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:目的观察1,25(OH)2D3对高糖诱导牛视网膜血管内皮细胞(BRECs)中血管内皮生长因子(VEGF)的表达水平变化及对细胞凋亡水平的影响。方法将分离培养的BRECs分为三组,分别为正常糖组、高糖组和高糖处理组。正常对照组细胞培养液含5mmol/L葡萄糖,高糖组细胞培养液含30mmol/L葡萄糖,高糖处理组细胞培养液含30mmol/L葡萄糖和50nM,1,25(OH)2D3。培养48h后收集细胞蛋白。蛋白免疫印迹法检测细胞VEGF及细胞凋亡相关蛋白Bax和Bcl-2表达水平;PI/Hoechst双染色法检测细胞凋亡。结果相比于正常糖组,高糖组中VEFG水平和Bax/Bcl-2比值显著增加;而在高糖处理组中表达水平远远低于高糖组,差异均有统计学意义(P〈0.05)。高糖的细胞凋亡水平高于正常糖组,而经过1,25(OH)2D3处理后,其细胞凋亡水平则有所下降,差异均有统计学意义(P〈0.05)。结论1,25(OH)2D3可以抑制高糖诱导BRECs中VEGF表达增加及细胞凋亡。
简介:目的探讨小儿喉乳头状瘤(JLP)患儿外周血T淋巴细胞(简称T细胞)亚群的变化及脾氨肽对JLP患儿T细胞亚群的影响和疗效。方法60例JLP病例随机分为JLP联合治疗组(30例)和JLP手术组(30例),所有JLP患者均采用喉内镜下电动吸割器切除新生物,联合治疗组术后同时口服脾氨肽辅助治疗,手术组不加任何免疫治疗。以同期体检的20例健康儿童做对照组,对JLP病例组(未治疗前)、健康对照组、JLP联合治疗组和JLP手术组进行外周血T细胞亚群的检测,并对脾氨肽辅助治疗JLP的疗效进行分析。结果治疗前JLP病例组CD3+T细胞、CD4+T细胞百分率及CD4+/CD8+T细胞比值较健康对照组降低,差异有统计学意义(P〈0.05);治疗后JLP联合治疗组CD3+T细胞、CD4+T细胞百分率明显升高(P〈0.05),CD4+/CD8+T细胞比值恢复平衡;治疗后JLP联合治疗组较JLP手术组的疗效明显提高(P〈0.05)。结论JLP患儿存在T细胞介导的免疫功能障碍,脾氨肽可恢复JLP患儿T细胞亚群的正常比例,改善患儿的细胞免疫功能,对JLP有较好的辅助治疗作用。
简介:AIM:Toinvestigatethelevelsofserumsolubleintercellularadhesionmolecules-1(sICAM-1)andneutrophilicexpressionofCD18inpatientswithvariousstagesofdiabeticretinopathyandtodeterminetheirdifferentexpressionpatterninthedevelopmentofdiabeticretinopathy(DR).·METHODS:LevelsofserumsICAM-1andCD18onthesurfaceofneutrophileweremeasuredin41DRpatients,theywereclassifiedinthreesubgroupsaccordingtothestageofretinopathyasdeterminedbyfund’sophthalmoscopy;10controlsubjectswerealsostudied.sICAM-1weremeasuredbyenzyme-linkedimmunosorbentassayandCD18byflowcytometry.·RESULTS:TheneutrophilicCD18expressionandserumsICAM-1levelwereallsignificantlyelevatedinalldiabeticsubgroupscomparedtocontrolsubjects(P<0.01).ThedifferencesofCD18andsICAM-1amongthediabeticsubgroupsweresignificantinCD18butnotinsICAM-1.TheprogressionofretinopathywasassociatedwithanincreasebothinCD18andinsICAM-1levelsbysimplecorrelationanalysis(β=0.74,P<0.001;β=0.38,P<0.01,respectively).ButstepwisemultipleregressionanalysisrevealedthatonlyCD18wasindependentdeterminantofretinopathy(β=1.04,P<0.01).·CONCLUSION:OurresultsconfirmthecontributionofendothelialandneutrophilicactivationinthedevelopmentofDRasindicatedbyincreasedlevelsofCD18andsICAM-1.However,adirectimplicationofCD18andICAM-1intheprogressionofDRcanbesupportedonlyintheCD18butnotICAM-1.CD18andICAM-1mayplaydifferentroleinthedevelopmentofdiabeticretinopathy.
