简介：MicroRNAs(miRNAs)aredynamicallyregulatedduringneurodevelopment,yetfewreportshaveexaminedtheirroleinspinabifida.Inthisstudy,weusedanestablishedfetalratmodelofspinabifidainducedbyintragastricallyadministeringoliveoil-containingall-transretinoicacidtodamsonday10ofpregnancy.Damsthatreceivedintragastricadministrationofall-transretinoicacid-freeoliveoilservedascontrols.ThemiRNAexpressionprofileintheamnioticfluidofratsat20daysofpregnancywasanalyzedusinganmiRNAmicroarrayassay.Comparedwiththatincontrolfetuses,theexpressionofmiRNA-9,miRNA-124a,andmiRNA-138wassignificantlydecreased(>2-fold),whereastheexpressionofmiRNA-134wassignificantlyincreased(>4-fold)intheamnioticfluidofratswithfetusesmodelingspinabifida.Theseresultswerevalidatedusingreal-timequantitativereverse-transcriptionpolymerasechainreaction.HierarchicalclusteringanalysisofthemicroarraydatashowedthatthesedifferentiallyexpressedmiRNAscoulddistinguishfetusesmodelingspinabifidafromcontrolfetuses.OurbioinformaticsanalysissuggestedthatthesedifferentiallyexpressedmiRNAswereassociatedwithmanycytologicalpathways,includinganervoussystemdevelopmentsignalingpathway.ThesefindingsindicatethatfurtherstudiesarewarrantedexaminingtheroleofmiRNAsthroughtheirregulationofavarietyofcellfunctionalpathwaysinthepathogenesisofspinabifida.Suchstudiesmayprovidenoveltargetsfortheearlydiagnosisandtreatmentofspinabifida.

简介：Wepreviouslyperformedtranscriptomesequencingandfoundthatgenesformatrixmetalloproteinases(MMPs),suchasMMP7and12,seemtobehighlyupregulatedfollowingperipheralnerveinjury,andmaybeinvolvedinnerverepair.Inthepresentstudy,wesystematicallydeterminedtheexpressionlevelsofMMPsandtheirregulatorsat1,4,7and14daysaftersciaticnervecrushinjury.Thenumberofdifferentiallyexpressedgeneswaselevatedat4and7daysafterinjury,butdecreasedat14daysafterinjury.Amongthedifferentiallyexpressedgenes,thosemostup-regulatedshowedfoldchangesofmorethan214,whilethosemostdown-regulatedexhibitedfoldchangesofmorethan2-10.Genesequencingshowedthat,atalltimepointsafterinjury,avarietyofMMPgenesinthe'InhibitionofMMPs'pathwaywereup-regulated,andtheirinhibitorgenesweredown-regulated.Expressionofkeyup-anddown-regulatedgeneswasverifiedbyquantitativereal-timepolymerasechainreactionanalysisandfoundtobeconsistentwithtranscriptomesequencing.TheseresultssuggestthatMMP-relatedgenesarestronglyinvolvedintheprocessofperipheralnerveregeneration.

简介：Thisstudyaimedtoexploretheroleofmechanicaltensioninhypertrophicscarsandthechangeinnervedensityusinghematoxylin-eosinstainingandS100immunohistochemistry,andtoobservetheexpressionofnervegrowthfactorbywesternblotanalysis.Theresultsdemonstratedthatmechanicaltensioncontributedtotheformationofahyperplasticscarinthebackskinofrats,inconjunctionwithincreasesinbothnervedensityandnervegrowthfactorexpressioninthescartissue.Theseexperimentalfindingsindicatethatthecutaneousnervoussystemplaysaroleinhypertrophicscarformationcausedbymechanicaltension.

