The with Electro-oculogram in Choroidal Melanoma Eyes

The electro-oculogram (EOG) in the involved eyes of 20 patients with a choroidal melanoma was compared to the EOG in the uninvolved eyes of these 20 patients and to the EOG in both eyes of 10 patients with a choroidal nevus in one eye and 19 normal control patients. A statistically significant reduction in the EOG occurred in eyes with melanomas compared to patients with nevi. No difference was found between the EOG of normals and the uninvolved eye of melanoma patients. Patients with nevi did not differ from normals. This observation was independent of tumor size and was not related to the extent of retinal detachment.     

脉络膜黑色素瘤与脉络膜脱离的超声鉴别诊断

分析了25例脉络膜黑色素瘤与75例(76眼)脉络膜脱离的超声显象特点。脉络膜黑色素瘤表现为半圆形或圆形、蘑菇形的实质性光团,肿瘤内有挖空现象,近半数有脉络膜凹陷,部分病例有声影。脉络膜脱离表现为1个至数个半球形光带或分叶的环形光带,多位于锯齿缘至赤道部之间,内部无回声,无脉络膜凹陷及声影。     

Extrascleral Extension of Choroidal Melanoma Diagnosed by Ultrasound

Four cases of choroidal melanoma with extrascleral extension are presented. In each case, contact B-scan ultrasonography identified extrascleral extension of choroidal melanoma as an area of relative echolucency immediately behind the globe. These ultrasonographic findings are contrasted with those demonstrated by computerized tomography. The importance of B-scan ultrasonography in the assessment of a patient with possible choroidal melanoma is discussed     

Syphilitic Scleritis and Choroidal Malignant Melanoma of the Same Eye

A 42-year-old Italian homosexual presented with a red painful eye associated with exudative retinal detachment that was subsequently attributed to syphilitic posterior scleritis. These findings all resolved with intravenous penicillin therapy. However, choroidal mass lesion persisted and subsequent ancillary imaging including B scan and ultrasonography confirmed the presence of a choroidal melanoma, which was treated with radioactive plaque therapy. This case report will describe the interesting findings of this unique presentation.     

Endoresection of Irradiated Choroidal Melanoma as a Treatment for Intractable Vitreous Hemorrhage and Secondary Blood-Induced Glaucoma

A 50-year-old man presented with uncontrolled erythroclastic glaucoma and recurrent vitreous hemorrhage from a previously irradiated choroidal melanoma. Despite trabeculectomy, intraocular pressure became uncontrolled due to increasing bleeding from the melanoma. The vitreous hemorrhage became voluminous and could not be stopped with multiple vitrectomies with endolaser, transpupillary thermotherapy, and transscleral laser. Endoresection of the regressed tumor and its bleeding vasculature resulted in immediate cessation of the bleeding and control of the intraocular pressure.     

In Vitro Study on Lethal Effect of Human Choroidal Melanoma OCM-1 Cell Line by Repeating-70℃Freeze Thawing

Objective: To investigate the effects of repeating-70℃freeze thawing on human choroidal melanoma cell line OCM-1. Methods: OCM-1 cells were frozen by repeating -70℃ freeze thawing with various durations and frequencies. Then the inhibit rate of cells was examined by MTT essay. The cell viability was measured by monoclonal formation assay. We also used the HE staining, immunohistochemistry staining and the laser-scanning confocal microscopy (LSCM) to investigate the morphological changes of the cells. Results: The growth of OCM-1 cells was inhibited by repeating -70℃ freeze thawing in time-dependent and frequency-dependent manners (P < 0.01). Different morphous including necrosis and apoptosis of the cells could be observed after -70℃ freeze thawing by the LSCM. Conclusion: Repeating -70℃ freeze thawing can not only kill cells directly and induce considerable cells to apoptosis, but also inhibit the growth of the survivals. The kill and wound ratio of the cells disposal with different times and frequencies present variance. And the distinction when treated with different frequencies during the same time is much more significant than different times with the same frequency, which guide clinical workers to choose repeating cryotherapy with short term method instead of single cryotherapy with long term in choroidal melanoma treatment.     

