简介：

简介：Inordertoinvestigatetheimmtmogenicityofthecontrolled-releasemicroencapsulatedhepatitisBvaccineinmice,polyethyleneglycol-poly-dl-lactide(PELA)microsphereswithentrappedHSsAgwerepreparedbydoubleemulsionW/O/Wbasedonsolventextractionmethods.BALB/cmicewereimmunizedwiththeencapsulatedvaccinebyoralfeedingorinjection.Bloodsampleswerecollectedat8^th,10^th,14^thand24^thweeks,respectively,andthelevelsofantibodyresponseweredetectedbyEI.ISA.Itwasfoundthatthescanningelectronmicroscopyshowedthepreparedmicrosphereshadsmoothandsphericalsurface,suitableforvaccinedelivery.Twogroupsofmiceorallyfedwiththeencapsulatedorconventionalrecombinantvaccines,respectively,theresereshowednoobviousdifferenceintheIgGlevels.At14^thweek,thegroupinjectedwithasingledoseofencapsulatedvaccinehadasimilarlevelofIgGresponsetothegroupinjectedwithtwodosesoftherecombinationvaccine.At24^thweek,theIgGlevelsofthegroupinjectedwithtwodosesofencapsulatedvaccinewerehigherthanthoseofthegroupinjectedwithtwodosesoftherecombinationvaccine.ItconcludesthatControlled-releasemicroencapsulatedhepatitisBvaccinepossessesthefeatureofslowlyreleasinginv/voandlongtimesimmtmogenicity.

简介：

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

简介：Toinvestigatethematernal-infantileinfectionwithhumanparvovirusB19,theIgGandIgMantibodiesagainsthumanparvovirusandtheB19-DNAinserumandperipheralbloodmononuclearcells(PBMC)ofpregnantwomenaswellastheserumIgMantibodyagainstB19andtheB19-DNAinserumandcordbloodnucleatedcells(CBNC)ofnewbornsweredeterminedbyELISAandnestedPCRrespectively.ItwasfoundthatthepositiverateoftheIgGantibodyagainsthumanparvovirusB19inseraof92pregnantwomenwas38.04%(35/92),andthatoftheIgMantibodyin720pregnantwomenwas9.03%(65/720).However,theIgMantibodyagainsthumanparvovirasB19wasnegativeinthecordbloodseraof95newborns.AstothehumanparvovirasB19DNA,noneof720pregnantwomenand95newbornswasprovedtobepositiveintheirsera,Nevertheless,thepositiverateoftheparvovirasB19DNAinPBMCwas3.06%(3/98)in98pregnantwomenand1.12%(1/89)inCBNCof89newborns.ItisconcludedthatthehistoryofinfectionwithhumanparvovirasB19existsincertainpregnantwomenwithasmallpercentageofpregnantwomeninfectedwithrecentoracuteinfectionsofB19virus.ThedetectionratesoftheB19viralDNAinPBMCofpregnantwomenandCBNCofnewbornswerehigherthanthoseinsera,indicatingthattheriskforverticaltransmissionisverylow.

简介：Thereversetranscriptase(RT)proteinofhepatitisBvirus(HBV)hasbeensuccessfullyexpressedbyrecombinanttechnologyinEschericahiacoli(E.coli).Inthisstudyweaimedtodevelopasemi-quantitativeassayforthestudyofHBVRTproteinusingthissystem.CompleteHBVpolymerasegenefromawildtypevirus(rt306P)andthepolymerasegenefromamutant,withrt306Psubstitutedbyserine(rtP306S)wereseparatelyfusedtothemaltosebindingprotein(MBP)geneandexpressedinE.colirespectively.TheexpressionlevelsofHBVpolymerasegenesfromthewildtypevirusanditscounterpartmutantatrt306werecompared.Whentheseproteinsweresemi-quantifiedbyWesternblottingusingrabbitanti-TPserum,thertP306SmutantshoweddecreasedexpressionofMBP-HBVpolymerase.Bythismethod,wehaveshownthattheexpressionlevelofHBVRTcouldbeaffectedbysubstitutionsinitsaminoacidsequences,andthismethodcouldbeusedtostudythecharacteristicsofHBVRTprotein.

