由于青光眼的早期诊断、发病机制、致病基因筛选及功能测定、视神经损伤与保护、滤过术后瘢痕调控、干细胞培养与自然动物模型形成、流行病学研究等基本问题尚处于探讨阶段，因此有必要回顾我国近5年来青光眼的临床与基础研究进展情况，分析青光眼研究领域存在的问题，明确今后青光眼的研究方向，进一步推动青光眼的临床与基础研究深入发展。

目的：探讨体外诱导小鼠胚胎干细胞（ESCs）分化为神经干细胞（NSCs）的可能性。方法：ESCs经胚胎体阶段，以视黄酸及视网膜müller细胞诱导，在NSCs选择性培养基中筛选培养扩增7d，观察形态变化，采用RT-PCR法检测nestin、glutaminase、Brn-3基因表达及免疫细胞化学法检测nestin等NSCs特异性标志物，并对其扩增及分化能力进行观察。结果：ESCs经初级诱导，NSCs选择性培养基筛选培养7d后，形成大量神经球样结构，可扩增传代。绝大部分神经球样结构呈nestin抗原阳性，整合BrdU，表达nestin、glutaminase、Brn-3基因，且可分化为神经元及神经胶质样细胞。结论：视黄酸及müller细胞诱导的ESCs，经选择性培养基筛选培养可获得NSCs，有望为青光眼等神经变性疾病提供新的治疗途径。

目的：建立人眼小梁细胞体外滤膜培养体系及通透阻力测定的模型。方法：将永生化的人眼小梁细胞株用酶消化法传代于细胞培养池的PET膜上，观察细胞的生长情况，同时分别在细胞融合后3、5d和1、2周，应用内皮细胞电阻测量仪（EVOM）测定滤膜上单层小梁细胞电阻，客观评价其通透阻力。结果：人眼小梁细胞在滤膜上生长良好，单层融合后3、5d和1、2周的平均电阻分别为（36.4±1.4）Ω、（35.0±1.7）Ω、（36.1±2.9）Ω、（39.3±3.0）Ω，各时间点无显著统计学差异（P=0.305）。结论：人眼小梁细胞滤膜上培养单层融合后，在不同时间段其阻力基本稳定。滤膜培养体系及应用EVOM对其通透阻力的测定可作为今后对小梁细胞特性研究的一个新的较好模型。

目的探讨治疗原发性闭角型青光眼三种手术方式的适应症和初步临床疗效观察。方法采用非随机临床对照研究方法。拟订手术适应症，对临床收治的63例72眼原发性闭角型青光眼进行手术处理：单纯抗青光眼手术——复合式小梁切除术（Trabeculectomy,Trab）、青-白联合手术——复合式小梁切除联合超声乳化白内障吸除+晶体囊袋折叠式人工晶体植入术（Phacotrabeculectomy+IOL，PhacoTrab+IOL）和单纯白内障手术——超声乳化白内障吸除＋晶体囊袋折叠式人工晶体植入术（Phaco+IOL）。比较不同适应症下三种手术方式初步的临床疗效。包括眼压控制情况、前房深度和房水流畅系数（C值）的变化，超声生物显微镜（UBM）观察房角和滤过泡，以及用抗青光药物辅助情况及并发症等。随访时间平均（11.7±4.9）个月。结果均经统计学处理。结果三组术后在随访期内均能将眼压控制在理想范围，术后眼压较术前明显降低（三组P值均＜0.001），前房深度：PhacoTrab+IOL和Phaco+IOL组术后明显较术前加深，差异有统计学意义（P值均＜0.001），但Trab组术前后改变不明显（P＞0.05）；UBM观察房角：PhacoTrab+IOL和Phaco+IOL组明显较术前加宽或重新开放，相应组C值术后较术前提高，差异有统计学意义（P分别＜0.001和＜0.01），Trab组房角变化不明显，但术后C值较术前亦有改善（P＜0.001）；三组均未见严重术中并发症；术后早期炎症反应最轻者为Phaco+IOL组；远期并发症Trab组有5眼发生低眼压。最后对视力的变化也作了讨论。结论不同手术方式适合不同的病人情况。超声乳化白内障吸除术因其具有相对房角开放和C值改善以及视力提高等优势，可考虑作为原发性闭角型青光眼治疗的选择性手段之一，但要注意适应症的选择。

