简介：AIM:ToinvestigatetheroleofRho-associatedproteinkinase(ROCK)inhibitor,Y27632,inmediatingtheproductionofextracellularmatrix(ECM)componentsincludingfibronectin,matrixmetallo-proteinase-2(MMP-2)andtypeIcollagenasinducedbyconnectivetissuegrowthfactor(CTGF)ortransforminggrowthfactor-β(TGF-β)inahumanretinalpigmentepithelialcellline,ARPE-19.METHODS:TheeffectofY27632ontheCTGForTGF-βinducedphenotypeinARPE-19cellswasmeasuredwithimmunocytochemistryasthechangeinF-actin.ARPE-19cellsweretreatedwithCTGF(1,10,100ng/mL)andTGF-β(10ng/mL)inserumfreemedia,andanalyzedforfibronectin,laminin,andMMP-2andtypeIcollagenbyRT-qPCRandimmunocytochemistry.CellswerealsopretreatedwithanROCKinhibitor,Y27632,toanalyzethesignalingcontributingtoECMproduction.·RESULTS:TreatmentofARPE-19cellsinculturewithTGF-βorCTGFinducedanECMchangefromacobblestonemorphologytoamoreelongatedswirlpatternindicatingamesenchymalphenotype.RT-qPCRanalysisanddifferentgeneexpressionanalysisdemonstratedanupregulationinexpressionofgenesassociatedwithcytoskeletalstructureandmotility.CTGForTGF-βsignificantlyincreasedexpressionoffibronectinmRNA(P=0.006,P=0.003respectively),lamininmRNA(P=0.006,P=0.005),MMP-2mRNA(P=0.006,P=0.001),COL1A1mRNA(P=0.001,P=0.001),COL1A2mRNA(P=0.001,P=0.001).PreincubationofARPE-19withY27632(10mmol/L)significantlypreventedCTGForTGF-βinducedfibronectin(P=0.005,P=0.003respectively),MMP-2(P=0.003,P=0.002),COL1A1(P=0.006,P=0.003),andCOL1A2(P=0.006,P=0.004)geneexpression,butnotlaminin(P=0.375,P=0.516).CONCLUSION:OurstudydemonstratedthatbothTGF-βandCTGFupregulatetheexpressionofECMcomponentsincludingfibronectin,laminin,MMP-2andtypeIcollagenbyactivatingtheRhoA/ROCKsignalingpathway.Duringthisprocess,ARPE-19cellswereshowntochangefromanepithelialtoamesenchymalphenotypeinvi

简介：AIM:ToInvestigatetheeffectsoftransforminggrowthfactorβ2(TGF-β2)andconnectivetissuegrowthfactor(CTGF)ontransdifferentiationofhumanlensepithelialcells(HLECs)culturedinvitroandsynthesisofextracellularmatrix(ECM).METHODS:HLECsweretreatedwithTGF-β2(0,0.5,1.0,5,10μg/L)andCTGF(0,15,30,60,100μg/L)fordifferenttimes(0,24,48,72h)invitroandtheexpressionofα-smoothmuscleactin(α-SMA),themaincomponentoftheextracellularmatrixtypeⅠcollagen(Col-1)andfibronectin(Fn)weremeasuredbyusingreal-timepolymerasechainreaction(PCR)andwestern-blot.RESULTS:TGF-β2andCTGFsignificantlyincreasedexpressionofα-SMAmRNAandprotein(P<0.05,P<0.001),FnmRNAandprotein(P<0.001),Col-1mRNAandprotein(P<0.001).TGF-β2couldinduceHLECsexpressionofCTGFmRNAandproteinindosedependentmanner(P<0.05,P<0.001).TGF-β2andCTGFcouldinduceHLECstoexpressα-SMA,FnandCol-1intime-dependentmanner.EachtimeofTGF-β2andCTGFinducedHELCsexpressionofα-SMA,Fn,Col-1mRNAandproteinwassignificantincreasecomparedwithcontrol(P<0.05,P<0.001).CONCLUSION:TGF-β2andCTGFcouldinduceHLECsepithelialmesenchymaltransitionandECMsynthesis.

简介：AIM:Tomeasurecentralcornealthickness(CCT)andpre-cornealtearfilmthicknessusingtheGalileidualScheimpfluganalyzer(GSA)inNewZealandWhiterabbits.METHODS:TennormalNewZealandWhiterabbits(20eyes)wereincludedinthisstudy.Withtheassistanceof0.1%fluorescein,thepre-cornealtearfilmcanbewellvisualized.BotheyesofeachrabbitwerescannedoncewiththeGSApre-andpost-instillationof1μL0.1%fluorescein.ThedifferencebetweenthetwomeasurementsofCCT(4-mmdiameter)wasrecordedasthepachymetricvaluesofthecentraltearfilm.RESULTS:TheCCTofpre-andpost-instillationwas388.8±9.5μmand407.0±10.5μm,respectively.Afterapairedt-testanalysis,thecentralpre-cornealtearfilmthicknessof4mmdiameterwas18.2±5.31μmwitha95%confidenceintervalof(15.7,20.6)μm(P<0.001).CONCLUSION:GSAcanbeusedtomeasureCCTandanalyzecentraltearfilmthicknessofrabbitswiththehelpoffluorescein.

简介：AIM:Toevaluatetheaccuracyofsphericalequivalent(SE)estimatesofadouble-passsystemandtocompareitwithretinoscopy,subjectiverefractionandatablemountedautorefractor.METHODS:Non-cycloplegicrefractionwasperformedon125eyesof65healthyadults(age23.5±3.0years)fromOctober2010toJanuary2011usingretinoscopy,subjectiverefraction,autorefraction(AutokeratorefractometerTOPCONKR-8100,Japan)andadoublepasssystem(OpticalQualityAnalysisSystem,OQAS,VisiometricsS.L.,Spain).Nineconsecutivemeasurementswiththedouble-passsystemwereperformedonasubgroupof22eyestoassessrepeatability.ToevaluatethetruenessoftheOQASinstrument,theSElaboratorybiasbetweenthedoublepasssystemandtheothertechniqueswascalculated.RESULTS:TheSEmeancoefficientofrepeatabilityobtainedwas0.22D.SignificantcorrelationscouldbeestablishedbetweentheOQASandtheSEobtainedwithretinoscopy(r=0.956,P<0.001),subjectiverefraction(r=0.955,P<0.001)andautorefraction(r=0.957,P<0.001).ThedifferencesinSEbetweenthedouble-passsystemandtheothertechniquesweresignificant(P<0.001),butlackedclinicalrelevanceexceptforretinoscopy;Retinoscopygavemorehyperopicvaluesthanthedouble-passsystem-0.51±0.50Daswellasthesubjectiverefraction-0.23±0.50D;Moremyopicvalueswereachievedbymeansofautorefraction0.24±0.49D.CONCLUSION:Thedouble-passsystemprovidesaccurateandreliableestimatesoftheSEthatcanbeusedforclinicalstudies.Thistechniquecandeterminethecorrectfocuspositiontoassesstheocularopticalquality.However,ithasarelativelysmallmeasuringrangeincomparisonwithautorefractors(-8.00to+5.00D),andrequirespriorinformationontherefractivestateofthepatient.