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: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: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:目的探讨囊袋张力环植入下的晶状体半脱位超声乳化手术的安全性及有效性。方法晶状体半脱位患者26例(28眼)。做透明角膜切口,连续环撕前囊,水分离后,将张力环植入晶状体囊袋内,使囊袋稳定居中,以行晶状体乳化抽吸,然后囊袋内置人人工晶状体。结果26例(28眼)全部成功行超声乳化手术并囊袋内植入人工晶状体,术后随访最佳矫正视力较术前明显提高(P〈O.05),23只眼人工晶状体位置良好,3只眼人工晶状体轻度倾斜但无明显偏中心,2只眼人工晶状体轻度偏中心。无严重并发症发生。结论囊袋内张力环植入可稳定晶状体囊袋,保证超声乳化手术及人工晶状体囊袋内植入的顺利进行,并可维持人工晶状体的长期稳定性,该手术方式安全有效。
简介:目的:探讨超声乳化术中透明质酸酶对筋膜下麻醉的疗效。方法:该研究是一项双盲随机临床试验,在Nikookari眼科医院进行了长达5mo。行超声乳化术的候选患者中接受筋膜下麻醉的42眼随机分为均等的两组,分别应用20g/L的利多卡因溶液2mL+150U/mL透明质酸酶(LH),和单纯20g/L的利多卡因溶液2mL(L)。筋膜下注射15min后活动受抑制。患者和医生都非常满意,对于术后疼痛的抑制(视觉模拟评分,thevisualanalogueseale,VAS)也同样满意。统计学分析采用列联表(包括卡方检验或Fisher’s精确检验)和参数分析(独立样本t检验)。结果:活动抑制(33.3%'US4.8%,P=0.04)、患者满意度(85.7%掷57.1%,P=0.04)和医生满意度(87.5%US52.4%,P=0.02),LH组明显高于L组。VAS在相同组内明显较低(1.90±1.45傩3.00±1.55,P=0.04)。结论:利多卡因溶液+透明质酸酶的筋膜下麻醉明显使得眼部活动受限,提高了患者和医生的满意度,并减轻了术后疼痛。
简介:目的观察生物羊膜与丝裂霉素C(MMC),应用于青光眼小梁切除术中治疗青光眼的临床疗效。方法随机选择39例(48只眼),分为羊膜组(22只眼)和丝裂霉素组(26只眼)。羊膜组施行小梁切除术联合巩膜瓣下生物羊膜植入术,丝裂霉素组施行小梁切除术联合巩膜瓣下放置丝裂霉素C,浓度为0.25mg/ml,时间2分钟,术后随访3-9月,比较两种手术方式的临床疗效。结果术后与术前相比:两组眼压均明显降低。术后并发症:羊膜组副反应小,引起的并发症主要有术后浅前房;丝裂霉素组引起的并发症主要有滤过泡渗漏、低眼压等。结论小梁切除联合生物羊膜植入术可有效地减少瘢痕组织形成,且并发症较丝裂霉素少,是治疗青光眼安全、有效的手术方法。
简介:目的:通过对视网膜脱离患者术前、术后的血流动力学研究及视网膜功能的了解,判断视网膜脱离患者的术后恢复情况,并探讨两种检测方法的临床价值。方法:对我院62例孔源性视网膜脱离患者,通过彩色多普勒(CDI)检测视网膜中央动脉(CRA)的收缩期峰值流速(Vmax)、舒张末期流速(Vmin)、阻力指数(RI);多焦视网膜电图(mfERG)分别对视网膜脱离患者的术前、术后进行检测,并以对侧正常眼作对照组。结果:CDI术前患眼与对照眼的CRA各项指标比较,差异无统计学意义。术后2wk视网膜脱离眼CRA的Vmax,Vmin均较术前降低,RI增高,差异有统计学意义(P〈0.05)。视网膜脱离术后患者脱离区mfERG的a波、b波振幅密度均明显高于术前,潜伏期比手术前明显缩短,有统计学意义(P〈0.05)。结论:通过mfERG,CDI的联合检测能及时了解视网膜脱离术后的视网膜复位情况及硅胶海绵垫压后CRA的血流情况,从而避免视网膜脱离术后视网膜部分复位不良,巩膜因缺血引起局部坏死等情况发生。
简介:目的探讨运用示教与反示教在年龄相关性白内障患者滴眼药护理中的效果。方法将100例患者随机分为实验组和对照组各50例,对照组采用传统的健康教育。实验组采用示教与反示教进行健康教育。由培训过的责任护士对患者有计划地从入院到出院,给予全程、个体化的示教与反示教健康教育。结果实验组患者在滴眼药知识、技能掌握及对护理工作满意度方面优于对照组,差异有统计学意义(P0.05),术后一周并发症的发生率明显低于对照组(P〈O.05)。结论运用示教与反示教对患者实施健康教育,可达到提高患者滴眼药的技能,自我效能感增加,遵医行为增强,自护能力提高,减少术后并发症,提高护理满意度。
简介:目的:观察MoriaM2型90与110μm角膜刀制作角膜瓣在准分子激光角膜原位磨镶术(laserinsitukeratomileusis,LASIK)的疗效和并发症,探讨MoriaM2型90刀头在LASIK中应用的有效性、安全性和优点。方法:选取通过术前检查并自愿行LASIK手术的患者105例202眼,按随机数字表分成两组,使用MoriaM2型90刀头LASIK患者51例98眼,110刀头LASIK患者54例104眼做对照,术后即使用光学相干断层扫描仪(OCT)检测两组角膜瓣厚度,观察两组术后1d;1wk;1,3mo裸眼视力、矫正视力和角膜瓣形态、对合情况、并发症。结果:90刀头组术后角膜瓣厚度为118.3±15.2μm,110刀头组术后角膜瓣厚度为130.5±17.1μm,有显著性差异。90刀头组均未发现层间点状金属碎屑,110刀头组有层间点状金属碎屑个例(12例),有显著性差异。两组角膜瓣形态、对合情况、术后反应、术后裸眼视力相当。结论:应用90刀头LASIK的疗效及并发症和110刀头LASIK相当,但90刀头保留角膜基质床相对较厚,可矫治的屈光度更大,术后层间点状金属碎屑并发症更少,具有更好的安全性和更宽的适应范围。