简介:Thedamageofhumancornealcellsencounterwiththeproblemofavailabilityofcornealcellsforreplacement.Limitationofthesourceofcornealcellshasbeenrealized.Anattemptofdevelopmentofcornealepithelial-likecellsfromthehumanskin-derivedprecursor(hSKPs)hasbeenmadeinthisstudy.Combinationofthreeessentialgrowthfactors:epidermalgrowthfactor(EGF),keratinocytegrowthfactor(KGF)andhepatocytegrowthfactor(HGF)coulddemonstratesuccessfullyinductionofhSKPstodifferentiationintocornealcells.Theinducedcellsexpressedtheappearanceofmarkersofcornealepithelialcellsasshownbythepresenceofkeratin3(K3)byantibodylabelandWesternblotassay.TheK3geneexpressionofinducedhSKPscellsasshownbyreversetranscription-polymerasechainreaction(RT-PCR)technologywasalsodemonstrated.ThepresenceofthesemarkersatbothgeneandproteinlevelscouldleadtoourconclusionthatthedirectionaltransdifferentiationofhSKPscellsintocornealepithelialcellswassuccessfullydoneunderthiscellinductionprotocol.Thefindingshowsanewlyavailablestemcellsourcecanbeobtainedfromeasilyavailableskin.Cellsfromautologoushumanskinmightbeusedforcornealdisordertreatmentinfutureclinicalapplication.
简介:AIM:Toestablishanuntransfectedhumancornealstromal(HCS)celllineandcharacterizeitsbiocompatibilitytoacellularporcinecornealstroma(aPCS).·METHODS:PrimaryculturewasinitiatedwithapurepopulationofHCScellsinDMEM/F12media(pH7.2)containing20%fetalbovineserumandvariousnecessarygrowthfactors.Theestablishedcelllinewascharacterizedbygrowthproperty,chromosomeanalysis,tumorigenicityassay,expressionofmarkerproteinsandfunctionalproteins.Furthermore,thebiocompatibilityofHCScellswithaPCSwasexaminedthroughhistologicalandimmunocytochemistryanalysesandwithlight,electronmicroscopies.·RESULTS:HCScellsproliferatedtoconfluence2weekslaterinprimarycultureandhavebeensubculturedtopassage140sofar.AcontinuousuntransfectedHCScelllinewithapopulationdoublingtimeof41.44hoursatpassage80hasbeendetermined.Resultsofchromosomeanalysis,morphology,combinedwiththeresultsofexpressionofmarkerproteinandfunctionalproteinssuggestedthatthecellsretainedHCScellproperties.Furthermore,HCScellshavenotumorigenicity,andwithexcellentbiocompatibilitytoaPCS.·CONCLUSION:Anuntransfectedandnon-tumorigenicHCScelllinehasbeenestablished,andthecellsmaintainedpositiveexpressionofmarkerproteinsandfunctionalproteins.Thecellline,withexcellentbiocompatibilitytoaPCS,mightbeusedforinvitroreconstructionoftissue-engineeredHCS.
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:AIM:Todemonstratethemorphologyandstructureofinvitroreconstructedtissue-engineeredhumancornealepithelium(TE-HCEP)withseedercellsfromanuntransfectedHCEPcellline.·METHODS:TheTE-HCEPswerereconstructedinvitrowithseedercellsfromanuntransfectedHCEPcellline,andscaffoldcarriersofdenudedamnioticmembrane(dAM)inair-liquidinterfaceculturefor3,5,7and9days,respectively.Thespecimenswereexaminedwithhematoxylin-eosin(HE)stainingofparaffin-section,immunocytochemicalstaining,scanningandtransmissionelectronmicroscopy.·RESULTS:DuringinvitroreconstructionofTE-HCEP,HCEPcellsformeda3-4,6-7and8-10layersofanHCEP-likestructureondAMsinair-liquidinterfaceculturefor3,5and7days,respectively.Butthecellsdeceasedto5-6layersandthestructureofstraifiedepitheliumbecamelooseatday9.Andthecellsmaintainedpositiveexpressionofmarkerproteins(keratin3andkeratin12),cell-junctionproteins(zonulaoccludens-1,E-cadherin,connexin43andintegrinβ1)andmembranetransportproteinofNa+-K+ATPase.TheHCEPcellsinTE-HCEPwererichinmicrovillionapicalsurfaceandestablishednumerouscell-cellandcell-dAMjunctionsatday5.·CONCLUSION:ThemorphologyandstructureofthereconstructedTE-HCEPweresimilartothoseofHCEPinvivo.TheHCEPcellsinthereconstructedTE-HCEPmaintainedthepropertiesofHCEPcells,includingabilitiesofformingintercellularandcell-extracellularmatrixjunctionsandabilitiesofperformingmembranetransportation.TheuntransfectedHCEPcellsanddAMscouldpromisinglybeusedinreconstructionHCEPequivalentforclinicalcornealepitheliumtransplantation.
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.
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:·Thisisacasepresentationofaverybizarreopenglobetraumawithanteriorsegmentforeignbody-fishinghookstuckinthecorneaandiris.Complicationsduetothiskindofeyetraumamightbeveryhazardousandwithseriousimpactonvisualfunction.Wearerepresentingourapproachandexperienceofthreestepmanagementofthiskindofeyeinjury:first-extracttheforeignbody,closeandreconstructtheeyeball,second-fightinflammation,andthird-restorethevisualfunctionbycataractsurgery.·
简介:AIM:Tocomparetheeffectoftopicallyadministeredandsubconjunctivallyinjectedbevacizumabonexperimentalcornealneovascularizationinratsfortwoweeksaftertreatment.METHODS:Twenty-eightSprague-Dawleyratsweredividedintofourgroupsof7animals.Eachcornealcenterofrighteyewascauterizedwithsilver/potassiumnitratefor8s.Aftercornealburning,bevacizumab(12.5mg/mL)wastopicallyadministeredthreetimesperday(TBgroup)fortwoweeksorsubconjunctivallyinjectedondays2and4aftercauterization(0.02mL;SBgroup).Asnegativecontrols,ratsreceived0.9%salinetopicallythreetimesperday(TSgroup)orsubconjunctivallyondays2and4(0.02mL;SSgroup).Digitalphotographsofthecorneaweretaken1and2weeksaftertreatmentandanalyzedtodeterminetheareaofcorneacoveredbyneovascularizationasthepercentageofcornealneovascularization.RESULTS:Oneweekaftertreatment,thepercentageofcornealneovascularizationwassignificantlylowerintheTBandSBgroupsthanintheTSandSSgroups(allP<0.05).Twoweeksaftertreatment,thepercentageofcornealneovascularizationwassignificantlylowerintheTBgroupthanintheTSgroup(P<0.05).Inallgroups,thepercentageofneovascularizationwasdecreasingastimepassed(allP<0.05)CONCLUSION:Topicallyadministeredbevacizumabhaslongerstandinganti-angiogeniceffectthansubconjunctivallyinjectedbevacizumabincornealneovascularizationfollowingchemicalinjuryinrats.