简介：Itiswellestablishedthatstemcellscandifferentiateintocelltypesoftheorganinwhichthesearetransplanted.However,theprocessisveryslowduetolackofunderstandingofsignalsimportantfortheirsurvivalanddifferentiation,mostoptimalstemcellsandtheirplasticity.Limitationsandadvantagesofvariouscellsubtypeswillbedescribed.Therateofstemcellsmobilizationandtheirsurvivalintheischemicenvironmentaremajorobstaclesinengraftmentanddifferentiationofstemcellsformeaningfulrepairoftheinfarctedmyocardium.Manipulationofstemcellswithischemicpreconditioning,combinedgeneandcelltherapytogetherwithsimultaneousactivationofdiversesignalingpathwaysformassivestemcellmobilization®enerationhassignificantimpactontherepairprocessbystemcells.Theseandotherdifficultiesencounteredinefficientuseofvariousstemcellshaveresultedininventionofinducedpluripotentstemcellswhichcouldrevolutionizethestemcellbasedtherapyandtheirapplicationsforunderstandingofhumandiseaseanddrugscreeninginthenearfuture.ReprogrammingofadultcellsintoiPScellswithouttheuseofviralvectorsisamajorchallengetowardsgettingiPScellswithoutviralintegrationintocells.Tomeetthischallengewehaverepro-grammedskeletalmyoblastsintoiPScellswithhighefficiencyusingepigeneticmodifiers.TransplantationofiPScellsderivedpurecardiacprogenitorsintoinfarctedmyocardiumledtoextensiverepopulationofscarareawithfullydevelopedmyocyteswithouttumorformationandresultinginmarkedimprovementincardiacfunction.Reprogrammingwithpurechemicalmeanswillmaketherapeuticuseofthesecellsmoresafer.Targetingtheinducedpluripotentstemcellstowardscardiacprogenitorsandtheirapplicationtowardstransplantationisamajorstepforwardinenhancingthemyocardialrepaircapacitybythesecells.

简介：backgroundBonemarrowmesenchymalstemcells(BMSCs)canbeisolatedandculturedtomanypassages.However,StemcellsincludingBMSCsquicklyundergosenescenceinculture.Thecellsenescenceandmulti-directionaldifferentiationhavehamperedproducingBMSCsinquantitywiththeirundifferentiatedstate.Inthisstudywereportanaturalcompound,vitaminC(Vc),maintainsBMSCsstemproperty.MethodsHumanBMSCswereisolatedfrombonemarrowandpurifiedby1.073g/mLdensitygradientcentrifugation.50ng/mLVcwereaddedtoBMSCsfordifferenttimepoint.FlowcytometrywasusedtodetectcellsurfacemarkersofBMSCswithorwithoutVctreatment.BMSCsproliferationwasanalyzedbyMTTassay.PCR(polymerasechainreaction)andreal-timePCRwereusedfordetectingc-kit,nanog,andOct-4genesexpressionlevels.DNAmethyltransferase(Dnmt)1andDnmt3blevelswerealsodetectedbyreal-timePCR.ResultsFlowcytometryshowedthatafterVctreatmentfor6h,thesurfacemarkersofBMSCswerealmostunchanged.VcincreasedtheproliferationactivityofBMSCsfrom6hto24h.PCRshowedtheexpressionofc-kit,nanog,andoct-4geneswereobviouslyincreasedinVctreatedgroupthancontrolgroupat12h.Real-timePCRshowedthatthelevelofc-kit,nanog,andoct-4geneswereunregulatedfrom6hto12hcomparedwithcontrolgroup.VcalsoincreasedDnmt3bbutnotDnmt1geneexpression.ConclusionsOurresultsshowedVcactsatleastacceleratesBMSCsproliferationandmaintainsstemcellproperty.Inourstudy,wehighlightedamethodofimprovingthespeedofBMSCsgenerationandprovidedadditionalinsightsintothemechanisticbasisofpreventingBMSCssenescence.