简介：BACKGROUND:Studieshavedemonstratedthatthemechanismsunderlyingcellularapoptosissignaltransductionfocusontwopathways:intracellularmitochondriaandextracellulardeathreceptor.Thecurrentevidencesupportsthatsignaltransductionofcellularapoptosisalsoincludesendoplasmicreticulumstresssignaltransduction.OBJECTIVE:ToobserveCaspase-12expressionandcellularapoptosisfollowingischemiainratswithprogressivespinalcordcompression,andtoverifytheinfluenceofendoplasmicreticulumstressontheapoptosisinducedbyspinalcordinjury.DESIGN,TIMEANDSETTING:Arandomized,controlled,animaltrialwasperformedattheInstituteofNeuroscienceinChongqingMedicalUniversitybetweenJanuaryandOctoberin2006.MATERIALS:Immunohistochemicalkit,diaminobenzidine,andTUNELkitwerepurchasedfromBeijingZhongshanBiotechnology,China;rabbitanti-ratCaspase-12monoclonalantibodywasprovidedbySantaCruz,USA.METHODS:SixtyWistarrats,aged3-4months,wererandomlyassignedtoamodelgroup(n=50),whichunderwentspinalcordcompressionintheL_1segmentfollowingL_1laminectomyandarticularprocessexcisiontoestablishamodelofprogressivespinalcordcompression,andasham-surgerygroup(n=10),whichunderwentonlylaminectomy.Startingwiththefirstdayaftersurgery,theratswerelocallyanesthetized,theskinwasopened,andthescrewwasrotatedby1/4ofacycle,twiceweekly.MAINOUTCOMEMEASURES:At3,7,14,21,and28daysaftersurgery,ratsfromeachgroupwereanesthetized,andthespinalcordswereresected.Pathologicalchangesfollowingspinalcordcompressionweredeterminedusinghematoxylin-eosinstaining,Nissldye,andtransmissionelectronmicroscopy.TheTUNELmethodwasusedtoobserveneuronalapoptosisinthecompressedspinalcordsegments.ImmunohistochemistryandWesternblotwereutilizedtodetectCaspase-12expressioninthecompressedsegments.RESULTS:Cellularswelling,neuraldegeneration,andalteredendoplasmicreticulumstructureswereobservedat3days

简介：ThisstudysoughttoidentifydifferentiallyexpressedproteinsinSH-SY5Ycellstreatedwithvalproicacid,usingtwo-dimensionaldifferencegelelectrophoresisanalysis.Threeproteinswereunambi-guouslyidentified:theeukaryotictranslationinitiationfactor4Aisoform1andATP6V1B2proteinweredownregulated,whiletheheterogeneousnuclearribonucleoproteinKwasupregulated.Moreover,allthreeproteinsareassociatedwithalteredexpressionduetooxidativestress.Ma-trix-assistedlaserdesorption/ionization-timeofflightmassspectrometryandproteinimmunoblottingassayconfirmedthedifferentialexpressionofeukaryotictranslationinitiationfactor4Aisoform1.Theresultsindicatethatvalproicacidexertsanantioxidationeffectbyregulatingtheexpressionofeukaryotictranslationinitiationfactor4Aisoform1.

简介：

简介：NewZealandrabbitswererandomlydividedintoanischemiagroup(occlusionoftheabdominalaortafor60minutes),anischemia-reperfusiongroup(occlusionoftheabdominalaortafor60minutesfollowedby48hoursofreperfusion)andasham-surgerygroup.Two-dimensionalgelelectrophoresisdetected49differentiallyexpressedproteinsinspinalcordtissuefromtheischemiaandischemia/reperfusiongroupsand23ofthemwereidentifiedbymassspectrometry.Intheischemiagroup,theexpressionofeightproteinswasupregulated,andthatoftheremainingfourproteinswasdownregulated.Intheischemia/reperfusiongroup,theexpressionoffourproteinswasupregulated,andthatoftwoproteinswasdownregulated.Inthesham-surgerygroup,onlyoneproteinwasdetected.Intheischemiaandischemia/reperfusiongroups,fourproteinsoverlappedbetweengroupswiththesamedifferentialexpression,includingthreethatwereupregulatedandonedownregulated.Theseproteinswererelatedtoenergymetabolism,celldefense,inflammatorymechanismandcellsignaling.

