简介:视网膜色素上皮细胞(retinalpigmentepithelium,RPE)的上皮间充质转化(epithelial-mesenchymaltransition,EMT)是增殖性玻璃体视网膜疾病(proliferativevitreoretinopathy,PVR)的主要发病机制,也是治疗脉络膜新生血管(choroidalneovascularization,CNV)时,造成抗血管内皮生长因子(anti-vascularendothelialgrowthfactor,anti-VEGF)疗效降低的重要因素。除此之外,随着细胞替代治疗作为年龄相关性黄斑变性(age-relatedmaculardegeneration,ARMD)的新型治疗方式而兴起,人类胚胎干细胞(hESC-RPE)、体细胞诱导多能干细胞(iPSC-RPE)定向诱导分化的RPE细胞以及来自于流产胎儿的胎儿视网膜色素上皮细胞(fetalRPE,fRPE)逐渐受到了研究者们的关注。为了发挥最好的治疗效果,保证治疗用细胞的稳定增殖及高效分化是极其重要的。但是在传代扩增的过程中,由于上皮间充质转化的存在,导致这些细胞出现增殖困难以及再分化能力下降,从而影响治疗效果并阻碍了治疗人群的普及,使细胞替代治疗难以推广。所以,鉴于上皮间充质转化在视网膜疾病的发生和治疗中都造成了重要影响,抑制视网膜色素上皮细胞发生上皮间充质转化就显得尤为重要,为此,我们将阻止上皮间充质转化研究的最新进展综述如下。
简介:恶性肌上皮瘤临床罕见,又称肌上皮癌,来源于腺体组织,英文文献报道迄今只有40余例[1],大多数(60%~70%)发生于腮腺,其余为小唾液腺和颌下腺,也见于乳腺等器官.现报告1例发生于咽旁间隙的肌上皮癌.
简介:目的:探讨去上皮瓣机械法准分子激光角膜上皮瓣下磨镶术(Epi—LASIK)矫治高度近视的临床疗效。方法:对86例169眼高度近视患者施行去上皮瓣Epi-LASIK,术后随访6mo。观察术后刺激症状、角膜上皮愈合时间、视力、屈光度、角膜上皮下雾状混浊程度(haze)。结果:所有患者术后均未主诉明显疼痛,仅表现有轻中度的异物感、畏光、流泪。角膜上皮愈合时间3~4d。术后6mo,全部86例169眼术后等效球镜均在±1.00D以内,未见最佳矫正视力下降,23眼(13.6%)最佳矫正视力提高1行以上。术后1mo,142眼(84.0%)角膜haze为0级,27眼(16.0%)角膜haze为0.5级;术后3mo,2例4眼不明原因出现3级haze;术后6too,所有患者角膜haze均为0级。结论:去上皮瓣Epi—LASIK治疗高度近视安全、有效,具有良好的可预测性和稳定性。个别患者在术后发生不明原因的迟发性haze有待于进一步研究。
简介:0引言急性后部多灶性鳞状色素上皮病变(acuteposteriormultifocalplacoidpigmentepitheliopathy,APMPPE)由Gass于1968年首先报道。本病发病急,目前临床并不多见。本文报道1例双眼急性后部多灶性鳞状色素上皮病变的病例,就其发病原因、病情演变及治疗、愈后进行简要的分析总结。
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:探讨胎牛血清对小鼠角膜上皮细胞生长和分化的影响。方法:分别在无血清低钙培养基(KSFM)和含100mL/L胎牛血清的KSFM中培养小鼠角膜上皮细胞。比较两种培养基中角膜上皮细胞的群体倍增(PD)。通过RT-PCR和Westernblotting方法检测p63、角蛋白19以及involucrin的表达。结果:在KSFM中,小鼠角膜上皮细胞可稳定传代超过25代,但在含胎牛血清的培养基中,细胞仅能存活3代。在KSFM中,培养细胞呈典型铺路石样外观,RT-PCR和Westernblotting结果显示细胞表达p63、角蛋白19以及involucrin。当培养基中加入胎牛血清后,细胞形态明显增大,呈扁平状;involucrin表达显著增强。结论:胎牛血清可抑制小鼠角膜上皮细胞增生、诱导其分化。
