简介：Objective:Toexplorethecultureconditionsofhumanneuralstemcellsandtoinvestigatetheultrastructureofneurospheres.Methods:Thecellsfromtheembryonichumancorticesweremechanicallydissociated.N2mediumwasadaptedtocultureandexpandthecells.ThecellswereidentifiedbyimmunocytochemistryandEMwasappliedtoexaminetheultrastructureofneurospheres.Results:Theneuralstemcellsfromhumanembryonicbrainsweresuccessfullyculturedandformedtypicalneurospheresinsuspension,andmostofthecellsexpressedvimentin,whichwasamarkerforneuralprogenitorcells,andthecellscoulddifferentiateintoneurons,astrocytesandoligodendrocytes.Invitromyelinformationinneurosphereswereobservedatanearlystageofculture.Conclusions:Humanneuralstemcellscanbeculturedfromembryonicbrains,canformthetypicalneurospheresinsuspensioninvitroandhavetheabilityofmyelinating,andmaybepotentialsourcefortransplantationintreatingmyelindisorders.

简介：Objective:Toinvestigatetheproliferativeeffectofkeratinocytegrowthfactor(KGF-2)onhumanadultkeratinocytes.Methods:Thestandardmediumwaskeratinocytegrowthmediumwithoutbovinepituitaryextract(BPE),hydrocortisoneorepidermalgrowthfactor(EGF).Keratinocytesfroma48-year-oldsubjectwereculturedandseededondisheswithstandardmediumofEGFincelldensityof2×104/32mm2.After24hours,themediumwasreplacedbythestandardmediumwith0,4,16,125and500ng/mlKGF-2,respectively.ThestandardmediumwithEGFwasusedasthepositivecontrolandthestandardmediumwithoutEGForKGF-2wasusedasthenegativecontrols.Thegrowthofkeratinocyteswasmonitoredby3-(4,5-dimethythiazol-2-yl)-2,5dipheyltetrazoliumbromide(MTT)assayandbyphotographsondays3,5and7,respectively.Results:KGF-2inconcentrationsof4-500ng/mlshowedasignificantproliferativeeffectondays5and7ascomparedwiththatofthenegativecontrols(P<0.01).Onday3thecellswereproliferatedto1.5-2.5-fold,onday5to3-5-foldandonday7to3-12-foldinKGF-2mediumasthatofthenegativecontrols.TheoptimalresponseoccurredwhentheconcentrationofKGF-2was125ng/mlonday7.CellproliferationwasalsoconsistentlyhigherinallKGF-2concentrationsascomparedwiththatofthepositivecontrols.Conclusions:KGF-2hassignificanteffectsontheproliferationofadultkeratinocytes,whicharemoreeffectivethanthatofEGF.ThisstudysupportsKGF-2canimprovethehealingofchronicwoundsinadultsinclinic.

简介：客观：为了探索可行性构造遗传工程人，神经干细胞(hNSCs)由lentivirus调停了多表示基因以便为针的绳索损害(SCI)的进一步的研究提供接枝来源。方法：从人的流产胎的大脑外皮的人的神经干细胞被孤立并且有教养，然后，基因被lentivirus修改两个都表示绿荧光蛋白质(GFP)和老鼠neurotrophin-3(NT-3)；转基因的表示被荧光显微镜，胎儿的老鼠的背面的根中心和槽污点的方法检测。结果：遗传工程hNSCs成功地被构造。所有在荧光显微镜下面表示了明亮绿的荧光的遗传工程hNSCs被观察。转基因的hNSCs的调节媒介能导致从背面的根中心(DRG)挥舞长出的神经突。遗传工程hNSCs表示了高级NT-3which能被使用槽污点检测。结论：遗传工程hNSCs调停了bylentivirus能被构造多成功地表示基因。

