Complement factor H polymorphism in age-related maculopathy in the Chinese population: the Beijing Eye Study.

PURPOSE: The purpose of this study was to analyze the association between the Y402H polymorphism in the complement factor H and soft drusen of the macula as part of age-related maculopathy in the Chinese population. METHODS: In the population-based Beijing Eye Study, the participants underwent a detailed ophthalmic examination including fundus photography. All fundus photographs were graded using the Wisconsin Grading System. Of 515 subjects with soft drusen in the macula, 208 (40.4%) subjects had blood samples taken and were thus eligible for the present study. These subjects were compared with 140 randomly selected control subjects from the Beijing Eye Study matched for age, sex, and rural versus urban area with the study group. The analysis of the genotype was performed by allele-specific digestion of polymerase chain reaction products. RESULTS: Dividing the study group into subjects with bilateral soft drusen and unilateral soft drusen showed a significant association between the Y402H polymorphism in the complement factor H gene and the study group with bilateral soft drusen with an odds ratio of 2.29 (95% confidence interval, 1.06-4.95). CONCLUSION: Also in the Chinese population, soft drusen as part of age-related maculopathy are associated with the Y402H polymorphism in the complement factor H gene despite a markedly lower frequency of C allele in the Chinese population than in white populations.

[Short hairpin RNA targeting vascular endothelial growth factor effectively inhibits expression of vascular endothelial growth factor in human retinal pigment epithelium].

OBJECTIVE: To design shRNA targeted to human vascular endothelial growth factor (VEGF) and to evaluate the effect of VEGF. shRNA on expression of VEGF in human retinal pigment epithelium (RPE) cells in vitro. METHODS: Human RPE cells were isolated with enzyme-assisted microdissection. The cells were identified by immunohistochemical method with antibody to cytokeratin and S-100. Plasma DNA was identified via restriction enzyme EcoRI and SamI. shRNAs (P1, P2) specific for human VEGF were designed. DNA expression vector is pSilencer 4.1-CMV of Ambion company. P3 is negative control nonspecific shRNA. There are 5 groups. Group 1: VEGF in cultured human RPE exposed to 100 micromol/L CoCl2 30 h; Group 2: VEGF in cultured human RPE in normal culture medium; Group 3, 4, 5: VEGF in cultured human RPE exposed to 100 micromol/L CoCl2 30 h after P1, P2, P3 transfection, respectively. VEGF level in conditioned media was measured by Western blot. RESULTS: The cells in culture could be stained with both cytokeratin and S-100 antibodies. The length of two fragment was 3.3 kb and 1.6 kb, respectively, which indicated that the extraction and purification were successful. The expression of VEGF in RPE was increased significantly (P < 0. 001) in group 1 as compared with group 2. Hypoxia-induced upregulation of human VEGF is halted by siRNA application in vitro (P < 0. 001 and P < 0. 001 in group 3 and 4 compared with group 1, respectively). shRNAs targeted hVEGF effectively and specifically inhibited hypoxia-induced VEGF levels in human RPE. The level of VEGF was reduced 65.9% and 52.4% in groups 3 and 4, respectively. There was no difference between group 5 and 1 (P = 0. 147). There was no difference of beta-actin production in RPE cells among groups. CONCLUSIONS: Delivery of shRNA can be used in vitro to target specific RNAs of VEGF and to reduce the level of the specific protein product (VEGF) in the targeted cells (human RPE). This work established the basis for the using of RNA interference in studies of retinal biology and for the treatment of a variety of retinal angiogenic diseases, especially the choroidal neovascularization.

[The prevalence and its effective factors of primary angle-closure glaucoma in defined populations of rural and urban in Beijing].

