Objective To investigate the contribution of Th immune bias to Coxsackie virus type B3-induced myocarditis on two different mouse strains.Methods BALB/c and C57BL/6 mice were infected intraperitoneally with Coxsackie virus B3.The incidence of myocarditis was evaluated with serum CK activity,appearance and H.E section of the heart tissue.Virus infection and replication were compared between the two mouse strains both in vitro and in vivo.Th immune bias was determined by cytokine expression pattern,antibody isotype and transcription factor T-bet/Gata-3 expression.Results Coxsackie virus B3 infected myocytes from both BALB/c and C57BL/6 mice both in vitro and in vivo.However,only BALB/c mice developed typical myocarditis after infection.Furthermore,BALB/c mice elicited a Th1 type immune response against CVB3 infection while C57BL/6 mice displayed a Th2 type immune response.Conclusion CVB3 infection induced a different pattern of pathological changes in cardiac muscle tissue in two different strains of mice.This difference was correlated with the different Th immune bias induced in these two mouse strains. if(document.getElementById('ChDivSummary').innerHTML!=""){CutSpan('ChDivSummary',500);DisplaySpanDiv('ChDivSummary');ClearSummaryOnLoad('SummaryLinkChID','SummaryLinkEnID');}else{CutSpan('EnDivSummary',1000);DisplaySpanDiv('EnDivSummary');ClearSummaryOnLoad('SummaryLinkEnID','SummaryLinkChID');}

Objective To explore the characteristics of immune lymphocyte subpopulation and liver biochemical markers in patients with HIV/HCV co-infection.Methods Some related laboratory makers of peripheral blood in the study group(40 hemophiliac patients with HIV/HCV co-infection)and in the control group(12 hemophiliac patients with HIV infection only)were detected and analyzed.Results There were significant differences between results of the two groups(study and the control)were in following:NK cell(9.26±5.98)% vs(12.19±5.80)%,P<0.05.The rate of CD4/CD8 0.39±0.21 vs 0.53±0.24,P<0.05.CD3(74.99±10.33)% vs(68.42±8.82)%,P<0.05.CD8(52.28±12.59)% vs(43.58±10.99)%,P<0.05.ALT(48.59±40.21)U/L vs(26.87±27.23)U/L,P<0.001.AST(61.66±51.02)U/L vs(34.17±30.24)U/L,P<0.001.GGT(136.50±111.50)U/L vs(43.88±36.17)U/L,P<0.001.CG(683.41±962.22)μg/dL vs(250.96±290.81)μg/dL,P<0.001.Hyaluronic acid(180.94±196.69)ng/mL vs(64.68±32.74)ng/mL,P<0.001.Albumin/globin(1.35±0.24)vs(1.50 ±0.34),P<0.05.Comparing with the patients infected with HIV only,NK cell,rates of CD4/CD8 and albumin/globin were decreased significantly,but the level of AST,ALT,GGT,CG,and liver fibrosis marker HA were increased significantly in the patients co-infected with HIV/HCV.Conclusion Patients co-infected with HIV/HCV may aggravate the imbalance of lymphocyte subpopulations,worsen liver injury and occur fibrosis tendency.

Objective To explore the characteristic and reason for indeterminate HIV Western Blot(WB),the limitation of WB and the possible amendatory measures.Methods Summing up and analysis the distribution of the indeterminate HIV WB among the tested people,the laboratory test results,the follow-up and the final HIV antibody outcome.Results The relative healthy people including the people of voluntary counseling and testing,the blood donors and pregnant women make up 50% of the indeterminate HIV WB,the follow-up for the indeterminate HIV WB is difficult and the there are few people of the indeterminate HIV WB have HIV antibody test again.There is false-positive in WB test,especially the false-positive of P24 is serious.Conclusion The indeterminate HIV WB is related to the false positive of WB test,measures should be taken to reduce the indeterminate HIV WB and to interpret the result accurately.

Objective To clone and express the Leptospira interrogans(L.interrogans)vaccine candidate gene,LB061 and to analysis the immunogenicity and conservation of the recombinant protein in different serovars of L.interrogans.Methods The characters of LB061 gene were predicted by bioinformatics software.The prokaryotic expression system of the LB061 was constructed by routine molecular biological techniques.Expression of the recombinant LB061 protein was induced by IPTG,and confirmed by Western blot.Balb/C mice were immunized with ercombinant LB061 and immune serum was used to study the immunogenicity of LB061 and the conservation of expressed LB061 in sixteen different serovars.In addition,antibodies against LB061 in rabbit serum infected with L.interrogans strain Lai(56601)were examined by Western blot.Results LB061 is a putative outer membrane protein in L.interrogans as predicted by bioinformatical analysiss.The prokaryotic expression system of LB061 was constructed successfully.Serum titers of anti-recombinant LB061 antibodies in immunized BALB/c mice were 1:32000.LB061 was detected in sixteen different serovars of L.interrogans.LB061-specific antibodies were detected in rabbit serum infected wit L.interrogans strain Lai by Western blot.Conclusion The vaccine candidate gene,LB061,of L.interrogans serogroup,icterohemorrhagic serovar,Lai strain can elicit antibody responses in rabbit during infection with Leptospira interrogans.Our studies provide a basis for further investigation on the role of LB061 as a vaccine candidate gene.

