Objective To sequence and analyze the full-length genome of one HEV strain,W2-1 isolated from a sporadic hepatitis E patient hospitalized in 1999 in Xinjiang,China.Methods Nested RT-PCR assays with 4 sets primers were used to amplify the entire genome.The PCR products were purified and sequenced.The full-length genome was acquired by assembling the fragmental sequences using the DNAstar 5.01 software.The genome of W2-1 was analyzed by comparing with the reference HEVs from GenBank.Results The complete genome of W2-1 is 7212 nt in length,including three open reading frames (ORF1-3) with 5079,1980 and 345 nt respectively,27 nt 5'UTR and 83 nt 3'UTR,and a 3' poly A tail.Phylogenetic analysis based on full-length genome showed that W2-1 belonged to genotype 1,subtype 1b.W2-1 had high homology with the HEV strains isolated in the large hepatitis E epidemic in Xinjiang in 1987-1989,sharing 97.2％-98.5％ nucleotide identity in the full length genome.W2-1 also showed high homology with 1b strains isolated in China after 2000,with 97.6％-99.2％ nucleotide identity.The specific amino acid sites in ORF1-3 proteins that distinct between genotype 1 HEV and the potential zoonotic strains did not change in W2-1.Conclusion W2-1 belongs to subtype 1b.The study indicates subtype 1b HEV has been circulating in China in a long period after hepatitis E outbreak in Xinjiang in 1986-1989.The amino acids of ORF1-3 of subtype 1b are conserved.

Objective To observe the efficacy for the eradication of Helicobacter pylori (Hp) by using the 10-day sequential therapy consisting of omeprazle,amoxicillin,clarithromycin,and furazolidone,and to compare with the traditional triple-drug therapy.Methods 155 adult patients,who were Hp positive with peptic ulcer,chronic gastritis with mucosal atrophy or erosion,were selected and randomly divided into two groups.The patients in 10-day sequential therapy group received omeprazole and amoxicillin in the first 5 days followed by another 5 days of ome-prazole,clarithromycin and furazolidone.The patients in traditional triple-drug therapy group recieved omeprazole,amoxicillin,and clarithromycin for 7 days.Results The HP eradication rate of the 10-day sequential therapy group was 92.31% ,while the traditional triple-drug group was 72.73%.The difference between the two groups was statistically significant (P<0.01).Conclusion The 10-day sequential therapy consisting of omeprazole,amoxicillin,clarithromycin and furazolidone achieved higher eradication rate,and it was safe,and good compliance.

Marburg virus (MARV) is a lethal virus that causes fatal hemorrhagic fever.It belongs to the Filoviridae family which also includes Ebola virus.MARV is similar to Ebola virus in structure and infection mechanism.Moreover,the diseases caused by them have similar clinical symptoms.However,researches on MARV are less than those on Ebola virus.In this review,we focus on the viral structure,especially the structure of MARV glycoprotein (GP) which determines its infectivity,functions of MARV GP as well as protective antibodies and vaccines against this protein.

Innovative medical talents training in higher medical colleges and universities is the requirements of the era. Based on the medical education of Xi'an Jiaotong University,the article systematically introduces some thoughts and methods of talents training system,which includes three aspects such as three-dimensional teaching environment(teaching resource construction,platform construction and construction of teachers' team),experiment curriculum system reform and innovation mechanism,etc. Construction of three-dimensional teaching environment is emphasized.