简介：ObjectivesToanalyzethesix-minutewalktest(6MWT)andgasexchangeof5hearttransplantationpatientsandtoapproachthevariationtendencyofexercisetolerance,oxygenuptake(VO2)andheartratechronotropicresponse.Methods5casesofhearttransplantationpatients(age25～52years)wereundertaken6MWT6～30monthsafteroperation,synchronizinggasexchangingparametersweremeasuredbywirelessportableremotesensingK4B2gasanalyzer,51normalcontrolswerecompared.ResultsThesix-minutewalkdistance(6MWD)of5patientswere(592.6±26.7)m(558～625)m,theascendingtendencyduringexercisewasslower,themaximumheartrateswere80%±6%ofage-predictingmaximalheartrate,lowerthannormalcontrol(86%);theendpointVO2/kgwere(21.8±1.4)mL/min·kg(19.94～23.60)mL/min·kg.ConclusionsThe6WMDandVO2of5patientsreachednormalrange,buttheheartratechronotropicresponseandVO2ascendingtendencywereslowerthanthoseofnormalcontrols.

简介：Withimprovementsintheirsurgicalandmedicalmanagement,thenumberofpatientswithcongenitalheartdisease(CHD)reachingadulthoodhasincreasedoverthelastdecade.AsthepopulationofadultCHDpatientscontinuestorise,anincreasingnumberofthesepatientswillrequireevaluationforhearttransplantation.ItisimportanttorecognizeadvancedheartfailureandotherassociatedcomplicationsearlyinthiscohortofcomplexpatientsforearlyreferraltoanadultCHDspecialist.Asthesepatientspresentwithuniquechallengesbecauseoftheirmultiplecomorbiditiesandcomplexanatomy,thereneedstobeacarefulselectionprocessfortransplantationtooptimizetheutilizationofdonororgans.

简介：IntroductionRecently,bonemarrowmesenchymalstemcells(MSCs)havebeenreportedtorepairchronicallyinfractedmyocardiumwithdirectinjection.However,itisverydifficulttolocalizetheinjectedcellsontotheischemicareatoregeneratesufficientcardiacmassinthethinnedscararea.Toovercometheproblem,wehaveutilizedourcellsheettechnologybasedontemperature-responsiveculturedishes.Whentheculturetemperatureisreducedfrom37℃to20℃,allcellsconnectedviacell-celljunctionproteinsareharvestedasasinglesheetwithoutusingproteolyticenzymes.Thistechnologyallowsustotransplantstemcellsinvivofortreatmentheartdiseasewithouttheproblemsmentionintheprevious.MethodsMaleClawnminipigswereusedinthisstudy.Bonemarrow(5-7mL)wascollectedundergeneralanesthesia.Histopaqe-1077(15mL),wereaddedtobonemarrowandcentrifuged.Thecellswerecollectedandculturedfor7days.Weseededthebonemarrow-derivedMSCsattheconcentrationof(6×10~5/ml)on60mmdiametertemperature-responsivedishesfor7days.Astheculturetemperaturedecreasedfrom37℃to20℃,MSCsheetdetacheditselfspon-taneouslyandfloatedupintotheculturemedium.Triplelayerswerestackedtogetherrepeatedlyformingspecialmultiplayer.Myocardialinfarctionwascreatedbytheligationoftheleftanteriordescendingbranchoftheleftcoronaryartery.Acellsheetswastransplantedontotheischemiaarea.Theechocardiographywasperformedtwoandfourweeksaftertransplantation.Thehearttissuewithcellsheetswereremovedandfixedwith10%formalinforhistologicalanalysisonemonthafterthetransplantationofcellsheets.ResultsMostMSCsarepositiveforCD29,CD90,CD146andCD73.ThesemeantheculturecellsheetswerecomposedofundifferentiatedMSCsandremainedmultipotent.Monolayers(20-30μm)andmultilayer(120μm)cellsheetswereproduced,whichretainedallcell-to-cellcontaction.Histologicalanalysesshowthecellsheetsbecomecloselycontactedwiththehearttiss

简介：ObjectivesToinvestigatetheprotectiveeffectofthrombopoietin(TPO)onmyocardialcellsinvitro.MethodsH9C2celllinewasmaintainedinIscove’smodifiedDulbecco’smedium(IMDM)supplementedwith10%calfserum.Beatingcellsfromheartventriclesofneonatalheartwereculturedataninvitrosystem.Apoptosisofthecelllineabovewasinducedbytreatmentofdoxorubicin(DOX)andwasblockedbyTPO.CellsurvivalrateofH9C2cellwasmeasuredbytheMTTassay.Changesofbeatingrateofneonatalmyocardialcellswerecapturedbydigitalcameraandbeatingratewascalculated.Flowcytometrywasemployedtostudyanti-apoptoticeffectofTPObystainingJC-1proteintoH9C2cell.ResultsMTTassaydemonstratedthatdoxorubicinreducedcellsurvivalrateby73.8%±1.1%,50ng·mL-1and100ng·mL-1TPOincreasedcellsurvivalrateby84.6%±3.6%(P<0.05),86%±4%(P<0.01)atadose-dependentmanner.Beatingrateofprimaryneonatalmyocardialcellsalsodecreasedto15%±8%at48h,100ng·mL-1TPOimprovedbeatingrateto48%±11%(P<0.01).TPOdecreasedapoptoticratefrom19%±9%to11%±6%(P<0.05).ConclusionsTPOhasprotectiveeffectonmyocardialcellsinvitro.Anti-apoptosisisoneofthemechanismsbywhichTPOprotectsinjuredheart.