Objective To develop a rapid and sensitive assay system to detect the seven virulence factors of Streptococcus suis,using two seqarate multiplex PCR assays.Methods The seven virulence factors were detected based on the gene sequences of mrp,epf,sly gdh,gapdh,orf2 and fbps.PCR primer pairs were produced for each of the 7 factors and the assay method,was developed and optimized.These genes were tested in 29 bacterial isolates by the multiplex assay.Results All PCR products were analyzed by electrophoresis on 2% agarose gels.Detection of all 29 Streptococcus suis strains was confirmed by bacteriological examination.The assay was validated by the positive and negative controls.Conclusion The results demonstrate that the multiplex PCR assay is a highly specific and sensitive diagnostic tool for the detection of seven virulence factors of Streptococcus suis. if(document.getElementById('ChDivSummary').innerHTML!=""){CutSpan('ChDivSummary',500);DisplaySpanDiv('ChDivSummary');ClearSummaryOnLoad('SummaryLinkChID','SummaryLinkEnID');}else{CutSpan('EnDivSummary',1000);DisplaySpanDiv('EnDivSummary');ClearSummaryOnLoad('SummaryLinkEnID','SummaryLinkChID');}

Objective The existence of a viable but non-culturable(VBNC)state has been described for Campylobacter jejuni.This state can be induced and detected using established VeroE6,HeLa cell lines.Methods VBNC C.jejuni cell were induced by low temperature and starvation.Sensitization test of established HeLa,Vero cells model to CDT of viable but non-culturable Campylobacter jejuni showed that the cell was still sensitive,and the result was further confirmed by RT-PCR test.The experiment found that VBNC C.jejuni cell still had the virulence to cell cultures although it decreased.Total and active cells were counted by conventional culture method and after staining with 49,6 diamino-2-phenylindole(DAPI).Induced VBNC cell were obtained by freezed at-70 ℃.Results The rest VBNC cell were confirmed by different state cell count.C.jejuni decreased about one magnitude after refrigeration.About 70% cell can be induced into apotosis before refrigeration and only 40% cell die after C.jejuni refrigeration.Conclusion Low temperature and nutrient poor conditions can induce cell enter into VBNC state.Remarkable change of C.jejuni cell to produce virulence can be observed,and we confirmed this further at RNA level.VBNC C.jejuni cell can still produce virulence although the quantity decreased remarkably. if(document.getElementById('ChDivSummary').innerHTML!=""){CutSpan('ChDivSummary',500);DisplaySpanDiv('ChDivSummary');ClearSummaryOnLoad('SummaryLinkChID','SummaryLinkEnID');}else{CutSpan('EnDivSummary',1000);DisplaySpanDiv('EnDivSummary');ClearSummaryOnLoad('SummaryLinkEnID','SummaryLinkChID');}

Objective To develop a polymerase chair reaction-capillary electrophoresis(PCR-CE)based method for the detection of biovars one and two of U.urealyticum and to determine whether these two biovars of U.urealyticum are associated with the nongonococcal urethritis(NGU)in the male patients.Methods Common primers for two biovars of U.urealyticum were synthesized and used for the PCR.Then PCR products obtained for two biovars of U.urealyticum were detected using capillary electrophoresis(CE).Two biovars of U.urealyticum were detected by the above method in the urine samples from the different male populations.The sensitivity and specificity of this PCR-CE based method was further examined.Results This PCR-CE method had a sensitivity of detection at 10 copies /50 μl.This method specifically amplified biovar 1 and biovar 2 of U.urealyticum.The prevalence of U.urealyticum biovar 2 in NGU group and nonchlamydial NGU(NCNGU)group was higher than that in the control group(P<0.05)but the prevalence of U.urealyticum biovar 1 in NGU group and NCNGU group did not differ significantly from that in the control group(P>0.05).Conclusion A PCR-CE based method for the detection of two biovars of U.urealyticum has been developed,which has good sensitivity and specificity.U.urealyticum biovar 2 is associated with NGU in the male.U.urealyticum biovar 1 can not cause NGU.