简介:目的:探讨丝裂霉素C(mitomycinC,MMC)在准分子激光原位角膜磨镶术(laserinsitukeratomileusis,LASIK)术后角膜上皮植入手术治疗中应用的临床疗效。方法:在31例31眼LASIK术后角膜上皮植入患者手术治疗中应用0.2g/LMMC,观察术后裸眼视力及角膜瓣愈合情况。结果:术后6mo裸眼视力平均为0.82±0.19,较术前明显提高(P〈0.05)。绝大部分患者角膜瓣愈合良好,4例患者角膜瓣边缘部分融解,但中心视力无明显影响。结论:在LASIK术后角膜上皮植入手术治疗中应用0.2g/LMMC可以取得良好的临床疗效。
简介:目的:探讨miR-181在白内障晶状体组织中的表达情况,及其对人晶状体上皮细胞凋亡的调控机制。方法:利用Realtimeq-PCR方法,检测年龄相关性白内障患者晶状体前囊膜和人晶状体上皮细胞凋亡模型中miR-181的表达情况;利用Lipofectamine2000瞬时转染miR-181mimic和inhibitor调节人晶状体上皮细胞中miR-181的表达,利用Realtimeq-PCR方法验证转染效率,利用流式细胞仪检测细胞凋亡率的变化。结果:与对照组相比,年龄相关性白内障患者晶状体前囊膜组和人晶状体上皮细胞凋亡模型组,miR-181的表达显著升高;miR-181mimic转染组,miR-181的表达显著升高,细胞凋亡率显著升高;miR-181inhibitor转染组,miR-181的表达显著降低,细胞凋亡率显著降低,差异均有统计学意义(P〈0.01)。结论:miR-181在年龄相关性白内障晶状体组织中呈高表达,miR-181能够促进人晶状体上皮细胞凋亡,从而可能在年龄相关性白内障的发病过程中发挥一定作用,miR-181可能成为白内障非手术治疗的一种新途径,但具体机制有待进一步研究。
简介:目的分析常年性变应性鼻炎的狗、猫上皮变应原皮肤点刺试验的阳性结果,对其他气传变应原的反应性,并进一步探讨环境接触与其相关性。方法总结1022例常年性变应性鼻炎患者采用阿罗格叶7种气传变应原标准化试液进行的皮肤点刺试验;分析狗、猫上皮变应原的皮试结果及合并其他气传变应原阳性的比例;并根据是否有环境接触分类,比较各组在阳性率分布上的差异。结果常年性变应性鼻炎中狗上皮变应原皮试阳性占13.80%,猫上皮阳性占6.95%;且98%以上合并其他气传变应原阳性,尤以合并粉尘螨、屋尘螨阳性居多。其中21.93%和26.32%环境接触狗/猫的变应性鼻炎相应狗/猫上皮变应原皮试阳性;无环境接触组的阳性率分别为12.78%和6.58%,差异均有统计学意义(P〈0.01)。结论常年性变应性鼻炎的狗上皮及猫上皮的阳性率位居前列,其中绝大多数合并其他气传变应原阳性。鉴于环境接触对狗、猫上皮过敏的高危险性,变应性鼻炎患者应该尽可能地避免饲养、接触宠物。
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:目的比较自体角膜缘上皮移植术和翼状胬肉切除术联合应用丝裂霉素C的疗效观察.方法将翼状胬肉69例(74眼)随机分为A、B两组,A组33例(36眼)作单纯手术切除,并在术中、术后应用0.2mg/ml丝裂霉素C;B组36例(38眼)在胬肉切除的基础上行自体角膜缘上皮移植术.术后平均随访20个月,按Prabhasawat提出的标准进行疗效评价,比较两组的治愈率、复发率、视力及角膜散光度.结果A组36眼中31眼获得临床治愈,治愈率为86.11%,复发率为13.89%.B组35眼中33眼获得临床治愈,治愈率为94.74%,复发率为5.26%;两组的治愈率和复发率差异有显著性(P<0.05).在裸眼视力和角膜散光度方面,B组优于A组,差异有显著意义(P<0.05).结论自体角膜缘上皮移植有利于重建角膜缘屏障功能,利用该方法治疗翼状胬肉可提高治愈率,降低复发率.在减轻角膜散光,提高视力方面也优于手术联合丝裂霉素.
