简介：ThepurposeofthisinvestigationistodemonstratewhethertriptolidecaninhibitTNF-αselectivelyandtostudythemechanism.TheabilityoftriptolidetoinhibittheproductionofTNF-αandIFN-γstimulatedbylipopolysaccharide(LPS)andphytohemagglutinin(PHA)onperipheralbloodmononuclearcell(PBMC)fromhealthydonorswasmeasuredbyELISAassays.TheimmunologicalmechanismofactionoftriptolideonTNF-αwasinvestigatedbypre-treatmentwithtriptolideofPBMCandmonecytesfollowedbyanalysiswithFlowCytometry(FCM).TheinhibitionofTNF-αandIFN-γbytriptol-ideoccurredinadosedependentmannerandtheIC50wasequalto5-10ng/mlforTNF-aand0.1-1ng/mlforIFN-γ.TheconcentrationsofTNF-αmeasuredafterthedifferentpre-treatmentswithtriptolideonPBMCandmonecytesareconsistentwithitseffectsonapopulationofCD14^+/TNF-αmonecytesshownonFCM.Thetwomethodsofpre-treatmentswithtriptolidemaysuggestdifferentclinicalsignificances,ImmunophenotypinganalysiswithFCMrevealedthattriptolidemaycompetewithLPSforbindingtotheCD14receptor.Thestudyprovidesbasicevidencethattriptolidemaybeusedasananti-inflam-matoryreagentfortreatmentofleprosyreactions.

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简介：Klebsiellapneumoniae(K.pneumoniae)isoneofthemaingram-negativebacilliinclini-calpractice.NosocomialinfectionscausedbyK.pneumoniaeproducingextended-spectrumβ-lactama-ses(ESBLs)areverydifficulttotreat.ThispaperinvestigatedtheresistantcharacteristicsofK.pneu-moniaeproducingESBLsandtheiraminoglycoside-modifyingenzymegeneexpressionsincludingN-acetyhransferasesandO-adenyhransfemses.BacteriaidentificationandESBLsconfirmatorytestswereperformedbyPhoenix~(TM)-100system.Andminimuminhibitoryconcentrations(MICs)ofgentamicin,amikacin,kanamycin,tobramycin,netilmicinandneomycinin53K.pneumoniaeisolateswerede-tectedbyagardilution.Inaddition,sixaminoglycoside-modifyingenzymegeneswereamplifiedbypolymerasechainreaction(PCR)andverifiedbyDNAsequencer.Itwasfoundthatimipenemandmeropenemagainst120K.pneumoniaeisolatesproducedpowerfulantimicrobialactivities.Theresis-tantratesofgentamicinandamikacinwere55.0%and46.7%,respectively.Exceptneomycin,MIC_(50)andMIC_(90)ofgentamicin,amikacin,kanamycin,tobramycinandnetilmicinin53K.pneumoni-aewereall>128μg/ml,andtheresistantrateswere83.0%,52.3%,75.5%,81.1%and69.8%,respectively.However,neomycinwasonly39.6%.Inaddition,fivemodifyingenzymegenes,includingaac(3)-Ⅰ,aac(3)-Ⅱ,aac(6')-Ⅰb,ant(3″)-Ⅰ,ant(2″)-Ⅰgenes,werefoundin53isoahesexceptaac(6')-Ⅱ,andtheirpositiverateswere11.3%,67.9%,47.2%,1.9%and39.6%,respectively.Itwasalsoconfirmedbynucleotidesequenceanalysisthattheaboveresistantgenessharednearly100%identitieswithGenBankpublishedgenes.TheresultsobtainedinthepresentstudyindicatedthatK.pneumoniaeproducingESBLsstrainsarerapidlyspreadinginourhospital,andtheirresistancetoaminoglycosidesmaybeassociatedwithaminoglycoside-modifyingenz-ymegeneexpressions.

简介：TotesttheantigenicactivityofMprotein(Mcprotein)intheinnermembraneofSARS-CoV,SARS-CoVMcprotein'sbaseslocatinginsidethemembranewerecloned,theHis-fusionproteinwasexpressedinE.coliandanalyzedforitsantigenicactivity.Amongthose7clinicallydiagnosedpatients'sera,therewere5positiveand2negativeinreactionwithHis-fusionprotein.Allofthe20healthypersons'seraandrabbitanti-OC43and229EwereofnegativereactionwithHis-fusionprotein.TheanimalsimmunizedwithHis-fusionproteinhaveproducedmulti-clonalantibody.TheHis-fusionproteincouldspeciallyreactwithclinicallydiagnosedSARSpatients'seraandtheanimalsimmunizedwithHis-fusionproteincouldproducespecificallymulti-clonalantibody,butitcouldnotreactwiththeseraofhealthypersonsandtherabbitanti-OC43and229E.