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。
简介:我科收治的患眼无视功能患者25例,保留自体巩膜壳,行羟基磷灰石义眼台植入术,现报告如下。
简介:AIM:Toreportthecytologyresultsof25-gaugetransconjunctival(25G-TSV)diagnosticvitrectomyincasessuspiciousforintraocularlymphoma(IOL),andcomparetheresultstothosereportedintheliterature.METHODS:Clinicalandcytopathologicalrecordsof18vitreousbiopsyspecimensobtainedvia25G-TSVdiagnosticvitrectomyin12patientssuspiciousforIOLwerereviewedretrospectively.Areviewoftheliteratureinregardstothediagnosticyieldsofvitreousspecimensobtainedvia25-gaugeand20-gaugediagnosticvitrectomyinsuspectedcasesofIOLwasperformed.RESULTS:Eighteeneyesfrom12patientswithclinicalsuspicionofIOLunderwentdiagnostic25G-TSV.ThecytopathologicalinvestigationsdemonstratedIOLin15eyes(83.3%).Vitreousanalysiswasnon-diagnosticin3eyes(16.7%).CONCLUSION:Twenty-five-gaugediagnosticvitrectomyyieldsadequatesampleforcytologicalevaluationofthevitreousincasessuspiciousforIOL.Thediagnosticresultsofthe25G-TSVinthecurrentstudyaresuperiortothosereportedfor20-gaugevitrectomybutequivalenttothosereportedfor25G-TSVinthepublishedliterature.
简介:AIM:Tocomparetheclinicaloutcomesofcombined25-gaugeparsplanavitrectomy(PPV)andphacoemulsification/posteriorchamberintraocularlens(PC-IOL)implantationwithvitrectomyalonesurgeryinpatientswithvariousvitreoretinaldiseases.METHODS:Atotalof306eyes(145withPPValoneand161withphaco-vitrectomy)wereenrolledinthisretrospectiveanalysis.Thesurgicalapproachwas25-gaugePPVcombinedwithphacoemulsificationandPC-IOLimplantationatthesametimeineyesinphaco-vitrectomygroupandonlyPPVineyesinvitrectomyalonesurgerygroup.Themainoutcomemeasureswerepostoperativeclinicaloutcomesincludedanteriorchamberinflammation,changesinintraocularpressure(IOP)andbestcorrectedvisualacuity(BCVA).RESULTS:Themostcommonpostoperativecomplicationwasanteriorchamberreactionwhichhashigherincidenceinphaco-vitrectomygroup(P<0.001).Themeanpostoperative1stdayIOPofvitrectomyalonegroupwassignificantlylowerthanthatofphaco-vitrectomygroup(16.3±5.8mmHgvs17.8±8.1mmHg,respectively,P=0.02).Hypotony(IOP(8mmHg)wasnotdifferentbetweengroupsinthepostoperative1stday(P>0.05).Themeanpreoperativevisualacuitywasnotdifferentbetweengroups(1.6±0.9logMARvs1.8±0.9logMAR,respectively,P>0.05).However,themeanvisualacuitywasdecreasedinvitrectomyalonegroupatthefinalvisitcomparedtophaco-vitrectomygroup(1.2±0.8logMAR,0.9±0.7logMAR,respectivelyP<0.05).CONCLUSION:Twenty-fivegaugePPVcombinedwithphacoemulsificationsurgeryisasafeandefficientprocedure,whichcanbepreferredinphacicpatientswithavarietyofvitreoretinaldiseasescomparedtovitrectomyalone.Despiteimprovedoutcomes,thisapproachisnotfreeoflimitationsasanteriorchambercomplicationsespeciallywithcombinedsurgery.
