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.
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.
简介:Wehaveconfirmedefficientanti-tumoractivitiesoftheperipherallymphocytestransducedwithap185HER2-specificchimericT-cellreceptorgenebothinmurineandinhumaninourpreviousstudies.TofurthertestthefeasibilityofchimericT-cellreceptorinabonemarrowtransplantationmodel,wefirst,madetwomurinetumorcelllines:MT901andMCA-205,toexpresshumanp185HER2byretroviralgenetransduction.MurinebonemarrowcellswereretrovirallytransducedtoexpressthechimericT-cellreceptorandgene-modifiedbonemarrowcellsweretransplantedintolethallyirradiatedmouse.Sixmonthsposttransplantation,p185HER2-positivetumorcells:MT-901/HER2orMCA-205/HER2wassubcutaneouslyorintravenouslyinjectedtomakemousemodelssimulatingprimarybreastcancerorpulmonarymetastasis.Theinvivoanti-tumoreffectsweremonitoredbythesizeofthesubcutaneoustumororcountingthetumornodulesinthelungsafterIndiainkstaining.ThesizeofthesubcutaneoustumorwassignificantlyinhibitedandthenumberofpulmonarynodulesweresignificantlydecreasedinmouserecipientstransplantedwithchimericT-cellreceptormodifiedbonemarrowcellscomparedwiththecontrolgroup.Ourresultssuggesttheefficientinvivoanti-tumoractivitiesofchimericT-cellreceptorgenemodifiedbonemarrowcells.
简介:WehavedevelopedandtestedchimericT-cellreceptors(TCR)specificforp185HER2.Intheseexperiments,retroviralvectorsexpressingtheN297orN29ξreceptorswereconstructedinpRET6.AmphotropicviralproducercellswereestablishedintheGALV-basedPG13packagingcellline.Ficollpurifiedhumanperipheralbloodlymphocytes(PBL)werevitallytransducedusinganoptimizedprotocolincorporatingactivationwithimmobilizedanti-CD3/anti-CD28monoclonalantibodies,followedbyviralinfectioninthepresenceoffibronectinfragmentCH296.Transducedcellswereco-culturedwithhumantumorcelllinesthatoverexpress(SK-OV-3)orunderexpress(MCF7)p185HER2toassayforantigenspecificimmuneresponses.BothCD4^+andCD8^+T-cellstransducedwiththeN297orN29ξchTCRdemonstratedHER2-specificantigenresponses,asdeterminedbyreleaseofTh1likecytokines,andcellularcytotoxicityassays.OurresultssupportthefeasibilityofadoptiveimmunothempywithgeneticallymodifiedT-cellsexpressingachTCRspecificforp185HER2.
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。
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 。
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.
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。
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.