简介：Intherecentpast,bonemarrow(BM)-derivedcellshavebeenusedtoregeneratedamagedcardiovasculartissuespostmyocardialinfarction(MI).Recentclinicaltrialshaveshowncontroversialresultsinrecoveringdamagedcardiactissue.Newprogresshasshownthattheunderlyingmechanismsofcell-basedtherapyreliesmoreheavilyonhumoralandparacrineeffectsratherthanonnewtissuegeneration.However,studieshavealsoreportedthepotentialofnewendothelialcellgenerationfromBMcells.Thus,effortshavebeenmadetoidentifycellshavinghigherhumoralortherapeuticeffectsaswellastheirsurfacemarkers.Specifically,BM-derivedCD31~+cellswereisolatedbyasurfacemarkeranddemonstratedhighangio-vasculogeniceffects.IwillpresentrecentadvancesinthetherapeuticuseofBM-derivedcellsandtheusefulnessofCD31~+cellsasanextgenerationcelltherapy.

简介：Rats(Rattusnorvegicus)havemanyadvantagesovermiceinscientificstudies,forexample,theyaremorerelevanttohumaninphysiologicalandpharmacologicalresponses.Therefore,ratsarebroadlyusedinexperimentalstudies.Therecentbreakthroughinthegenerationofratembryonicstemcells(rESCs)opensthedoortoapplicationofgenetargetingtocreatemodelsforthestudyofhumandiseases.Inaddition,theinvitrodifferentiationofrESCsintoderivativesofthreegermlineswillserveasapowerfultoolandresourcefortheinvestigationofmammaliandevelopment,cellfunction,tissuerepair,anddrugdiscovery.However,thedistinctcultureconditionandsignalinhibitor-dependedmaintenanceofrESCsstandasaconsiderablechallengeforitsinvitrodifferentiation.Toaddressit,weinvestigatedwhetherrESCsarecapableofformingterminaldifferentiatedcardiomyocytes.Wefoundthattheembryoidbodies(EBs)-basedmethodusedinmouseESC(mESC)differentiationfailedtoworkinthecultivationofrESCs.WethenmodifiedthedifferentiationprotocolandsuccessfullydevelopedaninvitrodifferentiationsystemtodifferentiaterESCsintothreeembryonicgermlayers.Byusingthismethod,therESCsformspontaneousbeatingcardiomyocyteswiththepropertiessimilartothosederivedfromfetalratheartsandmESCs.Thisuniquecellularsystemwillprovideanewapproachtostudytheearlydevelopmentandcardiacfunctionaswellastoperformpharmacologicaltestandcelltherapystudy(Grants:theStateMajorResearchProgramofChina(2009ZX09503-024,2010CB945603)andCAS(XDA01030000).

简介：BackgroundPreviousstudieshavesuggestedthatpatientswithlowendothelialprogenitorcell(EPC)countsandimpairedendothelialcolonyformingactivityhaveahigherincidenceforcardiovasculareventscomparedtopatientswithhighEPCcountsandfavorablecolonyformingactivity.ThepathophysiologicalbasisforthisfindingmaybeaninsufficientendothelialcellrepairbyEPC.TheobjectiveofthisstudywastodeterminewhetherthenumberofEPCsinperipheralbloodwasassociatedwiththepresenceandseverityofangiographicstenosisinpatientsofthelatephaseafteracutemyocardialinfarction(AMI).MethodsOnehundredandonepatientsundergoingcardiaccatheterizationinourhospitalwereenrolledinthestudy.ThenumberofcirculatingEPCswasmeasuredbyafluorescent-activatedcellsorter(FACS).Patientswithacutecoronarysyndromeswereexcluded.ResultsComparedwithpatientswithnormalcoronaryartery,thenumberofcirculatingEPCswassignificantlyreducedamongpatientsinthelatephaseafterAMI(P<0.01).Wealsofoundthatcomparedwiththecontrolgroup,thenumberofEPCsofsingle-vesselstenosisgroupandmulti-vesselstenosisgroupweresignificantlyreduced(P=0.005;P=0.001).ConclusionsThenumberofEPCsintheperipheralbloodisdecreasedinpatientsofthelatephaseafterAMI.TheEPCsnumbercorrelatedwithangiographicstenosisseverity,whichsuggeststhatendothelialinjuryinthedeficientcirculatingEPCsmayaffecttheseverityoftheheartdisorderandtheclinicalpresentations.

