简介:AIM:Toinvestigatethemorphologicalalteringeffectoftransforminggrowthfactor-β2(TGF-β2)onuntransfectedhumancornealendothelialcells(HCECs)invitro.METHODS:AfteruntransfectedHCECsweretreatedwithTGF-β2atdifferentconcentrations,themorphology,cytoskeletondistribution,andtypeIVcollagenexpressionofthecellswereexaminedwithinvertedcontrastlightmicroscopy,fluorescencemicroscopy,immunofluorescenceorWesternBlot.RESULTS:TGF-β2attheconcentrationof3-15μg/LhadobviouslyalterativeeffectsonHCECsmorphologyindoseandtime-dependentmanner,and9μg/Lwasthepeakconcentration.TGF-β2(9μg/L)alteredHCEcellmorphologyaftertreatmentfor36h,increasedthemeanopticaldensity(P<0.01)andthelengthofF-actin,reducedthemeanopticaldensity(P<0.01)ofthecollagentypeIVinextracellularmatrix(ECM)andinducedtherearrangementofF-actin,microtubuleincytoplasmandcollagentypeIVinECMaftertreatmentfor72h.·CONCLUTION:TGF-β2hasobviouslyalterativeeffectonthemorphologyofHCECsfrompolygonalphenotypetoenlargedspindle-shapedphenotype,indoseandtime-dependencemannerbyinducingmore,elongationandalignmentofF-actin,rearrangementofmicrotubuleandlargerspreadareaofcollagentypeIV.
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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:目的本研究针对初发性翼状胬肉局部使用丝裂霉素C(MMC)一次性注射治疗,用免疫组织化学方法对经MMC治疗的翼状胬肉标本中血管内皮生长因子(VEGF)和转化生长因子(TGF-β_1)的表达进行分析研究,并与未使用MMC治疗者的翼状胬肉标本作对比。观察局部注射MMC对翼状胬肉组织中VEGF和TGF-β_1的表达的影响。方法需手术者共19例20眼,随机分为二组:A组直接切除胬肉,留标本作免疫组织化学检查;B组于翼状胬肉颈部进针,向胬肉组织内局部注射MMC,0.1~0.2ml(0.4mg/ml),3周至10周后手术切除胬肉,留标本作免疫组织化学检查。分别对A、B二组标本中VEGF和TGF-β_1的表达进行分析研究。结果A组VEGF、TGF-β_1的表达较B组高,差异有统计学意义(P〈0.05)。结论丝裂霉素C翼状胬肉局部注射可以抑制静止期翼状胬肉中VEGF、TGF-β_1因子的表达。MMC在低浓度、低剂量下使用未出现眼部严重并发症。
简介:病例介绍病例1:患者,男,47岁。因右眼突然视力下降伴下方黑幕遮挡4天入院。原双眼有高度近视-14.00DS,入院时全身体检(-)。视力:右眼无光感,陈旧性视网膜脱离;左眼0.05(矫正)。左眼前节(-),眼底见9:30~4:00视网膜脱离,隆起度较高,累及黄斑区,颞上方约1:00时钟见一2PD大小马蹄孔。入院后第二天局部麻醉下行左眼冷凝加外加压加环扎术,外加压块、环扎带均位于赤道区,无放液并发症,手术顺利。术后第一天查眼底见脱离的
简介:·AIM:Toobtainwideningofapotentiallyoccludableangle,inaccordingtoKanski’sindications,throughpreventiveNd:Yaglaseriridotomy.Theobservationalstudywasperformedbyusinggonioscopyfortheselectionandfollow-upof1165treatedeyesandexploitingShaffer-Etiennegonioscopicclassificationasaquality/quantitytestoftheanglerecession.·METHODS:BetweenSeptember2000andJuly2012,586patientswereselectedattheOutpatients’OphthalmologicalClinicofthePoliclinicoUmbertoIofRomeinordertoundergoNd:Yaglaseriridotomy.AGoldmanntypecontactlens,Q-switchedmode,2-3defocus,and7-9mJintensitywith2-3impulsedischargeswereusedforsurgery.·RESULTS:Fromasearlyasthefirstweek,awhole360°anglewideningwereevidentinthepatients,thusshowingthesuccessofNd:Yaglaseriridotomyinsolvingrelativepupilblock.Theangleremainednarrowby270°in14eyesonly,despiterepetitionsoffurthertreatmentwithlaseriridotomyinadifferentpartoftheiris,twicein10eyesandthreetimesin4eyes.·CONCLUSION:Nd:YaglaseriridotomyrevealeditselfasbeingasafeandeffectivetreatmentinwideningthosecriticalShaffer-Etiennegrade1and2potentiallyoccludableangles.
