角膜是基因治疗的理想靶器官。角膜碱烧伤、角膜新生血管、角膜移植术后排斥反应因其病理机制复杂，所牵涉的致病因素众多而治疗困难，疗效不佳。本文就基因治疗在上述疾病中的应用加以综述，以了解基因治疗应用于角膜病变的新进展 。

Cornea is an ideal target organ for gene therapy. Corneal alkali burn, cornealneovascularization and corneal graft rejection tend to be with poor treatment elicacydue to its complex pathogenesis. This article aims to update the recent progress of genetherapy on corneal diseases.

神经营养性角膜病变是一种与角膜神经退行性改变有关的疾病，角膜神经的知觉和营养功能受损，导致角膜上皮缺损、角膜溃疡甚至角膜穿孔。目前人工泪液、治疗性角膜绷带镜、泪点栓塞、羊膜移植，睑缘缝合等治疗措施仍是治疗神经营养性角膜病变的主要治疗方式，对于轻中度病变患者，具有较好的治疗效果，而对于重度病变患者，药物治疗及简单的手术干预治疗效果不佳，病情反复发作。由于重度神经营养性角膜病变患者的角膜神经完全消失，丧失角膜感觉，对未恢复角膜神经营养功能的角膜白斑或溃疡患者行角膜移植术，可能导致角膜移植术后上皮持续不愈合，因此恢复角膜神经营养功能是复明的重要保障手段。角膜神经移植术是重度神经营养性角膜病变患者恢复角膜神经营养功能，提高角膜知觉，改善角膜透明度的重要和有效的治疗方法。角膜神经移植术通过将具有正常功能的供体神经移植到麻痹眼角膜缘周围，使神经末梢重新长入角膜基质，恢复角膜知觉功能。随着角膜神经移植术的术式的不断改进，其良好的术后效果和优点已经渐渐突显。角膜神经移植术包括直接角膜神经移植和间接角膜神经移植，促使角膜神经重新生长，重建角膜神经的营养和知觉功能。角膜神经移植手术已有40年历史，1981年Samii等首次报告了角膜神经移植术，2009年Terzis等成功地实施了第1例直接角膜神经移植术，2014年Elbaz等进行了第1例以腓肠神经作为间置移植物的间接角膜神经移植。封面展示了神经营养性角膜病变患者未接受治疗前的和接受角膜神经移植术后的眼表角膜图像。由于角膜神经退行性改变，角膜失去神经支配，继而出现角膜上皮缺损，角膜缘新生血管形成，经角膜神经移植后，角膜上皮愈合，角膜透明度改善，同时角膜缘新生血管消退。

神经营养性角膜病变是一种与角膜神经退行性改变有关的疾病，角膜神经的知觉和营养功能受损，导致角膜上皮缺损、角膜溃疡甚至角膜穿孔。目前人工泪液、治疗性角膜绷带镜、泪点栓塞、羊膜移植，睑缘缝合等治疗措施仍是治疗神经营养性角膜病变的主要治疗方式，对于轻中度病变患者，具有较好的治疗效果，而对于重度病变患者，药物治疗及简单的手术干预治疗效果不佳，病情反复发作。由于重度神经营养性角膜病变患者的角膜神经完全消失，丧失角膜感觉，对未恢复角膜神经营养功能的角膜白斑或溃疡患者行角膜移植术，可能导致角膜移植术后上皮持续不愈合，因此恢复角膜神经营养功能是复明的重要保障手段。角膜神经移植术是重度神经营养性角膜病变患者恢复角膜神经营养功能，提高角膜知觉，改善角膜透明度的重要和有效的治疗方法。角膜神经移植术通过将具有正常功能的供体神经移植到麻痹眼角膜缘周围，使神经末梢重新长入角膜基质，恢复角膜知觉功能。随着角膜神经移植术的术式的不断改进，其良好的术后效果和优点已经渐渐突显。角膜神经移植术包括直接角膜神经移植和间接角膜神经移植，促使角膜神经重新生长，重建角膜神经的营养和知觉功能。角膜神经移植手术已有40年历史，1981年Samii等首次报告了角膜神经移植术，2009年Terzis等成功地实施了第1例直接角膜神经移植术，2014年Elbaz等进行了第1例以腓肠神经作为间置移植物的间接角膜神经移植。封面展示了神经营养性角膜病变患者未接受治疗前的和接受角膜神经移植术后的眼表角膜图像。由于角膜神经退行性改变，角膜失去神经支配，继而出现角膜上皮缺损，角膜缘新生血管形成，经角膜神经移植后，角膜上皮愈合，角膜透明度改善，同时角膜缘新生血管消退。