目的鉴定常染色体显性遗传家族性开角型青光眼家系（GZ11）致病基因并探讨其致病机制。方法应用全基因组扫描、连锁分析、核苷酸直接测序法筛查GZ11家系的致病性基因突变，进一步利用体外基因转染技术，观察突变型基因在小梁细胞中的表达、分布及对细胞凋亡的作用。结果GZ11家系所有患者均存在myocilin基因第三外显子Pro370Leu杂合突变（16/16），而家系正常人中均未检测到该突变（0/8）。小梁细胞体外转染野生型和突变型myocilin基因后，经免疫印迹法检测证实，培养上清液中突变型myocilin蛋白的细胞外分泌较野生型明显减少；荧光免疫组化检测结果证实野生型myocilin蛋白在小梁细胞内呈点状、弥漫分布；而Pro370Leu突变型myocilin蛋白以积聚体形式存在于胞质内；经流式细胞仪检测突变型转染组凋亡细胞百分比较野生型转染组及对照组轻度升高。结论突变型（Pro370Leu）myocilin基因为GZ11家系的致病基因，突变型（Pro370Leu）myocilin蛋白在小梁细胞内以积聚体形式存在，可能是由于蛋白质错误折叠所致，并可能诱导小梁细胞凋亡。

Purpose: Hypericin, a specific inhibitor of protein ki-base C, has been reported to have potential as a thera-peutic dru for proliferative vitreoretinopathy (PVR) in vitro and in vivo. In the present strdies, we analyzed the dynamic changesin Ca2+ in flux and freeintracellular Ca2+ concentration Ca2+ of cultured human retinal pigment epith (RPE) cells after stimulation with hypericin in an attempt to elucidate its mechnism as a therapeutic drug for PVR, Methods: RPE cells were plated in a special plastic dish and then stimulated with 100 nM phorbol-12myristate-13-acetate (PMA) and/of 6 hypericin con-centrations (0.5, 1, 2, 3, 4 and 5 μM), after which Ca2+ influx and Ca2+ were determined using the fluores-cence Ca2+ dye fluo-3 AM and laser scanning confocal microscopy. Results: The fluorescence in resting Rpe Cells was strong and distributed throughout the cells. The nucleus appeared more fluorescent than the cyto-plasm. After stimulation with 0.5μM hypoericin, no ob-vious change of Ca2+ influx and Ca2+ was observed. In contrast, stimulation with higher concentrations of hy-pericin (1-5μM) led to a rapid decrease in Ca2+influx and Ca2+ which was significantly different from those de-tected without hypericin (contro experiments). In addl-tion , no significant differences in Ca2+ were found between 1 and 5μM hypericin used. Stimulation with hypericin, which was applled immediately after preincu-bation with PMA for 24 h not further change Ca2+ influx and Ca2+ Conclusion: In Rpe cells, high concen-trations of hypericin (1-5μM) significantiy inhibit Ca2+ influx and induce a decrease in Ca2+. Therefore, hypericin has potential as a therapeutic drug for PVR maybethrough its inhibition of the Ca2+ influx pathway.

目的：揭示活化星形胶质细胞去抑制对小鼠视神经再生的作用。方法：将Bcl-2高表达转基因小鼠(Bcl-2tg)和GFAP/Vimentin双基因敲除小鼠（GFAP-/-/Vim-/-）交配获得三基因突变小鼠(Bcl-2tg/GFAP-/-/Vim-/-,3M)。分别选取出生后5d(P5)小鼠3只及出生后18d(P18)小鼠3只制作标准的完全性视神经嵌断模型。应用GAP43免疫荧光染色特异性检测再生轴突。结果：Bcl-2tg/GFAP-/-/Vim-/-小鼠在P5时获得视神经再生,再生轴突可以延伸达视交叉水平,而P20时仅获得局部出芽再生。结论：在Bcl-2转基因使视网膜节细胞重获内源性再生能力基础上,通过活化星形胶质细胞去抑制可以促进视神经轴突再生,提示活化星形胶质细胞是抑制视神经再生的重要因素,从而为青光眼等视神经病变的再生治疗提出了新的方向。

