三种准分子激光手术中角膜切削厚度的对比研究

目的 通过术中利用光学相干厚度测量的方法评估飞秒激光制瓣联合LASIK、LASIK与LASEK三种手术中角膜切削深度的差异。设计 前瞻性比较性病例系列。研究对象 2013年3月到2014年5月行阿玛仕准分子激光矫正手术的患者103例200眼,其中飞秒激光制瓣LASIK组50例100眼,角膜板层刀LASIK组30例57眼,LASEK组23例43眼。方法 术中使用实时角膜厚度测量（OCP）技术分别测量制瓣后激光切削前和激光切削后的中央角膜厚度,从而计算实际角膜基质切削深度。比较各组实际角膜切削深度与激光系统预期最大切削深度的差异及三种术式实际角膜切削深度与理论切削深度差值的组间差异。主要指标 理论角膜切削深度、实际角膜切削深度。结果 飞秒激光制瓣LASIK组OCP所测角膜切削深度（117.22±31.13）μm,准分子激光系统预计角膜切削深度（99.52±25.47）μm（t=-12.55,P<0.001）。角膜板层刀制瓣LASIK组OCP所测角膜切削深度（116.30±22.93）μm,预计角膜切削深度（94.26±16.37）μm （t=-12.44,P<0.001）。LASEK组OCP所测角膜切削深度（106.72±23.77）μm,预计角膜切削深度（99.31±20.46）μm（t=-4.44,P<0.001）。实际角膜切削深度与预计值比较,飞秒激光制瓣LASIK组为（17.69±14.10）μm,角膜板层刀制瓣LASIK组为（22.04±13.38）μm,LASEK组为（7.41±10.95）μm。飞秒激光制瓣及板层刀制瓣LASIK组实际角膜切削深度与理论切削深度差值较LASEK术式组切削深度差值较大（P均<0.001）。飞秒激光组与角膜板层刀组之间差异无统计学意义（P=0.15）。结论 术中实时光学相干厚度测量的方法所测三种准分子激光手术实际角膜切削深度较激光系统预计值大,术前预测角膜基质床厚度时需考虑到预计值与实际值差值存在。（眼科, 2014, 23： 305-307）     

三种不同方案角膜交联后角膜厚度减小量的比较

目的 评估三种不同方案角膜交联(corneal crosslinking,CXL)治疗进展期圆锥角膜后最薄角膜厚度(thinnest-point corneal thickness,TCT)减小量的差异.方法 回顾性临床病例研究.选取2010年8月至2015年11月在海军总医院眼科确诊并行CXL治疗的进展期圆锥角膜连续性病例85例(110眼).21例(25眼)行标准的去上皮CXL(standard epithelium-off corneal crosslinking,S-CXL)治疗;14例(22眼)行1g·L-1核黄素乳酸钠林格液离子导入CXL(iontophoresis-assisted CXL,I-CXLa)治疗;50例(63眼)行1g·L-1核黄素蒸馏水溶液离子导入CXL(iontophoresis-assisted CXL,I-CXLb)治疗.应用ALLEGRO眼前节分析仪测量术前、术后TCT,比较三种CXL方案术后TCT减小量的差异.结果 S-CXL术后3个月、6个月、12个月TCT与术前差值分别为(-14.93±27.16)μm、(-31.94±22.89)μm、(-27.71±26.01)μm;I-CXLa术后3个月、6个月、12个月TCT与术前差值分别为(-20.14±19.09)μm、(-10.10±24.28)μm、(-7.11±22.26)μm;I-CXLb术后3个月、6个月、12个月TCT与术前差值分别为(-28.08±26.14)μm、(-21.08±25.62)μm、(-15.91±19.19)μm.术后3个月时,三组TCT减小量差异没有统计学意义(P =0.188);术后6个月、12个月时,S-CXL组和I-CXLa组比较差异均有统计学意义(均为P＜0.05),I-CXLb组和S-CXL组、I-CXLb组和I-CXLa组比较差异均无统计学意义(均为P＞0.05).结论 三种CXL方案治疗术后6个月和12个月,TCT减小量与交联方案相关,反映了交联强度的大小.I-CXLb术后TCT减小量小于S-CXL,但差异不显著,两者交联强度差异亦不显著.     

