简介:目的:构建表达小鼠CD4+T细胞钙支架蛋白AHNAK1的短发夹RNA(shorthairpinRNA,shRNA)慢病毒载体,并研究其对小鼠甲状腺相关性眼病(thyroid-associatedophthalmopathy,TAO)的抑制效应。方法:设计并筛选对AHNAK1具有良好干扰效力的shRNA序列,慢病毒载体包装干扰序列,感染小鼠CD4+T细胞,检测AHNAK1静默对T细胞功能的抑制作用,采用实验动物模型观察AHNAK1体内抑制甲状腺相关性眼病的效果。结果:成功筛选出具有良好干扰效力的shRNA,并包装入慢病毒。病毒滴度为1.0伊106TU/mL,转染慢病毒的CD4+T细胞展现出失能倾向,抑制炎症免疫反应;在动物模型中抑制T细胞中AHNAK1表达可以有效控制甲状腺眼病的发生发展,显著降低治疗组T细胞中IL-2、IL-1茁和IFN-酌的表达。结论:成功构建了表达小鼠AHNAK1shRNA的慢病毒,具有抑制T细胞分泌IL-2、IL-1茁和IFN-酌的表达效应,能够有效抑制甲状腺眼病的发生发展。
简介:3型Stargardt病(STGD3,MIM600110)是一种常染色体显性遗传的早发性黄斑营养不良性疾病,是一种单基因遗传性疾病。目前为止,已知的STGD3致病基因为极长链脂肪酸延长酶4基因(ELOVL4)。ELOVIA是位于内质网上的一种调节极长链饱和及多不饱和脂肪酸生物合成的限速缩合反应中不可或缺的膜蛋白。作为一种遗传性疾病,STGD3目前仍无有效治疗方法。然而,饮食补充极长链多不饱和脂肪酸可能是治疗STGD3的一种可行疗法。对STGD3基因及致病机制的研究可能将有助于发现STGD3的有效治疗方法及进一步了解其他遗传性视网膜变性疾病和更复杂的年龄相关性黄斑变性。
简介:AIM:Toinvestigatethelevelsofserumsolubleintercellularadhesionmolecules-1(sICAM-1)andneutrophilicexpressionofCD18inpatientswithvariousstagesofdiabeticretinopathyandtodeterminetheirdifferentexpressionpatterninthedevelopmentofdiabeticretinopathy(DR).·METHODS:LevelsofserumsICAM-1andCD18onthesurfaceofneutrophileweremeasuredin41DRpatients,theywereclassifiedinthreesubgroupsaccordingtothestageofretinopathyasdeterminedbyfund’sophthalmoscopy;10controlsubjectswerealsostudied.sICAM-1weremeasuredbyenzyme-linkedimmunosorbentassayandCD18byflowcytometry.·RESULTS:TheneutrophilicCD18expressionandserumsICAM-1levelwereallsignificantlyelevatedinalldiabeticsubgroupscomparedtocontrolsubjects(P<0.01).ThedifferencesofCD18andsICAM-1amongthediabeticsubgroupsweresignificantinCD18butnotinsICAM-1.TheprogressionofretinopathywasassociatedwithanincreasebothinCD18andinsICAM-1levelsbysimplecorrelationanalysis(β=0.74,P<0.001;β=0.38,P<0.01,respectively).ButstepwisemultipleregressionanalysisrevealedthatonlyCD18wasindependentdeterminantofretinopathy(β=1.04,P<0.01).·CONCLUSION:OurresultsconfirmthecontributionofendothelialandneutrophilicactivationinthedevelopmentofDRasindicatedbyincreasedlevelsofCD18andsICAM-1.However,adirectimplicationofCD18andICAM-1intheprogressionofDRcanbesupportedonlyintheCD18butnotICAM-1.CD18andICAM-1mayplaydifferentroleinthedevelopmentofdiabeticretinopathy.
简介:AIM:ToexplorethemolecularmechanismsinlensdevelopmentandthepathogenesisofPetersanomalyinSmad4defectivemice.METHODS:Le-CretransgenicmouselinewasemployedtoinactivateSmad4inthesurfaceectodermselectively.PathologicaltechniqueswereusedtorevealthemorphologicalchangesoftheanteriorsegmentinSmad4defectiveeye.ImmunohistochemicalstainingwasemployedtoobservetheexpressionofE-cadherin,Ncadherinanda-SMAinanteriorsegmentofSmad4defectivemiceandcontrolmiceatembryonic(E)day16.5.Real-timequantitativepolymerasechainreaction(qPCR)wasperformedtodetecttheexpressionofSnail,Zeb1,Zeb2andTwist2inlensofSmad4defectivemiceandcontrolmiceatE16.5.RESULTS:ConditionaldeletionofSmad4oneyesurfaceectodermresultedincorneaidysplasia,iridocornealangleclosure,corneolenticularadhesionsandcataractresemblingPetersanomaly.LossofSmad4functioninhibitedE-cadherinexpressioninthelensepitheliumcellsandcorneaiepitheliumcellsinSmad4defectiveeye.ExpressionofN-cadherinwasupregulatedincorneaiepitheliumandcorneaistroma.BothE-cadherinandN-cadherinweredown-regulatedatthefuturetrabecularmeshworkregioninmutanteye.TheqPCRresultsshowedthattheexpressionofTwist2wasincreasedsignificantlyinthemutantlens(P<0.01).CONCLUSION:Smad4isessentialtoeyedevelopmentandlikelyacandidatepathogenicgenetoPetersanomalybyregulatingepithelial-mesenchymaltransition.Twist2canberegulatedbySmad4andplaysanessentialroleinlensdevelopment.