目的:通过观察对金黄色葡萄球菌性角膜溃疡的疗效,筛选克拉霉案眼用凝胶的合适浓度。

方法:角膜实质层接种法建立40只家兔右眼金黄色葡萄球菌性角膜溃疡模型,将模型随机分成5组,每组8只免(8只眼),各组分别给予空白基质、0.1%克拉素眼用凝胶、0.25%克拉霉素眼用凝胶、左氧氟沙星凝胶、0.25%克拉霉素眼用凝胶联合重组牛碱性成纤维细胞生长因子(Recombinant bovine basic fibroblast growth factor, Rb-bFGF)，每天4次，每次2滴，分别在第1、3、5、7、10、14 天观察角膜病变情况及溃疡面积大小。

结果:在相同的给药方法下，0.1%克拉霉素眼用凝胶、0.25%克拉素眼用凝胶、左氧沙星凝胶、0.25%克拉霉素眼用凝胶联合Rb-bFCF均能使金黄色葡萄球菌性角膜溃疡面积缩小角膜病变好转，与空白基质组相比有统计学差异(P < 0.05)：0.25%克拉素眼用凝胶组疗效明显优于0.1%克拉荐素眼用凝胶组(P < 0.05)。

结论:制备的 0.25%克拉霉素眼用凝胶对金黄色葡萄球菌性角膜溃疡疗效肯定，可以进一步开发应用于临床。

Purpose:To screen proper concentration of clarithromycin ophthalmic gel by observingthe efficacy of different concertrations of clarithromycin ophthalmic gel for treatingstaphylococcal corneal ulcers.
Methods:Corneal ulcer was induced in the right eye of 40 rabbits, 3.0 x 10°CFU/mlstaphylococcus aureus suspension was injected midstromally into the central cornel.These rabbits were divided randomly into $ groups ,each group received respectivelytopical blank matrix, clarithromycin ophthalmic gel 0.1%, clarithromycin ophthalmicgel 0.25%,levofloxacin ophthalmic gel, clarithromycin ophthalmic gel 0.25% andrecombinant bovine basic fibroblast growth factor (Rb-bFGF), 4 times every day, 2drops each time. The eyes were examined respectively with the slit lamp beforetreatment(day0), on day3, day5, day7, day 10, day 14 to observe theprogression of corneal ulceration. including the area of the corneal ulcer and mark of keratitis.

Resuls:Under the same way of giving medicine, experimental coreal ulcer studiesshowed a statistically significant decrease in all tratement groups on measurements ofthe area of the comeal ulcer and mark of keratitis(P<0.05), and clarithromycinophthalmic gel 0.25% had a better action than clarithromycin ophthalmic gel 0.1%against staphylococcus aureus corneal ulcer.
Conclusion:Clarithromycin ophthalmic gel 0,25% was proved to be an effective ocularmedication for the therapy of gram-positive bacterial corneal ulcer. 

目的: 研究 TGF-β1 短期眼部应用对兔角膜碱烧伤后整合素 β1 表达和角膜上皮愈合的影响,探求其对角膜碱烧伤的治疗作用。

方法: 制备大耳白家兔角膜碱烧伤模型, 一组给予 TGF-β1 (浓度为 200 ng /ml) 局部滴眼, 每日 3 次, 连续 7 日; 另一组给予 PBS 溶液代替, 处理相同。于角膜碱烧伤后每日观察角膜上皮愈合面积, 并于烧伤后 6 h、1 d、3 d、7 d 和 14 d 5 个时间点应用免疫组化方法检测 TGF-β1 实验组与 PBS 组角膜整合素 β1 表达情况。