Applications of hydrogel materials in different types of corneal wounds

Severe corneal injury can lead to a decrease in light transmission and even blindness. Currently, corneal transplantation has been applied as the primary treatment for corneal blindness; however, the worldwide shortage of suitable corneal donor tissue means that a large proportion of patients have no access to corneal transplants. This situation has contributed to the rapid development of various corneal substitutes. The development and optimization of novel hydrogels that aim to replace partial or full-thickness pathological corneas have advanced in the last decade. Meanwhile, with the help of 3D bioprinting technology, hydrogel materials can be molded to a refined and controllable shape, attracting many scientists to the field of corneal reconstruction research.Although hydrogels are not yet available as a substitute for traditional clinical methods of corneal diseases, their rapid development makes us confident that they will be in the near future. We summarize the application of hydrogel materials for various types of corneal injuries frequently encountered in clinical practice, especially focusing on animal experiments and preclinical studies. Finally, we discuss the development and achievements of 3D bioprinting in the treatment of corneal injury.     

三种不同设备行屈光手术时的角膜切削误差比较

目的调查2种不同的准分子激光设备和1种全飞秒激光手术设备用于准分子激光原位角膜磨镶术（LASIK）时对中央角膜厚度切削误差的影响,探索角膜切削误差的变化规律。方法前瞻性病例对照研究。使用A型超声角膜测厚仪测量分别应用3种不同设备行手术治疗的138例（274眼）患者术前及术后1个月的中央角膜厚度。其中51例（100眼）使用Esiris机器行LASIK手术（Esiris—LASIK）,50例（100眼）使用VisxS4-IR机器行LASIK手术（Visx—LASIK组）,37例（74眼）使用VisuMax机器行SMILE手术（VisuMax—SMILE组）。所有眼按屈光度分为低、中、高度3组,分别计算中央角膜厚度及角膜切削误差,并分析切削误差与不同设备、近视程度、散光程度、角膜切削直径及手术前角膜厚度的关系。采用单样本t检验、双因素方差分析及Pearson相关进行数据分析。结果3组病例中,近视程度、术前中央角膜厚度及术前散光度对角膜切削误差均没有显著影响,设备或手术方式对角膜切削误差有显著的影响。在Esiris—LASIK组中,角膜的实际切削厚度小于预期切削厚度,差异有统计学意义（低度近视组,t=4．672,P〈0．01;中度近视组,t=10．629,P〈0．01;高度近视组,t=11．021,P〈0．01）;Visx—LASIK组中,角膜的实际切削厚度大于预期切削厚度,差异有统计学意义（低度近视组,t=3．910,P〈O．01;中度近视组,t=4．922,P〈0．01;高度近视组,t=4．807,P〈0．01）;在VisuMax—SMILE组中,角膜的实际切削厚度与预期切削厚度差异无统计学意义（中度近视组,t=1．158,P〉O．05;高度近视组,t=0．836,P〉0．05）。结论不同的手术设备会显著影响屈光手术中中央角膜厚度的切削误差。     

Chemotactic and haptotactic activities of fibronectin for cultured rabbit corneal epithelial cells

A previous study reported that both fibronectin and epidermal growth factor (EGF) stimulated corneal epithelial resurfacing. Fibronectin appears in the cornea after injury, and corneal epithelial cells migrate over the temporary fibronectin matrix. To determine whether fibronectin serves as chemoattractant and haptoattractant for the directed movement of corneal epithelial cells, the directed migration of cultured rabbit corneal epithelial cells was measured in vitro using a Boyden chamber. Chemotactic and haptotactic migration were assayed separately. Fibronectin was found to stimulate attachment of corneal epithelial cells and to have chemotactic, haptotactic and chemokinetic activities for the corneal epithelial cells. In contrast, EGF had no chemotactic activity.     

