简介:ThisstudywasaimedtoobservetheexpressionofP70S6kinase(P70S6K)inoralaciniccellcarcinoma.PT0S6kinaseexpressionwasexaminedbymeansofWestern-blottestandActivityas-say.Specimenswerefrom30casesoforalaciniccellcarcinomaand15casesofnormaloraltissuewereusedascontrols.StatisticalanalysissoftwareSPSS10.0wasusedforttesttodeterminetherelationshipbetweengeneexpressionandclinicalfeatures.TheexpressionlevelofP70S6Kincreasedobviouslyinoralaciniccellcarcinomatissue(P<0.01).ActivityassaywasthesameastheWestemblottest(P<0.01).P70S6Kexpressionlevelandactivityplayedanimportantroleinthedevelopmentoforalaciniccellcarcinoma.Inconclusion,P70S6Kisamplifiedandoverexpressedinoralaciniccellcarci-nomatissue,whichsuggestsapotentialoncogenicfunction.P70S6KandotherpossibletargetsofmTORcontributesignificantlytotumordevelopmentandthatinhibitionoftheseproteinsmaybethera-peuticforcancerpatients.OverexpressionofP70S6Kmaybeinvolvedinthepathogenesisoforalacin-iccellcarcinoma.
简介:Mycoplasmagenitaliumisthemaincausativeagentfornon-gonococcalandnon-chlamydialurethritis.P32istheputativesurface-exposedmembraneproteinofM.genitaliumandithassubstaintialidentityinaminoacidsequencewithadhesinproteinP30fromM.pnewnoniae.SinceM.pneumoniaemutantslackingP30proteinisdefectiveincytadherence,P32proteinhasbeenproposedtobeanessentialadhesinimplicatedintheadherenceofM.genitaliumtohostcells.TheprokaryoticexpressionvectorpET-30(+)/p32wasconstructedinthepresentstudy,andtherecombinantproteinwasexpressedinE.coliandpurifiedunderdenaturingcondition.Asdemonstratedbytheimmunoblottinganalysis,therecombinantproteincouldreactwithrabbitantiseraagainstM.genitalium,andadherenceinhibitionassayswerepetformedwithantiseraagainstthisrecombinantprotein.ItwasdemonstratedthatP32proteinapperaredtobeanadhesionproteinofM.genitalium,thusprovidingtheexperimentalbasisforbetterunderstandingofthepathogenesisofM.genitaliuminfectionandforthedevelopmentoftherelatedvaccinesagainsttheinfection.
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:CytoehromeP450norgenewasclonedintotheexpressionvectorpET-28toyieldtherecombinantexpressionplasmidpET-P450nor,whichcoulddirectthesynthesisofaeukaryoticderivedproteininEscherichiacoliBL21.Thevectorallowsoverproductionandsingle-steppurificationof(His)6-taggedcytoehromeP450norbythefacifitationofmetal(Ni^2+)chelateafl]nitychromatography.TheexpressionlevelofcytoehromeP450norwashighat30℃afterIPTGinduction.SDSPAGEandWesternblotanalysisshowedaspecificband(about43kDa).TheoverproducedcytochromeP450norwaspurifiedtoeleetrophoretichomogeneitywithin2.5handabout20.8mgpurifiedproteinwasobtainedfrom2Lcellculture.TheproliferationofSSMC-7721celllinecouldbeinhibitedbycytoehromeP450nor.Rabbitpolyclonalantibodies(titerover64000)wereproducedagainstrecombinantcytoehromeP450norandprovedtobeveryusefulforimmunoblottingstudy.AvailabilityofthisexpressionsystemandspecificantibodiesshouldfacilitatecharacterizationoftheroleofcytochromeP450norinthemetabolismofNO.
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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:ThepurposeofthisstudywastoconstructaneukaryoticDNAvectorencodingamultipleepitopeantigen(MFC)ofhepatitisCvirus(HCV)andahepatitisBsurfaceantigen(HBsAg),andexploretheeffectofHBsAggeneontheimmunityofHCVmultiple-epitopeDNAconstructinvitroandinvivoinmice.AnHCVDNAvector(pVAX1-HBs-MFC)wasconstructedbyfusingHBsAggenetotheNterminalofanHCVmultiple-epitopeantigengene.ThepVAX1-HBs-MFCwastransfectedintoHEK293TcellsanditsexpressionwasmeasuredbyELISAandWesternblotting.BALB/cmicewereintramuscularlyimmunizedwiththepVAX1-HBs-MFC,andanELISAapproachwasappliedtodeterminethespecificantibodytitersandsubtypesinthemouseserum.Thecross-reactivityoftheantibodieswasalsocheckedwithtwosynthesizedHCVhypervariableregion1(HVR1)peptides.TheIFN-γproductionandcellproliferationofthemousespleencellswereevaluatedbyELISAandMTS(3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium,innersalt)assays,respectively.TheexpressionofpVAX1-HBs-MFCwasdetectableinthetransfectedHEK293Tcells.TheserumantibodyresponsewaseffectivelyelicitedinBALB/cmiceinjectedwithpVAX1-HBs-MFC.ThehighesttiterofantibodyagainstHCV(MFC)was1:1280,andtheratioofIgG2a/IgG1was1.50±0.12atthefifthweekafterfirstimmunization.Moreover,thecollectedmouseserumantibodyhadtheabilitytocross-reactwiththetwosynthesizedHCVHVR1peptides.ThestimulationindexofthemousesplenocytestoMFCwas1.79±0.07,andtheIFN-γlevelwas287±6pg/mlatweek21afterfirstimmunization.ThehighesttiteroftheantibodyincontrolBALB/cmiceimmunizedwithpVAX1-MFCwas1:320,andtheratioofIgG2a/IgG1was1.33±0.11atweek5post-immunization.Furthermore,thestimulationindexofthemousesplenocytescellstoMFCwas1.52+0.06,andtheIFN-γlevelwas225±9.3pg/mlatweek21post-immunization.TheHBsAggenecanenhancetheeffectsofanHCVmultiple-epitope
简介:【摘要】目的:探究B超检查联合p16/Ki-67检测对宫颈癌的应用优势及中医推拿与护理的联合治疗效果。方法:对比组采用HPV检测,并应用常规护理干预;实验组加用p16/Ki-67检测与B超检查,并应用中医推拿与护理干预。结果:实验组的诊断准确率(96.96%)比对比组的(81.81%)高,实验组中联合检测的灵敏度比对比组要高;实验组的护理满意度评分比对比组高,住院时长和住院时间均比对比组短,生活质量评分以及预后评分均比对比组高。(P<0.05)。结论:B超检查联合p16/Ki-67检测对宫颈癌在临床的应用能够大幅度提升宫颈癌的诊断准确率,而中医推拿与护理的联合治疗能够明显提升病人术后的生活质量,缩减住院时间,加快诊断效率,及早治疗。
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简介:【摘要】目的:探究p16/Ki-67检测与B超对宫颈癌的诊断价值与中医推拿结合护理的影响效果。方法:选取从2019年1月至2019年12月宫颈癌的100例患者作为本次调查对象,采用随机分配方式分为对照组与观察组,50例/组。对照患者采用HPV检测,观察组则在B超联合p16/Ki-67检测,比较两组患者诊断准确率。结果:两组患者分别采取不同的检测诊断方法后,对比和分析两组患者诊断结果准确率,对照组为76%,观察组为90,差异明显具有统计学意义(P<0.05)。结论:针对宫颈癌患者采用p16/Ki-67检测与B超共同诊断,提高了诊断准确率,为临床的进一步治疗提供有价值的临床依据。同时,为患者介入中医推拿结合护理方案,对患者生活质量也具有积极影响。
简介:【摘要】目的:分析化学发光法检测2型糖尿病患者胰岛素与血清C肽水平临床价值。方法:将2018年1月至2019年8月在本院实施诊治的59例2型糖尿病患者纳入患病组,将同期于本院健康体检的59例受检者划入体检组。为受检者均实施化学发光法检测,比较受检者的胰岛素与血清C肽水平。结果:患病组的空腹血糖值、餐后2h血糖值均高于体检组,差距比较有统计学意义(p<0.05)。患病组的胰岛素和C肽含量均低于体检组受检者,数据比较有统计学意义(p<0.05)。结论:化学发光法检测2型糖尿病患者胰岛素与血清C肽水平,临床诊断检出效果较好,操作简单,可明确患者血清与胰岛素水平,为患者对症治疗。
简介:【摘要】 目的 分析6s管理在物质管理中的应用价值。方法 抽选来自2017年4月至2018年4月期间我院未实施6s管理的物质管理作为对照组,将2018年5月至2019年5月期间我院采用实施6s管理的物质管理作为观察组 ,比较两组出现物品存放不合理例数、寻找物品消耗时间、清洁环境耗时、月盘点耗时。结果 观察组物品存放不合理例数显著低于对照组,差异具有统计学意义,P<0.05;观察组寻找物品消耗时间、清洁环境耗时、月盘点耗时显著短于对照组,差异具有统计学意义,P<0.05。结论 对物质管理中采用6s管理,能够避免不良事件,有助于物品核对,减少物品存放不合理例数,缩短寻找物品消耗时间、清洁环境耗时、月盘点耗时间,在临床上显示出卓越成效,值得进一步推广使用。
简介:Tosupportthescientificbasisforrapididentificationofpathogenicbacteriaandotherstud-ies,thesequencesofhsp60geneinmajor34speciesof16genusofpathogenicbacteriaweresearchoutinGenBankandaproperpairofuniversaldegenerateprimerwasdesignedbymeansofthemolecu-larbiologicalsoftwawePrimer5.0andOligo6.0.ThisprimerwasthenusedinthePCRamplification,andthehsp60genefragmentsoftheselectedpathogenicbacteriacouldbeamplifiedusingthisdegener-ateprimer.Bywayofbioinformationalanalysis,theconservation,variationandtheinterspeciesphylo-geneticrelationsofthehsp60genesequencewereanalysed.Fromtheresultsofthecomparativestudyonsequences,itwasdemonstratedthatthehsp60genewascharacterizedbyconservationandvaria-tion,inwhichtheconservedandmutantregionsco-existedandseparatelydistributedwithmanysmallmutantregionsdistributedamongtheconservedregions,justlikethemosaic.Thephylogenetictreeamongdifferentpathogenicbacteriadrawnfromthehsp60geneanalysiswasprovedtobeconsistentwiththosefrom16SrRNAand23SrRNA.Itisconcludedthatthesequencedistributionofhsp60genewouldprovideasolidbasisfortherapididentificationofpathogenicbacteriaandthedevelopmentofadiagnosticmicroarray.