简介:AIM:Toexaminetheα-Galgeneexpressionanddistributioninthedifferentspecies/genusanddevelopingphaseanimalocularsurfacetissue.METHODS:α-Galbindingassaywerecarriedoutonvariousanimaleyesections.Photograph,slit-lampobservationonvariouseyeshowednormalcornealtransparence.RESULTS:Astrongα-Galexpressionininvertebratesandsomevertebratesoculartissue,butnoα-Galbindinginbirds,fishandmammal.α-Galexpressionchangeinthedevelopmentofmiceocularsurfacetissue(exceptsclera)anddisplaygenusdependencyinthedifferentmurineocularsurfacetissue.CONCLUSION:Thisstudyidentifiedspecificα-Galepitopesbindingareaintheocularsurfaceofseveralspeciesandmaysolvetheproblemthatnaiveocularsurfacemaybeusedasnaturalα-Galgeneknockoutmodel/highriskimmunologicrejectionmodelorocularsurfacescaffoldmaterial.
简介:目的:利用DNA探针杂交技术,结合显色探针技术建立一种新型、高灵敏的免疫检测体系,用于早期先天性白内障的筛查。方法:选取3个常染色体显性遗传的先天性白内障家系中患者14例,取静脉血并提取mRNA,建立CRYAB的捕获探针及显色探针。利用DNA探针,通过碱基配对原则形成三明治结构(捕获探针-DNA探针-显色探针)检测入选者的血样。1家系6例患者静脉血利用酶联免疫吸附测定(ELISA)法检测琢B-晶状体蛋白。结果:最佳条件下,双特异探针技术可检测到最低浓度的先天性白内障晶状体蛋白的突变基因,各突变位点检测率为99.5%~99.7%;ELISA法检测样本琢B-晶状体蛋白上调,阳性率为85.9%。双特异探针技术敏感性更高,检测位点更多,ELISA法仅局限于蛋白检测水平,精确性不高。结论:双特异探针检测技术操作简单,灵敏度高,可重复性高,经济实惠,在临床上用于产前诊断、优生优育具有重要的应用价值。
简介:目的研究高眼压及其持续时间对大鼠视网膜caspase-3表达的影响。方法通过前房灌注平衡盐液建立大鼠急性高眼压模型:高眼压持续时间均为4h,依据眼内压的不同将sD大鼠60只随机分为正常对照组、40mmHg(1mmHg=0.133kPa)组、60mmHg组、80mmHg组、100mmHg组,每组12只。将眼内压为80mmHg的48只sD大鼠随机分为正常对照组、2h组、4h组、8h组,每组12只。正常对照组不给予前房灌注,其他各组分别给予不同的高眼压或高眼压持续不同时间。各组灌注结束后快速摘除眼球,分别采用Western印迹法和免疫组织化学法检测视网膜caspase-3的表达。结果与对照组相比,40mmHg组大鼠视网膜caspase-3的表达无增加(P〉0.05);60mmHg组、80mmHg组和100mmHg组大鼠视网膜caspase-3的表达均强于正常对照组(q值分别为4.87、5.28和6.71;P〈0.01),但各组之间差异无统计学意义(P〉0.05)。与对照组相比,2h组大鼠视网膜caspase-3的表达差异无统计学意义(P〉0.05),4h组和8h组大鼠视网膜caspase-3的表达均增加(q值分别为2.81和3.67;P〈0.01)。表达caspase-3的视网膜细胞主要位于神经节细胞层、外丛状层和内丛状层。结论采用前房灌注平衡盐液制作急性高眼压模型研究高眼压大鼠视网膜caspase-3的表达时,眼内压为80mmHg、高眼压持续4h即可。
简介:AIM:Toinvestigatewhether15-Lipoxygenase-1(15-LOX-1)playsanimportantroleintheregulationofangiogenesis,inhibitinghypoxia-inducedproliferationofretinalmicrovascularendothelialcells(RMVECs)andtheunderlyingmechanism.METHODS:PrimaryRMVECswereisolatedfromtheretinasofC57/BL6JmiceandidentifiedbyanevaluationforFITC-markedCD31.ThehypoxiamodelswereestablishedwiththeBio-bagandevaluatedwithablood-gasanalyzer.ExperimentswereperformedusingRMVECstreatedwithandwithouttransferAd-15-LOX-1orAd-vectorbothunderhypoxiaandnormoxiaconditionat12,24,48,72hours.Theefficacyofthegenetransferwasassessedbyimmunofluorescencestaining.CellsproliferationwasevaluatedbytheCCK-8method.RNAandproteinexpressionsof15-LOX-1,VEGF-A,VEGFR-2,eNOsandPPAR-rwereanalyzedbyreal-timereversetranscriptionpolymerasechainreaction(RT-PCR)andWesternblot.RESULTS:RoutineevaluationforFITC-markedCD31showedthatcellswerepure.Theresultsofblood-gasanalysisshowedthatwhenthecultureswereexposedtohypoxiaformorethan2hours,thePo2was4.5to5.4Kpa.WeverifiedRMVECscouldbeinfectedwithAd-15-LOX-1orAd-vectorviaFluorescencemicroscopy.CCK-8analysisrevealedthattheproliferativecapacitiesofRMVECsinhypoxicgroupweresignificantlyhigherateachtimepointthantheywereinnormoxicgroup(P<0.05).Inahypoxiccondition,theproliferativecapacitiesofRMVECsin15-LOX-1groupweresignificantlyinhibited(P<0.05).Real-timeRT-PCRanalysisrevealedthattheexpressionsofVEGF-A,VEGF-R2andeNOsmRNAincreasedinhypoxiagroupcomparedwithnormoxiagroup(P<0.01).However,theexpressionsof15-LOX-1,PPAR-rmRNAdecreasedinhypoxiagroupcomparedwithnormoxiagroup(P<0.01).Italsoshowedthatinahypoxiccondition,theexpressionsofVEGF-A,VEGF-R2andeNOsmRNAdecreasedsignificantlyin15-LOX-1groupcomparedwithhypoxiagroup(P<0.01).However,15-LOX-1andPPAR-rmRNAincreasedsigni
简介:AIMTo学习discoidin象我一样domaincontaining蛋白质3的效果(EDIL3)在人的透镜的增长和上皮间充质的转变(EMT)上的弄空上皮的房间(LEC).METHODSRNA干扰被用来在vitro在人的LEC禁止EDIL3的表示。房间的形态学用一台转换显微镜被观察。房间增长用EdU工具包被估计。房间移植用Transwell房间被调查,LEC的EMT用共焦的显微镜并且西方的弄污被估计。转变生长因素(TGF)小径用数据显示出的西方的blotting.RESULTSThe被调查那个silencingEDIL3表达式改变了LEC形态学并且压制了LEC增长(P<0.05)并且移植(P<0.01)。而且,西方的弄污的结果证明那EDIL3弄空减少了光滑的肌肉肌动朊的表示(-SMA)(P<0.001)并且vimentin(P<0.01),当增加时E-cadherin的表示(P<0.001)。EDIL3弄空能压制Smad2的phosphorylation(P<0.01)并且Smad3(P<0.01)并且exracellular信号的激活调整了kinase(英皇家空军之阶级最低之兵)(P<0.05).CONCLUSIONThe调查结果显示EDIL3可能经由TGF小径在LEC参予增长和EMT并且可以是为以后的囊opacification的处理的一个潜在的治疗学的目标。
简介:研究显示,趋化因子受体3(chemokinereceptor3,CCR3)在眼部主要分布于视网膜色素上皮细胞中,亦表达于脉络膜血管内皮细胞(CECs)中。CCR3的特异性高表达在湿性年龄相关性黄斑变性(age-relatedmaculardegeneration,AMD)中被发现,并被证明在湿性AMD患者脉络膜新生血管(choroidalneovascularization,CNV)的产生中具有重要作用。本文拟对CCR3的结构、功能、目前研究存在的问题及未来的研究方向做一综述。相信随着对CCR3研究的进一步深入,必将帮助我们寻找到一种湿性AMD诊断和治疗的新方法,同时也可能对其它CNV性疾病研究以及新的抗CNV药物提供重要参考。
简介:目的在视网膜缺血再灌注模型上,观察再灌注后视网膜内caspase-3的动态变化,探讨caspase-3与视网膜细胞凋亡的关系。方法结扎大鼠左侧颈总动脉1h,然后再灌注,检测再灌注后1、6、12、24、48、72h大鼠视网膜内caspase-3的水平及视网膜细胞凋亡平均发生率。结果再灌注后视网膜内caspase-3的表达出现在光感受器细胞层,在再灌注后1、6、12、24、48、72hcaspase-3平均光密度分别为0.067±0.004、0.923±0.045、1.962±0.377、3.793±0.860、2.039±0.427、1.332±0.109,细胞凋亡平均发生率(%)分别为1.8±0.1,7.1±0.2,18.2±1.4,34.7±2.1,22.6±0.9,16.3±0.4。结论再灌注后大鼠视网膜caspase-3表达与细胞凋亡呈现正相关,caspase-3可以促进视网膜细胞凋亡的发生。
