Retinal regional differences in photoreceptor cell death and regeneration in light-lesioned albino zebrafish

Teleost fish regenerate retinal cells from a population of inner nuclear layer (INL) stem cells. To characterize photoreceptor regeneration in zebrafish (Danio rerio), adult albino fish were subjected to constant intense light to cause photoreceptor cell death. Retinal morphometry was performed on histological sections of control and light-lesioned albino retinas to compare the extent of light damage in the ventral, central and dorsal retinal regions. In addition, opsin immunohistochemistry and TUNEL were used to compare photoreceptor cell death in these different retinal areas, while PCNA immunolabeling quantified the cell proliferation that precedes the photoreceptor regeneration. Transgenic albino; Tg(alpha1-tubulin:egfp) zebrafish were also exposed to the intense light in order to examine regeneration-related gene expression changes. The light-lesioned retinas are characterized by extensive rod and cone photoreceptor cell death in the central and dorsal regions. In contrast, many of the rods and cones survive in the ventral retina. The highest levels of INL cell proliferation, which occurs subsequent to photoreceptor death, correspond to the retinal regions that suffer the greatest levels of photoreceptor damage. In the ventral retina, where photoreceptor cell death is minimal, cell proliferation is confined to the ONL. In addition, EGFP expression from the alpha1-tubulin promoter is increased in Müller glial cells in the light-damaged central and dorsal retina, while transgene expression in the ventral retina is restricted to small, round INL cells. Furthermore, expression of the HuC/D neuronal antigen is detected in a subpopulation of the Müller cells in the light-damaged superior retinal region. These data demonstrate that adult albino zebrafish display retinal regional differences in photoreceptor cell death and in the regeneration-related INL cell proliferation response. The high levels of INL cell proliferation and alpha1-tubulin:egfp transgene expression in the Müller cells may be graded in response to the degree of photoreceptor cell death. This suggests that the levels of photoreceptor damage may directly influence cell responses in the underlying retinal layers.     

Socioeconomic and Ethnic Disparities in Periocular Cutaneous Malignancies

ABSTRACT

Cutaneous malignancies make up the majority of periocular tumors diagnosed and treated by ophthalmologists. In this review, we examine literature regarding ethnic and socioeconomic disparities in incidence and clinical outcomes of the three most common cutaneous periocular tumors: basal cell carcinoma, squamous cell carcinoma, and melanoma. In all three tumor types, the literature shows an increased incidence among two groups: those with lightly pigmented skin and those of higher socioeconomic status. While incidence is high in these groups, clinical outcomes for these patients tend to be good. Those with lower socioeconomic status and ethnic minorities, on the other hand, have a low incidence but are more likely to have poor clinical outcomes. These disparities are likely the result of both biologic and behavioral differences between patients and could provide opportunities for intervention to change risk perception and improve outcomes.     

角膜缘干细胞培养及研究进展

角膜缘干细胞是一种特殊类型的细胞,位于角膜缘基底上皮层,在角膜上皮更新和创伤愈合中起重要作用。对于角膜缘干细胞缺乏或功能障碍的疾病,培养角膜缘干细胞进行移植将是一种重要且有效的治疗方法。对角膜缘干细胞的生物学特性和解剖定位、影响培养的干细胞增殖的调控因素、培养的角膜缘于细胞移植、移植所需载体的选择等方面的研究进展作一综述。     

眼部疾病中上皮细胞转型相关信号通路的研究进展

上皮细胞间质转型（EMT）是一种基本的病理生理现象,参与胚胎发育、组织重构和肿瘤转移等过程,以上皮细胞表型的缺失及间质特性的获得为重要特征,主要表现为具有极性的上皮细胞转化成具有活动能力、能够在细胞基质间自由移动的间质细胞。研究发现,各种刺激通过多种不同的信号途径诱导上皮细胞发生EMT是许多眼部疾病,如视网膜母细胞瘤、创伤后白内障、视网膜新生血管形成等重要的病理变化过程。就眼部疾病中与EMT有关的信号通路进行综述。     