Effects of Atropine Treatment on Choroidal Thickness in Myopic Children

PurposeTo examine the changes in choroidal thickness (ChT) after 6 months of 1% or 0.01% atropine treatment and the independent factors associated with eye elongation.MethodsA total of 207 myopic children aged 6 to 12 years were recruited and randomly assigned to groups A and B in a ratio of 1:1. Participants in group A received 1% atropine once a day for 1 week, and then once a week for 23 weeks. Participants in group B received 0.01% atropine once a day for 6 months. ChT and internal axial length (IAL) were measured at baseline, 1 week, 3 months, and 6 months.ResultsIn group A, the ChT significantly increased after a 1-week loading dose of 1% atropine (26 ± 14 µm; P < 0.001) and the magnitude of increase stabilized throughout the following weekly treatment. The internal axial length did not significantly change at the 6-month visit (−0.01 ± 0.11 mm; P = 0.74). In contrast, a decreased ChT (−5 ± 17 µm; P < 0.001) and pronounced eye elongation (0.19 ± 0.12 mm; P < 0.001) were observed in group B after 6 months. Multivariable regression analysis showed that less increase in ChT at the 1-week visit (P = 0.03), younger age (P < 0.001), and presence of peripapillary atrophy (P = 0.001) were significantly associated with greater internal axial length increase over 6 months in group A.ConclusionsOne percent atropine could increase the ChT, whereas 0.01% atropine caused a decrease in ChT after 6 months of treatment. For participants receiving 1% atropine, the short-term increase in ChT was negatively associated with long-term eye elongation. Younger age and the presence of peripapillary atrophy were found to be risk factors for greater eye elongation.     

三氧化二砷抑制人眼脉络膜黑色素瘤裸鼠移植瘤生长的研究

目的探讨三氧化二砷（ATO）对人眼脉络膜黑色素瘤细胞系OCM-1细胞裸鼠移植瘤生长的作用。方法建立裸鼠腋周皮下异位移植肿瘤模型36只；采用不同浓度（0．05、0．5和2．5mmol／L）ATO液0．2ml瘤体内局部注射治疗。计算移植瘤的相对体积、重量及抑瘤率；取瘤体及肝肾组织进行组织病理学和电镜检查。结果不同浓度的ATO均可以使肿瘤生长变慢、重量减轻，与对照组比较有显著统计学差异（P〈0．05）；抑瘤率随ATO浓度的升高而增加。电镜结果显示与对照组相比，治疗组瘤体内的肿瘤细胞发生明显坏死和凋亡的形态学改变。结论一定剂量的ATO直接作用于裸鼠脉络膜黑色素瘤移植瘤，可以通过诱导细胞凋亡及直接导致细胞坏死来抑制肿瘤的生长。     

Effects of Monocular Light Deprivation on the Diurnal Rhythms in Retinal and Choroidal Thickness

PurposeTo determine the effects of monocular light deprivation on diurnal rhythms in retinal and choroidal thickness.MethodsTwenty participants, ages 22 to 45 years, underwent spectral domain optical coherence tomography imaging every three hours, from 8 AM to 8 PM, on two consecutive days. Participants wore an eye patch over the left eye starting at bedtime of day 1 until the end of the last measurement on day 2. Choroidal, total retinal, photoreceptor outer segment + retinal pigment epithelium (RPE), and photoreceptor inner segment thicknesses were determined.ResultsFor both eyes, significant diurnal variations were observed in choroidal, total retinal, outer segment + RPE, and inner segment thickness (P < 0.001). For light-deprived eyes, choroid diurnal variation persisted, although the choroid was significantly thinner at 8 AM and 11 AM (P < 0.01) on day 2 compared to day 1. On the other hand, diurnal variations in retinal thickness were eliminated in the light-deprived eye on day 2 when the eye was patched (P > 0.05). Total retinal and inner segment thicknesses significantly decreased (P < 0.001) and outer segment + RPE thickness significantly increased (P < 0.05) on day 2 compared to day 1.ConclusionsBlocking light exposure in one eye abolished the rhythms in retinal thickness, but not in choroidal thickness, of the deprived eye. Findings suggest that the rhythms in retinal thickness are, at least in part, driven by light exposure, whereas the rhythm in choroidal thickness is not impacted by short-term light deprivation.     

