was determined by ELISA (n=55).Results: A large fragment copy number amplification was revealed by aCGH at chromosome 9p13 region (including CNTNAP3 gene) in untreated CD patients.Real-time PCR confirmed that the copy number of CNTNAP3 gene was amplified in peripheral blood of CD patients, especially steroid-naive patients as compared with the normal controls (208 616.4±126 984.7 and 233 453.3±113 520.8 vs.161 750.2 ±53 940.3, P<0.05 and P<0.01).In the clinical parameters analyzed in this study, only smoking was significantly correlated with CNTNAP3 copy number amplification (P<0.05).However, there was no significant difference in plasma CNTNAP3 level between CD patients with amplified copy number and normal controls (P>0.05).Furthermore, the plasma CNTNAP3 level in CD patients with amplified copy number was not correlated with the simplified endoscopic score for CD (P>0.05).Conclusions: Copy number amplification of CNTNAP3 gene might be involved in the pathogenesis of CD in Chinese population.Glucocorticoid treatment and smoking might affect the copy number variation of CNTNAP3 gene.Plasma CNTNAP3 level cannot discriminate CD patients from healthy subjects.Conclusions of this study needs to be further demonstrated and discussed.