简介：BACKGROUND:Anaminoacidimbalancehasbeenconsideredtoberesponsibleforepilepsypathogenesis.Gamma-aminobutyricacid-Breceptor(GABA_BR)inhibitsvoltage-sensitivecalciumionchannelsandGABAorglutamicacid(Glu)neurotransmitterrelease,whichpromotesorinhibitsonsetanddevelopmentofepilepsy.OBJECTIVE:ToexploretheeffectofbaclofenonGBR1aandGBR2mRNAexpressioninthehippocampusofepilepticratsfollowingkainicacid(KA)induction,andtostudytheadaptabilityofGABA_BRsubunits.DESIGN,TIMEANDSETTING:Arandomized,controlled,animalexperimentbasedonmolecularbiologywasperformedattheLaboratoryResearchCenterofSecondHospitalAffiliatedtoSoochowUniversityfromNovember2005toMarch2006.MATERIALS:KAwasprovidedbySigma,USA.InsituhybridizationdetectionkitofGBR1aandGBR2wasprovidedbyWuhanBosterBiologicalTechnology,China.GABA_BRagonist(baclofen)wasprovidedbySigma,USA.METHODS:Forty-fourepilepticratswererandomlyallocatedtoepileptic(n=28)anddrugintervention(n=16)groups.Theepilepticgroupwasfurtherdividedintopost-epilepticsubgroupsatdifferenttimepoints:6,12hours,1,3,7,15,and30days(n=4).Thedruginterventiongroupwasfurtherdividedintointerventioncontrolssubgroupsatvarioustimepoints:6hours,1day,and3days(n=4).Fouradditionalratswereconsideredthenormalcontrolgroupandnotmodeled,butwereinjectedwithsalineinthehippocampalCA3region.MAINOUTCOMEMEASURES:GBR1aandGBRmRNAexpressionwasdetectedintherighthippocampalCA1,CA3,anddentategyrus(DG)areasofthecontrol,epileptic,andinterferencegroupsatvarioustimeintervalsaccordingtoinsituhybridizationresults.RESULTS:(1)Duringtheearlystageofepilepsy(6and12hours),GBR1aandGBR2mRNAexpressionwasdecreased,andexpressionwaslessthanthecontrolgroupatonedayafterKAinduction(P<0.05).mRNAexpressionwasincreasedintheDG,butwasgreaterthanthecontrolgroupatday3(P<0.05).ExpressioninthehippocampalCA1andCA3regi

简介：BACKGROUND:PreviousstudieshavefocusedonthecorrelationbetweenNogo-AexpressionandmultiplesclerosisorbetweenNogo-Areceptor(NgR)expressionandmultiplesclerosisinthecentralnervoussystem.ExpressionpatternsofNogo-AandNgRremainpoorlyunderstoodinratmodelsofexperimentalautoimmuneencephalomyelitis(EAE).OBJECTIVE:ToobservedynamicchangesinNogo-AandNgRproteinexpression,andtoverifythecorrelationbetweenNogo-AandNgRprotein,aswellasexpressionpatternsatvarioustimepoints,inperiventriculartissueofEAErats.DESIGN,TIMEANDSETTING:Aneuroimmunological,randomized,controlledexperimentwasperformedattheClinicalInstituteofHunanPeople'sHospitalofChinafromSeptembertoNovember2008.MATERIALS:Immunohistochemistry(streptavidin-biotin-peroxidasecomplexmethod)kitwaspurchasedfromBoster,China.METHODS:Atotalof60female,Wistarrats,aged6-8weeks,wererandomlyassignedtoEAEandcontrolgroups(n=30,respectively).Guineapigspinalcordhomogenate,self-madecompleteFreund'sadjuvant(0.2mL/100g),andpertussisvaccine(0.2mL)weresubcutaneouslyinjectedintothehindlimbfootpadofratsfromtheEAEgrouptocreateratmodelsofEAE.CompleteFreund'sadjuvant(0.2mL)wasinfusedintoratsfromthecontrolgroup.MAINOUTCOMEMEASURES:Nogo-AandNgRproteinexpressionwasdeterminedinperiventricularwhitematterusingimmunohistochemicalmethods.Neurologicalscoresweredeterminedinallrats.RESULTS:RatsfromtheEAEgroupdevelopedacute-onsetEAEfollowingimmunization.Thepathogeneticsymptomsreachedapeakonday15,andneurologicalscoreswerealsogreatestatthistimepoint.Neurologicalscoresdecreasedwithrecoveryoftheillness.Nogo-Awasshowntobeexpressedinneuronalcellsandoligodendrocytes,andexpressionincreased11daysafterimmunization(P<0.01),decreasedbyday13(P<0.01),andthenincreasedagainbyday15.Nogo-AexpressionremainedgreaterintheEAEgroupcomparedwiththecontrolgroupatday30(P<0.01).IntheEAEgroup