Effects of Y-27632, a Rho-associated Kinase Inhibitor,on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells

PurposeHuman corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitro effects of different concentrations of Y-27632 on the differentiated HCEPs.MethodsHCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated into HCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescence staining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs were assessed in the presence of different concentrations of Y-27632.ResultsY-27632 enabled the isolation and expansion of HCEPs from the corneal endothelium. The differentiated HCEPs showed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632 increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632 also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs.ConclusionsY-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migration of differentiated HCEPs.     

The Clinical Role of <Emphasis Type="Italic">N</Emphasis>-Isopropyl-<Emphasis Type="Italic">p</Emphasis>-[<Superscript>123</Superscript>I]-Iodoamphetamine Single Photon Emission Computed Tomography in the Follow-up of Choroidal Melanoma After Radiotherapy

Background Single photon emission computed tomography (SPECT) images with N-isopropyl-p-[¹²³I]-iodoamphetamine (¹²³I-IMP) have recently been used for the sensitive and specific detection of melanoma.Cases Using ¹²³I-IMP SPECT, we observed three patients over a period of 18 months in whom choroidal melanoma had been diagnosed. Two underwent radiotherapy (cyber knife) in our clinic; the other patient was referred to us after 8 months of proton beam irradiation at another clinic.Observations In two of the three cases, no metastasis or tumor recurrence has been observed up to the present time. In these individuals, the average ¹²³I-IMP uptake in the pathological eye gradually and progressively decreased after radiotherapy to levels seen in the fellow eye. One eye of the three patients examined here, however, exhibited tumor recurrence in the ciliary body as well as hepatic metastasis 12 months after radiation treatment. In the pathological eye of this patient, the average ¹²³I-IMP uptake gradually decreased, but never reached the levels in the fellow eye during the observation period.Conclusions ¹²³I-IMP SPECT is a useful examination method not only for diagnosis but also for the follow-up of patients with choroidal melanoma. Jpn J Ophthalmol 2004;48:54–58 © Japanese Ophthalmological Society 2004     

巨噬细胞刺激人成纤维细胞增生及去炎松和道诺霉素对其的抑制

采用离体培养的人皮肤成纤维细胞并加入巨噬细胞调理培养液（MCM），及3H-胸腺嘧啶掺入液体闪烁检测，观察了成纤维细胞受MCM刺激后的增殖及药物对其的抑制作用。结果，加入MCM组细胞增殖率较对照组明显增加（P＜0．01），150ng／ml道诺霉素及联合用药组的细胞抑制率最高达95％以上（P＜0．01），250μg／ml去炎松的抑制率在不加MCM组约为20％～40％，而在加MCM组约为40％～60％（P＜0．01）。证实去炎松除有直接抑制作用外，更能明显抑制巨噬细胞活性因子对成纤维细胞增生的刺激作用。     

In Vitro Effects of Preservative-free and Preserved Prostaglandin Analogs on Primary Cultured Human Conjunctival Fibroblast Cells

Purpose

Long-term use of topical medication is needed for glaucoma treatment. One of the most commonly prescribed classes of hypotensive agents are prostaglandin analogs (PGs) used as both first-line monotherapy; as well as in combination therapy with other hypotensive agents. Several side effects of eye drops can be caused by preservatives. The purpose of this study was to evaluate the effects of PGs with varying concentrations of benzalkonium chloride (BAC), alternative preservatives, or no preservatives on human conjunctival fibroblast cells.