Spreading of cultured corneal epithelial cells on fibronectin and other extracellular matrices

Previous investigations demonstrated that fibronectin is an essential adhesive glycoprotein for the corneal epithelial cells. Fibronectin mediates attachment and spreading of the corneal epithelial cells. However, it seems to be important to know the changes in cellular receptor activity in order to understand the interactions of corneal epithelial cells and underlying extracellular matrix. In this investigation, we studied the effects of various culture times and conditions on the receptor activity of rabbit corneal epithelial cells for fibronectin. The cultured cells were placed on tissue culture plates that were coated with various concentrations of four extracellular matrices: fibronectin, laminin and collagen types I and IV. Freshly isolated corneal epithelial cells without culture did not spread on fibronectin and laminin; they spread only on collagen types I and IV. When the epithelial cells were cultured for 12 h or more, they spread on fibronectin. However, the spreading of the cells on collagen types I and IV was the same regardless of the culture period (up to 20 h). Only a small number of epithelial cells spread on laminin at the highest concentration examined after culture for 12 h or more. Thus, the corneal epithelial cells responded differently to fibronectin, collagen types I and IV, and laminin. Perhaps the receptor for fibronectin appears when epithelial cells are cultured, but the receptor(s) for collagen types I and IV are always present on the cell surface of corneal epithelial cells.     

Topical fibronectin therapy in persistent corneal ulceration

Fifteen patients with 20 episodes of persistent corneal ulceration, resistant to conventional therapy, were treated with topical autologous fibronectin. Thirteen corneal ulcers (eight patients) developed following penetrating keratoplasty, three patients had mucous membrane pemphigoid, two patients had herpetic keratitis and one each had Sj?gren's syndrome and a trophic corneal ulcer. A standard protocol for fibronectin administration was followed. This therapy healed 16 of the 20 ulcers after a mean duration of treatment of 2.3 months. Corneal ulceration associated with mucous membrane pemphigoid failed to respond to fibronectin. Corneal ulcers which recurred after cessation of fibronectin responded to reintroduction of this therapy. Topical fibronectin is an effective therapy for refractory corneal ulceration and is free of major side effects.     

PRK术中角膜上皮的去除

目的：了解ＰＲＫ术中激光（Ｌ组）、激光加机械刮除（Ｌ＋Ｓ组）及机械刮除（Ｓ组）三种去除角膜上皮方法在操作时间、术后上皮修复、视力、角膜上皮下混浊（Ｈａｚｅ）、切削偏中心及角膜中央岛发生等方面的异同。方法：对上述三种方法去除角膜上皮的ＰＲＫ术后追踪超过半年的４９６例（９６４只眼）近视眼进行统计学分析。结果：Ｓ组刮除上皮的时间是其它组的２倍，其上皮修复也相对较慢，偏中心及过矫发生的比例及程度也较重；Ｌ组屈光欠矫及中央岛的发生较多；Ｌ＋Ｓ组视力最好。三组Ｈａｚｅ发生差异无显著性。结论：激光去除上皮快捷、中心位易确定、术后恢复较快，但有时因角膜表面水分积聚不均、患眼眼球转动、角膜厚度的个体差异等可能导致角膜上皮去除不彻底或小岛残余；机械刮除角膜上皮彻底、可减少过多激光脉冲叠加对角膜的热损伤、玻璃体的扰动及视网膜的震荡，但因耗时长，角膜干燥变薄，术后易产生过矫，同时术中如角膜变混，中心位也较难确定；激光加机械刮除兼有二者的优点，并可克服其缺点，应视为首选。但对超高度近视，尤其患眼有严重近视眼视网膜病变时，可考虑机械刮除角膜上皮     

Topical fibronectin treatment in persistent corneal epithelial defects and corneal ulcers

Topical fibronectin, autologous and homologous, was used to treat nine patients (eleven eyes) with persistent corneal epithelial defects and corneal ulcers that failed to improve with standard therapy. The fibronectin was purified from autologous and homologous plasma by gelatin-Sepharose 4B affinity chromatography and administered topically, 500 micrograms/ml five times a day, for three weeks. Complete or nearly complete reepithelialization was achieved in all patients regardless of the source of fibronectin, autologous or homologous. But healing times varied. The average healing time was 41.7 +/- 14.7 days (35.7 +/- 12.4 days for autologous, 50.8 +/- 14.4 days for homologous). Ocular symptoms were relieved significantly, and no side effects were observed. Over an average follow-up period of 5.2 months, no recurrences were noted. The results showed that homologous, as well as autologous, fibronectin was effective in patients with persistent corneal epithelial defects and corneal ulcers.     

