简介：

简介：

简介：

简介：

简介：

简介：ToanalyzethegenomicmolecularstructureandgenotypeofhumanastrovirusisolatedfrominfantinGuangzhouofChina,theprimersweredesignedbasedonthegenomicsequenceofastrovirusfromtheGenBankandthetargetsequencewereamplifiedbyRT-PCR.ThenthePCR-productswereclonedtoTvectorandsequenced.ThegenomicnucleotidesequenceswereanalyzedbytheprogramsCLUSTALWandDNASTAR.ItwasfoundthatthefullgenomiclengthofHASTVgz01strainwas6721bpandtheORFswere6558bp.The5'and3'UTRwere82and81nucleotides.Thegenomeincluded3openreadingframes(ORFs):ORF1a,ORF1bandORF2.The5'-terminalORF1astartedatnucleotide83andextendedtonucleotide2845.ORFlb(nt2785tont4332)overlapedORFlaby61nucleotides.The3'-terminalORF2beganatnucleotide4325andterminatedatnucleotide6640.ORF2had2316nucleotides.ComparedwithotherastrovirussequencesinGenBank,thehomologyoftheaminoacidsequenceofORF2ofHASTVgz01strainwiththatofserotype4was93%.Homologywithotherserotypesrangedfrom61%to70%.ThecompletenucleotidesequenceofastrovirusHASTVgz01strainisolatedfromGuangzhouinChinawas6721bpinlength,GenBankaccessionNO.DQ344027.ComparingtheORF2ofastrovirusHASTVgz01withtheknownsequencesoftypes1-8thehighesthomologywasserotype4(93%).ComparativesequenceanalysisoftheHASTVgz01ORF2withthereportedhumanastrovirussequencesrevealedthattheisolatedastrovirusbelongstogenotype(serotype)4.

简介：

简介：

简介：

简介：

简介：

简介：

简介：

简介：ToclarifytheroleofAPOBEC3G(A3G)incellulardefenseagainsthepatitisBvirus(HBV),theexpressionofA3GinnormalhumanliverandtheregulationoftheA3Gexpressioninhep-atomacellline(HuH-7)wereinvestigated.ExpressionlevelofAPOBEC3smRNAinhumanliverwasdeterminedbyRT-PCR.HuH-7andHepG2cellsweretreatedwithvariousconcentrationsofIFN-α(0U/ml,100U/ml,500U/ml,1000U/ml)for12h.ThemRNAlevelsweremeasuredbyaquantitativeRT-PCR,theresultswerenormalizedrelativetothespecimenswithoutIFN-αstimulation.TotalproteinofHuH-7cellstreatedwithvariousconcentrationsofIFN-αfor48hwassubjectedtoWesternblotanalysis.Forreportergeneassay,HuH-7cellsweretransfectedwiththereporterplasmidscontainingIRF-Esitesanditsmutantswithdifferentlengths.Thenthecellsweretreatedwithorwithout1200U/mlIFN-aforadditional12h(1000U/ml)after24hoftransfection,andthecelllysatewaspreparedandassayedforluciferaseactivity.ItwasfoundthatnormalhumanliverexpressedthemRNAofA3G.A3GmRNAexpressioninHuH-7andHepG2cellswereup-regulatedbyIFN-αstimulationinadose-depen-dentmanner.WesternblotanalysisindicatedthatA3GproteinexpressionwasalsoenhancedbyIFN-αstimulation.SequenceanalysisshowedtheexistenceofputativesitesofIFNregulatoryfactorelement(IRF-E)in5'regionofA3Ggeneupstreamtheinitiationcodon.IFN-αstimulationresultsin6-to8-foldincreaseinluciferaseactivityincellstransfectedwiththeplasmidcontainingIRF-Esitesofthe5'upstreamsequences,whereasluciferaseactivitydidnotchangeincellstransfectedwiththeplasmidcontainingmutantIRF-EsitesorwithoutIRF-Esites.Asaconclusion,A3Gareexpressedinnormalhumanliver.A3Gexpressionwasup-regulatedbyIFN-αstimulationinhepatomacellsandcouldbeinvolvedinhostdefensemechanismsagainstHBV.ERF-Esitein5'regionofAP0BEC3Ggeneupstreamtheinitiationcodonplaysanimportantroleinthisp

简介：

简介：

简介：TheexpressionofIL-4inaratmodelofchronicpulmonaryinfectionbiofilmformationinducedbyPseudomonasaeruginosawasinvestigated,inwhichSPFWisterratswereinfectedviatracheawith0.1mlP.aeruginosastrainPAO579(10~9CFU/ml)inalginatebeadsortheplanktonicformofthisbacterialstrain(10~9CFU/ml),andon3,7and14dafterinfection,thebacteriologicalandpathologicalchangeswereobservedaswellastheexpressionofthecytokineIL-4wasdetermined.ItwasdemonstratedthatthecountofCFUperlungtissueincaseofbacteriainalginatebeadswassignificantlyhigherthanthatofbacteriainplanktonicform,withmoreseveregrosspathologicchangesandinflammatoryreactionsinthealginatebeadgroupincomparisonwiththatoftheplanktonicforms(P=0.002,P=0.004andP=0.002,respectively).Inaddition,theexpressionofIL-4inthealginatebeadgroupwasalsohigherthanthatintheplanktonicform(P=0.02,P=0.02andP=0.022,respectively).Apositivecorre-lationbetweenthelevelofIL-4expressionandthegrosslungpathologyinalginatebeadgroupexistedasdemonstratedbysimpleregressionanalysis(r=0.78,P<0.02).Itisconcludedthatthechronicpul-monaryinfectionwithbiofilmformationinducedbyP.aeruginosatendstohavetheprioritytotheTh2immuneresponse.

简介：

简介：

简介：