简介：Objective:Toobservehumanneuronalapoptosissecondarytotraumaticbraininjury,andtoelucidateitsregulativemechanismandthechangeofexpressionofapoptosis-relatedgenes.Methods:Specimensofbrainwerecollectedfromcasesoftraumaticbraininjuryinhumans.Thehistologicalandcellularmorphologywasexaminedbylightandelectronmicroscopy.TheextentofDNAinjurytocorticalneuronswasdetectedbyusingTUNEL.ByinsituhybridisationandimmunohistochemistrythemRNAchangesandproteinexpressionofBcl-2,Bax,p53,andcaspase3p20subunitwereobserved.Results:Apoptoticneuronsappearedfollowingtraumaticbraininjury,peakedat24hoursandlastedfor7days.Innormalbraintissueactivatedcaspase3wasrare,butashorttimeaftertraumaitbecameactivated.Theactivitypeakedat20-28hoursandremainedhigherthannormalfor5-7days.TherewasnoexpressionofBcl-2mRNAandBcl-2proteininnormalbraintissuebut8hoursafterinjurytheirexpressionbecameevidentandthenincreased,peakedat2-3daysandremainedhigherthannormalfor5-7days.TheprimaryexpressionofBax-mRNAandBaxproteinwashighinnormalbraintissue.At20-28hourstheyincreasedandremainedhighfor2-3days;onthe7thdaystheyreturnedtoanormallevel.Innormalbraintissue,p53mRNAandP53wereminimallyexpressed.Increasedexpressionwasdetectedatthe8thhour,anddecreasedat20-28hoursbutstillremainedhigherthannormalonthe5thday.Conclusions:Followingtraumaticinjurytothehumanbrain,apoptoticneuronsappeararoundthefocusoftrauma.ThemRNAandproteinexpressionofBcl-2,Baxandp53andtheactivityofcaspase3enzymeareincreased.

简介：探索骨头的效果的目的有带hepatocyte生长因素的adenoviral向量的导出髓的间充质的干细胞(BMSC)transfected(HGF，Ad-HGF)在灼伤创伤愈合上。从男Wistar老鼠的方法BMSC用由密度坡度centrifugation中等、与包含20%胎儿的牛的浆液(FBS)的DMEM有教养的Percoll分开被分开并且净化。当时，BMSC是有在感染(MOI)的100复合的最佳的基因transduction效率的Ad-HGF的transfected。transfection和在暂停的HGF的表示的效率被流动cytometry检测，酶分别地连接了immunosorbent试金(ELISA)。32只雌老鼠受到90慥?灳瑩吗？

简介：Objective:Toinvestigatethedifferentiativecapabilityofadulthumanbonemarrowmesenchymalstemcells(BMSCs)intoSchwann-likecells.Methods:BonemarrowswereaspiratedfromhealthydonorsandmononuclearcellswereseparatedbyPercolllymphocytesseparationliquid(1.073g/ml)withcentrifugation,cellswereculturedinDMEM/F12(1:1)mediumcontaining10%fetalbovineserum(FBS),20ng/mlepidermalgrowthfactor(EGF)and20ng/mlbasicfibroblastgrowthfactor(bFGF).Cellsofpassage1wereidentifiedwithimmunocytochemistry.Conclusions:BonemarrowcontainsthestemcellswiththeabilityofdifferentiatingintoSchwann-likecells,whichmayrepresentanalternativestemcellsourcesforneuraltransplantation.

简介：ObjectToexploretileeffectoflipofectin-c-erbB2antisenseoligodeoxynucleotidesonradiosensitivityofhumanovariancancercellllne.MethodsTheexpressionofc-erbB2wasdetectedbymeansofRT-PCR,cellularresponsetoirradiationwasevaluatedbytilecolonyformingassay.ResultsLipofectin-c-erbB2antisenseoligodeoxynucleotides（AS-ODN)couldsuppresstheexpressionofc-erbB2,andsignificantlydecreasedthecolonyformingrateofhumanovariancancercellsafterionizingirradiation（P<0.01),whilenon-trean~entandthesenseoligodeoxynucleotides{S-ODN)groupsdidnotdecreaseontheradio-resistancelevelofSKOV3cellline{P>0.05).Condusionc-erbB2antisenseoligodeoxynueleotidessensitizedtheSKOV3toionizingirradiationthroughdecreasingtheexpressionofe-erbB2,whichmightbetheresultofthefactthatc-erbB2antisenseoligodeoxynueleotidesinhibittheeelluarsignaltransductionpathwayrelatingtotheradiation-resistantphenotype.