简介:目的:探讨线粒体膜电位(△ψm)、Caspase3在As2O3诱导ACC-2细胞凋亡中的作用。方法:进行ACC-2细胞培养,将As2O3建立不同药物浓度梯度(0,1.0,2.0,4.0,8.0μmol/L)分别作用于ACC-2细胞,用Rh123染色,流式细胞仪检测8.0μmol/LAs2O3作用前、后(24h),ACC-2细胞的线粒体膜电位(△ψm)变化;用多功能酶标仪进行Caspase3活性检测。结果:空白对照组ACC-2细胞内Rh123荧光强度最强,8.0μmol/LAs2O3处理组ACC-2细胞内Rh123荧光强度减弱,其差异有显著性(P〈0.05);随着As2O3药物浓度的增高(0,1,2,4,8μmol/L),ACC-2细胞的Caspase3酶活力单位逐渐增加。结论:As2O3作用于ACC-2细胞,可通过降低线粒体膜电位从而引起细胞凋亡。随着As2O3药物浓度的增高,ACC-2细胞的Caspase3酶活力单位逐渐增加,Caspase3被激活,细胞可发生不可逆转的凋亡过程。
简介:目的:探讨眼眶IgG4相关疾病(IgG4-RD)的临床病理特点。方法:收集整理23例35眼眼眶IgG4-RD患者的临床病理资料,对其进行组织学和免疫组织化学观察,总结其临床和病理特点。结果:眼眶IgG4-RD患者23例35眼,其中男8例9眼,女15例26眼,年龄28-72(平均52.1)岁。19例30眼来源于泪腺,4例5眼来源于眶内其他部位。以泪腺区肿胀或眼球突出就诊。单侧11例,双侧12例。病程1mo-10a,平均27mo。1例1眼6mo后复发。大体:灰白色结节状肿物,泪腺表面有很薄的纤维膜包绕。组织学特点:泪腺腺泡、导管组织严重萎缩甚至消失,被大量密集的淋巴细胞、浆细胞及淋巴滤泡替代,伴有不同程度的纤维化。免疫组织化学染色:23例35眼IgG4阳性浆细胞均〉50个/HPF,IgG4/IgG阳性浆细胞比值〉40%。结论:眼眶IgG4-RD主要发生于泪腺组织,通过组织学特点和免疫组织化学IgG4的表达可明确诊断。IgG4-RD应早期筛查、预防和治疗。
简介:AIM:Tocomparethespeedofvisualrecoveryfollowingmyopicthin-flapLASIKwithfourfemtosecondlasers.METHODS:Eighty-eighteyesof46patientswhowereconsecutivelyscheduledforbilateralLASIKwiththeIntraLaseFS60(Group1),FemtoLDVCrystalLine(Group2),WavelightFS200(Group3)andVisuMax(Group4)femtosecondlaserswereenrolledin.Monocularuncorrecteddistancevisualacuity(UDVA),best-correcteddistantvisualacuity(CDVA),refraction,contrastsensitivityandhigher-orderaberrations(HOAs)wereevaluatedat1,3d,1wkand1mopostoperatively.RESULTS:Sixteeneyes(72.7%)achieved20/16and8eyes(36.4%)were20/12.5at1dinGroup2,whichwassignificantlymorethanother3groups.At1wk,20eyes(90.9%)achieved20/16inGroups2and4.At1mo,20eyes(90.9%)achieved20/16inGroup2andGroup4,whichweresignificantlymorethanothertwogroups.Whileby1mo,thedifferenceoftheresidualsphericalequivalent(SE)wasnotstatisticallysignificantamong4groups(P=0.121).Theinductionofsphericalaberration(SA)weresignificantlylessforGroups2,3,4thanforGroup1onedayaftersurgery(P=0.015).Thedifferencesamong4groupswerenotstatisticallysignificantbeforeandaftersurgeryoneverytimepoints(allP>0.05).CONCLUSION:Thethin-flapLASIKprocedureusingtheFemtoLDVCrystalLineandVisuMaxfemtosecondlasershowfastervisualperformancerecovery.
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