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:生物力学是研究组织或器官的力量与功能之间关系的学科,对研究眼科疾病的预防、发生、发展、诊疗具有重要作用。眼球是一个封闭的近似球体的器官,眼球内外的每一组织都有一定张力,而且相互联系,互为影响。眼内外及球壁某一局部组织的改变,可能会影响眼球某一区域组织的生物力学或功能的改变。随着生物力学的方法增多,大家越来越重视对眼部组织生物力学的研究,达到预防和治疗眼部疾病的目的。如使用胶原交联增强角膜的强度,防治圆锥角膜或扩张性角膜疾病;又如后巩膜加固术增强巩膜力学的方法来治疗或预防高度近视等。对眼外肌、角膜、巩膜、虹膜、晶状体的生物力学的研究是目前热点,本文就目前眼部生物力学的研究热点做一综述。
简介:AIM:Todeterminethehistopathologicalchangesofrifampicinappliedintravitreallyonretinalganglioncellsbymeansofstereologicalandhistopathologicalmethods.METHODS:Forthisstudytwenty-fourNewZealandadultrabbitsweredividedintofourgroups(n=6foreachgroup).50μg/0.1mL(group1),100μg/0.1mL(group2),150μg/0.1mL(group3)and200μg/0.1mL(group4),rifampicinwereinjectedintothevitreousoftherighteyesofanimals,theirlefteyeswereusedascontrol(group5).Afterthe28thdayofapplication,animalswereanesthetisedwithxylazine(8mg/kg,IM)andthentheireyeswereenucleatedimmediately.Patternsweretakenawayandeyeswerepreparedforbothstereologicalandelectromicroscopicobservation.RESULTS:Dependingonthehighdoseofrifampicin,somehistopathologicalchangessuchascytoplasmicdilatationanddamagedmembranewereobservedontheelectromicroscopiclevel.Usingquantitativeexamination,whichwasdoneatthelightmicroscopiclevel,itwasshownthatthenumberofneuronsdecreasedlinearlyasrifampicindoseincreasedwhencomparedwiththecontrolgroup.CONCLUSION:Basedonthesefindings,low-doserifampicin(50μg/0.1mL)maybeusefulfortreatmentoftheoculardiseases.
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:目的:观察结膜松弛症松弛结膜组织和泪液中黏蛋白的表达变化,探讨结膜松弛症的发病机制。方法:收集结膜松弛症组38例38眼和正常对照组36例36眼的结膜组织和泪液标本,分别行免疫组织化学染色检测结膜组织中黏蛋白(MUC2、MUC4、MUC5AC、MUC16)表达情况,ELISA检测泪液中黏蛋白(MUC2、MUC4、MUC5AC、MUC16)A值,结果行统计学分析,比较两组之间的差异。结果:结膜松弛症组的结膜上皮细胞中MUC2、MUC4与正常对照组表达无统计学差异(P=0.315、0.156);结膜松弛症组的结膜上皮细胞中MUC5AC、MUC16阳性表达细胞较正常对照组明显减少,差异有统计学意义(P=0.016、〈0.01)。结膜松弛症组泪液中MUC2的A值与正常对照组无统计学差异(P=0.651),结膜松弛症组中MUC4、MUC5AC的A值较正常对照组明显降低,有显著的统计学差异(均P〈0.01),结膜松弛症组中MUC16的A值较正常对照组下降,差异有统计学意义(P=0.022)。结论:结膜松弛症患者结膜组织和泪液中MUC5AC和MUC16均下降,对其进一步研究可能揭示结膜松弛症的发病机制。
简介:目的通过检测Fas和FasL在鼻息肉中的表达,研究细胞凋亡和Fas/FasL系统在鼻息肉发病机制中的作用.方法用免疫组化方法检测Fas和FasL在35例鼻息肉和17例正常下鼻甲组织中的表达.对Fas和FasL阳性细胞的类型、分布特点进行分析.结果1.Fas和FasL在鼻息肉组织中的表达均较正常下鼻甲组织明显增强,差异有显著性.2.Fas在上皮细胞和间质细胞中都有阳性表达,主要表达于息肉表面的上皮细胞和间质内的炎性细胞中.结论1.Fas/FasL系统介导的凋亡在鼻息肉的发病机制中有重要作用,可能是鼻息肉长期保持良性增生的重要因素.2.FasL在鼻息肉上皮细胞中的高表达与上皮细胞的过度增生及Fas/FasL系统的免疫监视作用有关.