OBJECTIVE: To investigate the prevalence of primary angle-closure glaucoma (PACG) of urban and rural residents in Beijing, China. A quantitative comparison was made based on the data collected from this epidemiological survey. We also identify some of the most typical risk factors associated with PACG. METHODS: Glaucomatous screening examination was applied to specific age group populations (aged 40 and older) in the defined district of Beijing and its remote rural county, from June to October, 2001. There are 4451 subjects in all, 1980 rural subjects and 2471 urban subjects, 1939 males and 2512 females. The screen and diagnostic methods used in this survey included van Herick methods and gonioscopy examination to estimate the peripheral depth of the anterior chamber, visual acuity, intraocular pressure, refraction status, stereoscopic fundus photography, and threshold-related visual field tests. Subjects regarded as suspected glaucoma and glaucoma patients are reexamined with standard glaucomatous examination. RESULTS: The response rate of rural and urban population is 79.6% (1980/2488), 87.1% (2471/2836), respectively. The prevalence of PACG (in aged 40 years or older population) resulted from this survey was 1.2% (95% CI 0.9% - 1.5%). However, the prevalence was different between urban and rural residents, 1.1% (95% CI 0.8% - 1.4%) vs. 1.6% (95% CI 1.2% - 2.0%). The prevalence of PACG in female was more than that in male, 1.7% (95% CI 1.3% - 2.1%) vs.0.8% (95% CI 0.5% - 1.1%). A drastic increase in prevalence of PACG with age increase was identified in both survey sites, however, this increase in rural subjects (aged 60-69 years group) occurred ten years earlier than those from urban subjects (aged more than 70 years group). Compared to urban residents, rural residents showed higher prevalence of unilateral low vision (39.3% vs. 20.6%) and blindness (28.6% vs. 14.7%). This survey also confirmed that, as people aging, refraction status became hyperopia, the depth of peripheral anterior chamber became narrow. In the different age groups, female and male groups, the changes of refraction status and the depth of peripheral anterior chamber paralleled the prevalence of PACG. CONCLUSIONS: The prevalence of PACG was obviously different in different groups. This could due to several factors including gender, age, change of refraction status and chamber angle as well.

[Adeno-associated virus mediated delivery and expression of GDNF cDNA in cultured bovine iris pigment epithelial cells].

OBJECTIVES: To obtain transgenic bovine iris pigment epithelial cells (IPECs) by adeno-associated virus (AAV) mediated delivery of cDNA of glial cell-line derived neurotrophic factor (GDNF). METHODS: AAV-GDNF was titrated by slot blotting. Cultured bovine passage two IPECs were transfected using AAV-GDNF at dosage of MOI (multiplicity of infection) = 50, then were cultured in DMEM medium complemented with 3% FBS for 4 weeks with no change of medium. The expression of GDNF in culture medium was examined using ELISA test. By using the same methods, cultured passage two IPECs were transfected with AAV-GFP (green fluorescent protein) at dosage of MOI = 50, and then were cultured in DMEM medium complemented with 20% FBS. The expression of GFP in IPECs was examined using fluorescence microscope every 2 days after transfection. RESULTS: (1) GFP expression in IPECs could not be detected until 4 days after transfection. The positive GFP expression in IPECs reached fastigium in day 8 or 10. (2) According to the results of ELISA test, concentration of GDNF in the culture medium was (17.14 +/- 1.10) micro g/L. CONCLUSIONS: AAV-GDNF can effectively transfect cultured IPECs, and the transgenic cells show a high expression of GDNF.

[The prevalence and its screening methods of primary open angle glaucoma in defined population-based study of rural and urban in Beijing].

OBJECTIVE: The purpose of the study is to determine the prevalence of primary open-angle glaucoma (POAG) in persons aged 40 or above in Beijing, China. METHODS: From June 2001 to October 2001, the screening population was identified by a door-to-door census in five metropolitan resident areas in the north of Beijing and three villages in a county south to Beijing. The screening included visual acuity, frequency doubling perimetry (FDP, C-20 screening program), non-contact tonometry, slit lamp microscopy, anterior chamber depth (Van Herick method), and fundus photography. The suspect glaucoma and definite glaucoma patients were asked to have an examination of Octopus 1-2-3 perimetry (G1X TOP threshold program), repeat tonometry, gonioscopy and fundus stereo-photography at Beijing Tongren Hospital. RESULTS: There were 4451 subjects who were examined at the study sites. The response rates in rural and urban were 79.58% and 87.13%, respectively. In this 40 years-old or above population, the prevalences of POAG were 1.97% in rural men, 2.07% in urban men, 1.04% in rural women and 1.42% in urban women. In this study, 92.30% POAG patients in rural and 87.30% POAG patients in urban were new diagnosed cases. The prevalence of POAG increased with age and the change was exponential. In 50% POAG patients first IOP measurement was less than 21 mm Hg (1 mm Hg = 0.133 kPa). The prevalence of monocular eye blindness was 15.40% and 10.90% in rural and urban, respectively. CONCLUSIONS: Owing to use fundus photography and integrated evaluation of optic disc, it is possible to diagnose in earlier stage of POAG. This study identifies more patients with POAG than any previous population-based studies of China, and is similar to other studies of Asia such as in India and Singapore. The reason of lower POAG prevalence in rural women than in urban may be that the anterior chamber depth of rural women is shallower than that of urban women. It may cause difficult to differentiate the chronic primary angle closure glaucoma from POAG at screening sites, so the part POAG patients may be included in primary angle closure glaucoma patients.