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:AIM:Toinvestigatethediffusioncharacteristicsofwaterofopticnerveandopticradiationinhealthyadultsanditsrelatedfactorsbydiffusiontensorimaging(DTI)at3T.METHODS:Atotalof107healthyvolunteersperformedheadconventionalMRIandbilateralopticnerveandopticradiationDTI.TheprimarydataofDTIwasprocessedbypost-processingsoftwareofDTIstudio2.3,obtainingfractionalanisotropyvalue,meandiffusivityvalue,principalenginevalue,orthogonalenginevaluebymeasuring,andanalyzedbytheSPSS13.0statisticalsoftware.RESULTS:Thebilateralopticnerveandopticradiationfiberspresentedgreencolorindirectionalencodedcolor(DEC)mapsandpresentedhighsignalinfractionalanisotropy(FA)maps.TheFAvalueoftheleftopticnervewas0.598±0.069andtherightwas0.593±0.065;themeandiffusivity(MD)valueoftheleftopticnervewas(1.324±0.349)×10-3mm2/sandtherightwas(1.312±0.350)x10-3mm2/s;theprincipalenginevalue(λ?)oftheleftopticnervewas(2.297±0.522)×10-4mm2/sandtherightwas(2.277±0.526)×10-3mm2/s;theorthogonalenginevalue(λ⊥)oftheleftopticnervewas(0.838±0.285)×10-3mm2/sandtherightwas(0.830±0.280)×10-3mm2/s;theFAvalueoftheleftopticradiationwas0.636±0.057andtherightwas0.628±0.056;themeandiffusivity(MD)valueoftheleftopticradiationwas(0.907±0.103)×10-3mm2/sandtherightwas(0.889±0.125)×10-3mm2/s;theprincipaleigenvalue(λⅡ)oftheleftopticradiationwas(1.655±0.210)×10-3mm2/sandtherightwas(1.614±0.171)×10-3mn2/s;theorthogonalenginvalue(λ⊥)oftheleftopticradiationwas(0.531±0.103)×10-3mm2/sandtherightwas(0.524±0.152)×10-3m
简介:目的:对比分析伴虹膜异色的青光眼睫状体炎综合征与Fuchs综合征患者的临床表现及特点,对两种疾病的鉴别要点作出分析及总结,利于临床医生对这两种疾病进行鉴别诊断。方法:对本院自2000年至今收治住院的3例伴虹膜异色的青光眼睫状体炎综合征(Posner-Schlossman’ssyndrome,PSS)患者进行详细的临床检查及研究,根据其临床资料总结出该病的临床特点,并对伴虹膜异色的PSS与Fuchs综合征的鉴别要点作出分析及总结。结果:共收集了3例伴虹膜异色的PSS患者,均为单眼反复发病,病程均在10a以上,发作时眼压明显升高,可达30.00~60.00mmHg,持续时间一般为3~7d,可自行缓解或用药后缓解,间歇期眼压正常,该3例患者除了具有典型的PSS表现外,虹膜均轻度异色。总结了该病在发作期及间歇期的临床表现,从发病情况、眼压波动情况、KP形态、晶状体情况、眼底及视功能情况详细论述了伴虹膜异色的青光眼睫状体炎综合征与Fuchs综合征的区别,并针对两种疾病的治疗提出了不同建议。结论:结合我们所提出的鉴别要点,对患者进行详细的临床检查,可以对这两种疾病进行鉴别诊断。
简介:AIM:Toassesstheeffectofmyopiaonthethicknessofretinalnervefiberlayer(RNFL)measuredby3Dopticalcoherencetomography(3D-OCT)inagroupofnonglaucomatousChinesesubjects.METHODS:Twohundredandfifty-eighteyesof258healthyChinesemyopicindividualswererecruitedandfourgroupswereclassifiedaccordingtotheirsphericalequivalent(SE):lowmyopia(n=42,-0.50