简介:目的:构建表达小鼠CD4+T细胞钙支架蛋白AHNAK1的短发夹RNA(shorthairpinRNA,shRNA)慢病毒载体,并研究其对小鼠甲状腺相关性眼病(thyroid-associatedophthalmopathy,TAO)的抑制效应。方法:设计并筛选对AHNAK1具有良好干扰效力的shRNA序列,慢病毒载体包装干扰序列,感染小鼠CD4+T细胞,检测AHNAK1静默对T细胞功能的抑制作用,采用实验动物模型观察AHNAK1体内抑制甲状腺相关性眼病的效果。结果:成功筛选出具有良好干扰效力的shRNA,并包装入慢病毒。病毒滴度为1.0伊106TU/mL,转染慢病毒的CD4+T细胞展现出失能倾向,抑制炎症免疫反应;在动物模型中抑制T细胞中AHNAK1表达可以有效控制甲状腺眼病的发生发展,显著降低治疗组T细胞中IL-2、IL-1茁和IFN-酌的表达。结论:成功构建了表达小鼠AHNAK1shRNA的慢病毒,具有抑制T细胞分泌IL-2、IL-1茁和IFN-酌的表达效应,能够有效抑制甲状腺眼病的发生发展。
简介:例1,男52岁.因咽部不适、头痛睡眠不好2年,在内地医疗单位诊断为阻塞性睡眠呼吸暂停低通气综合征(obstructvesleepapneahypopneasyndrome,OSAHS).在全身麻醉下行悬壅垂腭咽成形术(uvulopalatopharyngoplasty,UPPP),术后20d返回高原.1wk后出现咽痛、咽干、吞咽困难,夜间症状加重.检查:咽部充血,咽腔宽大,黏膜干燥无光泽,咽后壁较多黄白色干痂贴附.
简介:AIM:ToinvestigatetheregulationofEaf2proteininmouselenscellsapoptosisinducedbyultraviolet(UV)radiation.METHODS:AneyeofEaf2geneknockoutmiceornormalcontrolmicewasexposedtoUVradiation,andtheotheronewasnon-exposed.AlloflenseswereanalyzedbyTUNELandcaspase3activityassaystodeterminethedifferenceoftheapoptosisinducedbyUVradiation.Inaddition,exposedandnon-exposedlenseswereanalyzedbyquantifiedp53expressionandreal-timereversetranscription-polymerasechainreaction(RT-PCR)ofBax,Bid,Apaf-1,PumaandNoxa,tocompareEaf2geneknockoutmiceandnormalcontrolmice.RESULTS:UVradiationcausedapoptosisoflenscellsinnormalcontrolmiceandEaf2knockoutmice.Activityofcaspase3wassignificantlyhigherinnormalcontrolmicethanEaf2knockoutmice.Expressionofp53proteinwassignificantlyhigherinlensesexposedtoUVradiationthannonexposedlenses,butwassimilarbetweenEaf2geneknockoutmiceandnormalcontrolmiceinthesameUVcondition.AfterexposingtoUVradiation,theanalysisofreal-timeRT-PCRdemonstratedthatmRNAlevelsofPumaandNoxaweresignificantlyhigherinlensesofnormalcontrolmicethanEaf2geneknockoutmice,andthatmRNAlevelsofBax,BidandApaf-1werenotsignificantlydifferentbetweengeneknockoutmiceandnormalcontrolmice.CONCLUSION:Eaf2increaseslenscellsapoptosisinducedbyultravioletradiation.AndEaf2up-regulatesexpressionofthePumaandtheNoxatoactonlenscellsapoptosisafterUVradiation.
