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  • 简介:InordertorevealvariationandrevolutionofNPgenesofhumanavianH_5N_1influenzavirusstrains,theNPgeneofahumanavianH_5N_1influenzavirusstraininGuangdongwassequencedandtheglobalNPgenesofstrainswereretrieved.ThesequenceswereanalyzedbyDNAStar5.0,andtheevolu-tionaryspeedwasstudiedwithreferencetotheepidemiologicaldata.ItwasfoundthatNPgenesof45strainsduring1997-2006werehomologicallyclassifiedintothreegroups:strainsin1997-1998,strainsin2004-2005andstrainsfrom2003to2006.Therewere35substitutionsinNPsinallstrainsaccountingforaratioof7.03%(35/498).Anadditionalglycoproteindomain(NGT_(430-432))wasfoundinNPgenesinthestrainsof2003-2006,themutationofN_(370)SinGD-01-06resultedinoccurrenceofonemoreglyco-proteindomain(NES_(368-370)).Inthesynonymousvariation,K_svaluesinNPwere2.03×10~(-5)-2.55×10~(-5)Nt/dandK_avaluesinNPwere1.58×10(-6)-3.10×10~(-6)Nt/d.Theredidn′texistobviouslyselectivepres-sure.Anadditionalglycoproteindomainineverystrainof2003-2006andonemoreinstrainGD-01-06mightchangetheantigenicityofhumanavianH_6N_1influenzavirus.ThevariationonhumanavianH_5N_1influenzastrainsoccurredfrequentlyinthenaturalworld,whichwouldresultinhighprobabilityofhu-man-humantransmissionalongwiththenaturalevolutionofthevirus.

  • 标签: NP基因 禽流感 H5N1病毒 基因表达
  • 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.

  • 标签: 疫苗 免疫机制 蛋白质 原核表达
  • 简介:ThegenomeoftheenterohemorrhagicEscherichiacoliO157:H7EDL933contains177“O”-islands(OIs).TostudytheirpotentialcontributiontotheO157-specificpathogenicity,wesurveyedthedistributionof22OIsbyPCRandDNAhybridizationin17isolatesofShigatoxinproducing(Stx-positive)E.coliO157:H7,andcomparedwiththeirdistributionin21isolatesofStx-negativeE.coliO157and21isolatesofnon-O157entericpathogens.Fourteenof22OIswerepresentinnon-O157entericpathogensanalyzed.Eightof22OIswerefoundonlyinthe17Shigatoxin-(Stx)positiveE.coliO157:H7isolates,buttheywereabsentfromthe21Stx-negativeE.coliO157:NMandO157Hundisolatestested.Amongthe8OIs,onlyOI43orOI48wereexclusivelydetectedinStx-positiveE.coliO157:H7,absentfromneitherofStx-negativeE.coliO157andnon-O157entericpathogens,suchasSalmonella,ShigeUa,Citrobacter,Vibriocholera,enteropathogen-icE.coli(EPEC),enteroadherentE.coli(EAEC),enteroinvasiveE.coli(E1EC)andenterotoxingenicE.coli(ETEC).TheOI43andOI48are83kbinsizeandidenticalinDNAsequences,whichencodegenesforurease,telluriteresistanceandadherence.ByanalyzingtheirjunctiongeneswithPCRandDNAhybridization,wefoundthat21ChineseisolateshaveOI48only.However,for7Japanesepatientisolates,4haveOI43and3haveOI48;forAmericanisolates,2havebothofO143andOI48,2haveOI48only.ThesedataconfirmedthehighlyplasticityofthepathogenicE.coligenome.TheuniquepresenceofOI43/OI48inStx-positiveE.coli0157:H7denotesitscriticalroleinthepathogenicityspecifictothispathogen.

  • 标签: O-岛 埃希氏菌属 大肠菌O157 肠病原体 E.大肠菌 O157:H7
  • 简介:【摘要】目的:探讨连续护理联合24h动态血压监测对老年原发性高血压病病人睡眠及血压控制的影响。方法:抽取本院2019.01至2019.12时段内诊疗的86例原发性高血压老年病人,划分为甲组(43例)和乙组(43例)。甲组仅为常规护理,乙组为连续护理联合24h动态血压监测,评估病人睡眠及血压控制效果。结果:干预1个月、2个月、3个月时,患者血压水平和睡眠质量均优于第2天,但乙组改善幅度明显超过甲组,数据比较有意义(P<0.05)。结论:针对原发性高血压老年病人,连续护理联合24h动态血压监测模式,既可增强病人睡眠质量,还可稳控血压,促进疾病恢复,可推广。

  • 标签: 连续护理 24h动态血压监测 老年病人 原发性高血压 睡眠 血压控制
  • 简介:目的:基于重组流感病毒载体的呼吸道合胞病毒(RSV)疫苗的构建及在小鼠体内的免疫保护效果评价。方法:构建并拯救表达RSV A2型G蛋白胞外结构域(Gecto)的重组甲型流感病毒,将其命名为PR8NAGecto/WSN。体外验证重组病毒G蛋白表达与病毒生长动力学后,单剂滴鼻免疫BALB/c小鼠,并评价体液免疫、黏膜免疫与细胞免疫。免疫4周后,分别用RSV A2与RSV B9320进行攻毒,通过小鼠体重变化、肺组织病毒滴度及病理评价免疫保护效果。结果:单剂滴鼻免疫PR8NAGecto/WSN能在小鼠体内产生较强的体液免疫、黏膜免疫及细胞免疫。与对照组比较,免疫组小鼠经RSV A2或B9320两个亚型病毒攻毒后肺病毒载量与肺组织病理均明显改善。结论:单剂滴鼻免疫重组PR8NAGecto/WSN疫苗可在小鼠体内诱导较强的RSV特异免疫应答与攻毒保护。本研究为新型RSV黏膜疫...

  • 标签: 呼吸道合胞病毒流感病毒病毒载体G蛋白
  • 简介:目的:基于重组流感病毒载体的呼吸道合胞病毒(RSV)疫苗的构建及在小鼠体内的免疫保护效果评价。方法:构建并拯救表达RSV A2型G蛋白胞外结构域(Gecto)的重组甲型流感病毒,将其命名为PR8NAGecto/WSN。体外验证重组病毒G蛋白表达与病毒生长动力学后,单剂滴鼻免疫BALB/c小鼠,并评价体液免疫、黏膜免疫与细胞免疫。免疫4周后,分别用RSV A2与RSV B9320进行攻毒,通过小鼠体重变化、肺组织病毒滴度及病理评价免疫保护效果。结果:单剂滴鼻免疫PR8NAGecto/WSN能在小鼠体内产生较强的体液免疫、黏膜免疫及细胞免疫。与对照组比较,免疫组小鼠经RSV A2或B9320两个亚型病毒攻毒后肺病毒载量与肺组织病理均明显改善。结论:单剂滴鼻免疫重组PR8NAGecto/WSN疫苗可在小鼠体内诱导较强的RSV特异免疫应答与攻毒保护。本研究为新型RSV黏膜疫

  • 标签: 呼吸道合胞病毒流感病毒病毒载体G蛋白