三种仪器测量中央角膜厚度的对比研究

目的探讨Pentacam眼前段分析仪、Orbscan-Ⅱ眼前节分析仪与A超角膜厚度测量仪三种仪器测量中央角膜厚度的差异。方法126例（252眼）欲做准分子激光手术的近视患者分别用Pentacam眼前段分析仪、A超角膜测厚仪、Orbscan-Ⅱ三种仪器测量中央角膜厚度。用SPSS11.0统计学软件对不同方法测量的结果进行配对t检验,并作Pearson相关性分析。结果三种仪器所测得中央角膜厚度测量结果分别为：Pentacam（546.3±33.6）μm,Orbscan-Ⅱ（550.5±38.5）μm,A超角膜测厚仪（538.6±35.4）μm。三种检查仪测量结果两两之间差异均有统计学意义（P=0.000）,且有高度的相关性。A超角膜测厚仪比其他两种测量仪测量所得的中央角膜厚度要薄。再以A超角膜测厚仪测量的角膜厚度为基准把患者分为三组：〈520μm,≥520μm且〈580μm,≥580μm,对三种仪器测量的结果进一步对比分析。第一组角膜厚度〈520μm,71只眼,A超角膜测厚仪测量结果比Pantacam和Orbscan-Ⅱ测量结果要薄,差异均有统计学意义（P=0.000,P =0.000）。而Pentacam与Orbscan-Ⅱ的测量值差异无统计学意义（P=0.143）。第二组角膜厚度≥520μm且〈580μm,154只眼,Orbscan-Ⅱ测量结果比Pantacam较厚,Pantacam比A超角膜测厚仪测量结果要厚,差异均有统计学意义（P=0.000,P=0.000）。第三组角膜厚度≥580μm,27只眼,Orbscan-Ⅱ测量结果比Pantacam和A超角膜测厚仪测量结果要厚,差异均有统计学意义（P=0.000,P=0.000）,而Pentacam与A超角膜测厚仪的测量值差异无统计学意义（P=0.747）。结论在对中央角膜厚度的测量中,Pentacam、Orbscan-Ⅱ与A超角膜测厚仪三种仪器之间是不能互相替换的。     

Morphological changes of corneal subepithelial nerve plexus in different types of herpetic keratitis

Purpose  

We investigated by in vivo confocal microscopy alterations in the subepithelial nerve plexus in different types of herpes simplex keratitis (HSK).     

不同病理类型的葡萄膜黑色素瘤中微小RNA差异表达谱分析

背景 目前研究证实微小RNA(miRNA)参与大多数人类肿瘤疾病的发生和发展,其作用类似于抑癌基因或癌基因.葡萄膜黑色素瘤(UM)是成人常见的眼部恶性肿瘤,其发生和转移机制仍未完全阐明.探讨UM组织中miRNA的差异表达情况有望为UM的靶向治疗提供依据. 目的 筛选不同病理类型的UM组织中特异性miRNA表达谱. 方法 收集于2013年3月至2015年10月在北京同仁医院手术局部切除并经常规组织病理学和免疫组织化学检测证实为梭型细胞型UM的标本4例和上皮细胞型UM标本4例,采用miRNA芯片分别检测2种UM组织中miRNA的表达,收集同期死于非肿瘤疾病的8个供体眼的正常葡萄膜组织作为对照,利用组间差异倍数筛选出差异≥2倍差异表达的miRNA;用在线软件预测差异表达miRNA的靶基因,采用生物信息学方法分析靶基因参与的信号功能通路.采用实时定量PCR法验证芯片检测结果.结果 收集的梭形细胞型和上皮细胞型UM标本经组织病理学检查均得到确诊,免疫组织化学检测梭形细胞型及上皮细胞型UM组织中HMB45、黑色素-A和S-100均呈阳性反应.与正常葡萄膜组织比较,在梭形细胞型UM组织中差异表达的miRNA有109个,其中29个上调,80个下调,上调的miRNA包括miR-146a-5p、miR-25-3p和miR-29b-1-5p,下调的miRNA包括miR-126-5 p、miR-183-5p和miR-96-5p;上皮细胞型UM中差异表达的miRNA有50个,其中23个上调,27个下调,上调的miRNA包括miR-155-5p、miR-210和miR-378 a-5p;下调的miRNA包括miR-199a-5p、miR-143-3p和miR-143-5p.在梭形细胞型和上皮细胞型UM组织中共同上调的miRNA为miR-132-3p、miR-21-5p、miR-34a-5p和miR-34b-5p,共同下调的miRNA为miR-125b-2-3p、miR-126-3p、miR-199a-3p和miR-214-3p.梭形细胞型和上皮细胞型UM组织中差异表达的miRNA所预测的靶基因分别参与癌症通路、丝裂原活化蛋白激酶(MAPK)信号通路、Wnt信号通路、细胞间黏附、胞吞作用、前列腺癌通路、结直肠癌通路和细胞黏附通路.结论 与正常葡萄膜组织相比,梭形细胞型UM和上皮细胞型UM组织中存在多种miRNA的差异表达,梭形细胞型UM和上皮细胞型UM组织之间也存在明显的miRNA差异表达,这些差异表达的miRNA可通过不同的信号转导通路参与调控UM的生物学行为.     

