血小板微粒子的检测与其临床意义分析

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1、中国协和医科大学硕士研究生学位论文英文摘要De．eftonofplatelet-derivedmicroparticlesusingflowcytometryanditsclinicalsignificance．伪bstract)ObjectiveToestablishaflowcytomemc(FCM)methodforthequantitationofplatelet．derivedmicroparticles(PMP)bymeallSof3I_tmand0．8p．mlatexbeadsthatwereusedaSinternalstanda

2、rdsanddiscusstherelationshipbetweenthevariationofPⅣ口concentrationindifferentdiseasesanditsclinicalsignificance．MethodsPMPsuspensions(platelet-poorplaSma,PPP)wereextractedfromwholebloodbysequentialcentrifugation．FirstwholebloodspecimenWaScentrifugedat160910raintogetplatelet—ri

3、chplasma(PAP)．AfterfixationthesuspensionWaScentrifugedat1500915minplus3500915minagain．PPPWaSextractedfromsuspension．whichcontainednoplateletsbutPMPThenthePPPspecimenWaSlabeledbyapanelofMoAbs．DuringFCManalysis，39mandO．8uIlllatexbeadswereusedasinternalstandardsforP^，【Pquantitio

4、nbaSedontheirdifferentsizes．Inalogarithmicforwardscatter(LOGFSC)versuslogarithmicsidescatter(LOGssc)dotplot．P～口wasgatedbyO．89mbeadsandthetimeofanalysisWaSdefinedbycounting3／．tmbeads．UsmgaLOGfluorescencevs．LOGSSCdotplot．PK口Wasdiscriminatedfromthebackgroundnoiseaccordingtotheir

5、positivefluorescenceintensity．Whenthenumberof39mreachedtO2x104，thecountingcyclewasstoppedandthenumberofPMPscontainedinlmlofplasmacouldbecalculated．BythisnewmethodwemeasuredthePMPunderdifierentcondition，suchasvariedcemrifugationsettings．variedconcentrationofthrombinasagonistan

6、dthetimecoursetodeterminethebestsampleproceedingprotocolandevaluatethereliabilityofthisnewmethod．InthestudyofPⅣ口’Sclinicalsignificance．30specimensfromhealthydonorsweremeasuredinordertocalculatereferencerange．Thenwegott11ePMPresultsfrom97patientswhosuffereddifferentdiseasesand

7、comparedthemwiththosegotfromhealthydonorsEspecially,adynamicPⅣ伊concentrationof2casesofidiopathicthrombocytopenicpurpura(ITP)wereobservedduringtheirdiagnosisandpost·steroidrecoveryperiod．第4页、中国协和医科大学硕士研究生学位论文ResultsTheinternalstandardmethodwasprovedaneffectiveapproachatouantit

8、ationofPMP．Alltheresultswereconsistentwithpublications．Weconfirmedth