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:目的:利用人脐静脉内皮细胞体外培养模型,探讨肾上腺素刺激状态下海带多糖L01对内皮细胞的保护作用,以及其对其分泌的组织型纤溶酶原激活物(t-PA)的影响,从内皮细胞抗血栓功能方面探讨海带多糖L01对心血管系统的保护作用。方法:体外培养人脐静脉内皮细胞系ECV-304,用肾上腺素刺激24h、48h、72h采样,ELISA法测定HUVECs培养上清液中的t-PA含量,逆转录多聚酶链反应(RT-PCR)检测HUVEC细胞内t-PAmRNA的表达,其扩增产物经电泳后密度扫描及半定量分析。结果:肾上腺素模型组HUVECs培养液中t-PA(24h、48h)含量明显增加,;单给海带多糖对HUVECst-PA分泌无影响。海带多糖L01在肾上腺素刺激下给药24h,48h后t-PA抗原分泌明显降低(P〈0.01),且具有一定的量效关系,但HUVECst-PAmRNA表达各组均无明显差异。结论:肾上腺素可刺激内皮细胞分泌t-PA抗原,海带多糖L01对血管内皮具有一定的保护作用,减少肾上腺素刺激下HUVECs分泌t-PA,而对t-PAmRNA表达无明显影响。