简介:目的构建大鼠神经营养因子3(NT-3)基因真核表达载体并观察其在真核细胞内的表达情况。方法采用逆转录聚合酶链式反应(RT-PCR)技术从大鼠脑组织总RNA中扩增NT-3基因cDNA序列,将其克隆到真核表达载体pcDNA3中,经酶切鉴定和序列分析后,以阳离子脂质体Li-pofectamine2000介导转染L929细胞,应用免疫细胞化学和westernblot鉴定NT-3在细胞内的表达。结果RT-PCR产物为822bp的特异片段,重组质粒pcDNA3/NT-3酶切后产生822bp和5.2kb的片段,DNA测序证实822bp片段的碱基序列与大鼠NT-3基因序列完全一致,成功构建了pcDNA3/NT-3重组质粒。将其转染真核细胞后,免疫细胞化学、westernblot结果表明NT-3能在真核细胞中正确表达。结论成功构建了重组真核表达质粒载体pcDNA3/NT-3,为后续的研究奠定基础。
简介:BACKGROUND:TheprogressivedegenerationofdopaminergicneuronsinParkinson’sdiseaseisassociatedwithanactivatedglialreaction,combinedwithaninflammatoryprocess.Theseresponsesleadtotheproductionofcytokines,suchasinterferon-γ,tumornecrosisfactor-α(TNF-α),andinterleukin-1β.Inaddition,14-3-3proteinisacomponentofLewybodiesinParkinson’sdisease.OBJECTIVE:Toobservetheexpressionof14-3-3γandζprotein,aswellasTNF-α,inmousemicroglia,aswellaschangesafterlipopolysaccharide(LPS)activation.Toinvestigatepossiblemechanismsofdopaminergicneuronalinjuryduetoactivatedmicroglia.ToandclarifytheimmuneresponsemechanismsofParkinson’sdisease.DESIGN:Randomizedcontrolledobservation,cellstudy.SETTING:LaboratoryofDepartmentofNeurology,theAffiliatedUnionHospitalofTongjiMedicalCollege,HuazhongUniversityofScienceandTechnology.MATERIALS:TheBV-2immortalizedmurinemicrogliacelllinewaspurchasedfromChinaUnitcellcenter.LPSwasprovidedbySigmaCompany.CellcultureswerepurchasedfromGibco.Phospho-(Ser)14-3-3bindingmotifantibodywaspurchasedfromSantaCruzBiotechnologies.FITCwasprovidedbyLinfeiBiotechnology,Wuhan,China.TNF-αELISAwasprovidedbyJingmeiBiotechCo,Wuhan,China.TheflowcytometerwasprovidedbyBectonDickinson,Canada.METHODS:ThepresentexperimentwasperformedattheLaboratoryofDepartmentofNeurology,theAffiliatedUnionHospitalofTongjiMedicalCollege,HuazhongUniversityofScienceandTechnologyfromApriltoDecember2006.Themicroglialcellline,BV-2,wasculturedinvitroandstimulatedwithLPSfor2,6,12,and24hours.BV-2cultureswithoutLPSwereusedascontrols.MAINOUTCOMEMEASURES:Expressionof14-3-3γproteinwasdetectedbyflowcytometry.14-3-3ζpercentageexpressionandthemeanfluorescenceintensitywasdetectedbyimmunofluorescence.TNF-αexpressionwasdetectedbyELISA.RESULTS:14-3-3γproteinexpressionanalysis:followi
简介:我院收治3例垂体腺瘤合并脑血管病,现报告如下:例1,男,45岁,因'视力下降、性欲减退2年,头痛2个月'入院.体检:神志清,胡须、阴毛稀少.视力:左0.8,右眼前手动.左眼颞侧、右眼周边视野缺损.双乳房可挤出稀白乳汁.血皮质醇:上午8时为422.29nmol/l,下午4时为313.68nmol/l;血泌乳素(prolactin,PRL)>9.1nmol/l;雌二醇(estradiol,E)<70pmol/l,睾酮(testosterone,T)为5.86nmol/l,其余正常.颅脑CT、MRI示3.7cm×2.8cm大小囊、实性肿块,位于鞍内及鞍上区,视交叉明显受压.行右翼点入路肿瘤切除术.术中见右侧颈内动脉床突上段有大小为3mm的动脉瘤,给予夹闭.肿瘤位于视交叉前下方,有囊性变,质软易吸除.病理报告:垂体腺瘤.诊断:垂体腺瘤并动脉瘤.恢复顺利出院.随访5年无复发.