转基因视网膜神经节细胞系RGC-5的研究进展

为研究青光眼性视网膜神经节细胞（RGCs）损害的细胞和分子机制，多种细胞培养模型已经建立，但是转基因的视网膜神经节细胞RGC-5细胞系在国内未见报道。RGC-5细胞系是应用鼠视网膜细胞通过转染技术建立的，除了其形态和电生理特性与RGCs有所不同外，细胞的生长条件以及细胞膜或者细胞内表达的物质与RGCs一致。应用RGC-5细胞系已经建立了多种凋亡模型，研究发现这些模型的细胞在凋亡时的基因变化以及对神经保护剂的反应都与RGCs一致。因此加强对RGC-5的研究和应用，对进一步阐明RGCs的损伤和保护机制将非常有意义。现就这个细胞系的建立及已做的相关研究做一综述。     

Bcl-xl、Bad在眼组织细胞凋亡中的作用

凋亡既是一种自然过程,同时又受很多基因调节,如bcl-2家族中的bcl-xl和bad基因,前者具有与bcl-2相似的抑制细胞凋亡的作用,bad则促进细胞凋亡,二者在眼组织很多部位如角膜、脉络膜上皮细胞、感光细胞、视网膜神经节细胞中均有表达。说明部分眼内组织细胞的生理、病理过程中均有细胞凋亡参与调节,这就为某些眼内疾病的基因治疗提供了新的思路。     

Limbal stem cell disease: Treatment and advances in technology

Anterior segment stem cell technology, due to its already well-defined corneal limbal stem cells with greater ease of evaluation, has been at the forefront of ophthalmic stem cell treatment and technology since 1997. This paper provides an overview of the current standard of care for treatment of limbal stem-cell deficient conditions and reviews recent treatment technologies using ex vivo expansion of cultivated limbal stem cells of the cornea.     

Lens Epithelial Cell Proliferation and Cell Density in Human Age—relat

Purpose: To discuss the potential effect of the lens epithelial cell proliferation inage-related cataract.Methods: In vitro cell proliferation was assayed by MTT method to evaluate the lensepithelial cell density, index, and proliferation capacity in normal lens and all kinds ofage-related cataract. Capsulotomy specimens from all kinds of patients who underwentcataract phacoemulsification extraction surgery were compared with the lens epithelialspecimens from non-cataract lenses of Eye Bank eyes.Results:Lens epithelial cell density of central anterior capsule(LECD) in female normallens was higher than that in male, LECD in nuclear cataract (> NⅢ) was higher thanthat in normal lens, but in the mature cortical cataract, LECD was lower. Mitotic indexof three kinds of age-related cataracts in vivo had no statistical difference, neither didcell proliferation capacity of cultivated cells in vitro.Conclusion: The individual difference of lens epithelial cell density and proliferationcapacity in vivo may be an     

应用视网膜铺片和神经元逆行标记技术测定神经节细胞密度分布

目的:为视网膜神经节细胞(retina ganglion cell,RGC)的准确定量研究提供形态学的指标依据.方法:联合应用Nissl染色法和神经元逆行标记技术,并通过计算机图像处理技术测定RGC的形态分布特征.结果:两种标记方法在视网膜组织中标记的细胞形态、大小和密度分布呈现明显的差异.Nissl染色可以使所有细胞着色,神经元逆行标记技术仅使RGC着色.通过两者的综合分析,在RGC等密度曲线图中可以观察到在视神经乳头下方形成一个沿鼻颞侧轴方向伸展的高密度区,即视条纹,由视条纹至周边部细胞密度递减.结论:联合应用视网膜铺片法和神经元逆行标记技术两者的优点,能够较准确地测定RGC的密度、大小及其分布等形态特征.     

细胞周期与视网膜色素上皮细胞的生长

目前玻璃体视网膜手术在治疗PVR方面取得了巨大成功但是手术并不能完全阻止甚至可能刺激RPE细胞的迁移和增殖,从而导致PVR的复发。RPE细胞是参与PVR形成的一种重要细胞,因此有关药物抑制RPE细胞增殖的研究受到重视,本文综述了近年来影响RPE细胞周期药物的研究进展.展望了通过调控RPE细胞周期来抑制细胞增殖以防治PVR的前景。     

Sequential processing in vision: The interaction of sensitivity regulation and temporal dynamics

