简介：ToobtaintherecombinantsolubleproteinoftheextracellularfragmentofhumanTRAILgeneandtoidentifyitsfunctionpreliminarily,thisgenefragmentwasamplifiedfromperipheralbloodmononu-clearcells(PBMC)byRT-PCRandclonedintovectorpGEM-T-Easyforsequenceanalysis.Theexpres-sionvectorpET-30a/TRAILwasthenconstructedbyDNArecombinationmethodwithaHis-taggeneatthefrontoftheTRAILfragment,andtherecombinantproteinwasexpressedinE.coliBL21(DE3).Meanwhile,theexpressedtargetproteinwaspurifiedwithNi-NTAchromatographycolumnandidentifiedbySDS-PAGEandWesternblotting.TheproliferationinhibitionactivityofTRAIL-HiswasdetectedbyMTTassay.PIstainingandWright-GiemsastainingwereusedtodetectthepresenceoftheTRAIL-in-ducedcellapoptosis.ItwasdemonstratedthatthetargetproteinexpressedinE.coliBL21showedthesamerelativemolecularmassasthattheproteinexpectedandcouldberecognizedbyboththeanti-TRAILpolyclonalantibodyandanti-Hismonoclonalantibody.Inaddition,thisproteincouldalsoinhibitprolif-erationofhumanlymphomacelllineJurkatcellsorinduceapoptosisofthiscellline.ItisapparentthatarecombinantsolubleTRAILproteinwithbiologicalactivityisobtainedandthisprospectivestudycanlaysolidfoundationforfurtherresearchonthebiologicalactivityandapplicationintheanti-tumortherapy.

简介：InordertocompareandevaluatethreeanimalmodelsforstudyingthepathogenicityofStaphylococcusepidermidisstrains,threeexperimentalanimalmodels,namely,murineintra-venousLD50,mouseforeignbodyinfectionandratcentralvenouscatheter(CVC)infectionmodelswereusedtoassesstherelativevirulenceoftwoS.epidermidisstrains,ATCC12228and97-337.Theresultsfromthreeanimalmodelswerecomparable,indicatingS.epidermidis97-337wasmorevirulentthanstrainATCC12228.TheratCVCinfectionmodelbestmimickedtheconditionsofclinicalpatientswithintmvenonscatheters,andmoreinformationcouldbeobtainedfromthismodel.Weconcludethatdifferentinvivomodelsservefordif-ferentpurposes,andtheratCVCinfectionmodelismostsuitableforstudyingspecificcharacteristicsofcatheterrelatedinfectionscausedbyS.epidermidisstratus.

简介：HepatitisBvaccine,asthefirsthigh-effectiverecombinantcommercialvaccine,wassuccessfullydevelopedintheearly1980s.Sincethen,differentopinionshaveoccurredonthequalityofvaccineswithrapiddevelopmentoftargetgeneselecting,antigenexpressionsystem,andqualityevaluation.DifferentantigensofhepatitisBvaccinesarederivedfromdifferentexpressionsystem,andtherearealsosomedifferencesonmanufactureprocedureorglycosylateddegreeofantigen.

简介：ThepurposeofthisstudywastotesttheeffectivenessofsourcevirusstrainforthemanufactureoftheinactivatedSARSvirusvaccine,andestablishanexperimentalmethodandpreliminarystandardforpotencyevaluation.MiceweredividedintogroupsforbeingimmunizedwithcorrespondingseriallydilutedexperimentalSARSvirusinactivatedvaccine.Andtherabbitswereimmunizedwithundilutedvaccine.ChallengeassaywasconductedwithaheterologousSARSvirus.Andtheneutralizationantibodywasdeterminedwithplaquereductionneutralizationtest(PRNT),towhichtheneutralizationantibodyintheconvalescentserumofSARSpatientswascompared.Theexperimentalvaccineviralstrainswereprovedtobesuitableformanufacturingthevaccine.Miceimmunizedbyvaccinesofserialdilutionswereabletoelicitneutralizingantibody.Theantibodytiterfrommiceimmunizedwiththeundilutedvaccinecouldreachupto1:495.2,whilethoseofrabbitsimmunizedwiththeundilutedvaccinecouldreachaGMTof55.0-79.9.ThecapabilityoftheantibodytoneutralizethevirusfromGuangdongismoreefficientthanthatfromBeijing.TheGMTofneutralizingantibodyinSARSconvalescentslivinginsouthandnorthChinarangedfrom50.12to54.95,andthetitersofconvalescentsfromnorthChinawerehigherthanthosefromsouthChina.Miceandrabbitsusedasthemodelforevaluationofpotencyareofsensitivity,andthetestisofreproducibility.Thecandidatechallengeviralstrainsshowedarelativelyconsistenteffectonevaluatingantibodiesproducedbyvariousbatchesanddifferentvaccine-sourcestrains,hencetheycanbeusedtoevaluatepotencyofthevaccine.Themethodfortestingthevaccinepotencyandtheevaluationstandardwasestablishedpreliminarily.