简介:目的:改良法建立大鼠后发性白内障动物模型并观察LEC在PCO过程中的动态变化。方法:SD大鼠50只在连续环形撕囊、水分离后行晶状体囊外摘除术(ECLE),娩核后使用无菌空气恢复前房。分别于术后0,3,7,14及28d对术眼进行裂隙灯检查并处死动物,摘除眼球行光镜观察,免疫组化检测LEC的α-SMA表达。结果:100%术眼后囊膜存在,84%的鼠眼可用于检测。术后0h于前囊下和赤道部的内表面观察到LEC。PCO在术后3d出现,出现囊膜皱缩,整个晶体囊膜出现纺锤形细胞。术后7d时后囊膜明显混浊,见较多纺锤形细胞分布。所有动物于术后14d出现明显后囊膜皱缩,可见新生晶体纤维。术后28d见明显后囊膜增厚,新生晶体纤维填充囊袋,后囊膜未见细胞。LEC形态及分布恢复到术后0h状态。免疫组化检测见术后3d时α-SMA阳性表达。结论:改良法成功建立大鼠PCO模型,LEC在PCO形成中出现形态和分布上的动态变化。其将为在分子水平上探索PCO的发病机制及防治方法提供合适研究载体。
简介:目的探讨准分子激光上皮下角膜磨镶术(LASEK)治疗高度近视术后半年疗效及其相关影响因素.方法对接受LASEK手术的109只术眼进行半年的跟踪观察,将术眼按术前屈光度分为三组,Ⅰ组(23眼):-6.00~-9.00D;Ⅱ组(56眼):-9.25~-12.00D;Ⅲ组(30眼):-12.00以上.对各组术后裸眼视力、HAZE、角膜曲率、回退率及眼压的变化进行了分析.结果1.术后视力各组均比术前有显著改善(P<0.001);2.术后HAZE发生率随度数增加而上升,且程度加重,但术后各组6个月时角膜混浊比第一个月有明显改善(P<0.01);3.术后各组回退率高,回退率随度数增加而上升;3.术后各组眼压与术前均有差异(P<0.001),术后各组第6个月时眼压有显著降低(P<0.01).结论LASEK手术对高度近视及超高度近视均有明显治疗作用,但对超高度近视预测性较差,HAZE发生率较高且回退率高.
简介:AIM:ToidentifythefunctionofST2andexploretheroleofIL-33/ST2signalinginregulatingthepro-allergiccytokineproductioninhumancornealepithelialcells(HCECs).METHODS:HumancornealtissuesandculturedprimaryHCECsweretreatedwithIL-33indifferentconcentrationswithoutorwithdifferentinhibitorstoevaluatetheexpression,locationandsignalingpathwaysofST2inregulatingproductionofpro-allergiccytokineandchemokine.TheexpressionofmRNAwasdeterminedbyreversetranscriptionandrealtimePCR,andproteinproductionwasmeasuredbyenzyme-linkedimmunosorbentassay(ELISA),immunohistochemicalandimmunofluorescentstaining.ST2proteinwasdetectedindonorcornealepithelium,andST2signalwasenhancedbyexposuretoIL-33.·RESULTS:IL-33significantlystimulatedproductionofpro-allergiccytokinesthymicstromallymphopoietin(TSLP)andchemokine(CCL2,CCL20,CCL22)inHCECsatbothmRNAandproteinlevels.Thesestimulatedproductionsofpro-allergicmediatorsbyIL-33wereblockedbyST2antibodyorsolubleST2protein(P<0.05).Interestingly,theIκB-αinhibitorBAY11-7082orNF-κBactivationinhibitorquinazolineblockedNF-κBp65proteinnucleartranslocation,andalsosuppressedtheproductionsofthesepro-allergiccytokinesandchemokineinducedbyIL-33.CONCLUSION:ThesefindingsdemonstratethatIL-33/ST2signalingplaysanimportantroleinregulatingIL-33inducedpro-allergicresponses.IL-33andST2couldbecomenovelmoleculartargetsfortheinterventionofallergicdiseasesinocularsurface.