简介：Directgenetransferintosomatictissueinvivoisadevelopingtechnologywithpotentialapplicationforcancergenetherapy.Retrovirusvector,whichwasaneffectivevehicle,stillhassomedisadvantagesingeneratinghightiterrecombinantvectorsandmanipulatingtomediateinvirogenetransfer.Inthispaper,recombinantvacciniavirusvectorencodinghuman

简介：Oncolysate,adebrisoftumorcells,hasbeenproventobeeffectiveintumoractiveimmunotherapy,itwasreportedthatthevacciniavirus,especiallyrecombinantvacciniavirusesencodinghumanIL-2(rVV-IL-2),enhancedtheimmunogenicityoftransfectedtumorcells.Inthisexperiment,themurinemelanomacellB16-F10oncolysatestransfectedbyrVV-IL-2(IL-2VBO)wereusedasvaccine.TheIL-2VBOorTK-VBOwaspreparedbyincubatingB16-F10cellswithrVV-IL-2orrVV-TKat

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简介：Objective:Toinvestigatethebiologicalfunctionofplatelet-derivedgrowthfactorB(PDGF-B)onthesurvivalandproliferationofcatcornealendothelialcellssoastoprovidebasesforfurtherstudiesofitsroleinwoundrepairanditsclinicalapplication.Methods:TotalRNAwasextractedfromtheplacentatissuesofhealthypregnantwomenundergoinghysterotokotomyandPDGFeDNAwasobtainedwithre-versetranscription-polymerasechainreaction(RT-PCR).TheprokaryoticexpressionvectorpET-PDGF-BwasconstructedandexpressedtherecombinantPDGF-BinEscherichiacoli(E.coli)BL21(DE3).AfterpurificationandrefoldingonNi2+-chelationaffinitychromatography(NTA)column,itwasusedtoculturecatcornealendothelialcells.Cellproliferationwastestedbymodifiedtertrazoliumsalt(MTT)andflowcytometer.Andthemorphologicchangeandtheultrastructurewereob-servedunderaninvertedphasecontrastmicroscope,ascan-ningelectronmicroscopeandatransmissionelectonmicroscope,respectively.Results:PDGF-Bchainpeptide(PDGF-BB)genewassuccessfullyinsertedintotheprokaryoticexpressionvector,pET-28a(+).ThepurifiedrecombinedproteinpET-PDGF-Bshowedasinglebandonsodiumdodecylsulfatepolyacry-lamidegelelectropheresis(SDS-PAGE)withthemolecularweightofabout27u,whichwasinagreementwiththede-ducedvalue.MTTandflowcytometryshowedthatPDGF-BBpromotedthesurvivalandproliferationofcatcornealen-dothelialcells.Conclusions:Theconstructionofrecombinantprokary-oticexpressionvectorpET-PDGF-BandthepreparationofPDGF-BBproteinprovideafoundationforfurtherstudyofthefunctionofPDGF-BBandproducingbiologicalPDGF-BBprotein.TheexpressedPDGF-BBpromotestheprolif-erationofculturedcatcornealendothelialcells.

简介：Objective:Toevaluatetheosteogenicpotentialofbonemorphogeneticprotein(BMP)-2genetransfectedgoatbonemarrow-derivedmesenchymalstemcells(MSCs).Methods:Goatbonemarrow-derivedMSCsweretransfectedbyAdv-humanbonemorphogeneticprotein(hBMP)-2gene(Group1),Adv-betagaltransfectedMSCs(Group2)anduninfectedMSCs(Group3).Westernblotanalysis,alkalinephosphatasestaining,VonKossastainingandtransmissionelectronmicroscopywereadoptedtodeterminethephenotypeofMSCs.Thenthecellswereinjectedintothighmusclesofthenudemice.Radiographicalandhistologicalevaluationswereperformedatdifferentintervals.Results:OnlyAdv-hBMP-2transfectedMSCsproducedhBMP-2.Thesecellswerepositiveforalkalinephosphatasestainingatthe12thdayandwerepositiveforVonKossastainingatthe16thdayaftergenetransfer.Electronmicroscopicobservationshowedthatthereweremoreroughendoplasmicreticulum,mitochondriaandlysosomesinAdv-hBMP-2transfectedMSCscomparedtoMSCsofothertwogroups.Atthe3rdand6thweeksaftercellinjection,ectopicboneswereobservedinmusclesofnudemiceofGroup1.Onlyfibroustissueoralittlebonewasfoundinothertwogroups.Conclusions:BMP-2genetransfectedMSCscandifferentiateintoosteoblastsinvitroandinduceboneformationinvivo.