Homologous serum-stimulated fibronectin synthesis in human retinal pigmented epithelial cells.

PURPOSE: Fibronectin expression has been recorded in subretinal membranes from patients with proliferative vitreoretinopathy (PVR). Retinal pigmented epithelial (RPE) cells are a major cellular component of PVR membranes and synthesize fibronectin. We investigated the effects of human serum (as a model of vascular leakage in vivo) on the expression of fibronectin by cultured human RPE cells and compared the responses to those stimulated by fetal bovine serum (FBS). METHODS: Human RPE cells were incubated in M199 with 1, 10 and 40% human adult serum or FBS for 72 h. RESULTS: Retinal pigmented epithelial cells expressed maximum extracellular matrix fibronectin when exposed to 40% human serum using immunohistochemistry. Using ELISA to quantify fibronectin, 10 and 40% human serum significantly increased the total fibronectin (P < 0.01; n = 4), whereas FBS did not affect fibronectin expression. CONCLUSIONS: The results show that human serum contains substances stimulating fibronectin synthesis in human RPE cells.     

角膜内皮细胞压力调控的研究进展

随着角膜内皮细胞体外培养技术的不断改进,体外培养角膜内皮细胞构建工程化角膜内皮细胞移植膜,置换病变或受损的角膜内皮,恢复角膜透明性已成为可能.正常压力对角膜内皮细胞具有正向调控作用;高压对角膜内皮细胞损伤的原因是多因素的,其具体机制有待进一步研究.     

人脂肪组织来源的干细胞体外诱导为角膜上皮样细胞的分化潜能

目的 初步探讨人脂肪组织来源的干细胞(human adipose-derived stem cells,hADSC)体外诱导为角膜上皮样细胞的分化潜能.方法 分离、纯化、体外扩增hADSC,流式细胞术鉴定所获得细胞的CD29+、CD34-、CD49d+、CD105+、CD106-的表达情况.成脂、成骨诱导鉴定其多向分化潜能.在DMEM/F12体系、KM体系及上述二者等体积混合的KM/DMEM/F12体系内添加不同梯度浓度的生长因子对hADSC进行体外诱导:0μg·L-1、10μg·L-1、20μg·L-1、30μg·L-1、40μg·L-1、50μg·L-1的表皮生长因子(epidermal growth factor,EGF),及50μg·L-1 EGF+10μg·L-1 碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)和50μg·L-1 EGF+20 μg·L-1 bFGF.连续诱导21 d,观察hADSC形态学的改变及第21天时角膜特异性蛋白AE5(CK3/CK12)的免疫化学表达情况,比较不同培养体系及不同浓度生长因子对hADSC诱导作用的差异.结果 流式细胞仪测定传3-5代的细胞CD34-、CD106-、CD29+、CD49d+、CD105+.成脂、成骨体外诱导14 d后.油红O、碱性磷酸酶染色阳性率分别为74.6%和29.3%.KM体系中加入终浓度为0～50μg·L-1 EGF诱导21d后,hADSC AE5阳性细胞率均占90%以上.其中,40μg·L-1EGF诱导下的AE5阳性细胞率为98.7%,50μg·L-1 EGF可达到100%.而加入bFGF的hADSC 则为AE5弱阳性.而另2个体系对各浓度ECF、bFGF诱导的hADSC的作用均为阴性.结论 人吸脂术废弃液中可获得大量高纯度脂肪组织来源的干细胞,经体外条件诱导具备向角膜上皮样细胞分化潜能.添加EGF的角质细胞培养液有利于其分化,并具有浓度依赖性,bFGF则对分化有抑制性作用.