简介:目的:探讨褪黑素对过氧化氢诱导晶状体上皮细胞氧化损伤的保护作用。方法:晶状体上皮细胞传代培养后,分别加入不同浓度褪黑素预处理12h后,加入100μmol/LH2O2继续孵育24h,MTT比色法检测褪黑素对H2O2诱导的晶状体上皮细胞活力的影响,流式细胞仪检测细胞凋亡率,比色法检测凋亡相关因子Caspase-3及Caspase-9的活性。结果:MTT结果显示褪黑素对晶状体上皮细胞活性无影响,该药物可以抑制过氧化氢诱导的细胞活性的下降,流式细胞计数结果显示褪黑素可以抑制过氧化氢诱导的细胞凋亡,此外,褪黑素还可以减少过氧化氢所致晶状体上皮细胞内Caspase-3及Caspase-9的活性,并且,伴随褪黑素作用时间的延长其活性呈下降趋势。结论:褪黑素可以明显抑制过氧化氢诱导的晶状体上皮细胞的凋亡,从而为寻求有效的防治白内障药物提供可靠的实验依据。
简介:AIM:Toinvestigatetheexpressionofdendriticcell-associatedC-typelectin-1(dectin-1)attheearlyperiodofAspergillusfumigatusinfectioninrat’scornealepithelium.·METHODS:Atotalof72Wistarratswererandomlydividedintothreegroups:A,BandC.Therighteyeswerechosenasexperimentaleyes.GroupAwascontrolgroup.RatsingroupBwerenotinoculatedwithAspergillusfumigatus.GroupCwastakenasAspergillusfumigatuskeratitismodel.RatsingroupBandC(sixfromeachgroup)wereexecutedrandomlyat4,8,16and24hoursafterexperimentalmodelbeingestablishedtoassesstheexpressionofdectin-1mRNAthroughreal-timePCR.AnothersixratsingroupBandCwereexecutedrandomlyat24hourstoassesstheexpressionofdectin-1proteinthroughimmunohistochemistry.·RESULTS:Theresultsofreal-timePCRindicatedthatdectin-1mRNAexpressionwaslowincornealepitheliumofnormalrats’.Therewasnosignificantlydifferenceofdectin-1mRNAexpressioningroupAandB(P>0.05).TheexpressionofAspergillusfumigatusinfectedcornealepitheliumincreasedgraduallyafter8hoursingroupC.ThesynchronousexpressionofgroupAandChadsignificantdifference(P<0.01).Immunohistochemistydiscoveredthatdectin-1receptorexistedinnormalrat’scornealepithelium.Dectin-1proteinincreasedafter24hoursingroupC.TherewasasignificantdifferenceofsynchronousexpressioningroupBandC(P<0.01).·CONCLUSION:Dectin-1existsinrat’scornealepitheliumanditsexpressionsignificantlyincreasesattheearlyperiodofAspergillusfumigatusinfection.Dectin-1isapatternrecognitionreceptorthatexpressesincornealepitheliumandinvolvesinimmuneresponsetoAspergillusfungalkeratitis.
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.