简介:Hypoxia-induciblefactor1(HIF-1)attenuatesamyloid-betaproteinneurotoxicityanddecreasesapoptosisinducedbyoxidativestressorhypoxiaincorticalneurons.Inthisstudy,weconstructedarecombinantadeno-associatedvirus(rAAV)vectorexpressingthehumanHIF-1αgene(rAAV-HIF-1α),andtestedtheassumptionthatrAAV-HIF-1αrepresseshippocampalneuronalapoptosisinducedbyamyloid-betaprotein.OurresultsconfirmedthatrAAV-HIF-1αsignificantlyreducesapoptosisinducedbyamyloid-betaproteininprimaryculturedhippocampalneurons.DirectintracerebralrAAV-HIF-1αadministrationalsoinducedrobustandprolongedHIF-1αproductioninrathippocampus.SinglerAAV-HIF-1αadministrationresultedindecreasedapoptosisofhippocampalneuronsinanAlzheimer’sdiseaseratmodelestablishedbyintracerebroventricularinjectionofaggregatedamyloid-betaprotein(25–35).OurinvitroandinvivofindingsdemonstratethatHIF-1haspotentialforattenuatinghippocampalneuronalapoptosisinducedbyamyloid-betaprotein,andprovidesexperimentalsupportfortreatmentofneurodegenerativediseasesusinggenetherapy.
简介:目的总结颅眶沟通黏膜相关淋巴组织结外边缘区B细胞(MALT)淋巴瘤的诊治经验。方法回顾性分析l例眼眶MALT淋巴瘤原位复发及颅内、颌面部转移患者的临床资料,结合文献复习,讨沦其转移方式、影像学特点及治疗措施。结果该患者行联合手术治疗颅眶占位病变,颌面部病变未处理,术后病理学结果为MALT淋巴瘤;术后复查MRI示右侧眼眶肿瘤全切,术后2周左眼视力较术前改善。结论眼眶MALT淋巴瘤转移至颅内罕见,需于术联合放疗或化疗,术后长期随访十分重要。
简介:GinsenosideRg1(Rg1)hasanti-agingandanti-neurodegenerativeeffects.However,themechanismsunderlyingtheseactionsremainunclear.TheaimofthepresentstudywastodeterminewhetherRg1affectshippocampalsurvivalandneuriteoutgrowthinvitroafterexposuretoamyloid-betapeptidefragment25–35(Aβ25–35),andtoexplorewhethertheextracellularsignal-regulatedkinase(ERK)andAktsignalingpathwaysareinvolvedinthesebiologicalprocesses.Weculturedhippocampalneuronsfromnewbornratsfor24hours,thenaddedRg1tothemediumforanother24hours,withorwithoutpharmacologicalinhibitorsofthemitogen-activatedproteinkinase(MAPK)familyorAktsignalingpathwaysforafurther24hours.Wethenimmunostainedtheneuronsforgrowthassociatedprotein-43,andmeasuredneuritelength.Inaseparateexperiment,weexposedculturedhippocampalneuronstoAβ25–35for30minutes,beforeaddingRg1for48hours,withorwithoutAktorMAPKinhibitors,andassessedneuronalsurvivalusingHoechst33258staining,andphosphorylationofERK1/2andAktbywesternblotanalysis.Rg1inducedneuriteoutgrowth,andthiseffectwasblockedbyAPI-2(Aktinhibitor)andPD98059(MAPK/ERKkinaseinhibitor),butnotbySP600125orSB203580(inhibitorsofc-JunN-terminalkinaseandp38MAPK,respectively).Consistentwiththiseffect,Rg1upregulatedthephosphorylationofAktandERK1/2;theseeffectswerereversedbyAPI-2andPD98059,respectively.Inaddition,Rg1significantlyreversedAβ25–35-inducedapoptosis;thiseffectwasblockedbyAPI-2andPD98059,butnotbySP600125orSB203580.Finally,Rg1significantlyreversedtheAβ25–35-induceddecreaseinAktandERK1/2phosphorylation,butAPI-2preventedthisreversal.OurresultsindicatethatRg1enhancesneuriteoutgrowthandprotectsagainstAβ25–35-induceddamage,andthatitsmechanismmayinvolvetheactivationofAktandERK1/2signaling.更多还原
简介:目的筛选有效的MRP1siRNA序列,为RNAi的体内外研究提供依据.方法利用Dharmacon公司的RNA在线工具,设计了4对针对MRP1基因不同区域的siRNA序列,分别转染至U251等三种肿瘤细胞,采用荧光转染试剂观察转染效率;采用RT-PCR和Westernblot检测MRP1mRNA和蛋白的表达.结果siRNA1、siRNA2、siRNA3、siRNA4四对序列分别转染上述三种肿瘤细胞后的RT-PCR实验结果说明siRNA2和siRNA4对MRPmRNA有较强的抑制作用.结论siRNA2、siRNA4是可有效抑制MRP基因表达的序列.
