简介:AIM:ToinvestigatetheroleofBrn-3bindifferentiationprocessofstemcellsderivedfromretinalMiillercellsintotheganglioncell.METHODS:ThepassageculturemethodofMiillercellsfromretinaofnewbornSpragueDawleyratswascarriedoutbyrepeatedincompletepancreaticenzymedigestionmethod.Thecellsweredetectedbyfluorescenceactivatedcellsorter(FACS),immunohistochemistrytechnologyandreversetranscription-polymerasechainreaction(RT-PCR)todeterminethepurity.Thethirdpassageofcellswasinducedintheserum-freededifferentiationmedium.TheexpressionofthespecificmarkersKi-67andnestinofretinalstemcellswasmeasuredbyRT-PCRandWesternblot.Thecellproliferationofretinalstemcellswasdetectedby5-Ethynyl-2’-deoxyuridine(Edu)staining.Thecellswererandomlydividedinto5groupsasfollows:groupA:Brn-3bsiRNAgroup;groupB:Brn-3bcontrolsiRNAgroup;groupC:pGC-Brn-3b-greenfluorescentprotein(GFP)group;groupD:pGC-GFPgroup;groupE:controlgroup(withoutanyhandling).ThepurifiedMullercellswereculturedfor3-7d,then,thepercentageofganglioncellswascountedbyimmunofluorescencestaining.RESULTS:FACSdemonstratedthepurityofretinalMullercellswasmore97.44%.Afewsphericalcellspheresappeared.Immunofluorescencestainingshowedthatstemcellswithinthesphereswerepositiveforretinalstemcell-specificmarkersnestin(redfluorescence,92.94%±6.48%)andKi-67(greenfluorescence,85.96%±6.04%).Meanwhile,RT-PCRanalysisshowedcellspheresintheculturetohaveexpressedabatteryoftranscriptscharacteristicofstemcellssuchasnestinandKi-67,whichwereabsentintheMullercells.WesternblotanalysisfurtherconfirmedtheexpressionofnestinandKi-67inthecellspheresbutnotintheMullercells.Edustainingshowedmostofthenucleiwithinthecellsphereswerestainedred(82.80%±6.65%),suggestingthenewcellsphereshadthecapacityforeffectiveproliferation.ThestatisticsresultshowedthedifferencebetweenBrn-3bsiRNAgroupand
简介:目的探讨桥小脑角疾病的相位平衡快速梯度回波(Balance-FastFieldEcho,B-FFE)序列MRI影像学特点,评价B-FFE序列对桥小脑角疾病的诊断价值.方法应用B-FFE序列对136例桥小脑角病变者(三叉神经痛131例、半面痉挛5例)进行MRI检查,观察桥小脑角神经、血管、肿瘤显像情况,并与手术中资料进行比较.结果131例三叉神经痛患者中,117例为原发性三叉神经痛,MRI显示有血管压迫者102例,术中证实有血管压迫者95例,行显微血管减压+神经梳理术;14例为继发性三叉神经痛,均为桥小脑角占位性病变,术后病理证实7例为胆脂瘤,2例为听神经瘤,5例为脑膜瘤;5例半面痉挛患者MRI显示有血管压迫,术中证实有血管压迫,并予以血管减压.结论B-FFE序列MRI成像能清楚显示桥小脑角区三叉神经、面神经与血管的关系及是否有占位性病变,对诊断、术前评估和指导治疗有重要意义.
简介:目的:研究αB-晶体蛋白对大鼠急性高眼压后视网膜组织中αB-晶体蛋白含量,视网膜神经节细胞(retinalganglioncells,RGCs)中生长相关蛋白-43(growthassociatedprotein-43,GAP-43)表达和全视野视网膜电流图(full-fieldERG,F-ERG)的b波振幅差异,探讨αB-晶体蛋白对急性高眼压后RGCs轴突再生的影响。方法:采用随机分组设计的实验研究。本实验选择120只健康、无眼疾SD大鼠,随机分为以下4组:αB晶体蛋白组(αB)30只,生理盐水组(S)30只,假手术组(P)30只,急性高眼压组(H)30只,均以右眼为实验眼,分别于术后7d和14d各取5只术眼的视网膜,行Western-blot法,观察急性高眼压后视网膜中αB-晶体蛋白的表达;术后7,14,21d各取5只术眼的视网膜,行免疫组织化学法,观察急性高眼压后RGCs的GAP-43表达;术前和术后1mo时应用全视野ERG的b波振幅变化测定视网膜功能。组间数据比较采用单因素方差分析(One-wayANOVA),组间两两比较采用SNK-q检验。结果:αB组视网膜中αB-晶体蛋白的表达于术后7d较高,14d时表达减弱(P=0.000),但同一时间点明显高于其他三组(P=0.006,P=0.024,P=0.007;P=0.006,P=0.008,P=0.010);αB组RGCs的GAP-43表达于术后7d达高峰,14d时表达减弱,21d时仍有少量表达(P=0.000),但各时间点明显高于其他三组(P=0.001,P=0.002,P=0.001;P=0.015,P=0.002,P=0.006;P=0.005,P=0.003,P=0.005);ERG-b波振幅于术后1mo较低(P=0.014,P=0.004,P=0.003,P=0.006),其中术前各组ERG-b波振幅无明显差异(P=0.993),术后1mo时αB组ERG-b波振幅明显高于其他三组(P=0.000,P=0.004,P=0.002)。结论:外源性αB-晶体蛋白能够提高视网膜αB-晶体蛋白的表达;αB-晶体蛋白通过促进RGCs中GAP-43的表达,从而促进RGCs的轴突再生;αB-晶体蛋白可促进视网膜功能的恢复,对大鼠视网膜无毒性作用。
