简介：OurpreliminarystudiesconfirmedthatanactiveprincipleregionofBuyangHuanwudecoction,comprisingalkaloid,polysaccharide,aglycon,glucosideandvolatileoil,caninducebonemarrowmesenchymalstemcelldifferentiationintoneurons.Mitogen-activatedproteinkinasesignalingwasidentifiedasoneofthekeypathwaysunderlyingthisdifferentiationprocess.Thepresentstudyshowsphosphorylatedextracellularsignal-regulatedproteinkinaseandphosphorylatedp38proteinexpressionwasincreasedafterdifferentiation.Cellularsignalingpathwayblockingagents,PD98059andSB203580,inhibitedextracellularsignal-regulatedproteinkinaseandp38inmitogen-activatedproteinkinasesignalingpathwaysrespectively.mRNAandproteinexpressionoftheneuronalmarker,neuronspecificenolase,andneuralstemcellmarker,nestin,weredecreasedinbonemarrowmesenchymalstemcellsaftertreatmentwiththeactiveprincipleregionofBuyangHuanwudecoction.Experimentalfindingsindicatethat,extracellularsignal-regulatedproteinkinaseandp38inmitogen-activatedproteinkinasesignalingpathwaysparticipateinbonemarrowmesenchymalstemcelldifferentiationintoneuron-likecells,inducedbytheactiveprincipleregionofBuyangHuanwudecoction.

简介：BACKGROUND:Stereotacticinjection(striatumorlateralventricle)andvascularinjection(tailveinorcarotidartery)arenowoftenusedincellulartherapyforcerebralinfarction.Stereotacticinjectioncanaccuratelydelivercellstotheinfarctarea,butrequiresastereotacticdeviceandcausessecondarytrauma;vascularinjectioniseasyandbetterforhostneurologicaldeficitrecovery,butcancausethrombosis.OBJECTIVE:Tocomparethetherapeuticpotentialofadultbonemarrow-derivedmesenchymalstemcells(BMSCs)transplantationbyintraperitonealversusintravenousadministrationtocerebralischemicrats.DESIGN,TIMEANDSETTING:ArandomizedcontrolledanimalexperimentwasperformedattheCellRoomandPathologyLaboratory,BrainHospitalAffiliatedtoNanjingMedicalUniversityfromNovember2007toSeptember2008.MATERIALS:BMSCswerederivedfrom20healthySprague-Dawleyratsaged4-6weeks.METHODS:Forty-fiveadultmiddlecerebralarteryocclusion(MCAO)ratswererandomlydividedintocontrol,intravenousandintraperitonealinjectiongroups,with15ratsineachgroup.At21daysaftermodeling,ratsinthecontrolgroupreceived1mLof0.01mol/Lphosphatebufferedsalineviatailveininjectionandeachexperimentalratreceived4×106BMSCslabeledbybromodeoxyuridine(BrdU)viaintravenousorintraperitonealinjection.MAINOUTCOMEMEASURES:Angiogeninexpressionandsurvivaloftransplantedcellsweremeasuredbyimmunohistochemicalstainingofbraintissueininfarctionhemisphereat7,14or21daysafterBMSCtransplantation.Co-expressionofBrdU/microtubule-associatedprotein2orBrdU/glialfibrillaryacidicproteinwasobservedbydouble-labeledimmunofluorescenceofcerebralcortex.Evaluationofnervefunctionusingtheneurologicalinjuryseverityscoreandtheadhesion-removaltestwasperformedonthe1stand21stdaybeforeandafterMCAO,andat3,7,14or21daysafterBMSCstreatment.RESULTS:Angiogenin-positivenewvesselsweredistributedinthebilateralstriatum,hippocampusandcerebral

简介：BACKGROUND:Alpha-actinin(α-actinin)playsakeyroleinneuronalgrowthconemigrationduringdirectionaldifferentiationfromneuralstemcells(NSCs)toneurons.OBJECTIVE:Todetectinsitumicrodistributionandquantitativeexpressionofα-actininduringdirectionaldifferentiationofNSCstoneuronsinthetemporallobecerebralcortexofneonatalrats.DESIGN,TIMEANDSETTING:BetweenJanuary2006andDecember2008,cultureanddirectionaldifferentiationofNSCswereperformedatDepartmentofHistologyandEmbryology,PreclinicalMedicalCollege,ChinaMedicalUniversity.ImmuneelectronmicroscopywasperformedatDepartmentofHistologyandEmbryologyandDepartmentofElectronMicrology,PreclinicalMedicalCollege,ChinaMedicalUniversity.SpectrumanalysiswasperformedatLaboratoryofElectronMicroscopy,MentalResearchInstitute,ChineseAcademyofSciences.MATERIALS:Basicfibroblastgrowthfactor,epidermalgrowthfactor,brain-derivednervegrowthfactor,type-1insulinlikegrowthfactor,andα-actininantibodywereprovidedbyGibcoBRL,USA;rabbit-anti-ratnestinmonoclonalantibody,rabbit-anti-ratneuronspecificenolasepolyclonalantibody,andEDAX-9100energydispersiveX-rayanalysiswereprovidedbyPHILIPSCompany,Netherlands.METHODS:NSCs,followingprimaryandpassageculture,weredifferentiatedwithserumculturemedium(DMEM/F_(12)+10%fetalbovineserum+2ng/mLbrain-derivednervegrowthfactor+2ng/mLtype-1insulinlikegrowthfactor).MAINOUTCOMEMEASURES:Expressionofα-actinininneuron-likecellswasquantitativelyandqualitativelydetectedwithimmunocytochemistryusingenergydispersiveX-rayanalysis.RESULTS:Immunocytochemistry,combinedwithelectronmicroscopy,indicatedthatpositiveα-actininexpressionwaslikeaspheroidparticlewithhighelectrondensity.Inaddition,theexpressionwasgraduallyconcentratedfromthenuclearedgetothecytoplasmandexpandedintodevelopingneurites,duringdifferentiationofneuralstemcellstoneurons.Conversely,energydispersive