结果: 烧伤后 4 d、10 d、11 d、12 d 和 14 d 实验组和对照组上皮愈合率比较, 差异有统计学意义(P < 0.05) , 两组随着上皮修复过程的进行, 整合素 β1 的表达均逐渐增加, 烧伤后 7 d、14 d两个时间点实验组和对照组整合素 β1 平均灰度值比较, 差异有显著性(P < 0.05) 。

结论: TGF-β1 在活体实验中能促进整合素 β1 的表达, 而后者的增加可以促进角膜上皮细胞向损伤区域的移行和粘附, 从而减少碱烧伤愈合过程中上皮再次脱落现象, 有利于创伤愈合。

Purpose: To observe the effect of TGF-β1 applied topically to the alkali-injured rabbit eye on corneal epithelial wound healing and expression of integrin β1 and its therapeutic action on corneal alkali burns.
Methods: Alkali burn was produced in 60 corneas from 30 rabbits. Two groups were randomly divided. One group was treated with TGF-β1 solution (200 ng /ml) topically 3 times one day within the first 7 days, the other group was treated with phosphate-buffered saline (PBS) solution. The injured eyes were photographed after the fluorescence staining with a digital camera and the pictures were analyzed with computer-aided picture analysis system to calculate the rate of corneal epithelial healing. The expression of integrin β1 was investigated in the point 6 h, 1 d, 3 d, 7 d, 14 d after the injury by means of immunohistochemical analysis.
Results: On the 4th, 10th, 11st, 12nd and 14th days after the burning, the rate of corneal epithelial healing of TGF-β1 groups was markedly higher than that of the PBS group (P < 0.05) . The expression of integrin β1 in the cornea epithelial cells gradually increased during the wound healing. On the 7th and 14th days after the burning, the expression of integrin β1 in the cornea epithelial cells of TGF-β1 group was remarkably  higher than that of the PBS group(P < 0.05) .

Conclusions: TGF-β1 could up-regulate integrin β1 in vivo corneal alkali burn model, which could stimulate the cornea epithelial cells to migrate and adhere to the cornea stroma, that can reduce the cases of the epithelial cells_detachment from the cornea stroma and sustain the corneal reepithelization. 

目的:探讨碱性成纤维细胞生长因子(Basic fibroblast growthfacfor, bFCF)，表皮细胞生长因子(Epidermal growth factor, EGF)和神经细胞生长因子(Nerve growth factor, NGF)对体外培养的人角膜内皮细胞的生长调控作用。

方法:将相同数量的人角膜内皮细胞接种于96孔板。加人浓度分别为0 ng/ml、1 ng/ml、3 ng/ml、10 ng/ml、30 ng/ml、100 ng/ml的 EGF、bFGF 和 NGF 进行培养。5 天后 MTT法用检测增殖情况。
结果:在0 ng/ml、1 ng/ml、3 ng/ml、10 ng/ml、30 ng/ml、100 ng/ml 浓度下 bFGF 组的平均 OD 值分别为： 0.224±0.045、0.239±0.040、0.262±0.0342、0.278±0.0319、0.281±0.0324、0.260±0.0310 EGF组的平均 0D 值分别为: 0.228±0.0304、0.245±0.0418、0.267±0.0454、0.275±0.0347、0.271±0.0449、0.250±0.0253。NGF 组的平均 OD 值分别为：0.216±0.0187、0.228±0.0226、0.231±0.0225、0.242±0.0279、0.245±0.0294、0.247±0.0349。
结论:bFGF在 30 ng/ml范围内对内皮细胞生长有促进作用,并具有剂量依赖性。高于100 ng/ml时促生长作用降低。EGF在10 ng/ml范围内对内皮细胞生长有促进作用，并具有剂量依赖性。高于30 ng/ml 时促生长作用降低。NGF本次实验剂量范围内对角膜内皮细胞生长无明显作用。

