PDT联合ranibizumab玻璃体腔注射治疗年龄相关性黄斑变性合并CNV

目的：观察 PDT 联合玻璃体腔注射 ranibizumab (雷珠单抗)治疗老年性黄斑变性脉络膜新生血管( choroidal neovascularization,CNV)的疗效。
　　方法：将符合纳入标准,经吲哚青绿脉络膜血管造影(indocyanine green angiography, ICGA)、光学相干断层扫描( optical coherence tomography, OCT)检查确诊为黄斑区脉络膜新生血管( CNV)患者27例27眼,经PDT治疗后3~7 d内行 ranibizumab 玻璃体腔注射。观察治疗后1,3,6 mo、末次随访时行最佳矫正视力、FFA、ICGA、OCT 检查及有无并发症发生情况。
　　结果：最佳矫正视力提高17眼(63%),最佳矫正视力稳定6眼(22%),最佳矫正视力下降4眼(15%)。27例27眼治疗前平均渗漏面积为1005.69±105.47μm,治疗后1,3mo后平均875.54±103.27,423.37±79.68μm,与治疗前比较差异有统计学意义(P<0.01),视网膜黄斑中央厚度27例27眼治疗前平均厚度为485.58±122.59μm,治疗后1,3mo后平均398.84±105.32,297.74±89.18μm,与治疗前比较差异有统计学意义(P<0.01)。
　　结论：PDT 封闭 CNV 后,联合玻璃体内腔内注射ranibizumab,有效阻断新生血管复发,减少PDT再次治疗次数和并发症,可提高治疗效果。     

糖尿病大鼠玻璃体腔注射缺氧诱导因子-1α siRNA对血管内皮生长因子蛋白表达的影响

目的：观察缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)小干扰RNA(siRNA)对糖尿病大鼠视网膜组织中血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白表达的抑制作用,及其用于糖尿病新生血管性疾病治疗的可行性。

方法：以pSilencer2.1-U6neo为质粒载体,构建HIF-1α.siRNA 重组质粒。选取雄性Sprague-Dawley(SD)大鼠54只,随机分为正常对照组(15只)和实验组(39只)。实验组采用尾静脉注射链尿佐菌素(STZ)的方法建立糖尿病大鼠模型,造模成功后将实验组随机分为糖尿病组(15只)、基因治疗组(12只)和空载体组(12只)。正常对照组及糖尿病组大鼠均不做转染; 基因治疗组和空载体组分别转染HIF-1α.siRNA 重组质粒和pSilencer空载体质粒。行苏木精-伊红染色(hematoxylin-eosin staining,HE染色)观察视网膜组织形态,并采用免疫组织化学法检测VEGF蛋白的表达。分别于干扰后24、48、72h,1wk时计算VEGF蛋白的抑制效率。采用单因素方差分析和LSD-t检验进行统计学分析。

结果：HIF-1α.siRNA 重组质粒经酶切、测序鉴定,确定为目的序列。HE染色显示：正常对照组大鼠视网膜各层细胞排列整齐,细胞形态基本正常。糖尿病大鼠视网膜各层细胞排列紊乱,内界膜不完整,新生血管芽、新生血管簇呈垂直状突破内界膜生长。免疫组织化学染色显示：VEGF阳性表达为细胞浆出现棕黄色颗粒,主要位于神经节细胞层。正常对照组VEGF蛋白呈弱阳性表达,而DR对照组和空载体组表达明显增强,基因治疗组较DR组和空载体组表达明显减少,差异有统计学意义(P<0.05)。VEGF蛋白抑制率24、48、72h和1wk时分别为：27.4%、40.6%、47.5%、64.5%。

结论：HIF-1α.siRNA重组质粒能够抑制糖尿病大鼠视网膜中VEGF蛋白的表达,可能成为一种治疗糖尿病性新生血管疾病的新方法。     

CCR7信号通路与视网膜新生血管

视网膜新生血管性疾病是致盲的主要原因。趋化因子受体7(C-C chemokine receptor type 7, CCR7)可通过细胞外信号调节激酶(extracellular signal regulate kinase,ERK)通路促进血管内皮生长因子(vascular endothelial growth factor, VEGF)的表达,导致血管渗漏、血管内皮细胞增生以及新生血管形成等改变。趋化因子受体7的检测可指导视网膜新生血管性疾病的诊治。     