简介:AIM:ToIntroduceanewspecializedvisualacuitychartforamblyopicchildrenaged3-5yearsoldanditsclinicalapplications.METHODS:ThenewvisualacuitychartandnotationsweredesignedbasedonWeber-Fechnerlaw.Theoptotypeswereredagainstawhitebackgroundandwerespeciallyshapedfourbasicgeometricsymbols:circle,square,triangle,andcross.Aregulargeometricprogressionoftheoptotypesizesanddistributionwasemployedtoarrangein14lines.Theprogressionrateoftheoptotypesizebetweentwolineswas1.2589andthetestingdistancewas3m.VisualacuityscorecouldberecordedaslogMARnotationordecimalnotation.Agestratifieddiagnosticcriteriaforamblyopiaestablishedbyconsensusstatementondiagnosisofamblyopia(2011)amongmembersoftheStrabismusandPediatricOphthalmologyGroup,OphthalmologySociety,ChineseMedicalAssociation(SPOGOSCMA)wereillustratedinthenewvisualacuitychart.RESULTS:Whenassessingvisualacuityinchildrenaged3-5yearsold,thisnewvisualacuitychartthatconsistsoffoursymmetricalshapes(triangle,square,cross,andcircle)overcameaninabilitytorecognizethelettersofthealphabetanddifficultiesindesignatingthedirectionofblackabstractsymbolssuchasthetumbling’E’orLandolt’C’,whichthesubjectswerepronetoloseinterestin.Thevisualacuityscoremayberecordedindifferentnotations:decimalacuityandlogMAR.Thesetwonotationscanbeeasilyconvertedeachotherintheneweyechart.Themeasurementsofthisnewchartnotonlyshowedasignificantcorrelationandagoodconsistencywiththeinternationalstandardlogarithmicvisualacuitychart(r=0.932,P<0.01),butalsoindicatedahightest-retestreliability(89%ofretestscoreswerewithin0.1logMARunitsoftheinitialtestscore).CONCLUSION:Theresultsofthisstudysupportthevalidityandreliabilityofdistancevisualacuitymeasurementsusingtheneweyechartinchildrenaged3to5yearsoverawiderangeofvisualacuities,andtheneweyechartisgreatforearl
简介:AIM:ToidentifythegeneticdefectinaChinesefamilywithbilateralprogressivechildhoodposteriorcataract.METHODS:Atwo-generationfamilywasrecruitedinthisstudy.Familyhistoryandclinicaldatawererecorded.AllreportedcandidategenesassociatedwithcongenitalposteriorcataractwerescreenedbydirectDNAsequencing.·RESULTS:Allaffectedindividualspresentedposterioropacitiesinthelens.Directsequencingofthecandidategenesshowedaheterozygousc.2668C>TvariationinEPHA2gene,whichresultedinthereplacementofargininebycysteineatcodon890(p.R890C).Thismutationwasfoundintwoaffectedindividuals,butwasnotobservedin200normalcontrols.·CONCLUSION:Wereportanovelmutation(p.R890C)intheEPHA2receptortyrosinekinasegene.ThefindingexpandsthemutationspectrumofEPHA2inassociationwithposteriorcataract.