The goal of this work was to describe the interaction of sensitivity regulation and temporal dynamics through the primate retina. A linear systems model was used to describe the temporal amplitude sensitivity at different retinal illuminances. Predictions for the primate H1 horizontal cell were taken as the starting point. The H1 model incorporated an early time-dependent stage of sensitivity regulation by the cones. It was adjusted to reduce the effects of gap junction input and then applied as input to a model describing temporal amplitude sensitivity of Parvocellular and Magnocellular pathway retinal ganglion cells. The ganglion cell model incorporated center-surround subtraction. The H1 based model required little modification to describe the Parvocellular data. The Magnocellular data required a further time-dependent stage of sensitivity regulation that resulted in Weber’s Law. Psychophysical data reflect the sensitivity regulation of the retinal ganglion cell pathways but show a decline in temporal resolution that is most pronounced for the post-retinal processing of Parvocellular signals.     

Mantle Cell Lymphoma in a Lacrimal Gland in a Female and a Review of the Literature

Purpose： To report a rare case of Mantle cell lymphoma in lacrimal gland and review of the literature Case report： We report a case of a 59-year-old female who presented with an upper eyelid mass in the right eye for 3 months, without pain and irrigation. A computerized tomog- raphy （CT） scan showed a mass in the bilateral lacrimal gland region, more significant in right eye. The patient underwent a lacrimal gland mass excision surgery and diagnosis of mantle cell lymphoma by histopathology. Immunochemistry for CD20, CD79a, CD5, and CyclinD1 was positive. She was recommended to the Shantou cancer hospital for chemothera- pY.     

外加直流电场对人晶状体上皮细胞移行和增殖活力的影响

目的:观察外加直流电场对人晶状体上皮细胞(lens epithelial cell,LEC)移行和增殖活力的影响.方法:将培养的人晶状体上皮细胞系HLE-B3细胞暴露于电场强度为100、200、400 mV/mm的直流电场中,未受电场暴露的细胞作为正常对照组.倒置显微镜观察并记录电场暴露前及暴露后的细胞图像,并计数细胞数目;流式细胞仪检测电场暴露前及暴露24 h后的细胞凋亡率和细胞周期.结果:强度为400 mV/mm的电场暴露3 h后,HLE-B3细胞呈现朝向电场阴极一侧的定向移行.电场持续暴露后,HLE-B3细胞数逐渐减少,在电场作用6h和12h时分别较正常对照组减少12.6%和18.6%,差异均具有统计学意义(P<0.05).100、200、400 mV/mm电场暴露24 h后,HLE-B3细胞凋亡率分别为(9.2±1.9)%、(23.9±2.6)%、(54±2.5)%,与正常对照组相比明显增加(P<0.05);细胞周期检测结果显示,HLE-B3细胞进入G2/M期的比例随电场强度增加逐渐上升,其中200 mV/mm和400mV/mm电场中G2/M期细胞比例分别为(13.8±2.2)%和(15.6±2.5)%,与正常对照组相比差异具有统计学意义(P<0.05).结论:外加直流电场可诱导HLE-B3细胞的定向移行,随电场作用时间的延长和电场强度增加,细胞生长受到抑制,同时伴随细胞凋亡增加和细胞周期G2/M期阻滞.     

Different Sensitivities to Apoptotic Induction by Camptothecin between Normal and Senescent Lens Epithelial Cells

PURPOSE: To investigate whether normal and senescent lens epithelial cells have different defense abilities to apoptotic induction factor in vitro. METHODS: Rabbit lens epithelial cells were cultured, passed. When reaching confluence, cells from the first and seventh passage were stained by x-gal staining to detect cell senescence. Cell apoptosis was detected by TUNEL(Roche). 10 micromol/L camptothecin was used to induce cell apoptosis from the lens epithelial cells of the first and seventh passage to distinguish different sensitivities to apoptotic induction factor between normal and senescent cells. RESULTS: The senescent cells (41.17% +/- 5.24%) were detected in the lens epithelial cell culture of the seventh passage, which are higher than those of the first passage (0.98% +/- 0.39%). There was no apoptotic cell detected in the cell cultures undisturbed. Exposure of the first passage cells to camptothecin resulted in death of approximately 23.87% +/- 3.45% of the cells during a 36 hour exposure period. In contrast, significantly more lens epithelial cells died through the apoptosis (38.29% +/- 4.01%) from the seventh passage. CONCLUSION: Senescent cells increased with cell passage. Senescence lens epithelial cells do not undergo apoptosis if they were not disturbed. But the vulnerabilities to apoptotic induction between health and senescence cells were different.     