简介：InordertoobserveseveralantibodiestoliverantigensinChinesepatientswithdifferentfiverdiseasesandtodiscussthecharacteristicsoftheautoantibodiesinautoimmuneliverdiseases,from1412patients,detectedbyindirectimmumofluoreseence(IIF)initially,230patientswithabnormalALTwerechosenanddividedinto5groups:①autoimmunediseasesgroup,42cases:18withautoimmtmehepatitis(AIH),21withprimarybiliarycirrhosis(PBC),3withprimarysclerosingcholangitis(PSC).②HAVgroup,23cases;③HBVgroup,70cases;④HCVgroup,35casesand⑤NonA-Egroup,60cases.First,ANA,AMA,SMA,liver-kidneymicrosomalantibody(LKM)andsoonweretestedby1/F.Then,LKM-1,fivercytosofic-1(LC-1),solubleliverantigen/fiverpancreas(SLA/LP)andsubtypeofAMA(M2)aswellasANAprofilesuchasSS-A,SS-BanddsDNAweretestedbyWesternblotandimmtmoblotstripsassay,respectively.Theresultswerethatamong1412cases,thosediagnosedasAIH,PBCandPSCaccotmtedfor12.7‰,14.9‰and2.1‰,respectively,ofthesamplesbeingtested.2/230withLKM-1and2/230withSLA/LPwereseeninindividualsinfectedwithAIHandHCV,respectively.AllpatientswithPBCshowedAMAandM2antibodies.NospecificANApatternwasseeninAIHby1/Fbutanti-actinwasonlyfoundinpatientswithAIH.InNonA-Egroup,fourcaseswerepositiveofAMAandM2;threehadhightiterofSMAandother4hadSS-A,SS-BordsDNAantibodies,etc.Itwasconcludedthatthedetectionofanti-fiverantigens,ANAprofileandAMAsubtypeswerehelpfulforthediagnosisofautoimmunefiverdiseasesandoverlapsyndromes.InpatientswithNonA-Ehepatitis,thediagnosisofPBCorAIHshouldbetakenintoconsideration.

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.

简介：Highlevelsoflowmolecularweight(LMW)IgMincertaindiseasesareassociatedwithclinicalandlaboratoryindiceswhichreflecttheseverityofthedisease.TheseassociationssuggestthatLMWIgMmayplayanimportantroleintheimmunopathogenesisofthesediseases.TofurtherapproachthequestionconcerningthefunctionalactivityofLMWIgMindisease,apanelofLMWIgMandhighmolecularweight(HMW)IgMpreparationswithorwithoutrheumatoidfactor(RF)activitywereusedtoinvestigatetheirantibodybindingactivityandtheireffectorfunction.ItwasfoundthatLMWIgM-RFandHMWIgM-RFhadasimilarbindingcapacitytoFcfragmentastherewasnosignificantdifferenceintheaffinityindexbetweenthem.ItfurthershowedthattherateofactivationandtotalamountofutilizationofcomplementbyLMWIgMandHMWIgMwassimilar,althoughthemeanfluorescenceofC3depositionbyIgM-RFandHMWIgM-RFwasslightlyhigherthanthatofLMWIgM-RFandothercontrolRFantibodies.However,thecurrentstudydemonstratedthatLMWIgMhadstrongneutrophilactivatingpropertieswhencomparedwithHMWIgM.ThesefindingssuggestthatonemechanismofLMWIgMcontributingtotheimmunopathogenesisofRAmaybeduetotheformationofcirculatingimmunecomplex(CIC)byLMWIgMwithsubsequentactivationofneutrophils.WhetherLMWIgMhasotherfunctionalactivityindiseaseisunclearandneedsfurtherinvestigation.