简介：Icariin,themajoractivecomponentofChinesemedicinalherbepimediumbrevicornummaxim,isusedwidelyintraditionalChinesemedicineforthetreatmentofneurologicaldiseases.However,theeffectsoficariinonmyelininhibitoryfactorsareasyetunclear.Inthepresentstudy,administrationoficariinat20mg/kgshowedamarkedreductioninneurologicaldeficitofmiddlecerebralarteryocclusionrats.Icariinexhibitedbetterinhibitoryeffectsonmyelininhibitoryfactors:Nogo-A,myelin-associatedglycoproteinandoligodendrocytemyelinglycoproteininischemiaregionsofmiddlecerebralarteryocclusionratscomparedwithmonosialotetrahexosylganglioside.Theseresultsindicatethaticariinexhibitspotentinhibitoryeffectsonexpressionofmyelininhibitorsaftermiddlecerebralarteryocclusion-inducedfocalcerebralischemiainvivo.Thiseffectmaybemediated,atleastinpart,bytheinhibitionofbothNogo-A,myelin-associatedglycoproteinandoligodendrocytemyelinglycoproteinactivation,followedbytheenhancementofaxonalsproutingandregeneration,resultinginneurologicalfunctionalrecovery.

简介：BACKGROUND:Brainischemiainvolvessecondaryinflammation,whichsignificantlycontributestotheoutcomeofischemicinsults.Vascularendothelialgrowthfactor(VEGF)mayplayanimportantroleinthevascularresponsetocerebralischemia,becauseischemiastimulatesVEGFexpressioninthebrain,andVEGFpromotesformationofnewcerebralbloodvessels.Minocycline,atetracyclinederivative,protectsagainstcerebralischemiaandreducesinflammation,oxidativestress,andapoptosis.OBJECTIVE:ToobservetheinfluenceofminocyclineonVEGF,interleukin-1beta(IL-1β),andtumornecrosisfactoralpha(TNF-α)expressioninWistarratswithfocalcerebralischemia/reperfusioninjury,andtostudytheneuroprotectionmechanismofminocyclineagainstfocalcerebralischemia/reperfusioninjury.DESIGN,TIMEANDSETTING:Randomized,controlledexperiment,whichwasperformedintheChongqingKeyLaboratoryofNeurologybetweenMarch2007andMarch2008.MATERIALS:Atotalof36female,Wistarratsunderwentsurgerytoinsertathreadintotheleftmiddlecerebralartery.Animalswererandomlydividedintosham-operation,minocyclinetreatment,andischemia/reperfusiongroups,with12ratsineachgroup.Minocycline(HuishiPharmaceuticalLimitedCompany,China)wasdissolvedto0.5g/Linnormalsaline.METHODS:A0.5-1.0cmthreadwasinsertedintoratsfromthesham-operationgroup.Ratsintheischemia/reperfusiongroupunderwentischemiaandreperfusion.Theminocyclinegroupreceivedminocycline(50mg/kg)12and24hoursfollowingischemiaandreperfusion,whereastheothergroupsreceivedsalineatthecorrespondingtimepoints.MAINOUTCOMEMEASURES:mRNAandproteinexpressionofIL-1βandTNF-αwasmeasuredbyreversetranscriptase-polymerasechainreaction(RT-PCR)andenzymelinkedimmunosorbentassay(ELISA),respectively.VEGFmRNAandproteinexpressionwasexaminedbyRT-PCR,Westernblot,andELISA.RESULTS:Minocyclinedecreasedthefocalinfarctvolume.VEGF,IL-1β,andTNF-αexpressionw