A case of nodular posterior scleritis mimicking choroidal mass

The aim of this study is to report clinical and imaging findings, and treatment outcomes of a patient with nodular posterior scleritis. A 41-year-old woman was diagnosed as nodular posterior scleritis in the light of clinical and imaging findings. At first admission best corrected visual acuity was 20/50 in her right eye. Fundus examination revealed an amelanotic subretinal mass under the superior temporal arcade associated with subretinal fluid surrounding it. B-scan ultrasonography, optical coherence tomography, fluorescein angiography, and indocyanine green angiography findings confirmed the diagnosis. As treatment, nepafenac eye drops 3 times a day, and flurbiprofen tablet 100 mg twice a day were prescribed. After 4 weeks of treatment, the ocular pain was relieved, BCVA improved to 20/20, and subretinal mass totally regressed. Although the diagnosis of nodular posterior scleritis may be confusing, it has to be kept in mind in patients with a subretinal/choroidal mass. Multimodal fundus imaging may be helpful in differential diagnosis. The condition is usually curable with non-steroidal anti-inflammatory drugs and/or systemic steroids.     

Effect of human recombinant Endostatin protein on human angiogenesis

Tumor growth and metastasis are dependent on the development of new blood vessels. Inhibitors of new vessel growth have been widely investigated as anti-tumor agents. Endostatin, a 20 kDa C-terminal fragment of collagen XVIII inhibits endothelial cell proliferation, induces endothelial cell apoptosis, and can both inhibit and reverse tumor growth in mice. However, human recombinant endostatin has had limited testing against human tissue targets. To investigate the effect of human endostatin on a human vessel target over a broad range of concentrations (10^–12–10^–4 M), human placental vein disks were grown for a period of 2 weeks in a 0.3% fibrin clot overlayed with growth medium. Disks from five individual placentas were tested. For each placenta utilized, a control (medium and 20% fetal bovine serum [FBS]) group and a group treated with heparin (300 g/ml) and hydrocortisone 21-phosphate (350 g/ml) (heparin-steroid) at a dose known to inhibit angiogenesis were included. Endostatin was tested at concentrations of 10^–12–10^–4 M in medium containing 20% FBS. The rate of initiation and the angiogenic growth index (on a visually graded semi-quantitative scale of 0–16) were determined for all experimental conditions. Endostatin inhibited angiogenesis in our model only in high concentrations. At 10^–5 M, endostatin did not alter the percent of wells that initiated an angiogenic response, but significantly inhibited subsequent vessel growth. At 10^–4 M, endostatin was able to inhibit both initiation and subsequent new vessel growth. Human endostatin can inhibit the initiation of a human angiogenic response and inhibit the subsequent proliferation of human neovessels when used at high doses in a continuous exposure model.     

Is choroidal vascularity index a useful marker in different stages of idiopathic epiretinal membranes?

BackgroundThis study aims to evaluate the choroidal vascularity index in patients with idiopathic epiretinal membrane at different stages.MethodsThis prospective study included 125 eyes of 125 patients with idiopathic epiretinal membrane and 62 eyes of 62 healthy control subjects. In this study, epiretinal membrane stages were defined based on the spectral-domain optical coherence tomography staging system. The choroidal vascularity index was measured as the ratio of the luminal area to the stromal area in the central 1500 μm after binarization on enhanced depth imaging optical coherence tomography images. Data on epiretinal membrane stages, choroidal vascularity index, and best-corrected visual acuity were noted.ResultsOf 125 eyes with epiretinal membrane, 38 (30.4 %) had stage 1, 32 (25.6 %) had stage 2, and 55 (44 %) had stage 3 disease. Visual acuity was better in eyes with stage 1 or 2 epiretinal membrane than those with stage 3 epiretinal membrane (p < 0.001). The mean choroidal vascularity index was 2.29 ± 1.02 in the control, 2.23 ± 0.98 in the stage 1 epiretinal membrane, 2.22 ± 0.91 in the stage 2 epiretinal membrane, and 2.23 ± 1.11 in the stage 3 epiretinal membrane group. There was no significant difference between epiretinal membrane subgroups and the control group regarding the choroidal vascularity index (p = 0.81).ConclusionFrom the results obtained in the present study, the choroidal vascularity index was not effected by either the development or the progression of idiopathic epiretinal membrane.     