简介：Theaimofthisstudyistoinvestigatewhetherthreemononucleotidepolymorphismsatthelocus-1082,-819and-592inthepromoterregionoftheIL-10geneareassociatedwithchronicseverehepatitis.TheIL-10-592andIL-10-1082polymorphismsweregenotypedbypolymerasechainreaction-restrictionfragmentlengthpolymorphismanalysis(PCR-RFLP)whilepolymerasechainreaction-se-quencespecificprimer(PCR-SSP)assaywasusedtotesttheIL-10-819polymorphism.Thepolymor-phismsofIL-10-1082,-819and-592genesweredetectedin98patientswithchronicseverehepatitis(CSH),478patientswithchronichepatitisB(CHB),223asymptomatic(chronic)HBVcarriers(ASC)and267patientswithself-restrictedHBV.Therewassignificantdifferenceofthepolymor-phismsofIL-10-1082,IL-10-819andIL-10-592genesbetweenCSHgroupandothergroups.Thefre-quencyofAAgenotypeatIL-10genepromoter-1082locusinchronicseverehepatitispatientswashigherthanthatinasymptomaticHBVcarriers(X~2=13.314,P=0.001),andself-restrictedHBVpatients(X~2=13.545,P=0.000);thefrequencyofCCandACgenotypeatIL-10genepromoter-592locusinchronicseverehepatitispatientswashigherthanthatinchronichepatitispatients(X~2=15.970,P=0.000)(X~2=20.414,P=0.000),asymptomaticHBVcarriers(X~2=21.283,P=0.000)(X~2=28.309,P=0.000)andself-restrictedHBVpatients(X~2=17.047,P=0.000)(X~2=16.528,P=0.000);thefrequencyofTCgenotypeatIL-10genepromoter-819locusinchronicseverehepatitispatientswashigherthanthatinchronichepatitispatients(X~2=58.961,P=0.000),asymptomaticHBVcarriers(X~2=53.255,P=0.001)andself-restrictedHBVpatients(X~2=39.616,P=0.001).Sointerleukine-10genepolymorphismwasassociatedwiththechronicsevereheoatitis.

简介：TostudytheexpressioncharacteristicofJapaneseencephalitisvirus(JEV)prMEandEproteinsandtheefficacyofDNAimmunizationbydifferentrecombinantplasmidscontainingJEVprME(2001bp)andE(1500bp)genes,tworecombinants(pJMEandpJE)containingJEVprMEandEgenesfusedwithFLAGwereconstructedandthentransfectedintoHepG2andCOS-1cellsbylipnsomefusion.TheexpressionfeatureofFLAG-prME(about72kDa)andFLAG-E(about54kDa)proteinsintransfectedcellswereanalyzedbyWesternblotandtwoantibodysystems(anti-FLAGandanti-E).BALB/cmicewereimmunizedwith100μgoftwokindsofrecombinantsbyintramuscularinjection,andJEVJaGAr-01strains(10^5PFU/100μl)weregiventoBALB/cmicebyintraperionealinjection3wkaftertwiceDNAimmunizationbyalethalviruschallenge.BALB/cmicewereobservedfor21daysafterchallenge.80%plaquereductionneutralizationtestwasperformedtotitrateneutralizationantibodybeforeandafterviralchallenge.ItwasfoundthattheexpressionofproteinsassociatedwithpJMEandpJEwasdeterminedintransfectedcellswithanti-FLAGandanewproteinof11kDawasdetectedinHepG2andCOS-1cellstransfectedwithpJME.OnlyE(53kDa)proteinwasidentifiedastransfectedwithpJMEusingantiE.HigherlevelofneutralizationantibodiesandtheefficacyofprotectiveimmunitywereinducedwithpJMEimmunization,andweresimilartothoseinducedbyinactivatedJapaneseencephalitisvaccine,butwerebetterthanthoseinducedwithpJE.ItconcludesthattheexpressionlevelfromprMtoEproteinsofJEVisdifferentinvitro,andtheinvitroexpressionefficiencyofpJMEwasbetterthanthatofpiE.FLAG-prMEproteinexpressedbypJMEcouldbecleavedbypeptidasefromhost.TheefficacyofDNAimmunizationiscorrelatedtotheexpressioncharacterizationofrelatedproteinsexpressedinvitro.