Purpose: To investigate effect of bFGF, EGF and NGF on growth of cultured humancorneal endothelial cells.
Methods: Cultured human corneal endothelial cells were seeded into individual wellsof 96-well tissue culture plate with the same culture media containing separately bFCF, EGF or NGF with a serial of concentrations of 0 ng/ml、1 ng/ml 、3 ng/ml、10 ng/ml 、30 ng/ml and 100 ng/ml and then cultured for 5 days. Then MTT method wasused to detect the growth of the cells.
Results: The averaged OD values of the cell wells containing bFCF with a serial of concentrations of 0 ng/ml、1 ng/ml、3 ng/ml、10 ng/ml、30 ng/ml and 100 ng/ml were 0.224±0.045, 0.239±0.040, 0.262±0.0342, 0.278±0.0319, 0.281±0.0324, 0.260±0.0310. The averaged OD values of EGF group and NGF group were separately 0.228±0.0304，0.245±0.0418, 0.267±0.0454, 0.275±0.0347, 0.271±0.0449, 0.250±0.0253 and 0.216±0.0187, 0.228±0.0226, 0.231±0.0225, 0.242±0.0279, 0.245±0.0294，0.247±0.0349.
Conclusion: bGFC can promote the growth of human corneal endothelial cells in adose dependent manner while with concentration lower than 30 ng/ml. bFGF withconcentration that is higher than 100ng/ml will weaken this effect. EGF can alsopromote proliferation of human corneal endothelial cells demonstrating a linear dosedependent effect when its concentration is lower than 10 ng/ml and this effect decreasedwhen its concentration was higher than 30 ng/ml, NGF showed no effect on the growthof human cornea endothelial cells in this study.

目的：分析有意愿接受准分子激光角膜屈光手术治疗近视的患者在术前检查后未行手术的原因。方法：选取2015年1月至2017年6月在安徽医科大学附属第一医院眼科激光中心检查的近视患者，分析其中未行激光手术的原因。结果：共2 875例患者准备接受准分子激光角膜屈光手术，其中821例(28.6%)检查后未进行手术，男462例(56.3%)，女359例(43.7%)，年龄18~51(23.69±5.85)岁。821例中社会心理因素影响484例(59.0%)，角膜薄或角膜厚度不足182例(22.2%)，超高度近视(>10.00 D)100例(12.2%)，高眼压19例(2.3%)，弱视13例(1.6%)，角膜曲率异常11例(1.3%)，眼底病变7例(0.9%)，角膜变性3例(0.4%)，其他2例(0.2%)。结论：近视治疗术前检查后未进行准分子激光角膜屈光手术的原因多种多样，其中社会心理因素及角膜厚度不足是最主要的原因。术前详细检查，严格掌握手术适应证和禁忌证以及充分医患沟通是手术安全的保障。

Objective: To analyze the reasons for not performing laser refractive keratomileusis surgery among refractive surgery candidates under regular preoperative examination. Methods: The patients with myopia examined in the Center of Ophthalmology in the First Affiliated Hospital of Anhui Medical University Laser between January 2015 and June 2017 were selected, the reasons for not performing laser refractive keraomileusis after regular preoperative examination were analyzed. Results: A total of 2 875 patients requested refractive surgery and 2 054 (71.4%) of them received refractive surgery. Among 821 (28.6%) patients who did not get laser refractive keratomileusis, 462 (56.3%) were male and 359 (43.7%) were female, aged 18–51 years (range, 23.69±5.85 years). The most common reason for not offering refractive surgery were social psychological factors (59.0%), low central corneal thickness (22.2%), high myopia (12.2%), high intraocular pressure (2.3%), poor corrected visual acuity (1.6%), corneal topography anomaly (1.3%), retinal disease (0.9%), corneal (0.4%) and other diseases (0.2%). Conclusion: Reasons for not performing refractive surgery are quite diverse. Social psychological factors and inadequate corneal thickness were the most common reasons in the present study. Careful preoperative examination, strictly mastering indications and contraindications and full doctor-patient communication should be done for the safety of surgery.