Clinicopathological Analysis of Malignant Eyelid Tumor Cases at Yamagata University Hospital: Statistical Comparison of Tumor Incidence in Japan and in Other Countries

Purpose

To report the clinical and histopathological features of malignant eyelid tumor cases treated in our clinic. We also compared the differences in the frequency of malignant eyelid tumor in various regions of Japan and worldwide.

Methods

Retrospectively, we studied the records of the 38 cases of malignant eyelid tumor treated in Yamagata University Hospital over the last 17 years. The statistical comparison with various countries was based on reports of case studies in those countries.

Results

Data from our clinic: Among the total of 38 cases, 15 cases (39.5%) were diagnosed as basal cell carcinoma, 11 cases (28.9%) as sebaceous gland carcinoma, and 4 cases (10.5%) as squamous cell carcinoma. In addition, three cases were malignant melanoma, two adenocarcinoma, one Merkel cell carcinoma, one malignant peripheral nerve sheath tumor, and one malignant lymphoma. The ages of patients ranged from 45 to 92 years (mean, 72.0 ± 12.4 years). Most of the cases were treated by complete resection of the tumors and eyelid reconstruction. Radiation or cryotherapy were added when required. The prognosis of the cases with basal cell carcinoma and squamous cell carcinoma was good, and that of the other tumors was relatively poor. During the same period, in Caucasians, basal cell carcinoma constituted about 80%–90% of the malignant eyelid tumors, whereas in Japan and Asian countries, basal cell carcinoma, sebaceous gland carcinoma, and squamous cell carcinoma each constituted about 20%–40%.

Conclusions

A racial difference in the incidence of basal cell carcinoma, sebaceous gland carcinoma, and squamous cell carcinoma can be considered in making a diagnosis. Jpn J Ophthalmol 2005;49:349–354 © Japanese Ophthalmological Society 2005     

干细胞移植在视网膜神经节细胞损伤修复中的应用

进行性视网膜神经节细胞损伤在一些致盲性眼病中屡见不鲜。目前临床上缺乏有效的损伤修复方法,然而最近研究显示干细胞移植为受损视网膜神经节细胞的保护和替代治疗提供了新思路。本文将就干细胞移植为基础的研究进展进行综述。     

HMGA1在葡萄膜黑色素瘤中的表达及对细胞增殖和侵袭的影响

目的：探讨高迁移率族蛋白A1(HMGA1)蛋白在葡萄膜黑色素瘤(UM)组织中的表达,以及抑制该基因表达对细胞增殖和侵袭的影响。

方法：选取2014-02/2019-08在我院接受手术治疗的UM患者53例53眼,同期留取因外伤摘除眼球的正常葡萄膜组织34例34眼。采用免疫组织化学法检测组织中HMGA1蛋白表达。将体外培养的人UM细胞系M23分为HMGA1下调组、阴性对照组和空白组,分别转染HMGA1干扰序列、阴性对照序列和不作任何处理,采用实时荧光定量PCR术检测HMGA1 mRNA表达,CCK-8法检测细胞增殖能力,Transwell法检测细胞迁移和侵袭能力。

结果：UM组织中HMGA1蛋白阳性表达率为77%,高于正常葡萄膜组织中的29%(P<0.001); 与未发生巩膜浸润、未累及睫状体和未发生眼外生长相比,HMGA1蛋白在发生巩膜浸润、累及睫状体和发生眼外生长的组织中阳性表达率升高(均P<0.05)。与阴性对照组和空白组相比,HMGA1 mRNA在HMGA1下调组细胞中相对表达量降低,且HMGA1下调组细胞培养24、48、72、96h时吸光度OD值降低,迁移细胞数和侵袭细胞数均明显减少(均P<0.05)。