Objective To establish HIV-1 seroconversion panels with the samples collected by National Institutes for Food and Drug Control ( NIFDC) and to evaluate the window periods of HIV enzyme immunoassay ( EIA) diagnostic kits used for blood screening with them. Methods Serum specimens were collected from different plasma donation stations in China. All suspected HIV infection specimens were screened for HIV by using the nucleic acid amplification testing (NAT), Western blot confirmatory assay and P24 quantitative detection assay. The HIV env gene sequences were amplified by RT-PCR for further confirmation of HIV infection. The PCR products were sequenced and genotyped. The confirmed seroconver-sion panels were used to evaluate the early detection capabilities of the 4th and 3rd generation HIV EIA diag-nostic kits used in blood screening in china. Results A total of 8 sets of HIV seroconversion panels com-prised of 36 samples were confirmed in this study, including 6 sets of AE subtype, 1 set of B subtype and 1 set of unknown genotype. Those seroconversion panels were tested with HIV diagnostic kits produced by 19 different manufacturers. For the early detection of HIV infection, the 4th generation HIV diagnostic kits with a score of 9. 4 points were better than the 3rd generation HIV diagnostic kits whose score was 3. 6 points (P<0. 01, t=8. 547). Some of the domestic 4th generation HIV diagnostic kits were similar to the imported kits in the early detection of HIV infection. In terms of the diagnosis of HIV infection, the HIV-1 NAT was at least 2 weeks earlier than the HIV EIA diagnostic kits. The sensitivity of confirmatory assay was lower than that of the diagnostic kits. Four out of five 4th generation HIV diagnostic kits showed declined signal to cut off ( S/CO ) ratio , indicating the probability of false detection during the second window period . Conclusion Eight sets of NIFDC HIV-1 seroconversion panels were established in this study. With those panels we found that there were differences in the window period between different EIA diagnostic kits used for HIV blood screening.

ObjectiveTo investigate the characteristics of left ventricular structural damage in patients with primary aldosteronism(PA).MethodsA total of 438 inpatients with hypertension from January 2007 to June 2010 were screened for PA,and diagnosis was made in 213 PA patients and 225 patients with essential hypertension (EH).The left ventricular structure of all subjects was evaluated according to the results of echocardiographic measurements.Results The duration of hypertension and plasma aldosterone level in patients with PA were significantly higher (P＜ 0.01 ),while the serum potassium and plasma renin activity were significantly lower (P＜0.01 ) than those in patients with EH.The interventricular septum thickness,left ventricular end-diastolic dimension,left ventricular mass index (LVMI),left ventricular end-diastolic volume,and stroke volume in patients with PA were significantly higher than those in EH patients( P＜0.01 ) after the duration of hypertension was corrected.In patients with PA,the prevalence of left ventricular hypertrophy (LVH) was higher than that in EH patients ( 53.1％ vs 33.8 ％,x2 =16.57,P ＜ 0.01 ). Normal left ventricular geometry ( NG ),concentric remodeling ( CCR ),concentric hypertrophy( CCH),and eccentric hypertrophy (ECH) were found respectively in 24.9％,22.1％,22.1％,and 30.9％ of patients with PA.Multiple stepwise regression analysis showed that the seated plasma aldosterone level (β=0.43,P ＜ 0.01 ) and systolic blood pressure (β =0.45,P ＜ 0.01 ) were the main factors that influenced LVMI.The course was the main parameter that influenced relative wall thickness(β=0.011,P＜0.05 ).Conclusion The prevalence of LVH is higher in patients with PA than that in EH patients.The eccentric hypertrophy is the most common left ventricular geometrical pattern in patients with PA.

Objective To explore the effects of inducible co-stimulator (ICOS) gene on the cytotoxic activity of cytokine-induced killer (CIK) cells against cholangiocarcinoma cells. Methods CIK-ICOS cells were obtained by stable transfecting ICOS genes into CIK cells through the adenovirus vector whereas untransfected and EGFP-transfected CIK cells were treated as controls. The proliferation and apoptosis of different CIK cells, as well as their cytotoxicity against cholangiocarcinoma cells in the three groups were detected. The expressions of IFN-T, IL-2 and TNF-α in the supernatant of different CIK cells were measured by ELISA. SCID mice with cholangiocarcinoma were randomly divided into CIK group, CIK-EGFP group, CIK-ICOS group and normal saline group. The cytotoxic activity of CIK-ICOS cells against cholangiocarcinoma cells in vivo was observed. Results CIK-ICOS cells displayed better proliferation than CIK cells and CIK-EGFP cells. At day 20 and 23 of culture, the apoptosis rate of CIK-ICOS cells was 0.69% and 0.89%, respectively, while that of the CIK cells was 2.90% and 4.92%. The cytotoxic effect of CIK-ICOS cells at different E: T ratio against cholangiocarcinoma cells was significantly stronger than that of CIK cells and CIK-EGFP cells (F=13.37, 6.46, 25.51, P<0.05). The concentration of IFN-γ in CIK-ICOS cultured supernatant was (49.50±4.73)μg/L, which was significantly higher than that in the cultured supernatant of CIK cells [(30.53±3.73)μg/L] and CIK-EGFP cells [(30.12±2.64)μg/L](F=38.89, P<0.05). The growth of cholangiocarcinoma was significantly slower in CIK-ICOS group than that in CIK group and CIK-EGFP group, whereas the necrosis area of tumor was larger and the CIK cells in CIK-ICOS group was more than those in the other two groups. Conclusions CIK cells had the function of killing cholangiocarcinoma cells in vitro and in vivo. After ICOS genes were transfected into CIK cells, the survival time of CIK cells in vitro was prolonged and the proliferation of CIK cells was enhanced, as well as the secretion of IFN-γ was increased so that the cytotoxicity of CIK cells against cholangiocarcinoma cells in vitro and in vivo was enhanced.