简介:目的:探讨眼附属器B细胞非霍杰金淋巴瘤(B—cellnon-Hodgkinlymphoma,NHL)中Skp2,p27和PTEN的表达。方法:收集1995年到2011年青岛大学附属医院眼科石蜡包埋标本,用免疫组化法分别检测眼附属器B细胞NHL(n=30)标本中Skp2,p27和PTEN的表达,以眼部反应性淋巴组织增生(n=10)作为对照组。以患者的年龄、性别、发病部位,病理类型作为眼附属器B细胞NHL的的分类标准。结果:Skp2,p27和PTEN的表达与患者的年龄、性别、发病部位无关,而与病例类型有关。眼附属器B细胞NHLSkp2表达率与眼部反应性淋巴组织增生相比显著增高。p27,PTEN表达率与反应性淋巴组织增生相比显著降低。随眼附属器B细胞NHL病理分级的提高,Skp2的表达显著增高,p27和PTEN的表达显著降低。在黏膜相关淋巴组织结(mucosa—associatedlymphoidtissue,MALT)外边缘区B细胞淋巴瘤(diffuselargeB—celllymphoma,DLBCL)中,Skp2分别与p27,VFEN成负相关,p27和PTEN成正相关。结论:Skp2的表达升高,p27,PTEN蛋白的缺失以及可能与眼附属器B细胞NHL的发生有关;其中在MALT外边缘区DLBCL中,三种蛋白存在相关性。联合三种蛋白的检测眼附属器B细胞NHL的不同病理类型有重要意义。
简介:AIM:Todeterminewhethersinglenucleotidepolymorphism(SNP)rs641153isassociatedwiththeriskofage-relatedmaculardegeneration(AMD),weperformedasystematicmeta-analysisof15eligiblestudies.SNPinthecomplementfactorB(CFB)geneisconsideredtohavesignificantassociationwithAMDsusceptibility,butthereisgreatdiscrepancyintheseresults.METHODS:TheeligiblestudieswereidentifiedbysearchingthedatabasesofPubMed,EMBASE,andWebofScience.Oddsratios(ORs)with95%confidenceintervals(CIs)wereusedtoassesstheassociation.AlldatawereanalyzedusingStatasoftware.RESULTS:Theassociationbetweenrs641153andAMDriskwasstatisticallysignificantunderthehomozygousmodel(AAvsGG:OR=0.26,95%CI=0.15-0.45,P_h=0.973,/~2=0.0%,fixedeffects),dominantmodel(AA+GAvsGG:OR=0.49,95%CI=0.40-0.59,P_h=0.004,/~2=56.4%,randomeffects)andrecessivemodel(AAvsGA+GG:OR=0.30,95%CI=0.17-0.51,R_n=0.983,I~2=0.0%,fixedeffects).Thesameresultswerealsoobservedinthestratifiedanalysesbyethnicity,sourceofcontrolandsamplesize.CONCLUSION:Ourmeta-analysissuggeststhatrs641153intheCFBgenemayplayaprotectiveroleinAMDsusceptibility,thelateAMDinparticular,bothinCaucasiansandinAsians.
简介:AIM:Anaerobicbacteriacancauseocularinfections.WetestedtheOxyPlateTMAnaerobicSystem(OXY)toisolatepertinentanaerobicbacteriathatcancauseoculardisease.METHODS:OXY,whichdoesnotrequiredirectanaerobicconditions(i.e.bags,jars),wascomparedtoconventionalisolationofincubatingculturemediainanaerobicbags.Standardcoloniescountswereperformedonanaerobicocularbacterialisolatesunderaerobicandanaerobicconditions(anaerobicbags)usingagarmedia:1)OXY(aerobiconly),2)5%sheepblood(SB),3)Chocolate,and4)Schaedler.Thebacteriatestedwerede-identifiedocularisolatesculturedfromendophthalmitisanddacryocystitisthatinclude10Propionibacteriumacnesand3Actinomycesspecies.Thecolonycountsforeachbacteriaisolate,oneachculturingcondition,wererankedfromlargesttosmallest,andnon-parametricallycomparedtodeterminethebestculturingcondition.RESULTS:Allanaerobicconditionswerepositiveforalloftheanaerobicisolates.SBandSchaedler’sagarunderaerobicconditionsdidnotsupportthegrowthofanaerobicbacteria.SparsegrowthwasnotedonchocolateagarwithPropionibacteriumacnes.Asananaerobicsystem,SBinananaerobicbagisolatedhighercolonycountsthanOXY(P=0.0028)andchocolateagar(P=0.0028).CONCLUSION:AlthoughOXYdidnottesttobemoreefficientthanotheranaerobicsystems,itappearstobeareasonablealternativeforisolatinganaerobicbacteriafromocularsites.Theuseofanagarmediuminaspeciallydesignedplate,withouttherequirementofananaerobicbag,renderedOXYasanadvantageoverotheranaerobicsystems.
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