简介：Acupunctureandmoxibustiontherapynon-specificallyraisesthepainthreshold,relievesregionalmusculartension,improveslocalbloodcirculation,accelerateseliminationofalgogenicsubstancessuchasacidmetabolites,andpromotesneuralregenerationandfunctionalrehabilitation.However,theeffectsofacupuncturearesyntheticallyinfluencedbymultiplefactors,whichvarywithacu-puncturepoint,intensity,andmanipulation.ThepresentstudyexploredtheanalgesiceffectsandmechanismsofspecificSancaiacupuncturemanipulationforsciatica.ResultsrevealedthatSancaiacupuncturemanipulationsignificantlyreducedinflammatoryfactorinterleukin-6expressioninthebloodserumofratswithsciatica,exhibitedanti-inflammatoryeffectsandpromotedrehabilitationofinjurednerves.Inaddition,Sancaiacupuncturemanipulationexhibitedsuperiorcurativeeffectsoverconventionalacupuncturemanipulation.

简介：MitochondrialK+-ATP(mito-KATP)channelsplayanimportantroleincellularfunctionandsurvivalfollowingischemicstress.Thepresentresultsrevealedthatinterventionwithdiazoxide,amito-KATPchannelopener,ledtoanincreaseinBcl-2expressioninthecerebralcortexofratssubjectedtocerebralischemiareperfusioninjury.Inaddition,theinterventionalsoledtoclearimprovementsinneuronalmitochondrialmorphologyandconsciousnesspost-injury.Glibenclamide,amito-KATPchannelblocker,exhibitedtheconverseeffects.Bothdiazoxideandglibenclamideexerteddose-dependenteffects(inparticular,at18mg/kgdiazoxideand25mg/kgglibenclamide).Thesefindingssuggestthatdiazoxideexertsaneuroprotectiveeffectoncerebralischemiareperfusioninjurybyopeningmito-KATPchannelsandupregulatingBcl-2expression.

简介：Previousstudieshaveshownthatbaicalinpreventedironaccumulationaftersubstantianigrainjury,reduceddivalentmetaltransporter1expression,andincreasedferroportin1expressioninthesubstantianigraofrotenone-inducedParkinson’sdiseaserats.Inthecurrentstudy,weinvestigatedtherelationshipbetweenironaccumulationandtransferrinexpressioninC6cells,toexplorethemechanismsoftheinhibitoryeffectofbaicalinonironaccumulationobservedinParkinson’sdiseaserats.Ironcontentwasdetectedusinginductivelycoupledplasma-atomicemissionspectroscopy.Resultsshowedthatironcontentdecreased41%afterblockingdivalentmetaltransporter1andferroportin1proteins.Aftertreatmentwithferricammoniumcitrateofdifferingconcentrations(10,50,100,400μg/mL)inC6gliomacells,cellsurvivalrateandferroportin1expressionwerenegativelycorrelatedwithferricammoniumcitrateconcentration,butdivalentmetaltransporter1expressionpositivelycorrelatedwithferricammoniumcitrateconcentration.Baicalinordeferoxaminereduceddivalentmetaltransporter1expression,butincreasedferroportin1expressioninthe100μg/mLferricammoniumcitrate-loadedC6cells.Theseresultsindicatethatbaicalindown-regulatedironconcentration,whichpositivelyregulateddivalentmetaltransporter1expressionandnegativelyregulatedferroportin1expression,anddecreasedironaccumulationinthesubstantianigra.