简介：Toinvestigatetheprevalenceandgenotypeofextendedspectrumbeta-lactamases(ESBLs)mediatedbyplasmidinGram-negativebacteriafoundinsouthernChina,atotalof1184clinicalisolatesofnon-repetitivestrainsofGram-negativebacteriawerecollectedin2001from5differentcitiesinsouthernChina.TheESBLs-producingisolatesweredistinguishedbymeansofthephenotypeconfimatorytestbasedontheNCCLScriteriaandweresubjectedtoplasmidconjugationandelectroporationexperiments.Thoseclinicalisolatessucceededinplasmidtransfershadundergoneplasmidconjugationandelectro-transformation,plasmidDNAextractionandPstⅠdigestlinger-printinganalysis,aswellasthetmiversalprimerPCRamplificationoftheTEM,SHV,CTX-M,VEB,PERandSFOgenesandtheDNAsequencinginordertodeterminethegenotypesofESBLsandtheirplasmidlocations.ItwasfoundthattheincidenceoftheESBLs-producingstrainsofGram-negativebacteriawas14.6%(173/1184)with67strainsoftransconjugantsand11strainsofelectro-transformants,inwhichCTX-M-14typewas33.3%(26/78);CTX-M-3typewas23.1%(18/78);CTX-M-9typewas14.1%(11/78);CTX-M-5typewas6.4%(5/78);CTX-M-13typewas2.6%(2/78);SHV-5typewas7.7%(6/78);SHV-12typewas5.1%(4/78),SHV-2atypewas2.6%(2/78)andunidentifiedtypewas5.1%(4/78).29.5%ofthewildstrainsalsocarriedbroad-spectrumbeta-lactamasesTEM-1andSHV-1types.TheabovementionedESBLsgeneswerelocatedontransferableplasmidswithvariablesizes(from35to190kb).TheCTX-MtypeESBLswascharacterizedbyhigh-levelofresistancetocefotaxime.ItconcludedthattheCTX-M-typewasthemostprevalentgenotypeinclinicalisolatesofGram-negativebacteriainsouthernChina,andtheSHVtyperanksinthesecondplace.TEM-,VEB-,Toho-andPER-typeswerenotfoundintheseisolates.

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简介：TheaimofthisstudyistoinvestigatethefeasibilityandmechanismofhIL-2-preSDNAvaccineaspreventionandtherapeuticapproachagainstHepatitisB.EukaryonexpressionvectorinvolvinghIL-2andpreSgenewasconstructedwithrecombinanttechniqueandtransferredintonormalBALB/cmiceandHBVtransgenicmice(Tg-Mice)respectively.Tnenaseriesofdetectionwereperformed:detectionofanti-preS2,HBsantibodyandHBsAginBALB/cmiceandTg-micewithELISA,quantificationofHBVDNAcopiesinHBVTg-miceserumwithreal-timePCR,determinationofhepatitisdegreewithimmunopathologicalHEstaininganddetectionofliverfunction.Anti-preS1canbedetectedat4^th,6^thand10^thweekininoculatedBALB/cmice.Injectionwithgenegungainedanadvantageovermuscularandsubcutaneousinjectionsinceitacquiredjust1/10inoculationquantity(10μg/mouse).HighestexpressionofIgG2aat4^thweeksuggestedThl-mediatedimmuneresponse,whichfacilitatedHBVcleaning.OfallinoculatedHBVTg-mice,80%ofthemshowedanfi-preS2,HBsantibodypositiveandHBVDNAdecreased,and20%showednegativeforHBsAg.HEstainingtohepatictissueshowedobviousinfiltrationofinflammatorycells,swellingandgranulardegenerationofhepatocytes.Inourstudy,IL-2-preSDNAvaccinewhichcanprovokethehumoralandcellularimmuneresponseandbreaktheimmunetolerancesupportsthedesignationandconstructionofnewvaccineagainstHBVandspecificimmuneremedyforHBVcontinuousinfection.

简介：Inordertoelucidatethemolecularandimmunologicalmechanismsaswellasthepathogenesisofhemorrhagicfeverwithrenalsyndrome(HFRS),theCD8^+cytotoxicTlymphecytes(CTL)clonewasestablisheddirectlyfromperipheralbloodmononuclearcells(PBMC)ofpatientswithHFRS.TheactivitiesofCTLweredetectedasusualwithEBV-transformedlymphoblastoidcellline(BLCL)astargetcells.TheresultsshowedthattheCTLclonecouldrecognizedandkilledthetargetcellswithspecificityofnucleocapsidproteinofHantaanvirus(HTNVNP)withthecytotoxicitypercentagesof50.2%,25.4%and39.0%respectively.TheseresultsdemonstratedthattheantigenicepitopesofHTNVNPmainlylocatedontheC-temainaloftheviralnucleocapsidprotein.