目的：探讨大泡性角膜病变(bullous keratopathy，BK)患者行飞秒激光辅助的角膜内皮移植术(endothelium keratoplasty，EK)的围手术期护理。方法：回顾性分析在南京总医院眼科行飞秒激光辅助角膜内皮移植的BK20例患者，分析并总结患者术前的心理疏导、术前准备、眼科检查、术中如何配合医生、术后特殊体位、高眼压的护理及如何正确用药和出院健康教育等。结果：患者得到及时、有效的护理，术后视力均有所提高，内皮贴伏好，无排斥反应及其他并发症的发生。结论：通过对患者围手术期的护理，可提高护理质量，缩短患者住院时间。

Objective: To observe the perioperative nursing of femtosecond laser-assisted corneal endothelial transplantation in the treatment of patients with bullous keratopathy. Methods: A retrospective analysis of 20 patients with bullous keratopathy with femtosecond laser-assisted corneal endothelial transplantation in our department. The preoperative psychological counseling, preoperative preparation, ocular examination, how to cooperate with the doctors in operation, nursing of special position, high intraocular pressure after surgery, how to use eye drops correctly and health education after discharge were summarized. Results: Nursing were performed to the patients timely and effectively. Postoperative visual acuity of all the patients were improved, the endothelial and corneal stroma bed are well combined and no graft rejection and other complications occurred in the patients postoperatively. Conclusion: We concluded that the quality of nursing was improved and the time of hospitalization was shortened after perioperative nursing.

角膜屈光手术是目前屈光手术的主流术式，随着全飞秒、全激光手术方式的发展，手术变得更加安全精准，不仅角膜创伤小，术后恢复时间也进一步缩短。角膜具有屈光特性和典型的生物软组织力学特性，角膜力学特性不仅参与维持角膜形态，影响角膜手术尤其屈光手术的效果及预后，而且还与部分角膜疾病的发生和发展密切相关。近年来生物力学研究发展迅速，其在眼部疾病的诊疗中发挥着越来越重要的作用。角膜生物力学的变化与术前角膜的形态、不同手术方式的选择、术后角膜厚度的改变等多种因素相关，但手术导致的角膜自身形态改变是不可逆的，若术后角膜生物力学的变化较大，可能会引起医源性角膜扩张、继发性圆锥角膜等并发症的发生。为了规避术后角膜扩张风险和指导个性化的术式选择，了解角膜生物力学特性的影响至关重要。文章对角膜的基础结构、角膜生物力学特性、生物力学测量方法和不同术式及不同角膜瓣厚度术后生物力学变化的研究进展进行综述，为近视患者的个性化精准治疗提供理论指导。

Corneal refractive surgery is currently main stream of refractive surgery. With the development of femtosecond and laser surgery, the surgery has become safer and more accurate, resulting in less corneal trauma and a shorter postoperative recovery time. In recent years, biomechanics research has rapidly progressed, and its clinical application has gradually increased. The cornea not only possesses refractive properties but also exhibits typical biological soft tissue mechanical properties. Corneal mechanical properties not only play a role in maintaining corneal morphology but also influence the outcome and prognosis of corneal surgery, especially refractive surgery, and are closely related to the occurrence and development of some corneal diseases. Corneal refractive surgery involves cutting the cornea according to the patient's diopter, which disrupts the integrity of the cornea and inevitably affects its biomechanical stability. Changes in corneal biomechanics are associated with various factors, such as preoperative corneal morphology, the selection of different surgical methods, and postoperative changes in corneal thickness. However, the self-morphology changes caused by surgery are irreversible. If the postoperative changes in corneal biomechanics are significant, it may lead to complications such as postoperative corneal dilation and secondary keratoconus. To avoid postoperative iatrogenic corneal dilation and guide personalized surgical choice, it is crucial to understand the limits of influence of corneal biomechanical properties. This article reviews the research progress regarding corneal biomechanical properties and changes associated with corneal refractive surgery.

角膜缘的细胞，特别是角膜缘干细胞，对于维持角膜的透明和健康至关重要。基于影像技术对角膜缘进行高精度可视化评价是相关疾病诊疗的重要手段。眼科临床使用的裂隙灯显微镜、共聚焦显微镜、眼前节光学相干断层扫描仪(optical coherence tomography,OCT)等成像技术，因低分辨、低对比度、侵入性等原因，限制了其在角膜缘细胞结构及功能影像评估中的应用。本团队创新研发新型双模态全视场光学相干断层扫描仪(full-field OCT，FFOCT)，成功实现了无标记的角膜缘细胞级分辨率结构及功能成像。FFOCT基于空间非相干光平面干涉原理提取组织内部散射光，获得微米级分辨率三维结构成像；通过FFOCT原始相干信号的高时空分辨率采集及动态特征解析，实现源于活细胞新陈代谢运动的无标记细胞功能影像可视化。双模态FFOCT创新性地整合了高分辨率、无标记的结构及功能成像模态，不仅清晰获取角膜缘组织的高精结构特征如Vogt栅栏、角膜缘隐窝、血管壁等，同时还能捕捉不同角膜缘细胞内的代谢活性动态变化，无需使用外源荧光染料或标记剂，为角膜缘生物学及疾病机制研究提供全新细胞水平结构及功能成像方法，具有广泛应用前景。