简介:目的观察1,25(OH)2D3对高糖诱导牛视网膜血管内皮细胞(BRECs)中血管内皮生长因子(VEGF)的表达水平变化及对细胞凋亡水平的影响。方法将分离培养的BRECs分为三组,分别为正常糖组、高糖组和高糖处理组。正常对照组细胞培养液含5mmol/L葡萄糖,高糖组细胞培养液含30mmol/L葡萄糖,高糖处理组细胞培养液含30mmol/L葡萄糖和50nM,1,25(OH)2D3。培养48h后收集细胞蛋白。蛋白免疫印迹法检测细胞VEGF及细胞凋亡相关蛋白Bax和Bcl-2表达水平;PI/Hoechst双染色法检测细胞凋亡。结果相比于正常糖组,高糖组中VEFG水平和Bax/Bcl-2比值显著增加;而在高糖处理组中表达水平远远低于高糖组,差异均有统计学意义(P〈0.05)。高糖的细胞凋亡水平高于正常糖组,而经过1,25(OH)2D3处理后,其细胞凋亡水平则有所下降,差异均有统计学意义(P〈0.05)。结论1,25(OH)2D3可以抑制高糖诱导BRECs中VEGF表达增加及细胞凋亡。
简介:AIM:Topresenttheoutcomeofmodifiedgridlaserphotocoagulation(GLP)indiffusediabeticmacularedema(DDME)ineyeswithoutextrafovealand/orvitreofovealtraction.METHODS:InclusioncriteriafortheretrospectivestudywereDDMEeyesofpatientswithtypeⅡdiabetesmellitusthathad≥4monthsoffollow-upfollowingGLP.Onlyoneeyeperpatientwasanalyzed.Using3-Dspectral-domainopticalcoherencetomography(3-DSDOCT),eyesthathadeitherextrafovealorvitreofovealtraction,orhadbeenpreviouslytreatedbyanintravitrealmedication(s)wereexcluded.TreatedDDMEeyesweredividedinto4groups:A)'Classic'DDMEthatinvolvedthecentralmacula;B)edemadidnotinvolvethemacularcenter;C)eyesassociatedwithcentralepiretinalmembrane(ERM);D)DDMEthatwasassociatedwithmacularcapillarydropout≥2disc-diameter(DD).RESULTS:GLPoutcomein35DDMEeyesafter4-24(mean,13.1±6.9)monthswasasfollows:GroupA)18eyeswith'classic'DDME.Followingoneor2(mean,1.2)GLPtreatments,best-correctedvisualacuity(BCVA)improvedby1-2Snellenlinesin44.4%(8/18)ofeyes,andworsenedby1linein11.1%(2/18).Centralmacularthickness(CMT)improvedby7%-49%(mean,26.6%)in77.8%(14/18)ofeyes.CausesofCMTworsening(n=4)werecommonlyexplainable,predominantly(n=3)associatedwithemergenceofextrafovealtraction,5-9monthspost-GLP.GroupB)GLP(s)inDDMEthatdidnotinvolvethemacularcenter(n=6)resultedinimprovedBCVAby1-2linesin2eyes.However,thecentralmaculabecameinvolvedintheedemaprocessaftertheGLPin3(50%)eyes,associatedwithanemergenceofextrafovealtractioninoneoftheseeyes4monthsfollowingtheGLP.GroupC)GLPfailedinall5eyesassociatedwithcentralERM.GroupD)GLPwasofpartialbenefitin2of6treatedeyeswithmacularcapillarydropout≥2DD.CONCLUSION:EyeswithDDMEthatinvolvedthemacularcenterwerefoundtoachievefavourableoutcomesafterGLP(s)duringmid-termfollow-up,unlesscomplicatedpre-GLPorpost-GLPbyvltreoretinalinterfaceabnormalities,oftenextrafovealtra