简介：CpGoligodeoxynucleotides(CpGODN)asadjuvanthavebeenextensivelystudiedinrecentyears.PhosphodiesterCpGODN(POCpGODN)canperfectlymimicbacterialDNAinenhancingimmuneresponsebutarevulnerabletonucleasesinvivo.ThisstudyaimedtoevaluatetheimmunostimulatorypotentialandsafetyofphosphodiesterCpGODNencapsulatedinnonphospholipidliposomes.BALB/cmicewereimmunizedintramuscularlywithdifferentformulationsofliposomes,CpGODNandhepatitisBsurfaceantigen(HBsAg).TheresultsdemonstratedthattheencapsulatedPOCpGODNwereprotectedagainstrapiddegradationinvivoandretainedtheiradjuvantactivity.POCpGODNencapsulatedwithHBsAginliposomesinducedstrongTh1-biasedorTh1/Th2mixedhumoralimmuneresponseinmicewiththemagnitudesimilartotheirphosphothioateequivalentinthesameformulation.HighIFN-gammaproductioninducedbythisformulationconfirmedthegenerationofstrongcellularimmuneresponse.Additionally,co-deliveryofHBsAgandPOCpGODNimprovedtheimmuneresponseoverthatobtainedwithseparatedelivery.Safetyexperimentshowedthatliposome-encapsulaedPOCpGODNandHBsAgcausedmildsystemicandmoderatelocaladversereaction.Inconclusion,ourdatashowsthatPOCpGODNencapsulatedinliposomesfullyexhibittheirTh1-typeadjuvantactivityandactasapotentialadjuvantforvaccines.

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简介：ToinvestigatetheHLA-A,-BallelepolymorphisminHanpopulationofShandongprovinceandtoexplorethepossibilitytofindouttheHLA-A,-B-matchedcordblooddonorsforstemcelltransplantationtobeusedinotherareainChina,5844umbilicalcordbloodsamplesweretakenfromHanpopulationdonorsofShandongprovince,andassayedwithPCR-sequence-oligonucleotide(PCR-SSO)assay.InShandongHandonors,20allelesatHLA-Alocusand46allelesatHLA-Blocuscouldbedetectedasrevealedinthepresentstudy.Amongthe20allelesatHLA-Alocus,themostprevalentfiveallelesincludedA*02(0.3041),A*11(0.1443),A*24(0.1434),A*30(0.0975)andA*33(0.0859),while,thealleleswithlowergenefrequenciesincludedA*34(0.0006),A*25(0.0005),A*66(0.0005),A*74(0.0004)andA*(0.0001).Ofthe46HLA-Ballelesdetected,themostprevalentfivealleleswereB*13(0.1348),B*51(0.0713),B*62(0.0712),B*61(0.0676)andB*60(0.0642);whilealleleswithlowergenefrequenciesincludedB*77(0.0001),B*76(0.0002),B*47(0.0003),B*42(0.0003)andB*72(0.0004).IncomparisonwiththoseoftheotherHanpopulationinChina,theHLA-A,-BgenefrequenciesintheumbilicalcordbloodofShandongprovincepossessuniquedistributionfeaturesamongtheinvestigatedpopulationsfromvariousregionsofthesameraceorigin,andthedifferencesinvariousregionsofthesameracewerelessthanthoseamongthedifferentrace.ItisevidentthattheHLA-A,-BallelesoftheumbilicalcordbloodtakeninShangdongprovinceshowhighdegreeofpolymorphism,anditmightbepartofthoseofNorthernHanpopulationinChina.So,itisreasonableforpatientsofNorthernChinesetoreceiveHLAclassI-matchtransplantofcordbloodstemcellsfortissueandorgantransplantationfromShangdongumbilicalcordbloodbank.

简介：ThepurposeofthisstudywastoconstructaneukaryoticDNAvectorencodingamultipleepitopeantigen(MFC)ofhepatitisCvirus(HCV)andahepatitisBsurfaceantigen(HBsAg),andexploretheeffectofHBsAggeneontheimmunityofHCVmultiple-epitopeDNAconstructinvitroandinvivoinmice.AnHCVDNAvector(pVAX1-HBs-MFC)wasconstructedbyfusingHBsAggenetotheNterminalofanHCVmultiple-epitopeantigengene.ThepVAX1-HBs-MFCwastransfectedintoHEK293TcellsanditsexpressionwasmeasuredbyELISAandWesternblotting.BALB/cmicewereintramuscularlyimmunizedwiththepVAX1-HBs-MFC,andanELISAapproachwasappliedtodeterminethespecificantibodytitersandsubtypesinthemouseserum.Thecross-reactivityoftheantibodieswasalsocheckedwithtwosynthesizedHCVhypervariableregion1(HVR1)peptides.TheIFN-γproductionandcellproliferationofthemousespleencellswereevaluatedbyELISAandMTS(3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium,innersalt)assays,respectively.TheexpressionofpVAX1-HBs-MFCwasdetectableinthetransfectedHEK293Tcells.TheserumantibodyresponsewaseffectivelyelicitedinBALB/cmiceinjectedwithpVAX1-HBs-MFC.ThehighesttiterofantibodyagainstHCV(MFC)was1:1280,andtheratioofIgG2a/IgG1was1.50±0.12atthefifthweekafterfirstimmunization.Moreover,thecollectedmouseserumantibodyhadtheabilitytocross-reactwiththetwosynthesizedHCVHVR1peptides.ThestimulationindexofthemousesplenocytestoMFCwas1.79±0.07,andtheIFN-γlevelwas287±6pg/mlatweek21afterfirstimmunization.ThehighesttiteroftheantibodyincontrolBALB/cmiceimmunizedwithpVAX1-MFCwas1:320,andtheratioofIgG2a/IgG1was1.33±0.11atweek5post-immunization.Furthermore,thestimulationindexofthemousesplenocytescellstoMFCwas1.52+0.06,andtheIFN-γlevelwas225±9.3pg/mlatweek21post-immunization.TheHBsAggenecanenhancetheeffectsofanHCVmultiple-epitope

