简介：TofindSchistosomajaponicum(S.j)newantigengenethusprovidemoreusefulvaccinecandidates,thecDNAlibraryofS.jadultwormwasscreenedwithseraofrabbitsimmtmizedwiththemembraneantigensofSchistosomajaponicumhepato-portalschistosomula(SjHmAg).ThepositivecloneswereamplifiedbyPCRandsequenced,thenthesequencesofcloneswerecomparedwithallsequencesinGenBankdatabaseusingBlastprocess.ThenewclonesweresubmittedtoGenBankforaccessionnumbers.Fifteenpositivecloneswereobtainedafterthreeroundsofimmunoscreening.ThesizeofS.jcDNAfragmentsinpositiveclonesrangedfrom0.7kb-3.0kbafterautomaticallyexcisedwiththehelperphage.SequenceanalysisrevealedthatpartialsequenceofcloneM5hadsignificanthomologywithS.jmitochondriarnRNA,theotherpositivecloneswerenewS.jgenes.M2clonesequence(GenBankaccessionnumberAF502579)was730bplongithada117bpopenreadingframe(ORF).ThesequenceofM15(GenBankaccessionnumberAF502582)hasnotransmembraneregionandencodes92aminoacids,anditsproteincontainsaferredoxinsiron-sulfurbindingregionsignatureandtwoVWFCsignalregions.ThesizeofM1,M8,M9,M12(GenBankaccessionnumbers:AF502578,AF502580,AF500622,AF502581)rangesfrom402bpto766bp.ItconcludedthattheserafromrabbitimmunizedwithSjHmAgcouldrecognizeS.jspecificantigensmolecules,andtheseantigensmayinducetheprotectiveimmunityagainstS.jinfection.

简介：HumanprimaryepithelialcellsofrenalpelviswasestablishedtoinvestigatetheadherenceofuropathogenicEscherichiacoli(UPEC)tothiscellline,inwhichtheprimarycellculturewasperformedbyusingcultivationofthenormalepitheliumofrenalpelvisinkeratinocyteserumfreemedium(K-SFM)withepidermalgrowthfactor(EGF)andbovinepituitaryextract(BPE).BothUPEC132obtainedfromurinespecimenofpatientswithpyelonephritisandthepilus-freerepresentativestrainE.coliK-12p678-54wereusedtostudytheadherenceofthesestrainsonhumanprimaryepithelialcellsofrenalpelvis.TheUPECadherencewasperformedwithobservationonthemorphologicalchangesoftheadheredcells,whiletheadhesionratesandindiceswerecalculatedindifferenttimesofexperiment.Inaddition,thevirulencegeneshlyandcnf1ofUPEC132weredetectedbymultiplexPCRassay.Inthisstudy,thehumanprimaryepithelialcellsofrenalpelviswasfoundtoexhibitthecharacterofthetransitionalepithelialcells.Comparedwiththecontrolgroup,theadhesionratesandindicesbegantoincreasefrom15minoftheexperimenttimeandreacheditspeakin120min.TheadhesionrateandindexofUPEC132tohumanprimaryepithelialcellsofrenalpelviswere74.4%and34.0respectively.ManymicroscopicchangesintheprimarycellsadheredwithUPEC132couldbedetected,suchasroundingorirregularityinshape,unevennessinstainingandthecytoplasmicandnuclearchanges.ItsuggeststhathumanprimaryepithelialcellsofrenalpelviscanbeusedfortheexperimentonUPECadhesion,thusprovidingabasisforthefurtherstudyonthepathogenesisofUPEC.

简介：WehavedevelopedandtestedchimericT-cellreceptors(TCR)specificforp185HER2.Intheseexperiments,retroviralvectorsexpressingtheN297orN29ξreceptorswereconstructedinpRET6.AmphotropicviralproducercellswereestablishedintheGALV-basedPG13packagingcellline.Ficollpurifiedhumanperipheralbloodlymphocytes(PBL)werevitallytransducedusinganoptimizedprotocolincorporatingactivationwithimmobilizedanti-CD3/anti-CD28monoclonalantibodies,followedbyviralinfectioninthepresenceoffibronectinfragmentCH296.Transducedcellswereco-culturedwithhumantumorcelllinesthatoverexpress(SK-OV-3)orunderexpress(MCF7)p185HER2toassayforantigenspecificimmuneresponses.BothCD4^+andCD8^+T-cellstransducedwiththeN297orN29ξchTCRdemonstratedHER2-specificantigenresponses,asdeterminedbyreleaseofTh1likecytokines,andcellularcytotoxicityassays.OurresultssupportthefeasibilityofadoptiveimmunothempywithgeneticallymodifiedT-cellsexpressingachTCRspecificforp185HER2.

简介：TheaimofthisstudyistoinvestigatecyclinE,pl6inkdaandki67aspossiblediagnosticbiomarkersforcervicalpreneoplasiausingcervicalexfoliated-cellspecimens,andevaluatethesignificanceforscreeningpatientsathighriskofdevelopingcervicalcarcinoma.TheexpressionofcyclinE,pl6inkdaandki67wasexaminatedin78cervicalexfoliatedepithelialspecimensdiagnosedasatypicalsquamouscellsofundeterminedsignificance(ASCUS)(12cases),cervicalintraepithelialneoplasia(CIN)oftype1(17cases),CIN2_3(38cases)andinvasivecarcinoma(11cases)usingimmunohistochemicalanalysis,andsimultaneously,theDNAstatusofhumanpapillomavims(HPY)type16/18wasdetectedbypolymerasechainreaction(PCR)usingtypespecificprimers,cyclinE,pl6inkdaandki67werealloverexpressedinCINsandinvasivecarcinoma,comparedwithlittleexpressioninASCUS(P<0.005).OverexpressionofcyclinEwasobservedinCIN1(94.1%,X^2=21.16,P<0.01),andp16inkdaandki67wereoverexpressedininvasivecarcinoma(100%and90.9%respectively).Thedegreeofpl6inkdaandki67expressioncorrelatedwellwiththatofepitheliallesions(P<0.005).HPV16/18infectionwasassessedinC1Nsandinvasivecarcinomasamples,andrevealedasignificantrelationshipwiththedegreeofcervicalepitheliallession.Theexpressionlevelofpl6inkdaandki67seemedmorecloselyassociatedwithHPVI6infectionthanthatofcyclinE(rs=1.0vsrs=0.4).Only1caseinCINIanddcasesinCIN2-3ofHPV18positivesamplesweredetected.Thereforenostatisticalsignificancewasfoundbystatisticalanalysis.OverexpressionofcyclinE,pl6inkdaandki67inCINsandinvasivecarcinomacellsdemonstratesthepotentialuseofcyclinE,pl6inkdaandki67asdiagnosticbiomarkersforHPV-relatedcervicalneoplasticlesions.Inaddition,thistechniquecanbeusedforscreeningpatientsathighriskofdevelopingcervicalcarcinoma.