简介：Toinvestigatetheprotectiveeffectofepigallocatechingallate(EGCG)ontheimmunefunc-tionofdendriticcells(DCs)afterultravioletBirradiation(UVB)anditsunderlyingmechanisms,themonocyteswereisolatedfromperipheralbloodandcultivatedintoDCswithcytokines,suchasGM-CSFandIL-4.DCswereharvestedaftercultivationfor7dandsubjectedtoirradiationwithdifferentdosagesofUVB.Then,200μg/mlofEGCGwereaddedincertaingroupsimmediatelyafterirradiation.DCssimplytreatedwithUVBortreatedwithbothUVBandEGCGwereco-culturedwithlymphocytes,andMTFassaywasusedtodetecttheabilityofDCstostimulateproliferationoflymphocytes.SurfacemarkersCDS0,CD86,HLA-DRandCD40weredetectedbyflowcytometry,andthelevelsofIL-10andIL-12secretedfromDCs24haftercultivationweremeasuredbyELISA.ItwasdemonstratedthatUVBirradiationcouldinhibittheabilityofDCstostimulatetheproliferationoflymphocytesandsurfaceexpressionsofCDS0,CD86,HLA-DRandCD4onDCsinadose-dependentmanner.TheinhibitionrateofDCswasimprovedtosomeextentaftertreatmentwith200μg/mlofEGCG.Whentheconcentra-tionofEGCGexceeded100μg/ml,theenhancingeffectofEGCGontheexpressionoftheco-stimulat-ingmoleculesonDCscouldbedemonstratedinadose-dependentmanner.UVBshowednosignificantinfluenceonthesecretionofIL-10andIL-12fromDCs,whileEGCGcoulddown-regulatethesecretionlevelofIL-12andup-regulatethatofIL-10.ItisconcludedthatEGCGcanantagonizetheinhibitoryeffectonDCsinducedbyUVBirradiation.Thisfunctionhassomerelationshipwithitsprotectingeffectoftheexpressionoftheco-stimulatingmoleculeonthesurfaceofDCsandthesecretionlevelofIL-10andIL-12.

简介：AbstractToinvestigatetheroleofCD4^+helperT(Th)cellsinthememoryCTL-mediatedanti-tumorimmunity,theRAG-1geneknockoutmicewereadoptivelytransferredwithOT-1cellstogeneratethememoryCTL,theC57B1/6miceimmunizedwiththeepitopepeptideofOVAspecificThcellsandwithdifferentadjuvantswereadopfivelytransferredwiththesememory-CTLs,andthentheanimalswerechallengedwithtumorcellsEGT.ItwasfoundthatalthoughthesimpleimmunizationofmicewiththeepitopepeptideoftheOVAspecificThcellscouldgeneratemoreeffectCTL,butthiseffectwasnotsostrongenoughtoresistcompletelythechallengeswithtumorcells.Nevertheless,thememoryCTL-mediatedanti-tumorimmuneeffectrequiredthehelpsofTh1andTh2cells.Thecross-regulationbetweenThlandTh2cellsseemedtobebeneficialforthehosttogeneratemoreeffectorCTLformountinganefficientanti-tumorresponse.ItconcludedthattheinteractionbetweenThlandTh2cellsmightbemoreimportantthanthesinglesubsetofThcellsinthememoryCTL-mediatedanti-tumorimmuneresponse.Moreattentionshouldbepaidinthisregardforthefuturestudies.

简介：RapiddifferentialidentificationofMycobacteriumspeciesisessentialforeffectivediagnosisandmanagementofmycobacteriosis.Theaimofthisstudywastodevelopanovelmultiplexprobearraybasedonthe16S-23SrRNAgeneinternaltranscribedspacersequenceforthegenotypingofmycobacteriatothespecieslevel.Apairofprimersandasetofgenus-andspecies-specificprobesweredesignedfromtheconservedandpolymorphicregionsofthe16SrRNAgene,internaltranscribedspacer,and23SrRNAgenesequencesofmycobacteria.Weusedanovelmultiplexprobearrayforidentificationof266clinicalspecimensobtainedfrompatientswithmycobaterialinfection.Theresultsshowedthattheoverallspecificityandsensitivityofournovelprobearraywereboth100%forthegenus-specificprobeandMycobacteriumtuberculosiscomplex-specificprobe.Therewere79.3%(23/29)ofnontuberculousmycobacteriawhichcouldbeidentifiedtothespeciesleveldirectlyinthespecimensfromChina.SomeintraspeciesheterogeneityinM.avium,M.intracellulare,M.chelonaeandM.abscessuswasobserved.Withtheincreaseofsequencesofinternaltranscribedspacerandnumbersofwholemicrobialgenomes,andfurtheroptimizationofprobes,themultiplexprobearraywillbecomeapromisingtoolfortherapidandaccurateidentificationofmycobacteriainordinaryclinicallaboratories.

简介：During2004,atotalof124batchesofHIVantibodyELISAsfromdomesticandoverseasmanufacturers,comprisingapproximately60milliontests,weretestedforqualityandreleasedforscreeningbloodinChina.Theinter-andintra-batchvariation,specificity,andsensitivitywereevaluatedusingalaboratorypanelandclinicalsamples.Theinter-batchvariationwaslessthan15%andonly2of12assayshadintra-batchvariationoflessthan20%for4dilutionsofacontrolspecimen.257samplesconfirmedpositiveforHIVantibodyand4826negativesamplesfromdifferentregionsinChinawereusedtoevaluatethesensitivityandspecificityoftheassays.Theresultsshowedthatthesensitivityisintherangefrom93.7%to100%forassayssampleddirectlyfromthemanufacturers,and91.4%-99.6%forthoseretrievedfromtheconsumers;thespecificitywasintherangefrom97.88%to99.97%.ThetestingenvironmentmayvaryindifferentregionsofChina.Therefore,manufacturersshouldproviderobustassaystosatisfytherequirementsofthesediverseenvironments,andespeciallyreducetheintra-assayvariationandimprovethestabilityofthekits.