简介：ObjectivesTodetectionofchlamydiapneumoniae(Cpn)DNAinthecirculatingmononuclearcellfractionsofcoronaryheartdiseaseandtoinvestigatetheassociationbetweeninfectionwithchlamydiapneumoniaeandcoronaryheartdisease(CHD)andprospectivelywhetherblood-basednestedpolymerasechainreaction(nPCR)isusefulinidentifyingCpninfection.MethodsTheperipheralbloodmononuclearcell(PBMC)CpnDNAwasexaminedusingnPCRtechniqueandconfirmedbyelectrophoresisin150patientswithCHD.Select55patientswithclinicalsuspectedCHDbutangiographyresultarenormalascontrolgroup(CG).Thenweconductedaprospective,randomized,double-blind,placebo-controlledstudyof6monthsofazithromycinandplacebotreatmentinCHDgroup.PatientswithCpnDNApositivewerethenrandomizedtoreceiveazithromycinorplacebo.Aftertreatmentbloodsamplewerecollectedforrepeatedmeasurement.ResultsChlamydiapneumoniaeDNAwasdetectedin49(32.7%)of150personswithCHDandin1(1.8%)of55personswithcontrolgroup,oddsratio26.2,95%confidenceinterva13.52-194.98.ThepositivityratesofnPCRinCHDgroupswerehigherthanthoseincontrolgroup.16cases(29.1%)inlatentcoronaryheartdiseases(LCHD)group,19cases(39.6%)inunstableangina(UAP)group,and14cases(29.9%)inacutemyocardialinfarction(AMI)groupwereCpnpositivebynPCR.TherewerenosignificantdifferenceamonginAMIUAPandLCHDgroup.ThereweresignificiantdifferenceinCpnDNAnegativeratesaftertheazithromycinandtheplacebotreatment.ConclusionsChlamydiapneumoniaeispresentinPBMCofasignificantproportionofpersonswithCHD.Thepotentialroleofchlamydiapneumoniaeincoronaryatherosclerosismaythereforebemorerelatedtoaccelerationofdiseaseorsystemiceffectsbypersistentinfectionthantosuddeninitiationofprogressivecoronaryarterydiseasebyacuteinfection.ThedetectionofCpnDNAinPBMCwithnPCRmaybeofgreatvalueforidentifyingCpncarriersandfo

简介：ObjectivesToinvestigatetheanti-apoptoticeffectsofmesenchymalstemcells(MSCs)onhypoxicinjuredcardiacmyocytesinvitro.MethodsMSCswereisolatedfrombonemarrowofSprague-Dawley(SD)rats,andcardiacmyocytesfromneonatalrats.Theratcardiacmyocyteswereco-culturedwithMSCsorMSC-conditionedmediainanoxia(95%N2+5%CO2)for72hours.CellapoptosiswasmeasuredbyHoechst33258staining.TheexpressionofBcl-2andBaxincardiacmyocyteswastestedbyWesternBlot.ResultsTheapoptoticratewas51.6%±2.4%whencardiacmyocyteswereculturedincontinuoushypoxiaandwassignificantlydecreasedwhencardiacmyocyteswerecoculturedwithMSCsorMSC-conditionedmedia(15.1%±5.4%and24.0%±4.2%respectively,P<0.001).ThedecreasedexpressionofBaxinthecardiacmyocyteswasgreatlyrelatedtothedecreasingofapoptosis,buttherewasnodifferenceinBcl-2expressionamongthesegroups.ConclusionsCo-culturedMSCsshowedsignificantanti-apoptoticeffectsoncardiacmyocytesincontinuoushypoxia.ThemechanismmaybetheinteractofcelltocellandparacrineofcytokineswhicheffectedtheexpressionofBaxinthecardiacmyocytes.

