简介:AIM:TOinvestigatetheimmunogenicityofcandidateDNAvaccineagainsthepatitisCvirus(HCV)deliveredbytwoplasmidsexpressingHCVenvelopeprotein1(El)andenvelopeprotein2(E2)antigensrespectivelyandtostudytheeffectofCpGadjuvantonthiscandidatevaccine.METHODS:RecombinantplasmJdsexpressingHCVEIandE2antigensrespectivelywereusedtosimultaneouslyinoculatemicewithorwithoutCpGadjuvant.AntiserawerethencollectedandtJtersofantJ-HCVantibodieswereanalyzedbyELISA.Onemonthafterthelastinjection,animalsweresacrificedtopreparesingle-cellsuspensionofsplenocytes.ThesecellsweresubjectedtoHCVantigenspecificproliferaionassaysandcytokinesecretionassaystoevaluatethecellularimmuneresponsesofthevaccinatedanimals.RESULTS:AntibodyresponsestoHCVEIandE2antigensweredetectedinvaccinatedanimals.AnimalsreceivingCpGadjuvanthadslightlylowertitersofanti-HCVantibodiesinthesera,whilethesplenocytesfromtheseanimalsshowedhigherHCV-antigenspecificproliferation.Analysisofcytokinesecretionfromthesplenocyteswasconsistentwiththeaboveresults.Whilenoantigen-specificIL-4secretionwasdetectedforallvaccinatedanimals,HCVantigen-specificINF-γ,secretionwasdetectedforthesplenocytesofvaccinatedanimals.CpGadjuvantenhancedthesecretionofINF-γ,butdidnotchangetheprofileofIL-4secretion.CONCLUSION:VaccinationofmicewithplasmidsencodingHCVE1andE2antigensinduceshumoralandcellularimmuneresponses.CpGadjuvantsignificantlyenhancesthecellularimmuneresponse.
简介:摘要慢性阻塞性肺疾病(COPD)是一种年龄和吸烟相关的进行性肺疾病,由于慢性支气管炎和肺气肿的影响,呈现出不可逆性气流的受限。COPD的发病率和死亡率不断增加,但是并没有有效的治疗策略。DNA甲基化是目前研究中备受关注的表观遗传学调控机制,与COPD的发生与发展密切相关。因此,针对DNA甲基化进行治疗可能存在重要的临床价值。
简介:巨噬细胞在免疫和动态平衡起一个重要作用。在经由特定的受体的病原体识别之上,他们很快导致煽动性的回答。这个过程紧在transcriptional水平被控制。DNA有约束力的锌手指蛋白质CCCTC有约束力的因素(Ctcf)是远程的染色质相互作用的一个关键管理者并且协调在抄写因素和基因表达式进程之间的特定的通讯。在这研究,Ctcf基因被使用转基因的Cre-LoxP系统明确地在myeloid房间删除。在myeloid房间的Ctcf基因的有条件的删除在vivo导致了温和显型。显著地展出的Ctcf缺乏的老鼠减少了主要histocompatibility的表示在肝的建筑群(MHC)班II。Ctcf缺乏的巨噬细胞表明了正常表面显型和吞噬作用能力。在像使用费的受体(TLR)之上刺激,他们生产了支持inflammatorycytokinesIL-12和IL-6的正常层次,但是表明了一个强烈损害的能力生产肿瘤坏死因素(TNF)和IL-10,以及表示IL-10家庭成员IL-19,IL-20和IL-24。一起拿,我们的数据表明涉及巨噬细胞功能调整的Ctcf的一个角色。
简介:目的:进一步了解乙肝产妇的母乳中HBVDNA含量及其传染性,更好地指导母乳喂养。方法:利用荧光定时定量PCR检测技术,对45例乙肝血清学阳性的产妇进行血浆及乳汁中HBVDNA的定量检测。结果:45例产妇血浆和乳汁中HBVDNA的检出率分别为73%(33、45)和67%(30、45),其HBVDNA含量的对数值具有良好的相关性,(r=0.837,n=45,P<0.01),HBeAg(+)与HBeAb(+)两组间比较血浆及乳汁HBVDNA的含量,两者存在显著性差异(P<0.01)。HBeAg(+)组含量高于HBeAb(+)组。结论:乳汁和血浆中HBVDNA含量具有较好的相关性,血浆中HBVDNA高拷贝的产妇最好不要进行母乳喂养。产妇血浆中HBeAg与乳汁及血浆中HBVDNA含量都具有明显关系,它的存在提示血浆及乳汁中的HBVDNA含量较高。
简介:AIM:NitrativeandoxidativeDNAdamagesuchas8-nitroguanineand8-oxo-7,8-dihydro-2'-deoxyguanosine(8-oxodG)formationhasbeenimplicatedininitiationand/orpromotionofinflammation-mediatedcarcinogenesis.TheaimofthisstudyistoclarifywhethertheseDNAlesionsparticipateintheprogressionofintrahepaticcholangiocarcinoma.METHODS:Weinvestigatedtherelationoftheformationof8-nitroguanineand8-oxodGandtheexpressionofhypoxia-induciblefactor-1α(HIF-1α)withtumorinvasionin37patientswithintra-hepaticcholangiocarcinoma.RESULTS:Immunohistochemicalanalysesrevealedthat8-nitroguanineand8-oxodGformationoccurredtoamuchgreaterextentincanceroustissuesthaninnon-canceroustissues.HIF-1αcouldbedetectedincanceroustissuesinallpatients,suggestinglowoxygentensioninthetumors.HIF-1αexpressionwascorrelatedwithinduciblenitricoxidesynthase(iNOS)expression(r=0.369andP=0.025)and8-oxodGformation(r=0.398andP=0.015).DoubleimmunofluorescencestudyrevealedthatiNOSandHIF-1αco-localizedincanceroustissues.Notably,theformationof8-oxodGwascorrelatedsignificantlywithlymphaticinvasion(r=0.386andP=0.018).Moreover,8-nitroguanineand8-oxodGinnon-canceroustissueswereassociatedsignificantlywithneuralinvasion(P=0.042andP=0.026,respectively).TheseresultssuggestthatreciprocalactivationbetweenHIF-1αandiNOSmediatespersistentDNAdamage,whichinducestumorinvasivenessviamutations,resultinginpoorprognosis.CONCLUSION:Theformationof8-nitroguanineand8-oxodGplaysanimportantroleinmultiplestepsofgeneticchangesleadingtotumorprogression,includinginvasiveness.