Objective To establish a high throughput phenotypic test for the detection of drug re-sistance in human immunodeficiency virus(HIV)strains. Methods The gene encoding luciferase was in-activated through restriction enzyme digestion and ligation. LacZ gene was used to replace the genes encoding original protease and reverse transcriptase. pol genes were amplified from pSG3△env plasmid and cloned in-to a new backbone plasmid through infusion. The factors that might affect the results of the test were opti-mized. Results The parental backbone plasmid pNL4-3. Lac was constructed,of which the gene encoding luciferase was inactivated and bearing the LacZ gene instead of genes encoding protease and reverse tran-scriptase. Several influential factors including cell numbers(10 000 / well),virus inoculation(200 TCID50 /well)and the concentration of DEAE-dextran(15 μg/ ml)were optimized. The reproducibility of this test was confirmed by testing 12 anti-HIV drugs against 2 pseudovirus strains 8 times,presenting the coefficient of variations(CVs)from 4. 32% to 28. 46% . Six types of pseudovirus were constructed and tested against the 12 anti-HIV drugs,the results of which were compared with those by using the pSG3△env-based pseud-ovirus test. The results of the two tests presented good consistency. Conclusion The high throughput phe-notypic test based on pNL4-3. Lac plasmid,combining the advantages of pSG3△env and pNL4-3 systems, could be used to analyze the drug resistance patterns of HIV-1 infectors and screen new drugs for antiretrovi-ral therapy in a rapid and effective way.

Objective To study the in vivo expression and biodistribution of Ad5-Fluc (Adenovirus carrying firefly luciferase genes) in mice.Methods The recombinant Ad5-Fluc virus was constructed and infected to BALB/c or nude mice through three different routes.The protein expression level,tissue distribution and the characteristics of infection were analyzed by in vivo bioluminescence imaging technology.Results Compared to other two routes,the BALB/c mice infected through muscular route had the longest expression cycle (over 60 days) and the highest expression level,while the virus was transferred into the liver and spleen after infection.The nude mice had a significantly extended expression cycle than BALB/c mice.Moreover,the characteristic of liver tropism was eliminated after Ad5 F35 infection in mice,while maintained similar expression efficiency.Conclusion Due to the highest expression efficiency,the muscular route would be the optimal route for Ad5 vector based vaccination.In addition,Ad5F35 virus could become an ideal alternative vaccine vector for eliminating the liver tropism.

Objective:To conjugate IVIG and MTX to produce a specific cytotoxicity upon phagocytes.Methods :MTX was conjugated with IVIG by indirect conjugating methods. HSA was used as an intermedi-ary to conjugate MTX with IVIG. The indirect immunofluorescence was adopted to test the binding abilityof Fc fragment. MTT assay was used to measure the cytotoxicity of conjugation on phagocytes. Results:Conjugation showed stronger cytotoxicity upon target cells than free MTX,and it showed only less cyto-toxic effect on Fc receptor negative cells compared with the positive ones. The specific cytotoxicity of IVIG-HSA-MTX was significantly stronger than that of MTX. Conclusion: In vitro the conjugation showed ahighly specific cytotoxicity upon phagocytes.