MicroRNAs in laser-induced choroidal neovascularization in mice and rats: their expression and potential therapeutic targets

Choroidal neovascularization characterizes wet age-related macular degeneration. Choroidal neovascularization formation involves a primarily angiogenic process that is combined with both inflammation and proteolysis. A primary cause of choroidal neovascularization pathogenesis is alterations in pro-and anti-angiogenic factors derived from the retinal pigment epithelium, with vascular endothelium growth factor being mainly responsible for both clinical and experimental choroidal neovascularization. MicroRNAs(miRNAs) which are short, non-coding, endogenous RNA molecules have a major role in regulating various pathological processes, including inflammation and angiogenesis. A review of recent studies with the mouse laser-induced choroidal neovascularization model has shown alterations in miRNA expression in choroidal neovascularization tissues and could be potential therapeutic targets for wet age-related macular degeneration. Upregulation of miR-505(days 1 and 3 post-laser), miR-155(day 14) occurred in retina; miR-342-5 p(days 3 and 7), miR-126-3 p(day 14) in choroid; miR-23 a, miR-24, miR-27 a(day 7) in retina/choroid; miR-505(days 1 and 3) in retinal pigment epithelium/choroid; downregulation of miR-155(days 1 and 3), miR-29 a, miR-29 b, miR-29 c(day 5), miR-93(day 14), miR-126(day 14) occurred in retinal pigment epithelium/choroid. Therapies using miRNA mimics or inhibitors were found to decrease choroidal neovascularization lesions. Choroidal neovascularization development was reduced by overexpression of miR-155, miR-188-5 p, miR-(5,B,7), miR-126-3 p, miR-342-5 p, miR-93, miR-126, miR-195 a-3 p, miR-24, miR-21, miR-31, miR-150, and miR-184, or suppression of miR-505, miR-126-3 p, miR-155, and miR-23/27. Further studies are warranted to determine miRNA expression in mouse laser-induced choroidal neovascularization models in order to validate and extend the reported findings. Important experimental variables need to be standardized; these include the strain and age of animals, gender, number and position of laser burns to the eye, laser parameters to induce choroidal neovascularization lesions including wavelength, power, spot size, and duration.     

Early Contribution of Pericytes to Angiogenic Sprouting and Tube Formation

Immunostaining with endothelial and pericyte markers was used to evaluate the cellular composition of angiogenic sprouts in several types of tumors and in the developing retina. Confocal microscopy revealed that, in addition to conventional endothelial tubes heavily invested by pericytes, all tissues contained small populations of endothelium-free pericyte tubes in which nerve/glial antigen 2 (NG2) positive, platelet-derived growth factor beta (PDGF beta ) receptor-positive perivascular cells formed the lumen of the microvessel. Perfusion of tumor-bearing mice with FITC-dextran, followed by immunohistochemical staining of tumor vasculature, demonstrated direct apposition of pericytes to FITC-dextran in the lumen, confirming functional connection of the pericyte tube to the circulation. Transplantation of prostate and mammary tumor fragments into NG2-null mice led to the formation of tumor microvasculature that was invariably NG2-negative, demonstrating that pericytes associated with tumor microvessels are derived from the host rather than from the conversion of tumor cells to a pericyte phenotype. The existence of pericyte tubes reflects the early participation of pericytes in the process of angiogenic sprouting. The ability to study these precocious contributions of pericytes to neovascularization depends heavily on the use of NG2 and PDGF beta -receptor as reliable early markers for activated pericytes.