简介:AIM:Toinvestigatewhethertheresponseofacentralhexagonalelementcorrespondingtothemacularareainconventionalmultifocalelectroretinography(mfERG)testswasthesameasthatofexperimentalmfERGusingsinglecentralhexagonalelementstimulation.METHODS:Prospective,observationalstudy.Thirtyhealthysubjectswereincludedinthisstudy.mfERGrecordingswereperformedaccordingtotwoprotocols:stimuluswith37hexagonalelements(protocol1),andstimuluswithasinglecentralelementcreatedbydeactivatingtheother36hexagonalelements(protocol2).Wecompareddifferencesbetweenring1parametersineachprotocol.RESULTS:Inprotocol1,thefirstpositivecomponent(P1)implicittimeandP1amplitudewere37.8±1.8msand6.3±2.7μV.Aftersingleelementstimulation(protocol2),doublepositivewavesappeared.TheimplicittimeandamplitudeofP1were40.7±2.4ms(P<0.001)and9.1±3.3μV(P=0.001),respectively.Theimplicittimeandamplitudeofthesecondpositivecomponent(P2)were68.0±4.5ms(P<0.001,comparedwithP1inprotocol1)and12.3±4.7μV(P<0.001,comparedwithP1inprotocol1),respectively.TheamplitudeofP2inprotocol2wasabouttwotimeshigherthanthatofP1inprotocol1.CONCLUSION:mfERGresponsesofacentralhexagonalelementinasingleelementstimulationprotocolaredifferentfromthoseofmultipleelementstimulation.Thepositivewaveismoreenhancedcomparedtothatoftheconventionalprotocolanditelongatedintotwowavelets.
简介:目的:探讨白内障合并晶状体脱位范围〉2个象限的患者,Ⅰ期行白内障囊内摘除术+前部玻璃体切割术,术后3mo矫正视力〉0.3者,Ⅱ期行小切口两点固定人工晶状体悬吊术的临床疗效。方法:对2014-07/2016-12我院白内障科就诊的34例34眼白内障合并晶状体脱位范围〉2个象限的患者,Ⅰ期行白内障囊内摘除术+前部玻璃体切割术,3mo后矫正视力〉0.3者,Ⅱ期行小切口两点固定人工晶状体悬吊术,分别观察患者术后1wk,1、3mo裸眼视力、最佳矫正视力、眼压、角膜散光度、术后并发症情况。结果:随着术后恢复时间的延长,患者各期的裸眼视力和最佳矫正视力均较术前有明显提高。术后3mo最佳矫正视力0.1~〈0.3者1眼,0.3~〈0.5者8眼,0.5~〈0.7者16眼,〉0.7者9眼,达到或接近术前的最佳矫正视力。术后1wk,1、3mo眼压处于正常范围内。手术并没有明显增加角膜的散光度。结论:对于白内障合并晶状体脱位范围〉2个象限的患者,Ⅰ期行白内障囊内摘除术+前部玻璃体切割术,3mo后矫正视力〉0.3者,Ⅱ期行小切口两点固定人工晶状体悬吊术能有效确切地提高视力,稳定眼压,术后并发症少,是较为安全可靠的治疗方式。
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: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。
简介:研究内源性大麻素N-花生四氢酸氨基乙醇(anan-damide,AEA)对体外培养的牛眼小梁细胞细胞骨架及PGE2表达的影响。方法:对牛眼小梁细胞进行原代及传代培养,并行鉴定。对传3代的牛眼小梁细胞分别施加AEA浓度分别为0,0.1,1.0,10/μmol/L的无血清培养液,作用24h后提取细胞上清液,并将细胞置于倒置显微镜下摄像,计算机图像分析系统计算细胞面积。用ELISA法检测PGE2浓度的变化。结果:AEA可以产生明显的细胞舒张效应,呈浓度依赖性。而且在一定范围内可以成剂量依赖性的促进PGE2的表达。与对照组均有显著性差异(P〈0.05)。结论:AEA可引起小梁细胞的舒张和促进PGE2的释放。