Methods

Primary human conjunctival fibroblast cells were used in these experiments. Cells were exposed to the following drugs: BAC at different concentrations, bimatoprost 0.01% (with BAC 0.02%), latanoprost 0.005% (with BAC 0.02%), tafluprost 0.0015% with/without 0.001% BAC and travoprost 0.004% (with 0.001% Polyquad) for 15 and 30 minutes. Cell cytotoxicity was evaluated by phase-contrast microscopy to monitor morphological changes of cells, Counting Kit-8 (CCK-8) assay to cell viability, and fluorescent activated cell sorting (FACS) analysis to measure apoptosis.

Results

BAC caused cell shrinkage and detachment from the plate in a dose-dependent manner. Morphological changes were observed in cells treated with bimatoprost 0.01% and latanoprost 0.005%. However, mild cell shrinkage was noted in cells treated with tafluprost 0.0015%, while a non-toxic effect was noted with travoprost 0.004% and preservative-free tafluprost 0.0015%. CCK-8 assay and FACS analysis showed all groups had a significantly decreased cell viability and higher apoptosis rate compared with the control group. However, travoprost 0.004% and preservative-free tafluprost 0.0015% showed lower cytotoxicity and apoptosis rate than other drugs.

Conclusions

This in vitro study revealed that BAC-induced cytotoxicity is dose-dependent, although it is important to emphasize that the clinical significance of toxicity differences observed among the different PGs formulations has not yet been firmly established. Alternatively preserved or preservative-free glaucoma medications seem to be a reasonable and viable alternative to those preserved with BAC.     

雪旺细胞源性神经营养活性物质对培养视网膜节细胞正常及缺气损伤环境中存活及生长的作用

目的：探讨雪旺细胞（Schwann cells,SC)源营养神经活性物质（SC derived neurotrophic activity,SCNA)对体外培养视网膜节细胞正常及缺血损伤环境中存活及生长相关蛋白（GAP）43表达的作用。方法：培养乳鼠SC，制备不同蛋白浓度的SCNA，加入原代培养的视网膜细胞中，MTT法检测SCNA活性培养荧光金逆行标记的新生3d的SD乳鼠视网膜细胞，接种于24孔板中。培养第2d时SCNA作用组培养液中加入300mg/L SCNA,对照组不加；培养第5d缺气损伤组在培养液表面加入液体石蜡造成缺气损伤。12h后去除液体石蜡，观察细胞形态和计数荧光金逆行标记的视网膜节细胞（retinal ganglion cell,RGC).Western Blot法分析SCNA对视网膜细胞GAP43表达的影响。结果：SCNA能促进培养视网膜细胞的存活，并有蛋白浓度依赖关系。加SCNA后，视网膜细胞生长明显旺盛，悬浮的死细胞较少，RGC数量明显多于视网膜细胞单纯培养组（F＝62．89，P＜0．01）。缺气损伤组视网膜细胞出现肿胀改变，加有SCNA缺气损伤组细胞形态大致正常，RGC数与对照组相比有显著性差异（F＝49．27，P＜0．01）。GAP43的表达在视网膜细胞正常及缺气损伤SCNA作用组上调。结论：SCNA对培养RGC具有明显营养作用，并上调GAP43的表达。在培养液中加入SCNA，可以减轻“缺气”造成的损伤，提高体外培养RGC在损伤环境中的存活。     