简介:目的:探讨HSP60(热休克蛋白60)在大鼠急性青光眼模型视网膜组织中的表达及其与血清中相应抗体的关系。方法:将SD大鼠70只随机分为高眼压组60只,正常对照组10只。大鼠全身及表面麻醉后,将一盛有等渗的生理盐水的储容器相连的7号针头于3:00位的角膜缘处刺入前房,提升储容器的高度,使眼内压达到110mmHg,但不超过150mmHg,观察大鼠眼前段变白,且视网膜色白,未见红色反光时即为视网膜缺血,缺血1h后再灌注,并于再灌注后2,6,12,24,72,168h将大鼠麻醉过量致死,处死前测眼压,抽血2mL,供酶联免疫吸附测定使用,分析视网膜组织内HSP60抗原产生的血清中相应抗体水平。并立即取出眼球,同时对鼠眼视网膜组织进行石蜡切片,采用免疫组化法检测视网膜组织中HSP60的表达及分布情况,并对检测结果进行统计学分析。结果:急性高眼压诱导的视网膜缺血/再灌注组眼压明显升高,视网膜组织中的HSP60阳性表达率在术后各时间点,高眼压组与正常对照组比较,差异有统计学意义(F=97.21,40.72,83.85,95.82,48.63及44.37,均P〈0.01)。神经节细胞(retinalganglioncell,RGC)中HSP60阳性表达随着眼压升高及高眼压持续时间延长逐渐增强,且视网膜神经纤维层中也出现较明显的HSP60阳性表达。结论:HSP60表达增强可能在急性高眼压所致的视神经病变中具有重要作用。
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:检测DNA氧化损伤标志物8-羟基脱氧鸟苷(8-OHdG)在翼状胬肉和正常结膜组织中的表达,探讨DNA氧化损伤在翼状胬肉发病机制中的作用。方法:收集在我科行翼状胬肉切除手术的原发性翼状胬肉组织标本35例,并收集术眼颞上方正常球结膜标本5例作对照。采用免疫组织化学法检测翼状胬肉标本中8-OHdG的表达,并与正常球结膜组织的标本进行对照。结果:在35例翼状胬肉组织中有24例呈阳性表达,阳性表达率为69%,而正常结膜组织中无8-OHdG的表达,其阳性表达率的差异具有显著统计学意义(P=0.007)。8-OHdG的阳性表达位于胬肉组织上皮细胞的细胞核,呈棕黄色着染,上皮下的纤维血管组织及正常结膜组织无表达。结论:在翼状胬肉组织中8-OHdG呈阳性表达,而正常结膜组织中不表达,提示DNA氧化损伤在翼状胬肉的发病机制中发挥重要作用。
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:AIM:Tocomparetheregularityandaccuracyoflaserinsitukeratomileusis(LASIK)flapscreatedbytheZiemerFEMTOLDV'Classic'(Ziemer'Classic')andZiemerFEMTOLDVCrystalLinefemtosecondlaser(ZiemerCrystalLine).METHODS:Fourier-domainopticalcoherencetomography(RTVueOCT)wasusedtomeasurethemorphologyof200LASIKflapsof100consecutivepatientscreatedwiththeZiemerClassic(100flaps)ortheZiemerCrystalLine(100flaps)atoneweekpostoperatively.Flapthicknesswasevaluatedat36specifiedmeasurementpointsoneachflap.Forallprocedureswithbothlasers,thenominalflapthicknesswas110μm.RESULTS:ThemeanflapthicknessoftheZiemerCrystalLinegroup(102.49±2.68μm)wasthinnerthanthatoftheZiemerClassicgroup(107.65±5.09μm)(P<0.01).Averagethicknessofallflapswasuniformwithin4μmatallmeasurementpoints.TheflapsintheZiemerCrystalLinegroupweremoreregularthanthoseintheZiemerClassicgroupwhenmeasuredfromthecentertotheperiphery.Themaximumdeviationfromthenominal110μmof36measurementswas8μmintheZiemerClassicgroup,whileintheZiemerCrystalLinegroupitwas9μm.Withinthe3600measurementsonthe100eyes,differencesgreaterthan20μmwereobserved0.14%intheZiemerClassicgroup,and0.04%intheZiemerCrystalLinegroup.CONCLUSION:TheflapscreatedwiththeZiemerFEMTOLDVCrystalLinefemtosecondlaseraremoreuniformandthinnerthanthosecreatedbytheZiemerFEMTOLDVClassicfemtosecondlaser.