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  • 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
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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
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  • 简介:摘要:国际糖尿病联盟(IDF)提出糖尿病的治疗目标不能仅局限于控制血糖水平,还要将提高病人的生活质量作为重要指标。1DF提出的五项基本措施,即饮食治疗、体育锻炼、药物治疗、糖尿病教育和自我血糖监测,在21世纪仍然需要对糖尿病进行综合治疗。此外,还应注意同时积极处理心脑血管疾病危险因子如高血压、脂代谢紊乱等。

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  • 简介:【摘要】目的:分析对 老年 2型糖尿病 (T2DM)患者采用利拉鲁肽治疗的 临床效果。方法:选取我院 2018年 2月 -2019年 4月收治的 110例 老年 2型糖尿病 (T2DM)患者为分析对象,给予所有患者利拉鲁肽进行治疗,经治疗 8 周后,对患者治疗前与治疗后的空腹血糖、餐后 2h 血糖、体重指数以及胰岛 β 细胞 功能指数进行对比。 结果:经所有患者 8 周治疗后所有相关临床指标均优于治疗前,治疗前后指标对比差异存在显著临床可比性 ( P<0.05)。 结论:临床上对 老年 2型糖尿病 (T2DM)患者采用利拉鲁肽治疗效果显著,在改善患者胰岛功能以及血糖时具有较高的安全性,值得在临床推广应用 。

  • 标签: 利拉鲁肽 老年 2 型糖尿病 (T2DM) 治疗效果
  • 简介:【摘要】康复新液是一种通利血脉,养阴生肌的纯中药制剂。临床可辅助胃黏膜修复,但其副作用尚不清楚。我们将康复新液用于2例由于痛风长期服用非甾体消炎药所致的胃溃疡患者,偶然间发现康复新液诱发了痛风性关节炎的急性发作。回顾病史及查阅文献,我们猜测可能是由于其昆虫成分增加血尿酸有关。本病案记载了康复新液的潜在不良反应,为临床其相关禁忌症提供参考。

  • 标签: 痛风 胃溃疡上消化道出血 康复新液 高尿酸血症 嘌呤
  • 简介:Analysisofthefrequencyofantigen-specificcytotoxicTlymphocytes(CTLs)exvivoislargelydependentontheuseofMHC/peptidetetramers.However,thelatterreagentshavenotbeenwidelyavailable,mostlikelybecauseoftheircostlyandtime-consumingproduction.InthisreportweutilizedaneconomicstrategytoconstructHLA/peptidetetramerswithrecombinantpeptide-linkedβ2microglobulin(β2m).TheHLA-A2-restricted,melanomaantigenMARTl-derivedpeptideMARTI27-35(AAGIGILTV)wasfusedtotheNterminusofhumanβ2mthrougha15-aminoacid(aa)-longlinkerbeforebeingrefoldedwiththerecombinantbiotinylatedHLA-A2heavychainectodomain.Theresulted2-component(2C)monomerwasthentetramerizedwithphycoerythin-labeledstreptavidin.Theexperimentalresultshowedthatthe2CHLA-A2/MARTI27-35monomerwasshowntobindtotheHLAclassⅠcomplex-specificmonoclonalantibodyW6/32andtheHLA-A2/MARTI27-35complex-specificsinglechainantibodyfragment(scFv)8.3,suggestingthecorrectnessofitsspecificity.Furthermore,the2CHLA-A2/MARTI27-35tetramerdetectedaspecificCD8^+TcellpopulationinHLA-A2-restrictedmelanomainfiltratinglymphocytesastheconventional3CHLA-A2/MARTI27-35tetramer.Theyieldof2CHLA-A2/MARTI27-35monomerwas2.5timesmorethanthatoftheconventional3Cmonomer.Takentogether,thesedataindicatethattheHLA-A2/MARTI27-35tetramercanbegeneratedconvenientlythroughtheuseofMARTI27-35peptide-β2mfusionproteins,whichcanfacilitatethemonitoringofHLA-A2-restricted,MARTl-specificCTLresponsesinpatientswithmelanoma.

  • 标签: 细胞毒素 T淋巴细胞 特异性抗原 缩氨酸四聚物 Β2微球蛋白
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  • 简介:【摘要】目的:分析综合护理干预在 2型糖尿病肾病护理中所取得的护理价值。方法:选择我院在 2017年 4月 ~2019年 5月诊治的 2型糖尿病肾病患者 84例作为护理对象,将患者均分为观察组 42例和对照组 42例,观察组应用综合护理干预,对照组应用常规护理,对比两组患者护理后血糖和肾功能指标、患者护理前后情绪状态。结果:观察组患者的血糖和肾功能指标均优于对照组, P<0.05;在护理前,两组患者负面情绪评分相似,经过护理后,观察组患者负面情绪评分降低,数据对比有意义。结论:在 2型糖尿病肾病患者的护理中应用综合护理干预可显著改善病情,缓解不良情绪,取得了积极的护理价值,值得推广。

  • 标签: 2型糖尿病肾病 护理干预 价值
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  • 简介:ToexploretheantiviraleffectandmechanismofpolysaccharidefromSpirulinaplatensis(PSP)onherpessimplexvimstype2(HSV-2),astandardstrainofHSV-2(333strain)wasusedtoinvestigatetheantiviraleffectofPSPinvitro.PSPinvariousconcentrationswasappliedtodifferentstagesofHSV-2replicationcycle.Finally,thevirusinfectivity(TCID50),cytopathiceffect(CPE),andMTTstainingmethodforviablecells(MTTassay)wereusedasmarkerstoevaluatetheeffectofPSPonHSV-2.ThequantityofHSV-DNAwasdetectedbyreal-timefluorescencequantitativePCR(FQ-PCR).TheHSV-2infectedVerocellultrastructureswereobservedbytransmissionelectronmicroscopy(TEM).TheresultsshowedthatPSPhadlittlecytotoxiceffectonVerocells,itcouldnotdirectlyinactivateHSV-2infectivity.PSPnotonlyinterferedinadsorptionofHSV-2toVerocellsbutalsoinllibitedHSV-2biosynthesisinthecells.FQ-PCRresultsshowedthattheinhibitoryrateonHSV-DNAalsoincreasedinadose-dependentandtime-dependentmanner.TEMalsoconfirmedthatPSPexhibitedpronouncedinhibitoryeffectonHSV-2.Inconclusion,theantiviraleffectofPSPonHSV-2maybeattributedtotheinhibitionofvimsadsorption,vimsreplicationandsynthesisincells.

  • 标签: 抗滤过性病原体 多醣 病毒 免疫机制
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  • 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