角膜缘的细胞，特别是角膜缘干细胞，对于维持角膜的透明和健康至关重要。基于影像技术对角膜缘进行高精度可视化评价是相关疾病诊疗的重要手段。眼科临床使用的裂隙灯显微镜、共聚焦显微镜、眼前节光学相干断层扫描仪(optical coherence tomography,OCT)等成像技术，因低分辨、低对比度、侵入性等原因，限制了其在角膜缘细胞结构及功能影像评估中的应用。本团队创新研发新型双模态全视场光学相干断层扫描仪(full-field OCT，FFOCT)，成功实现了无标记的角膜缘细胞级分辨率结构及功能成像。FFOCT基于空间非相干光平面干涉原理提取组织内部散射光，获得微米级分辨率三维结构成像；通过FFOCT原始相干信号的高时空分辨率采集及动态特征解析，实现源于活细胞新陈代谢运动的无标记细胞功能影像可视化。双模态FFOCT创新性地整合了高分辨率、无标记的结构及功能成像模态，不仅清晰获取角膜缘组织的高精结构特征如Vogt栅栏、角膜缘隐窝、血管壁等，同时还能捕捉不同角膜缘细胞内的代谢活性动态变化，无需使用外源荧光染料或标记剂，为角膜缘生物学及疾病机制研究提供全新细胞水平结构及功能成像方法，具有广泛应用前景。

圆锥角膜（KC）是一种典型的扩张性眼病，以角膜扩张变薄并向前锥形突起为特征，严重时可致盲。KC三联征之一铁锈色Fleischer环，主要由上皮细胞基底膜周围的铁离子沉积组成。近年来，越来越多研究表明，铁稳态失衡可能与KC的发生和发展密切相关。KC患者泪液中铁相关蛋白的异常表达，提示铁稳态失衡可能是诱发KC的潜在致病机制。此外，角膜上皮细胞内铁稳态失衡导致细胞内铁离子异常积聚，进而引发活性氧和脂质过氧化物的大量生成，最终可能触发细胞铁死亡。从恢复铁稳态角度出发，螯合过量的铁离子和调控铁死亡过程关键靶点可能是未来KC潜在的治疗方法。目前关于铁稳态失衡导致KC发病的具体机制仍存在诸多谜团。随着相关研究的不断深入，有望通过改善角膜铁稳态失衡，为KC临床治疗带来新的思路和突破，也为KC患者提供更精准和个体化的治疗策略。

圆锥角膜（KC）是一种典型的扩张性眼病，以角膜扩张变薄并向前锥形突起为特征，严重时可致盲。KC三联征之一铁锈色Fleischer环，主要由上皮细胞基底膜周围的铁离子沉积组成。近年来，越来越多研究表明，铁稳态失衡可能与KC的发生和发展密切相关。KC患者泪液中铁相关蛋白的异常表达，提示铁稳态失衡可能是诱发KC的潜在致病机制。此外，角膜上皮细胞内铁稳态失衡导致细胞内铁离子异常积聚，进而引发活性氧和脂质过氧化物的大量生成，最终可能触发细胞铁死亡。从恢复铁稳态角度出发，螯合过量的铁离子和调控铁死亡过程关键靶点可能是未来KC潜在的治疗方法。目前关于铁稳态失衡导致KC发病的具体机制仍存在诸多谜团。随着相关研究的不断深入，有望通过改善角膜铁稳态失衡，为KC临床治疗带来新的思路和突破，也为KC患者提供更精准和个体化的治疗策略。