简介：AIM:Toconstructanon-resistantandattenuatedSalmonellatyphimurium(S.typhimurium)strainwhichexpressesconservativeregionofadhesionABofHelicobacterpylori(Hpylon)andevaluateitsimmunogenicity.METHODS:TheABgeneamplifiedbyPCRwasinsertedintotheexpressionvectorpYA248containingasdgeneandthroughtwotransformationsintroducedintothedeltaCya1deltaCrp1deltaAsdattenuatedSalmonellatyphimuriumstrain,constructingbalancedlethalattenuatedSalmonellatyphimuriumstrainsX4072(pYA248-AB).BridgedELISAmethodwasusedtomeasuretheexpressionofABantigeninsonicateandculturesupematant.AccordingtothemethoddescribedbyMeacock,stabilityoftherecombinantwasevaluated.Semi-lethalcapacitytestwasusedtoevaluatethesafetyofrecombinant.Theimmunogenicityofrecombinantwasevaluatedwithanimalexperiments.RESULTS:TheattenuatedS.typhimuriumX4072(pYA248-AB)whichexpressesABwassuccessfullyconstructed.Furthermore,bridgedELISAassayshowedthatthecontentofABinrecombinantX4072(pYA248-AB)culturesupematantwashigherthanthatwasinthalluslyticliquor.AndafterrecombinantX4072(pYA248-AB)wasculturedfor100generationswithoutselectionpressure,bheentirerecombinantbacteriaselectedrandomlycouldgrow,andtheABantigenwasdefectedpositivebyELISA.ThegrowthcurveoftherecombinantbacteriashowedthatthegrowthstatesofX4072(pYA248)andX4072(pYA248-AB)werebasicallyconsistent.ThesurvivalrateofC57BL/6wasstill100%,at30daftermicetakingX4072(pYA248-AB)1.0×l0^10cfuorally.OralimmunizationofmicewithX4072(pYA248-AB)inducedaspecificimmuneresponse.CONCLUSION:Invitrorecombinantplasmidappearstobestableandexperimentsonanimalsshowedthattherecombinantstrainsweresafeandimmunogenicinvitro,whichprovidinganewliveoralvaccinecandidateforprotectionandcareofHpyloriinfection.

简介：AIM:RecombinedplasmidpETNF-P16wasconstructedtoinvestigateitsexpressionpropertiesinesophagealsquamouscarcinomacelllineEC9706inducedbyX-rayirradiationandthefeasibilityofgene-radiotherapyforesophagealcarcinoma.METHODS:RecombinedplasmidpETNF-P16wasconstructedandtransfectedintoEC9706cellswithlipofectamine.ELISA,Westernblot,andimmunocytochemistrywereperformedtodeterminetheexpressionpropertiesofpETNF-P16inEC9706aftertransfectioninducedbyX-rayirradiation.RESULTS:EukaryoticexpressionvectorpETNF-P16wassuccessfullyconstructedandtransfectedintoEC9706cells.TNFαexpressionsweresignificantlyincreasedinthetransfectedcellsafterdifferentdosesofX-rayirradiationthaninthoseafter0Gyirradiation(1192.330-2026.518pg/mL,P<0.05-0.01),andtheTNFαexpressionsandP16weresignificantlyhigher6-48hafter2GyX-rayirradiation(358.963-585.571pg/mL,P<0.05-0.001).NoP16expressionwasdetectedinnormalEC9706cells.However,therewasstrongexpressioninthetransfectedandirradiationgroups.CONCLUSION:X-rayirradiationinductioncouldsignificantlyenhanceTNFαandP16expressioninEC9706cellstransfectedwithpETNF-P16plasmid.Theseresultsmayprovideimportantexperimentaldataandtherapeuticpotentialforgene-radiotherapyofesophagealcarcinoma.