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：ToconstructandexpressthefusionproteinStx2B-IntiminC300ofEHEC0157:H7,andtofurtherinvestigateitsimmunoprophyiacticpotential,thegeneofStx2B(stx2b)fromEHEC0157:H7chromosomewasclonedintopMD18-Tvector.Thereafter,theamplifiedgenewasclonedintoprokaryoticexpressionplasmidpET-28a(+)-eaeC300,whichwasconstructedpreviously.TherecombinantpasmidpET-28a(+)-stx2b-eaeC300wastransformedintoE.coliBL21(DE3).Afterinducement,theproteinStx2B-IntiminC300wassuccessfullyexpressedandanalyzedwithsodiumdodecylsulfatepolyacrylamidegelelectrophoresis(SDS-PAGE),WesternblottingandN-terminalaminoacidresidualsequencing.Toevaluateitsimmunoprophyiacticpotential,itwasprimarilypurifiedbyion-exchangechromatographyandinjectedinto30BALB/cmicewithAl(OH)3inthesubscapularregion.Tendaysafterthelastboostervaccination,20micewereattackedwithEHEC0157:H7lysateandtheprotectiveefficacywasobserved.Inthepresentstudy,thegeneofStx2B-IntiminC300wassuccessfullyclonedintopET-28a(+)vector.TheresultsofSDS-PAGEandWesternblottingassayshowedthatthefusionproteinwassuccessfullyexpressedintheinclusionbodyform,accountingfor25%oftotalexpressionproducts,anditsmolecularweightwasabout43kDa.TheresultoftheN-terminalaminoacidresidualsequencingshowedthatitwasidenticaltothatofthemoleculardesigned.Thepuritywasabout75%afterprimarypurification.AnimaltestsrevealedthatthefusionproteinStx2B-IntiminC300haselicitedhightiterofprotectiveantibodyrelatively.TheseresultsdemonstratethatthefusionproteinStx2B-IntiminC300issuccessfullyexpressedinprokaryoticexpressionsystemandshowscertainimmunoprophyiacticpotential.

简介：

简介：InordertoinvestigatewhetherlipoxinA4(LXA4)hasanantagonisticeffectonlipopolysaccharide(LPS)-inducedsynthesisofinterleukin(IL)-1β,IL-6andIL-8inratpulmonarymicrovascularendothelialcells(PMVEC),andtoexplorethemolecularmechanismsofsignalpathwayinLXA4actions,culturedPMVECweretreatedwithLPS,withorwithoutpreincubationwithLXA4.ProteinsofIL-1β,IL-6andIL-8insupernatantwereanalyzedbyenzyme-linkedimmunosorbentassay(ELISA).ExpressionsofmRNAofIL-1β,IL-6andIL-8weredeterminedbyRT-PCR.Expressionsofphosphorylationofphosphoinositide3-kinase(PI3-K)andmyeloiddifferentiationfactor88(MyD88)wereanalyzedbyWesternblot.ActivitiesofDNA-bindingofnuclearfactor-kappaB(NF-κB)andactivatorprotein-1(AP-1)weremeasuredbyelectrophoreticmobilityshiftassay(EMSA).TheresultsshowedthatLPSinducedproductionofIL-1β,IL-6andIL-8inratPMVECviaMyD88/PI3-K/NF-κBandAP-1pathway-dependentsignaltransduction.LPS-stimulatedexpressionofPI3-K,activitiesofNFκBandAP-1,secretionofproteinandexpressionofmRNAofIL-1β,IL-6andIL-8butnotMyD88expressioninPMVECwereinhibitedbyLXA4inadose-dependentmanner.Inconclusion,LXA4inhibitssynthesisofIL-1β,IL-6andIL-8bydown-regulationofPI3-K/NF-κBandAP-1signalpathwayinPMVEC.