简介：BACKGROUND:Matrixmetalloproteinase-9(MMP-9)expressionincreaseswithintracerebralhemorrhage,andparticipatesinthepathophysiologicalprocessesofsecondarybraininjuryafterintracerebralhemorrhage.OBJECTIVE:ToinvestigatetheeffectsofmildhypothermiaonMMP-9expressionandbrainedemaintheperihematomalregionofexperimentalintracerebralhemorrhagerats.DESIGN,TIMEANDSETTING:Therandomized,controlledexperimentwasperformedattheCentralLaboratoryofShandongProvincialHospitalbetweenMayandSeptember2007.MATERIALS:Seventy-two,Wistar,malerats,12-weeksold,wereusedforthisstudy.Rabbitanti-MMP-9primaryantibodywaspurchasedfromBoster,China.METHODS:Wistarratswereequallyandrandomlydividedintonormothermiaandmildhypothermiagroups.Thetwogroupseachcomprisedcontrol,6-hourintracerebralhemorrhage,24-hourintracerebralhemorrhage,48-hourintracerebralhemorrhage,72-hourintracerebralhemorrhage,and1-weekintracerebralhemorrhagesubgroups,withsixratsineachsubgroup.Ratmodelsofintracerebralhemorrhagewereestablishedbyinjecting100μLofautologousbloodintotheratcaudatenucleus.Ratsinthemildhypothermiagroupreceivedfourhoursoflocalmildhypothermiaimmediatelyfollowingtheinjection.Intracerebraltemperaturewasmaintainedat(33±0.5)℃.Subsequently,intracerebraltemperaturewasspontaneouslyrecoveredat25℃.Ratsinthecontrolsubgroupwerenotinjectedwithautologousbloodandreceivedonlywithintracerebralhemorrhage.MAINOUTCOMEMEASURES:BrainwatercontentandMMP-9expressionsurroundingthehematomaregion.RESULTS:MMP-9expressionincreasedat6hours,andbrainedemareachedapeakat48hoursafterintracerebralhemorrhage.MMP-9expressionwassignificantlydecreasedinthemildhypothermiagroupcomparedwiththenormothermiagroupateachtimepoint(P<0.05).CONCLUSION:MildhypothermiacansignificantlyinhibitMMP-9overexpressionandrelievebrainedemafollowingintracerebralhemo

简介：BACKGROUND:Previousstudieshaveshownthatthemitochondrialstructureandfunctionaredamagedinanimalmodelsofepilepsy.Inaddition,theBcl-2proteiniscapableofregulatingmitochondrialstability.OBJECTIVE:ToobserveandvalidatechangesinmitochondrialstructureandBcl-2expression,andtoanalyzethesecharacteristicsinthehippocampalCA3regionofratmodelsofepilepsy.DESIGN,TIMEANDSETTING:Thisrandomized,controlled,animalexperimentwasperformedattheLaboratoryofElectronMicroscopyandDepartmentofHistologyandEmbryology,LuzhouMedicalCollegebetween2007and2008.MATERIALS:CoriamyrtinwasprovidedbythePharmacyFactoryofWestChinaUniversityofMedicalSciences.TheprimaryandsecondaryantibodieswereprovidedbyZhongshanGoldenbridgeBiotechnology,Beijing.METHODS:Atotalof44adult,male,SpragueDawleyratswererandomlydividedintocontrol(n=11)andepilepsy(n=33)groups.Ratsintheepilepsygroupwereinducedbycoriamyrtin(50μg/kg),whichwasinjectedintothelateralventricles.Theratswerethenobservedat3,6,and24hoursafterepilepsyinduction,with11ratsateachtimepoint.Epilepsywasnotinducedinratsfromthecontrolgroup.MAINOUTCOMEMEASURES:PathologicalchangesinthehippocampalCA3regionwereobservedbylightmicroscopy;Bcl-2expressionwasanalyzedbyimmunohistochemistry;andmitochondrialchangesinthehippocampuswereobservedundertransmissionelectronmicroscopy.RESULTS:(1)ThecontrolgroupdisplayedverylittleBcl-2proteinexpressioninthehippocampalCA3region.However,after3hoursofepilepsy,expressionwasvisible.By6hours,expressionpeakedandthensubsequentlydecreasedafter24hours,butremainedhigherthanthecontrolgroup(P<0.05).(2)Mitochondriaweredamagedtovaryingdegreesintheepilepsygroups.Forexample,mitochondriaedema,cristaespaceincrease,anddisappearanceofmitochondriawereapparent.Moreover,mitochondrialdamageoccurredpriortopathologicalchangesintheneuronsandnucleolus.CONCLUSION:

简介：Inthisstudy,wesoughttoelucidatetheeffectsofmelatoninonlearningandmemoryaswellasapoptosisandexpressionoftheBaxorBcl-2proteinsinthesubgranularzoneofthedentategyrusinpinealectomizedrats.UsingtheMorriswatermazeandtheolfactorymemorytests,wefoundthattheaverageescapelatencyinpinealectomizedratswasclearlyincreasedcomparedwithsham-operatedrats.Moreover,theaverageescapelatencyinthemelatonin-treatedandpinealectomizedratswaslongerthanthatinthesham-operatedratsandshorterthanthatinthepinealectomizedanduntreatedrats.Immunohistochemistryandterminal-deoxynucleoitidyltransferasemediatednickendlabeling(TUNEL)showedthattherewerefewerBaximmunoreactivecellsandTUNEL-positive(apoptotic)cellsbutmoreBcl-2immunoreactivecellsinthemelatonin-treatedratscomparedwiththepinealectomizedrats.Thesham-operatedratsshowednumbersofthesecellssimilartothemelatonin-treatedrats.TheseexperimentalfindingsdemonstratethatmelatonintreatmentmayreduceabnormalapoptosisbypromotinggeneexpressionofBaxandsuppressinggeneexpressionofBcl-2inthesubgranularzoneofthedentategyrusinpinealectomizedrats.Theseeffectsappeartoresultintheinhibitionofcellularapoptosisandtheimprovementofspatiallearningandmemoryinpinealectomizedrats.

简介：BACKGROUND:Studieshavesuggestedthatfibronectinleucine-richtransmembraneprotein3(FLRT3)isrelatedtoinjuryandregenerationofthenervoussystem.However,theexpressionandbiologicalcharacteristicsoftheseproteinsremainpoorlyunderstood.OBJECTIVE:ToobtainFLRT3C-terminalgenefragments,toeffectivelyexpressandpurifythetargetproteins.DESIGN,TIMEANDSETTING:AnobservationalstudyofcellularandmolecularbiologywasperformedatthelaboratoryofHistologyandEmbryologyinXiangyaSchoolofMedicine,CentralSouthUniversitybetweenOctober2007andJune2008.MATERIALS:ThreeSpragueDawleyadultratswereusedtoextracttotalRNAfromratbrains.ThepGEX4T3andEscherichiacoli(E.coli)JM109werepurchasedfromPromega.E.coliBL21wasprovidedbyNovagen.METHODS:FLRT3proteincodingC-terminalDNAfragments,atalengthof786bp,wereamplifiedusingRT-PCRtechniquefromrattotalRNA.TheamplifiedproductswereclonedintotheexpressionvectorpGEX4T3.ArecombinantexpressionvectorwasthenconstructedandintroducedintoE.coliBL21.IsopropyI-D-thiogalactopyranosidewasappliedtoinduceexpressionofrecombinantGSTfusionproteins,followedbyisolation,purification,andrenaturationofinclusionbodiesthatcomprisedrecombinantproteins.Finally,thepurifiedrecombinantproteinwasobtained.MAINOUTCOMEMEASURES:DeterminationofFLRT3C-terminalDNAsequence;expressionoftargetproteinswasassayedbySDS-PAGEelectrophoresis;purifiedrecombinantproteinwasidentifiedwithWesternblotmethods.RESULTS:FLRT3proteincodingC-terminalDNAfragments,atalengthof786bp,weresuccessfullyharvestedthroughRT-PCRamplification,andwerethenclonesintotheprokaryoticexpressionvectorpGEX4T3.Theresultsofthesequencewereconsistentwiththeknowngenesequence.SDS-PAGEanalysisdemonstratedthattherewasaspecificproteinbandintherecombinantGSTfusionproteinsatarelativemolecularmassof56,600.Therecombinantproteinwasobservedintheinclusionbody,andh

