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1、OnlineAppendixforthefollowingJACCarticleTITLE:AMicro-RibonucleicAcidSignatureAssociatedWithRecoveryFromAssistDeviceSupportinTwoGroupsofPatientsWithSevereHeartFailureAUTHORS:RaviRamani,MD, DeborahVela,MD, AnaSegura,MD,DennisMcNamara,MD,BonnieLemster,MPH, VishnupriyaS
2、amarendra,BS,RobertKormos,MD, YoshiyaToyoda,MD, ChristianBermudez,MD,O.H.Frazier,MD, ChristineS.Moravec,PhD,JohnGorcsanIII,MD, HeinrichTaegtmeyer,MD,DPhil,CharlesF.McTiernan,PhDAPPENDIXMethodsCardiacfunctionalcharacterization.ForthefunctionalcharacterizationoftheTes
3、tcohort,weusedechocardiographicautomatedborderdetectionfrommid-ventricularshort-axisimagesandnoninvasivearterialpressuretomeasurebeat-to-beatresponsesin2-to5-minutetrialsofdecreaseddeviceflow.Anincreaseinstrokearea,>40%increaseinfractionalareachange,orapreloadadjust
4、edmaximalpower>4.0mW/cm4withlowdeviceflowwereassociatedwithsuccessfuldeviceremoval.Additionalevaluationofrecoveryinvolvedexercisephysiologywithgasexchangeanalysisandinvasivehemodynamicsbyrightheartcatheterization,bothwiththeVADratelowered.Rightheartcatheterizationwa
5、sperformedusingthesameVADflowreductionprotocolasforechocardiography(1,2).Theclinicalassessmentofheartfailureetiologyforeachcohortwasequallydistributedineachgroup,aspresentedinSupplementaryTable1.Tissuecollectionandprocessing.LVfreewallsampleswerecollectedintheoperat
6、ingroom,immediatelyimmersedincoldcardioplegia,flashfrozenwithin15minutes,andstoredat-80oC.AsubsetofTestcohortsampleshadtissuefixed(2%paraformaldehyde)andfrozenforfuturemicroscopicanalyses(3).AllValidationcohortsampleshadtissuefixedinformalin(4).Screeningreal-timePCR
7、array.PCRarraywasperformedonVADcoresamplesobtainedfrom6patientsineachgroupoftheTestcohort.RNAwasisolated(QiagenmiREasy),enrichmentforsmallRNAs(SABiosciencesRT2qPCR-GrademiRNAIsolationKit),copiedtocDNA,andanalyzedwithamicroRNAmicroarrayplatform(SABiosciencesMAH-3100,
8、Genome-WideArray)onanAppliedBiosystems7900HTinstrumentonsequentialdays.Arraydataanalysis.FluorescencedataacquiredwithAppliedBiosystemsSequ