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：TheaimofthisstudyistoexplorethegenomicmolecularorganizationandgenogroupofhumannorovirusfrominfectedinfantsinGuangzhouofChina.PrimersweredesignedaccordingtothegenomicsequenceofnorovirusintheGenBank,andthenorovirusgenomewasamplifiedbyRT-PCR.ThePCR-productswereclonedintoTvectorandsequenced,andthegenomicnucleotidesequenceswereanalyzedwiththeprogramsCLUSTALW/X,DNASTARandRAT(RecombinationAnalysisTool).TheNVgz01straingenomeis7558bpinlengthandencodesthreeopenreadingframes(GenBankaccessionNo.isDQ369797).ThegenomicsequencesofNVgz01werecomparedwiththoseofnomvirusinGenBank,whichrevealedthatthehomologywithgenogroupⅡrangesbetween76%-90%,andgenogroupⅠbe-tween43%-44%.TheORF1regionshared94%and88%identitywithMc37andFarmingtonstrains,respectively;thecapsidregion(ORF2)shared65%and94%identitywithMc37andFarmingtonstrains,respectively.Phylogenetictreeswerereconstructedbytheneighbor-joiningmethod.ComparativecompletesequenceanalysisoftheNVgz01withreportedhumannorovimsgenomicsequencesrevealedthatthisisolatebelongstogenogmupⅡ.TheORF1andORF2regionsshareddifferentidentitywithMc37andFarmingtonstrains,suggestingNVgz01couldbearecombinantvires.

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简介：Toinvestigatetheroleofnegative-regulatoryfactorsA20,IRF-4andTRAF4ofthetoll-likereceptor(TLR)signalpathwaysinimmunologicalpathogenesisofKawasakidisease(KD),48childrenwithKawasakidisease,16childrenwithinfectiousdisease(ID)and16age-matchedhealthychildrenwerestudied.Reverse-transcriptionPCR(RT-PCR)andreal-timePCRwereusedtoevaluatetheexpres-sionlevelsofnegative-regulatoryandeffectivefactorsintoll-likereceptor4(TLR4)signalpathwaysandproinflammatoryfactorsinperipheralbloodmonocyte/macrophage(MC).Inthisstudy,expressionlevelsofTLR4,MD-2,MyD88,IRAK-4,TRAF6,TAK1,andTAB2mRNAinKDgroupweredetectedtobeelevatedsignificantlyduringacutephaseofKD.Transcriptionlevelsofnegative-regulatoryfactorsA20,IRF-4andTRAF4mRNAinKDorIDpatientsincreasedremarkably.However,expressionsofIRF-4andTRAF4inKDpatientsweredetectedtobelowerthanthatinIDpatients,exceptthattran-scriptionlevelsofA20werefoundtobehigherthanthatinIDpatients.Simultaneously,expressionsofproinflammatorycytokinessuchasL-1β,IL-6andTNF-αinKDpatientsweresignificantlyelevatedcom-paredwiththoseinIDpatients.Furthermore,itwasfoundthatstimulationoflipopolysaccharide(LPS)remarkablyup-regulatedtheexpressionsofnegative-regulatoryfactorsA20,IRF-4andTRAF4inKDpa-tientsorhealthyvolunteers.ThemRNAlevelsofallthethreefactorsinKDpatientswerefoundtobelowerthanthatinthelatter.Inaddition,transcriptionlevelsofIRF-4andTRAF4inKDpatientswithcoronaryarterylesion(KD-CAL~+)weredetectedtobelowerthanthoseinKDpatientswithoutcoronaryarterylesion(KD-CAL~-)duringacutephase,whilethatofA20inKD-CAL~+groupwerelowerthanthatinthelatter.AndthelevelsofexpressionsofproinflammatorycytokinesinKD-CAL~+groupwerefoundtobehigherthanthoseinKD-CAL-group(P<0.01).Thesefindingssuggestthataberrantexpressionofnegative-regulatoryfactorsofTLRssignalpathwaysmaybeinvolved

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