简介:AIM:Todeterminetheproliferativepotentialandthemaintenanceofstemcellactivityinstoredhumanlimbaltissues,andcorrelatethiswiththepreservationtime,cellviabilityandtheexpressionofstemcellmarkers.METHODS:Thirtylimbalrimsweresplitinto4partsandstoredincornealpreservationmediumat4℃for0,1,4,or7days.ThelimbalstemcellandmitoticmarkersP63,CK19,proliferatingcellnuclearantigen(PCNA),andKi67weredeterminedbyimmunohistochemicalstaining.Theproliferativepotentialoflimbalepithelialcellswasassessedbycellviability,theabilityofgeneratingstratifiedepithelium,andcolonyformingassay.RESULTS:Thestoredtissuesmaintainedlimbalstratifiedstructureto7daysandexhibitedcomparableexpressionlevelofstemcellandmitoticmarkers.Theproportionofviablecellsdecreasedwiththeprolongedpreservationtime,whilecolonyformingefficiencydecreasedfromthe1stdayanddisappearedatthe4thday.Wheninoculatedonamnioticmembrane,thecellspreservedfor1dayformedastratifiedepithelium,whilethecellsfrom4days’preservationformedadiscontinuouslayer.CONCLUSION:Thecolonyformingefficiencyoflimbalepithelialstem/progenitorcellsdecreasedrapidlywiththeincreasingpreservationtime,whiletheexpressionlevelofmarkersandcapacityofformingepithelialmonolayeronamnioticmembranedecreasedgradually.Thelimbalepithelialstemcellslosttheirfunctionearlierthanthelostexpressionlevelofstemcellmarkers.Thismayhelpustobetterchoosetheappropriatepreservationgraftsforfuturelimbalstemcelltransplantation.
简介:AIM:Topresentretinalmicrostructure,metabolismandfunctionabnormalitiesinthecourseofmultipleevanescentwhitedotsyndrome(MEWDS)byHeidelbergspectralismodalityimagingplatformandobserveitsoutcomebyEDI-SD-OCTandtwowavelengthautofluorescence.METHODS:Acaseofmultipleevanescentwhitedotsyndromeina23-year-oldfemalepresentedinitiallywitha15-dayhistoryoffloatersandacentralscotomaintherighteye.Toestablishthediagnosis,multimodalityimagingwasperformed,namely,bluelight-fundusautofluorescence(BL-FAF,excitation488nm,emission>500nm),near-infraredfundusautofluorescence(NIR-FAF,excitation787nm,emission>800nm)usingaconfocalscanninglaserophthalmoscope,fundusfluoresceinangiography(FFA),indocyaninegreenangiography(ICGA),spectrum-domainenhancedepthimagingopticalcoherencetomography(SD-EDI-OCT),multifocalelectroretinography(mf-ERG)andfundusphotograghwereperformedandfollowedupattheeighthmonthafterinitiallyvisiting.RESULTS:Opticalcoherencetomography(OCT)showedatransientdisruptionofthefovealphotoreceptoroutersegmentsincorrespondencetofovealgranularity.NIR-FAFshowedhypoautofluorescentareas,≤40μminsize,mostlyconcentratedaroundtheposteriorpoleanditstemporalsidelessthanthatinBL-FAF.Mf-ERGshowpinnacledisappearedinfoveaandmaculaandresponsesdecreasedmarkedlycomparedwiththefolloweye.Attheeighthmonthfollowup,hyperfluorescenceinBL-FAFweredisappear,while,NIR-FAFHypofluorescentspotsinearlystageofsuchlesionwerereduced.ButOCTdemonstratedthestructurewasrecoveredinresidualHypofluorescentareainNIR-FAF.Thesubfovealchoroidalthicknesswasdecreasedfrom372μmto307μmslightlyandcostlinewasrecovered.CONCLUSION:MEWDSisabenignself-healingdiseaseandthereisnopathologicalevidencetoinvestigatethenaturalcourseofsuchdisease.SD-OCTallowshighlydetailedimagesapproachinghistopathologytocertifythemicrostructura
简介:AIM:ToinvestigatetheinterferingeffectofY-27632,aROCK-Iselectiveinhibitor,onthesignaltransductionpathwayoftransforminggrowthfactor-β1(TGF-β1)inocularTenoncapsulefibroblasts(OTFS)invitro.METHODS:AfterOTFSfrompassages4to6invitrowereinducedbyTGF-β1andthentreatedbyY-27632,thechangesoftheOTFScellcycleswereanalyzedviaflowcytometry,andtheproteinsexpressionoftheα-smoothmuscularactin(α-SMA),connectivetissuegrowthfactor(CTGF),collagenIwerecalculatedbyWesternblot.AfterOTFStreatedbythedifferentconcentrationsofY-27632,theexpressionlevelsoftheα-SMA,CTGFandcollagenImRNAwereassayedbyRT-PCR.RESULTS:Y-27632hadnomarkedlyeffectontheOTFScellcycles.AftertreatedbyTGF-β1,OTFSinG1periodsignificantlyincreased.ThecellcyclesdistributionbybothTGF-β1andY-27632hadnoremarkabledifferencefromthatincontrolgroup.Y-27632significantlyinhibitedtheproteinsexpressionsofbothα-SMAandCTGF,whiletosomeextentinhibitedthatofcollagenI.TGF-β1significantlypromotedtheproteinsexpressionsofα-SMA,CTGFandcollagenI.AfterOTFStreatedbybothTGF-β1andY-27632,ofα-SMA,theproteinexpressionwassimilarwiththatincontrolgroup(P=0.066>0.05),buttheproteinexpressionofCTGForcollagenI,respectively,wassignificantlydifferentfromthatincontrolgroup(P=0.000<0.01).Thedifferencesofexpressionsoftheα-SMA,CTGFandcollagenImRNAin30,150,750μmol/LY-27632groupwerestatisticallysignificant,comparedwiththoseincontrolgroup,respectively(α-SMA,P=0.002,0.000,0.000;CTGF,P=0.014,0.002,0.001;collagenI,P=0.003,0.002,0.000).CONCLUSION:BlockingtheRho/ROCKsignalingpathwaybyusingofY-27632couldinhibitthecellularproliferationandtheexpressionofbothCTGFandα-SMAwhateverOTFSinducedbyTGF-β1ornot.Y-27632suppressedtheexpressionofcollagenImRNAwithoutinduction.