简介:目的探讨可调节缝线技术应用于小梁切除术的临床疗效。方法对92例(107只眼)原发性闭角型青光眼,随机分为两组;观察组(A组)48例56只眼行可调节缝线小梁切除术;对照组(B组)44例51只眼行传统小梁切除术。观察比较术后前房、眼压、住院日等情况。结果术后一周内浅前房的发生率观察组为16.1%(9只眼),对照组为31.4%(16只眼);术后半年,眼压的控制两组无明显差异(P〉0.05);平均住院日观察组小于对照组,差异有显著性(P〈0.05)。结论小梁切除术联合应用可调节缝线技术,可减少术后早期浅前房的发生,增加手术安全性,缩短平均住院日,但对眼压的长期控制并不优于传统的小梁切除术。
简介:目的:观察角膜缘干细胞培养液对血管内皮细胞增殖的影响。方法:分别培养角膜缘干细胞及球结膜上皮细胞,并通过免疫组化检测AE-5蛋白鉴别角膜缘干细胞。搜集两组培养细胞的上清液加入培养的人脐静脉血管内皮细胞中,并设立对照组。培养24h后通过MTT法检测细胞增殖情况并进行统计学分析。结果:角膜缘干细胞AE-5染色呈阴性,而球结膜上皮细胞为阳性。加入角膜缘干细球结膜上皮细胞和对照组培养液的血管内皮细胞MTT值分别为2.097±0.079,1.981±0.034和1.990±0.044。三组间有显著性差异(F=9.169,P=0.000)。加入角膜缘干细胞培养上清液的血管内皮细胞增殖活性明显高于其他两组(P=0.005和P=0.007)。结论:角膜缘干细胞培养液能够促进血管内皮细胞的增殖,这可能是角膜缘干细胞的特征。本研究从功能学角度为角膜缘干细胞理论提供更多的依据。
简介:目的观察上调白细胞介素10(IL-10)的表达对家兔慢性细菌性鼻窦炎动物模型上颌窦黏膜创伤修复的影响。方法以慢病毒(LV)为表达载体,上调IL-10的表达(LV-IL-10)。实验分3组:生理盐水注射组、LV-IL-10治疗组和LV-GFP注射组(GFP为绿色荧光蛋白)。制作家兔慢性细菌性鼻窦炎模型。在创伤前3d,各组上颌窦内侧壁黏膜下分别注射生理盐水、LV-IL-10和LV-GFP。于创伤后第3天和第10天取再生的上颌窦内侧壁黏膜,用实时聚合酶链反应和酶联免疫吸附试验分别在mRNA水平和蛋白水平检测相关细胞因子的表达。苏木素-伊红(HE)染色和Masson三染色法观察创伤后第10天再生黏膜的形态特征。结果在创伤后第3天和第10天再生的上颌窦黏膜中,LV-IL-10治疗组的胶原沉积明显减少,炎症反应较轻,且IL-6以及Ⅰ型和Ⅲ型胶原的mRNA表达水平明显下降。IL-6、IL-8以及Ⅰ型和Ⅲ型胶原的蛋白表达水平也明显下降,但3组间再生的上颌窦黏膜中转化生长因子β(TGF-β)mRNA的表达无明显差别。结论在家兔慢性细菌性鼻窦炎创伤修复的动物模型中,上调IL-10的表达可以抑制再生黏膜中的炎症反应,减少胶原沉积,改善再生黏膜的组织重塑。
简介:·AIM:Toinvestigatechangesinimmunogenicityofcryopreservedlimbalstemcells.·METHODS:Cryopreservedlimbalstemcells,freshprimarylimbalstemcellsandblankcontrolswereinoculatedsubcutaneouslyinC57BL-6miceandthepercentageofCD25cellsinlimbalexplantswasdeterminedbyflowcytometryatday21postinoculation.Morphologicalstudieswereperformedbylightandelectronmicroscopyoflimbalexplantsections.·RESULTS:Thenumberofregionalandsystemiclymphocytesderivedfromcryopreservedlimbalstemcellswaslowerthanthatfromfreshprimarylimbalstemcells.·CONCLUSION:Lymphocytesderivedfromcryopreservedlimbalstemcellsshowedchangesinimmunogenicity,butthesignificanceisunknown.Thecryopreservationandthawingmethodsawaitfurtherstudy.·