简介：Toexploretheroleofnuclearfactor-κB(NF-κB)inthesignalpathwayofproteinkinaseC(PKC)regulatingtheproliferationandapoptosisofTlymphocytesinasthma.Tlymphocyteswereisolatedfromtheasthmaticmodelofguineapigsandtheasthmaticpatients.EithertheTcellsstimulatedwithPMAaloneorthosestimulatedwithPMAtogetherwithpyrrolidinedithiocarbamate(PDTC)wereincubatedfor1and24h.TheproliferationofandthepresenceofNF-κBinthecellsincubatedfor1hwereobservedbyMTTandimmunohistochemicalstaining,respectivelyAndthecellsincubatedfor24hwereobservedfortheapoptosisbyTUNEL.Alltheassayswereparalleledwithcontrols,andallthedatawereanalyzedstatisticallywiththesoftwareSAS.ThepercentageofcellsofnuclearpositivestainingofNF-scBandtheproliferationofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithPMAweresignificantlyhigherthanthoseofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithoutPMArespectively(P<0.01)andthoseofTlymphocytesfromnormalcontrolguineapigsandnormalcontrolpersonsstimulatedwithPMArespectively(P<0.01),andweresignificantlyreducedbyPDTC(P<0.01).TheapoptosisindexofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithPMAweresignificantlylowerthanthoseofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithoutPMArespectively(P<0.01)andthoseofTlymphocytesfromnormalcontrolguineapigsandnormalcontrolpersonsstimulatedwithPMArespectively(P<0.01),andweresignificantlyinducedbyPDTC(P<0.01).ThereweregoodpositivecorrelationbetweenthepercentageofcellsofnuclearstainingofNF-κBofTlymphocytesandtheproliferationofTlymphocytes(r=0.51-0.72,P<0.001),andalsogoodnegativecorrelationbetweenthepercentageofcellsofnuclearstainingofNF-scBandtheapoptosisindexofTlymphocytes(r=-0.55-0.71,P

简介：Theaimofthisstudywastoanalyzethepointmutationoftheexon1atcodon54ofthemannose(ormannan)-bindinglectin(MBL)geneinhealthyindividualsofChineseHansandMongolianpopulation,andtofindoutanyassociationbetweentheplasmalevelsofMBLandthegenemutationfrequencyinbethgroupsofindividuals.Bloodsampleswerecollectedrandomlyfrom56healthyindividualsofChineseHansand37Mongolian.ThedetectionofthepointmutationsoftheMBLgenewasperformedbypolymerasechainreaction-restrictionfragmentlengthpolymorphism(PCR-RFLP)anddetectionsforplasmalevelsofMBLweredeterminedbyusingMBLELISAkits.AMBLPCRmethodofassaywasestablishedwithhighspecificity,andgoodreproducibility.ByoptimizingthePCRcondition,theoptimalannealingtemperaturewas55℃,andthelowestdetec-tionlimitwas160pg.Nobandswerefoundinnon-specificitysamples(HAV,HBV,HCVandTB),andthesequencesofPCRproductswerethesameastheexpectedones.AlsoaMBLPCR-RFIPwasestablished.Uponelectrophoresisofthedigestedproductsin3%agarosegel,therewere3patterns:inwhich2bandscorrespondtomoleculeweight232bpand93bp;1band,correspondstomoleculeweight325bpand3bandscorrespondtomoleculeweight325bp,232bpand93bp,respectively.Threebandsof325bp,232bpand93bpofpointmutationswerefoundatcedon54ofMBLcedinggene.FrequenciesinhealthyHanandMongolianpopulationwere0.2321and0.1757respectively.TheaverageplasmaMBLconcentrationwas1998.750μg/L,withSDof1505.152in56healthyHanpopulationand2525.676μg/L,withSDof1955.188in37Mongolian.AnegativecorrelationbetweenMBLconcentrationandgenemutationfrequencywasfoundinhealthyHanpopulation.Frequencyofpointmutationwas1.00whentheMBLconcentrationswerebelow100μg/L;frequencyofpointmutationwas0.4524whentheconcentrationwas100μg/Lto1000μg/L;andthefrequencyofpointmutationwas0.0156whentheconcentrationwaso

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