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简介：ObjectivesToob-servetheeffectofdifferentestrogenlevelsonthesecretoryfunctionofvascularendothelialcellsoffemalerats,andstudytheeffectofmodulationofestrogenlevelontheexpressionofvascularcelladhesionmolecule-1andtheconcentrationofestrogenreceptorinvascularendothelialcells.MethodsRadioim-munologywasusedtomeasuretheserumconcentrationofendothelinandPGI2,andcopper-cadmiumreductionwasemployedtomeasuretheserumcontentofnitrogenmonoxide.Radioligandbindingandflowcy-tometrywereusedtomeasuretheexpressionofestrogenreceptorandvascularcelladhesionmolecule(VCAM-1)ofvascularendothelialcellsrespectively.Results1.TheserumconcentrationofnitricoxideandPGI2decreasedwhentheovariesoffemaleratswereremoved.Inovariectomizedrats,givenestrogen,theconcentrationrose(P<0.05),buttheplasmaconcentrationofendothelinwasadversetoit.2.Theconcentrationofestrogenreceptorofvascularendothelialcel

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简介：ObjectivesToconstructarecombinantplasmidcarryingenhancedgreenfluorescentprotein(EGFP)andhumanvascularendothelialgrowthfactor(VEGF)121geneanddetectitsexpressioninratmesenchymalstemcells(MSCs).MethodsHumanVEGF121cDNAwasamplifiedwithpolymerasechainreaction(PCR)frompCD/hVEGF121andwasinsertedintotheeukaryoticexpressionvectorpEGFPC1.AfterbeingidentifiedwithPCR,doubleenzymedigestionandDNAsequencing.TherecombinantplasmidpEGFP/hVEGF121wastransferredintoratMSCswithlipofectamine.TheexpressionofEGFP/VEGF121fusionproteinweredetectedwithfluorescencemicroscopeandimmunocytochemicalstainingrespectively.ResultsTherecombinantplasmidwasconfirmedwithPCR,doubleenzymedigestionandDNAsequencing.ThefluorescencemicroscopeandimmunocytochemicalstainingresultsshowedthattheEGFPandVEGF121proteinwereexpressedinMSCs48haftertransfection.ConclusionsTherecombinantplasmidcarryingEGFPandhumanVEGFwassuccessfullyconstructedandexpressedpositivelyinratMSCs.ItoffersapromisetoolforfurtherresearchondifferentiationofMSCsandVEGFgenetherapyforischemialcardiovasculardisease.

简介：BackgroundOurpreviousstudyshowedthe150mg/mLfetalcardiacsupernatant(FCS)couldinducedifferentiationofBMSCsintocardiomyocye-likecellswithoutcardiomyocytetouch,butdifferentiationefficiencyisnothighenough.Inhibitionofglycogensynthasekinase-3enhancedtheproliferationandsurvivesofstemcells.Wetestedif6-bromoindirubin-3-oxime(BIO,glycogensynthasekinase-3inhibitor)enhancestheeffectsofFCSondifferentiationofBMSCsandexplorethegrowthfactorsinFCS.MethodsBMSCswereisolatedfromthefemurandtibiaoffour-week-oldmaleSprague-Dawleyratsandco-culturedwithFCS(150mg/mL)thatwasmadefromfetalheartsfromnineteen-daypregnantWistarrats.BIOwithdifferentconcentration(0,1,10,and100nM)wasintroducedinculturedishes.Transforminggrowthfactorbeta1(TGF-β1),bonemorphogeneticprotein2(BMP-2)andAktincardiacsupernatantandculturemediumwereassayedwithELISAmethods.ResultsAfterco-culturingwithFCS,beatingmyotubeswereobservedin25.9%BMSCsdishesafter1to2weeks’culture.ThelevelsofTGF-β1andBMP-2inFCSconcentrationswerenomorethanthatinyoungandadultcardiacsupernatant.AllBIOgroupssignificantlyenhancedtheeffectsofFCSondifferentiationofBMSCsintothecardiomyocyte-likecells(1nM,83%;10nM,73%;100nM,100%).AktlevelswerehigherinBMSCsculturalmediumwithFCS.ConclusionsFCScouldinducethedifferentiationofBMSCsintothecardiomyocyte-likecells.TGF-β1andBMP-2mightnotplayaroleinthedifferentiationofBMSCsinducedbyFCS.BIOenhancedtheeffectsofFCSonthedifferentiationofBMSCsintocardiomyocyte-likecells,whichmightinvolvetheAktpathway.