[Prevalence of low vision and blindness in defined populations in rural and urban areas in Beijing].

OBJECTIVE: To investigate the prevalence and causes of low vision and blindness in Beijing residents aged 40 and over. METHODS: 4,451 residents aged 40 and over in 3 rural communities and 5 urban communities in Beijing underwent eye examination, including examination of distant and near visual acuity (VA), best corrected distant and near VA, pinhole VA, and visual field, slit lamp biomicroscopy, and dilated ocular examination in the form of in-home survey by defined population-based sampling. The medical history was surveyed too. The data were analyzed based on the criteria of the World Health Organization. RESULTS: The general prevalence rates of low vision and blindness were 0.99% (95% CI: 0.70-1.28) and 0.39% (95% CI: 0.21-0.57) respectively. The prevalence rate of low vision in females was 1.45%, 2.23 times that of males (0.65%) (OR: 1.97, 95% CI: 1.00-3.95). The prevalence rate of low vision of rural residents was 1.76%, 2.89 times that of urban residents (0.61%) (OR: 2.93, 95% CI: 1.43-6.11). The prevalence rate of blindness in females was 0.64% and 0.37% in males. The prevalence rate of blindness of rural residents was 1.06%, 2.04 times that of the urban residents (0.52%) (OR: 3.77, 95% CI: 1.41-10.62). The 3 major causes of blindness were cataract (37.50%), glaucoma (29.20%), and high myopic macular degeneration (8.30%). The prevalence of blindness increased with age. CONCLUSION: The prevalence rates of low vision and blindness are higher in the rural areas. Cataract, glaucoma, and high myopic macular degeneration are the major causes of blindness. The prevalence of low vision and blindness are influenced by age, sex; area, health care level, educational level, and environmental factors.

[The application of digital photography with retroillumination for lens in cataract study].

OBJECTIVE: To evaluate the reliability of digital photography with retroillumination for lens as a tool to assess the degree of lens opacity, and to study the morphological features of some lens diseases using the digital images. METHODS: The cataract screener (Neitz CT-S) was connected to a computer capable of acquiring digital images of lens. The digital lens retroillumination and slit lamp images were collected in 64 eyes. Three independent separate examiners took three sets of different CT-S digital photographs in four cataract subjects. The area of lens opacification in the images was measured and analyzed using threshold adjustment in PhotoShop software and a customized program in Matlab software. RESULTS: The distinct digital images of lens cortical opacity, posterior subcapsular opacity and other lens diseases were obtained with CT-S computer digital photography, which images were much better than slit lamp sectional digital images. The sensitivity and specificity of measuring lens opacification with CT-S digital images were 90% and 94% respectively. However, the digital lens retroillumination images were not as sensitive as the slit lamp images for nuclear cataracts. The variation coefficients of lens opacity area in the three sets of CT-S digital images in the same subject, which were taken by the independent examiners, were from 2.23% to 10.86%. The cortical opacification in CT-S digital images was linear, cuniform or clustered aggregate shadow configuration. The posterior subcapsular opacification was discal vacuolar or granular appearance. CONCLUSIONS: The reproducibility of the CT-S digital images taken by the independent examiners is excellent. This technique can objectively show cortical or posterior subcapsular opacities, but nuclear cataracts.