目的研究单纯疱疹病毒胸腺激酶（HSV-tk）基因系统对人眼球筋膜囊成纤维细胞（HTFs）增殖的抑制作用及其作用机制。方法　构建含HSV2tk基因的逆转录病毒载体pL（tk）SN并转入包装细胞系PT67进行病毒包装，G418筛选抗性克隆并扩大培养收集病毒上清，测定病毒滴度。透射电镜观察鉴定病毒颗粒;聚合酶链反应(PCR)技术鉴定tk基因在病毒基因组中的整合。用病毒感染HTFs细胞，G418筛选并扩大培养，逆转录聚合酶链反应（RT2PCR）技术和免疫印迹法检测tk基因的表达；加不同浓度前药更昔洛韦（GCV）后，以MTT法检测tk基因对HTFs细胞增殖的抑制作用；采用Hoechst33258染色及流式细胞仪检测凋亡细胞；电镜观察细胞形态学改变。结果酶切鉴定结果显示成功构建了逆转录病毒载体pL（tk）SN，电镜下包装细胞产生的病毒颗粒呈椭圆形,直径约100nm；病毒基因组中，PCR扩增出了tk基因；测得病毒滴度为4×107cfu/ml；被转染的HTFs细胞，用RT2PCR及免疫印迹法检测到tk基因在mRNA与蛋白水平均有表达；HSV-tk-GCV系统对HTFs细胞增殖的抑制作用表现为剂量依赖性，5×10-3g/L的GCV作用5d可将细胞全部杀死,IC50为6×10-4g/L,HSV2tk2GCV系统对HTFs细胞的杀伤机制表现为坏死与凋亡,凋亡率随GCV浓度的升高而增加。结论　HSV2tk基因系统能有效抑制人眼球筋膜囊成纤维细胞的增殖,作用机制表现为细胞的坏死与凋亡。(中华眼科杂志,2006,42:2122217)

PURPOSE. To investigate whether photorefractive keratectomy (PRK) performed in infant primates can modify emmetropization and therefore could be used to study mechanisms of refractive error development. METHODS. Six healthy rhesus monkeys ranging in age from 2 to 3 months were randomly divided into two groups (n=3 each). Anisometropia was induced in each animal by performing PRK on one eye. Hyperopic anisometropia was induced in group A monkeys by flattening the cornea of the right eye, whereas myopic anisometropia was produced in group B monkeys by steepening the cornea of the right eye. Corneal morphology and topography, refractive status, and axial growth were evaluated over a 5-month observation period. RESULTS. All the PRK-treated corneas were re-epithelialized and transparent within 3 days after surgery. Subsequently, all the surgically treated eyes exhibited interocular alterations in axial growth rate that were appropriate to compensate for the PRK-induced anisometropia. Specifically, vitreous chamber elongation rates were faster in the eyes with induced hyperopias than in their fellow eyes (0.63

Objective To inve stigate the genetic ba sis of the pathogene sis of a Guangzhou (GZ11) pedigree with primary open-angle glaucoma (POAG) 1 Methods DNA fragment s of the trabecular me shwork inducible glucocorticoid re sponse protein (TIGR) gene from 4 typical POAG patient s and 2 normal subjects were amplified by polymera se chain reaction ( PCR) 1 The amplified PCR fragment wa s cloned into a pT2Adv vector , and direct sequencing wa s carried out on an ABI2373 automated DNA sequencer using dye-terminator chemistry to detect the mutation1 Re s ult s 　The TIGR gene mutation wa s identified in the selected subject s of this pedigree1 This mutation is a “C2to2T”transition at po sition 370 , different from that of we stern countrie s and equivalent to the po sition change found in J apane se patient s with familial POAG1 No mutation wa s found in the TIGR gene fragment in 2 normal subject s of the pedigree1 Co nclus io ns 　The se preliminary re sult s provide insight s into the pathogene sis of POAG by the TIGR gene mutation , and into the underlying action of the different mutations in oriental and we stern peoples.