结论：UM组织中HMGA1蛋白阳性表达率升高,下调M23细胞中HMGA1表达可减少细胞增殖,抑制细胞迁移和侵袭。     

干细胞与角膜眼表重建研究进展

角膜是眼表的重要防护屏障和光学系统的重要组成部分,其完整性和透明性是实现有效视功能的重要前提。各种原因造成的眼表疾病未经及时有效治疗都有可能发展到终末阶段———角膜缘干细胞功能障碍,包括角膜缘干细胞数目的减少和微环境的病理改变。自体角膜移植无疑是治疗角膜缘干细胞功能障碍最有效的方法。但是供体角膜来源十分有限,且切除健眼组织行角膜移植可能会对健眼造成长期损害。随着干细胞研究的不断深入和组织工程技术的兴起,组织工程角膜应运而生,并且迅速发展。其基本原理为选用生物性能良好的支架材料,体外模拟角膜缘干细胞微环境,诱导各类种子干细胞分化为角膜类上皮,然后再行角膜眼表重建。常用种子细胞包括角膜缘干细胞、胚胎干细胞、诱导多能干细胞、骨髓间充质干细胞、皮肤干细胞、口腔黏膜干细胞。本文就上述干细胞在角膜眼表重建中的应用作一综述。     

Mechanisms of inhibition of elemene on human lens epithelial cell proliferation in vitro

AIM: To study the effects of elemene (Ele) on proliferation and cell cycle of human lens epithelial cells B3 (HLE-B3) and the mechanisms of its signal transduction. METHODS: Recombinant human basic fibroblast growth factor (rhbFGF) was used to induce proliferation of HLE-B3 cells, which were incubated with 80mg/L Ele for 24 hours. The inhibitory effects of Ele on the proliferation of HLE-B3 cells were evaluated by MTT method. The effect of Ele on HLE-B3 cell cycle was analyzed by flow cytometry(FCM). The expressions of protein kinase A (PKA) and protein kinase G (PKG) of HLE-B3 were also analyzed by FCM. RESULTS: Ele altered the cell cycle of HLE-B3 and effectively inhibited HLE-B3 cell proliferation induced by rhbFGF. Ele up-regulated PKA and down-regulated the expression of PKG in HLE-B3 cell. CONCLUSION: Ele inhibits HLE-B3 proliferation, making it an attractive potential agent in regimens to treat after- cataracts.     

牛眼小梁细胞培养,形态和骨架超微结构及动力学研究

OBJECTIVE: To establish the method of culturing calf trabecular cells (CTCs) in vitro, to understand the morphology and function of CTCs, to probe into the effect of resistance of aqueous outflow in the pathogenesis and mechanism about primary open angle glaucoma(POAG). To direct the clinical using of drugs and to open up new antiglaucomatous medicine by pharmacological studies of CTCs. METHODS: Trabecular meshwork was collected from twenty eyeballs of calf donors after slaughter. The tiusse was primarily cultured and cells were subcultured. The growing characteritics and morphological features of cultured primary and passaged cells were observed by light and electron microscopes. Cell kinetics of the third and tenth passaged cells were analysed using autoradiography and flow cytometry. The influence of the antiglaucomatous drugs 0.25 mg.ml-1 epinephrine (EPI) and 0.025 mg.ml-1 dipivalyl epinephrine (DPE) on cell kinetics of the third passaged cells was studied. RESULTS: The growing characteritics and morphological features of cultured CTCs were as same as those of human trabecular cells. Growing types of CTCs included most of epitheial cell and few of fibroblast. The amount of cellular microfilaments was reduced, DNA synthesis time(Ts) and cell cycle time(Tc) were obviously prolonged with passaged increasing. Antiglaucomatous drugs-EPI (0.25 mg.ml-1) and DPE (0.025 mg.ml-1) made microfilaments dissolving, Ts and Tc obviously prolonging. CONCLUSION: Establishing the method of culturing CTCs in vitro and understanding their morphology, function and pharmacological effects provided an important information for studying human trabecular cells and probing into the effect of resistance of aqueous outflow in the pathogenesis and mechanism about POAG. These studies indicated that antiglaucomatous drugs-EPI (0.25 mg.ml-1) and DPE (0.025 mg.ml-1) influenced obviously microfilaments and cell kinetics of the third passaged cells and suggested that it is not to be ignored that 1% EPI and 0.1% DPE may make CTCs' microfilaments dissolving and may inhibit CTCs' division and proliferation when they are used in clinical therapy.