银杏叶提取物对激光诱导的大鼠脉络膜新生血管的疗效

目的:观察银杏叶提取物对激光诱导的大鼠脉络膜新生血管(choroidal neovascularization,CNV)的疗效。方法:选取60只大鼠,随机分为4组,分别为正常组、模型组、实验组、生理盐水组,每组15只大鼠。除正常组外其余各组均行氪激光光凝制作CNV模型,模型组不做任何处理,实验组均在光凝后每日行腹腔注射银杏叶提取物(Extract of Ginkgo Biloba,EGb761),生理盐水组在光凝后每日腹腔注射生理盐水。于光凝后7,14,21d后对各组大鼠行眼底造影检查,观察CNV的渗漏情况,然后立即处死各组动物,行脉络膜视网膜切片,HE染色后观察各组视网膜及脉络膜的结构和CNV的情况。结果:荧光造影结果显示正常大鼠脉络膜无渗漏,7d时各组大鼠脉络膜渗漏均较轻微,14,21d时模型组和生理盐水组大鼠脉络膜渗漏明显,实验组大鼠脉络膜荧光素渗漏点数明显要少于对应的模型组和生理盐水组差异具有统计学意义(P<0.05)。实验组大鼠的视网膜、脉络膜结构保存比模型组更好。结论:腹腔注射EGb761可以明显减少大鼠CNV的形成,用药时间越长,疗效越好。     

冬凌草甲素协同TRAIL诱导OCM-1A细胞凋亡的作用机制

目的：通过实验研究考察肿瘤坏死因子相关的凋亡诱导配体（TRAIL）与冬凌草甲素联合使用时对脉络膜黑色素瘤细胞株OCM-1A细胞和C918细胞的抑制作用。方法：实验研究。以体外培养OCM-1A细胞和C918 细胞为研究对象,加入不同浓度的冬凌草甲素（终浓度分别为0.625、1.25、2.5、5、10、20 μmol/L）和TRAIL（终浓度分别为6.25、12.5、25、50、100、200 ng/ml）,利用MTT法检测细胞增殖活性,用PI单染结合流式细胞技术检测细胞周期分布,Annexin-V/PI双染检测细胞凋亡,用免疫印迹检测凋亡相关蛋白如死亡受体DR5、A-Caspase-3 蛋白、XIAP蛋白表达量变化。数据采用单因素方差分析和独立样本t检验进行分析。结果：TRAIL单独使用时,对OCM-1A细胞有一定程度的抑制,TRAIL浓度为200 ng/ml时抑制率最高为36.5%;冬凌草甲素单独作用时,对OCM-1A细胞的抑制不明显;当二者联合使用时,对OCM-1A的抑制作用明显增强,TRAIL 200 ng/ml联合冬凌草甲素20μmol/L作用于OCM-1A时抑制作用最强,抑制率为87%,而对C918的抑制增强效果不明显。流式细胞仪检测结果发现处于G2 期细胞显著增多,提示细胞被阻滞在G2/M期,免疫印迹结果显示DR5、A-Caspase-3 表达增强,而凋亡抑制蛋白XIAP表达减弱。TRAIL与冬凌草甲素联合使用后对C918细胞的诱导凋亡作用不明显,差异没有统计学意义。结论：冬凌草甲素和TRAIL共同作用于OCM-1A细胞时,明显增强各自单独作用时候诱导的凋亡作用。     

人角膜上皮细胞体外培养的生长曲线及几种抗生素的影响

目的通过人角膜上皮细胞培养研究细胞的生长曲线和抗生素的影响作用。方法组织块接种后在标准条件下培养，传代细胞进行细胞计数和３Ｈ－ＴｄＲ测定，并加入抗生素观察细胞的生长情况。结果角膜上皮细胞呈膜状扩展增殖，形成单层时间为１３～１５天，群体倍增水平５．９４，具有上皮细胞的特征。传代细胞接种后第４天进入指数生长期，３Ｈ－ＴｄＲ渗入增加。结论加入不同浓度的三种抗生素表明庆大霉素超过１．０ｍｇ／ｍｌ，妥布霉素超过２．０ｍｇ／ｍｌ浓度具有明显的细胞毒性作用，氯霉素介于二者之间。