简介:目的探讨术前三维稳态构成干扰(3D-CISS)序列和三维时间飞跃(3D-TOF)序列MRI在原发性三叉神经痛责任血管判断中价值。方法回顾性分析2016年1~12月首次行微血管减压术(MVD)治疗的79例原发性三叉神经痛的临床资料。术前均行3D-CISS和3D-TOF序列MRI检查。以术中发现为判断责任血管的标准。结果术中发现单一责任血管56例(70.8%),2支及以上责任血管19例(24.1%),无责任血管4例(5.1%);责任血管中包含至少一条动脉73例(92.4%),单纯静脉2例(2.5%)。术前影像学检查阳性75例,阴性4例。75例阳性中,术中发现责任血管73例,未发现责任血管2例;4例阴性中,术中发现责任血管2例,2例未发现责任血管。术前3D-CISS和3D-TOF序列MRI判断责任血管的灵敏性为97.3%(73/75),特异性为50.0%(2/4),准确率为94.9%[(73+2)/79];判断责任血管包含动脉的灵敏性为98.6%(72/73),特异性为50.0%(3/6),准确率为94.9%[(72+3)/79]。结论术前3D-CISS联合3D-TOF序列MRI可用于判断原发性三叉神经痛责任血管,尤其对于包含动脉的责任病变,准确率较高,为MVD提供参考信息。
简介:目的:评价高分辨率3D-FIESTA+c成像及图像处理技术显示脑神经及其病变的价值。方法采用3D-FIESTA+c对20例健康志愿者和20例临床疑是因血管等原因压迫相应脑神经具有临床症状的患者进行扫描及图像后处理。由2名神经放射学医师根据20名健康志愿者480支脑神经显示的清晰程度分为清晰、较清晰、不清晰3个等级,清晰和较清晰定义为显示,不清晰定义为未显示;临床病例中,脑神经与血管关系分为无接触、接触、压迫。结果12对脑神经显示率分别为:嗅神经84.3%,视神经100%,动眼神经100%,滑车神经43.8%,三叉神经100%,外展神经100%,面神经100%,前庭蜗神经100%,舌咽神经、迷走神经及副神经复合体100%,舌下神经47.1%。20例脑神经症状患者,16例确诊为脑神经与周围血管接触或压迫,且均被临床治疗证实。结论高分辨3D-FIESTA+c成像与图像后处理技术相结合可显示脑神经及其病变,能准确定位血管走向及其与脑神经的关系,为临床医生提供准确、全面的影像学资料。
简介:目的研究GOLPH3在胶质瘤组织中的表达及其临床意义.方法选取山东大学齐鲁医院和聊城市脑科医院神经外科自2008年7月至2009年12月间手术切除并经病理证实的人脑胶质瘤标本76例,其中Ⅰ级13例,Ⅱ27级例,Ⅲ级25例,胶质母细胞瘤11例.另取9例因脑创伤行内减压术患者的正常脑组织标本作为对照.应用RT-PCR、Westernblotting分别检测各标本中GOLPH3mRNA和GOLPH3蛋白的表达.结果RT-PCR与Westernblotting检测结果显示正常脑组织中GOLPH3mRNA和GOLPH3蛋白不表达,不同级别胶质瘤组织GOLPH3mRNA、GOLPH3蛋白的阳性表达率差异无统计学意义(P>0.05),而其表达值差异有统计学意义(P<0.05),且随着肿瘤病理级别的增高,胶质瘤组织中GOLPH3mRNA、GOLPH3蛋白表达值增高,差异有统计学意义(P<0.05).相关性检验显示GOLPH3蛋白的表达值与肿瘤的病理分级呈正相关(rs=0.961,P=0.000).结论GOLPH3在不同级别的胶质瘤组织中均有表达,其表达值与肿瘤的病理分级呈正相关,可能是胶质瘤发生发展的重要机制之一.
简介:Anumberofpreviousstudiesofacupunctureacupointspecificityhaveusedshamacupoints,shamacupunctureormeridianacupointsatagreatdistancefromeachotherascontrolsinfunctionalMRI(fMRI)experiments.However,fewstudieshavecompareddifferentmeridianacupointswithinthesamesegment,whichareassociatedwithsimilarlyintenseneedlesensations.WeperformedfMRIon12healthyyoungvolunteersandobserveddifferencesinbrainactivationelicitedbyacupunctureoftheTaixi(KI3)andQiuxu(GB40)acupoints.AcupuncturewasappliedattheTaixiandQiuxuacupoints,usingamultiple-blockfMRIdesignwiththreeblocks,involvingthreealternationsofrestingandtaskphases.Afterscanning,needlesensationwasassessed.ThebehavioralresultsrevealedthatthesubjectiveneedlesensationwassimilarbetweentheTaixiandQiuxuacupoints.ThefMRIresultsrevealedthatacupunctureattherightTaixiacupointactivatedtherightsuperiortemporalgyrus(BA22),leftmiddlefrontalgyrus(BA46)andinferiorfrontalgyrus(BA45),bilateralparietallobepostcentralgyrus(BA2),rightparietallobe(BA3),andleftparietallobe(BA40).AcupunctureattherightQiuxuacupointactivatedtheleftsuperiortemporalgyrus(BA42),rightparietallobepostcentralgyrus(BA40,BA43),rightinferiorfrontalgyrus(BA47),bilateralsuperiortemporalgyrus(BA22),andrightinsulaBA13.TheseresultssuggestthattherightTaixiandQiuxuacupointsactivateddifferentbrainareas.
简介:Inthepresentstudy,weconstructedalentivirus,FIV-CMV-GFP-miR-7-3,containingthemicroRNA-7-3geneandthegreenfluorescentproteingene,andusedittotransfecthumangliomaU251cells.Fluorescencemicroscopyshowedthat80%ofU251cellsexpressedgreenfluorescence.Real-timereversetranscriptionPCRshowedthatmicroRNA-7-3RNAexpressioninU251cellswassignificantlyincreased.ProliferationwasslowedintransfectedU251cells,andmostcellswereintheG1phaseofthecellcycle.Inaddition,theexpressionoftheserine/threonineproteinkinase2wasdecreased.ResultssuggestedthattransfectionwithalentiviruscarryingmicroRNA-7-3caneffectivelysuppressepidermalgrowthfactorreceptorpathwayactivityinU251cells,arrestcellcycletransitionfromG1phasetoSphaseandinhibitgliomacellgrowth.