简介：BACKGROUND:ThedetectionofdifferentialgeneexpressioninbrainispossiblebycDNAmicroarraytechnology,andthescreeningofdifferentiallyexpressedgenesmightprovideabiologicalbasisforgene-targetedtherapyfortumors.OBJECTIVE:TodetectthedifferentialexpressionofgenesamongastrocytomaSHG-44(WHOgradeIV),CHG-5(WHOgradeII),andATRA-treatedSHG-44celllinesbycDNAmicroarray.DESIGN:Laboratoryexperimentsinvitro.SETTING:DepartmentofNeurobiology,theThirdMilitaryMedicalUniversity.MATERIALS:TheexperimentwasperformedattheDepartmentofNeurobiologyintheThirdMilitaryMedicalUniversityoftheChinesePLAfromJanuarytoOctober2007.TheSHG-44cellline(WHOgradeⅣ)wasestablishedbyProf.ZiweiDu,andtheCHG-5cellline(WHOgradeII)wassetupbyProf.XiuwuBianfromtheThirdMilitaryMedicalUniversityoftheChinesePLA.ThecDNAmicroarraycontaining9182knowngeneswaspreparedandprovidedbyDr.YangZhongattheCityUniversityofHongKong.METHODS:ToscreendifferentiallyexpressedgenesfromthegeneexpressionprofilesdetectedbycDNAmicroarraycomparisonsweremadebetweenCHG-5andSHG-44cellsandbetweenSHG-44cellswithorwithouttreatmentwith10μmol/LATRA.SomedifferentiallyexpressedgeneswereselectedrandomlyforNorthernBlotanalysistoconfirmtheresultsofthemicroarray.Thedeterminationcriteriafordifferentialgeneexpressionwereasfollows.①TheratioofCy5signaltoCy3wasgreaterthan2.0orlessthan0.5.②Theresultsofthetriplicatemicroarrayhybridizationsshowedthesametrendinthreeexperiments.③Ageneappearedatleasttwotimesonthetriplicatemicroarrayhybridizations,andthe3rdvaluedidnotshowacontradictorytrend.AnormalizedratioofCy5intensitytoCy3greaterthan2.0orlessthan0.5wasconsideredtorepresentup-regulatedordown-regulatedgeneexpression,respectively.MAINOUTCOMEMEASURES:Theidentificationofgenesthatweresimilarlyregulated(overlapping)dur

简介：BACKGROUND:Epidemiologicstudieshaveindicatedthattheincidenceofstrokeinpremenopausalfemalesislowerthaninmalesatthesameage,butitsignificantlyrisesinpostmenopausalfemales.Estrogenisusedclinicallytoalleviateinjurycausedbycerebralischemia.Ithasbeenhypothesizedthattheneuroprotectiveroleofestrogenrelatestoangiopoietin(Angpt),whichplaysanimportantroleinvascularization,vascularremodelingandmaturation.OBJECTIVE:Toobserveandvalidatetheeffectofestradiolonangiopoietin-1(Angpt1)mRNAexpressioninovariectomizedratswithfocalcerebralischemiaafterreperfusion,soastoexplorethemolecularmechanismsofestradiol-mediatedprotectionfromcerebralischemicdamage.DESIGN,TIMEANDSETTING:Randomized,controlled,molecularbiology,prospectiveanimalstudy.TheexperimentwasperformedattheCentralLaboratoryofChongqingMedicalUniversityfromSeptembertoDecember2005.MATERIALS:Fiftyhealthyfemalewildtype(WT)ratsaged6monthsandfiftyfemaleratsaged6monthswithknockoutoftheestrogen-alphareceptorgene(ERKO).METHODS:WTratsandERKOratsweredividedintoestradiolandcontrolgroups(n=25),andinjectedin-tramuscularlywithestradiolbenzoate(100μg/kgperday)orcornoil(1mL/kgperday)for7days,30daysafterbilateralovariectomy.Ratmodelsofcerebralischemia/reperfusionwereestablishedwiththemiddlecerebralarteryocclusionmethod.After30minutesofmiddlecerebralarteryocclusion,ratsfromtheestradiolandcontrolgroupswereinjectedintramuscularlywithestradiolbenzoateorcornoilattheabovedose.MAINOUTCOMEMEASURES:Weusedradio-immunityanalysisandlaser-Dopplerflowmetrytomeasureplasmaestradiollevelsandchangesincerebralbloodflow.WeusedimmunohistochemicalstainingofCD34epitopestomeasurechangesinthecapillarydensityinbrainfollowingcerebralischemia/reperfusion,andquantitativeRT-PCRanalysistoassessmRNAexpressionlevelsofAngpt1,Angpt2,Tie2,vascu