简介:目的通过对荷瘤裸小鼠进行异种基质金属蛋白酶-2(c-MMP-2)DNA疫苗联合伽玛刀(γ-刀)治疗胶质瘤,观察二者的协同治疗作用,并对其作用机制进行初步探讨。方法制备纯化c-MMP-2DNA疫苗,裸鼠腋下注入大鼠C6胶质瘤细胞株1×10^6/只,建立裸鼠胶质瘤动物模型,待肿瘤长至约1cm(接种后第16d)开始治疗。γ-刀治疗以50%的等剂量曲线覆盖,边缘剂量13Gy;异种MMP-2DNA疫苗治疗组小鼠自荷瘤后次日开始后肢肌肉注射c-MMP-2DNA疫苗(1mg/ml),100μl/次,每周一次,连续4W。观察瘤体大小、测量肿瘤重量;免疫组化法测肿瘤组织微血管密度;末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口标记法(TUNEL)检测各组肿瘤细胞凋亡指数;HE染色观察重要器官组织坏死情况。结果异种MMP-2DNA疫苗联合γ-刀治疗胶质瘤能明显抑制肿瘤组织的生长,并延长荷瘤小鼠的生存期,与其他3组比较,联合治疗组免疫组化显示小鼠的肿瘤组织中微血管密度明显减少;TUNEL显示凋亡指数明显增多。结论c-MMP-2DNA疫苗联合γ-刀治疗能够抑制肿瘤生长、抑制肿瘤血管生成,二者的联合应用具有协同效应,是一种独特的抗肿瘤治疗途径。
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:目的探讨基质金属蛋白酶2(MMP-2)及抑癌基因PTEN在人脑星形瘤中的表达及二者与人脑星形细胞瘤侵袭性的关系。方法用免疫组织化学SABC法检测50例人脑星形细胞瘤组织和10例正常人脑组织中的MMP-2和PTEN蛋白的表达,并且分析二者与人脑星形细胞瘤临床病理分级的关系。结果MMP-2和PTEN在低度恶性星形细胞瘤和高度恶性星形细胞瘤组织中表达差别有统计学意义(p〈0.05)。随着星形细胞瘤恶性度增高,MMP-2的表达强度呈上升趋势而PTEN表达强度逐渐下降;Spearman等级相关分析表明人脑星形细胞瘤中MMP-2和PTEN之间呈负相关(Rs=-0.518,P〈0.01)。结论MMP-2和PTEN是人脑星形细胞瘤分化程度和转移的潜在生物学指标,联合检测MMP-2和PTEN更有利于判断星形细胞瘤生物学行为和病理分级。
简介:目的探讨以最近发展起来的核转染技术直接将编码绿色荧光蛋白DNA质粒转染到兔原代骨髓基质细胞的细胞核内进行基因修饰的可行性。方法从兔股骨抽取骨髓,密度梯度离心法获取原代骨髓基质细胞。以Nucleolector^TM技术转染pEGFP-C2(EGFP组),以同期培养未转染的细胞作为对照组。测定细胞的活力、贴壁率、生长曲线以及转染的效率。结果在转染后24h成功发现EGFP的表达。两组细胞具有相似的形态学变化、贴壁率以及生长曲线。EGFP的表达逐渐增强,至第6天达到最高峰(47.8%),观察一个月未发现表达减弱。结论pEGFP-C2基因核转染对兔原代骨髓基质细胞的体外增殖无明显影响;EGFP可以作为兔骨髓基质细胞有效的基因表达标记;Nucleofector^TM技术是一种简易而高效的转染兔原代骨髓基质细胞的方法。