简介:摘要目的探讨血清学HBsAg阳性产妇乳汁中HBV-DNA检出率,以指导母乳喂养。方法运用3种不同处理方法处理754例HBsAg阳性产妇乳汁,再用荧光定量PCR法检测其HBV-DNA含量。结果754例HBsAg阳性产妇乳汁中HBV-DNA检出率方法1为44.4%(335/754)方法2为33.3%(251/754),方法3为51.9%(391/754)(P<0.01);病毒载量的对数值分别为3.87±0.79;3.65±0.68;4.35±0.84(P<0.05)。结论运用方法3处理乳汁标本HBV-DNA检出率及病毒含量均高于其它两种方法,运用方法3筛查出的阴性结果更可靠,乳汁HBV-DNA阴性的哺乳期妇女可以进行母乳喂养。
简介:Rat-1cellsweretransfectedwithDNAfromhumanesophagealcancer2K,4K,6K,7K.8K.Thetransformingfociwereobtainedandthetransformingcelllineswereestablished.Thecelllinescanformlargercolonyinsoftagar.Thosenudemiceinjectedsubcutaneouslywiththecellssufferedfromlargerfibroussarcoma.Thisindicatesthatthecelllineshavecarcinogenicity.TheexperimentalresultssuggestthathumanDNAsequenceandhumanHa-rasspecial616Kb(BamHI)bandarepresentintheDNAofthetransformingcells.Theover-expressionofrasgeneproductsP21werefoundinthetissuesofexophagealcancer,thetissuesadjacenttotumorandthetransformingcells.
简介:Inordertoanalyzethesequencesoftheinternaltranscribedspacer(ITS)includingthe5.8SribosomalDNA(rDNA)ofcommondermatophytes,soastoobtainarapidandaccuratemethodtoidentifythespeciesofdermatophytesandtoestablishthephylogenetictreeofthesespeciestounderstandtheirrelationship,16strainsofdermatophyteswerecollectedandpreliminarilyidentifiedbymorphologicalcharacteristics.GeneralprimersforfungiITS1andITS4wereusedtoamplifytheITSrDNAofeachstrainswithPCR.ThePCRproductsafterpurificationweresequenceddirectlyandwereanalyzedthroughinternet.Intheresults,11strainswereidentifiedbymeansofmorphologicalfeatures,amongwhich5strainswereTrichophyton,5strainswereMicrosporumand1wasEpidermaphyton,whichwasconsistentwiththeresultsbymolecularbiology.Inthe5unidentifiablestrains,1strainwasprovedtobeChrysosporiumbymolecularbiology.Thesestrainsstudiedcouldbedividedinto3differentclassesasindicatedintheanalysisofthephylogenetictreeofthesequencesinITS,whichwerequitedifferentfromthoseofmorphologicalclassification.ItisevidentfromtheaboveobservationsthatthemolecularmethodofanalysisontheITSsequencesisarapid,highlysensitiveandaccurateapproachforthedetectionofdematophytespecies,however,itstillexhibitssomelimitationsneedingthesupplementationwithmorphologicalidentification.
简介:目的研究乙型肝炎免疫球蛋白(HBIG)对肝细胞的跨膜转运作用和对乙型肝炎病毒(HBV)感染细胞模型的乙型肝炎表面抗原(HBsAg)、HBVDNA分泌的抑制作用。方法用含HBIG的培养基培养QSG7701细胞,在不同时间点定量检测培养上清的抗HBs,计算HBIG的透过率;用含HBIG的培养基培养HepG2.2.15细胞数天,或先以,定浓度的HBIG共培养,第3天后更换为不含HBIG的培养基继续培养细胞。于不同时间点定量检测培养上清的HBsAg、HBVDNA;用MTT比色试验观察药物的细胞毒性。结果浓度为0.1~0.4IU/mL的HBIG与QSG7701细胞共孵育48h时,细胞内约有38%~46%的HBIG。浓度为0.1~10.0IU/mL的HBIG作用第3、6.9天时能抑制HepG2,2.15细胞培养上清中HBsAg、HBVDNA的分泌(P〈0.01)。住尤药物继续培养的第5、7天,培养卜清中HBsAg浓度较有药物培养的第3天低且继续下降(P〈0.01),第9~11天逐步增高;培养上清HBVDNA水平在第5天时也较有药物培养的第3天低并继续下降(P〈0.01),第7~11大时逐步增高。结论在体外细胞实验中,HBIG能跨膜转运入肝细胞内,细胞内外的HBIG能抑制HBsAg、HBVDNA的分泌。