Association Study of ARMC9 Gene Variants with Vogt-Koyanagi-Harada Disease in Japanese Patients

Purpose: To investigate whether variants in the ARMC9 gene encoding KU-MEL-1 are associated with Vogt-Koyanagi-Harada (VKH) disease in a Japanese population.

Methods: We recruited 380 Japanese patients with VKH disease and 744 Japanese healthy controls to genotype seven single-nucleotide polymorphisms (SNPs) in ARMC9. We also performed imputation analysis of the ARMC9 region and 195 imputed SNPs were included in the statistical analysis.

Results: We observed an increased frequency of the A allele of rs28690417 in patients compared with controls (P = 0.0097, odds ratio (OR) = 1.46). The A allele had a dominant effect on VKH disease risk (P = 0.011, OR = 1.51). However, these significant differences disappeared after Bonferroni correction (corrected P > 0.05). The remaining 201 SNPs did not show any significant association with disease risk.

Conclusions: Our study suggests that ARMC9 variants do not play a critical role in the development of VKH disease.     

Investigation of Association between Autophagy-Related Gene Polymorphisms and Pseudoexfoliation Syndrome and Pseudoexfoliation Glaucoma in a Spanish Population

Purpose: Cellular stress conditions are important mechanisms implicated in the pathogenesis of pseudoexfoliation syndrome. One of the potential cellular responses to these stress conditions is induction of autophagy. The purpose of this study was to evaluate whether genetic variants in three critical genes of autophagy (ATG16L, ATG2B, ATG5) may be involved in the development of pseudoexfoliation syndrome (XFS) and pseudoexfoliation glaucoma (XFG) in a Spanish population. Methods: 108 patients (64 XFS, 44XFG) and 118 healthy controls were evaluated. The analysis of genetic polymorphisms was performed by standard TaqMan allelic discrimination technique. Results: No significant differences in either genotype distributions or allelic frequencies of the tested polymorphisms were found between patients with XFS/XFG and control subjects. Conclusions: Our results suggest that these three genes that are critical components of the autophagy pathway (ATG16L, ATG2B, ATG5) are not significant risk factors among Spanish patients with either XFS or XFG.     

Association Analysis of Polymorphisms in Genes Related to Oxidative Stress in South Indian Type 2 Diabetic Patients with Retinopathy

Background: Diabetic Retinopathy (DR) is one of the most common microvascular complications of type 2 diabetes mellitus (T2DM) and is polygenic with a multitude of genes contributing to disease susceptibility. The present study aimed at exploring the association between DR and seven polymorphisms in oxidative stress-related genes, i.e. ACE, eNOS, p22phox subunit of NAD(P)H oxidase, PARP-1 and XRCC1 in South Indian T2DM subjects.

Materials and methods: The study included 149 T2DM subjects with DR (diagnosed through funduscopic examination) and 162 T2DM patients with no evidence of DR. The selected polymorphisms were genotyped by polymerase chain reaction (PCR) and Taqman allele discrimination assay.

Results: There was no significant difference in the genotype and allele distribution of ACE ins/del, eNOS-786T>C, 894G>T, 4a4b and p22phox 242C>T polymorphisms between T2DM groups with and without DR. Contrastingly, there appeared to be a significant association of PARP-1 Val762Ala and XRCC1 Arg399Gln polymorphisms with DR, wherein 762Ala allele seemed to confer significant protection against DR (p?=?0.01; OR?=?0.51 [0.3–0.86]), while the presence of 399Gln allele was associated with an enhanced risk for DR (p?=?0.02; OR?=?1.52 [1.07–2.15]). Multiple logistic regression analysis revealed a significant and independent association of Val762Ala and Arg399Gln polymorphisms and other putative risk factors with DR in T2DM individuals.

Conclusions: The polymorphisms in the DNA repair genes PARP-1 and XRCC1 tended to associate significantly with DR. While Val762Ala polymorphism was associated with reduced susceptibility to DR, the Arg399Gln polymorphism contributed an elevated to risk for DR in South-Indian T2DM individuals.     

HMGB1和TOLL样受体9在糖尿病大鼠视网膜组织中的表达

目的 探讨HMGB1和TOLL样受体(Toll-like receptors,TLR)9在糖尿病大鼠视网膜中的表达.方法 采用链脲佐菌素腹腔注射的方法制作糖尿病大鼠模型,于注药后4周、8周、12周处死大鼠,取视网膜进行HE染色和免疫组织化学染色检测,并与正常对照组大鼠作对比.结果 HMGB1和TLR9在正常对照组和糖尿病4周、8周、12周时的光密度值分别为0.103 7±0.001 1、0.132 3±0.0005、0.145 0±0.002 0、0.155 7±0.001 5和0.084 3±0.0047、0.1164±0.0094、0.132 1±0.000 1、0.139 0±0.0042,在糖尿病大鼠视网膜中的表达呈时间依赖性增加,主要表达在神经节细胞层、内核层和外核层,与正常对照组相比差异均有统计学意义(均为P＜0.01).结论 HMGB1和TLR9在糖尿病大鼠视网膜中表达增加,HMGB1-TLR9信号通路可能参与了糖尿病视网膜病变的发生发展.     

Vascular Endothelial Growth Factor Gene Polymorphisms and Vitreous Proteome Changes in Diabetic Retinopathy

Abstract

Ischemic retinal diseases, particularly diabetic retinopathy, continue to significantly impact vision and remain a leading cause of vision loss in working-aged adults. Identifying specific genetic risk factors for ischemic-driven pathways that increase susceptibility to developing diabetic retinopathy is a priority to allow development of accurate risk assessment algorithms, employ earlier intervention, and design novel treatment strategies to reduce the associated visual complications. Single nucleotide polymorphisms (SNPs) in the VEGF gene have been shown to influence the expression of the VEGF protein. Several studies suggest that SNPs in the VEGF gene mediate genetic predisposition to diabetic retinopathy. In addition, alterations in the vitreous proteome, including carbonic anhydrase mediated vascular permeability, have been found to be associated with sight-threatening proliferative diabetic retinopathy and macular edema. Inhibition of these factors could provide new therapeutic opportunities for the treatment of diabetic retinopathy.     

RASGRF1基因遗传多态性与甘肃地区近视易感性的相关性

目的：分析RASGRF1基因顺式调控元件中单核苷酸多态性与甘肃地区近视人群的相关性。方法：系 列病例对照研究。选取2018 年1 月至2019 年1 月就诊于甘肃省人民医院眼视光学中心的高度近视患 者166 例（332 眼）和中低度近视患者92 例（184 眼）分别作为高度近视组和中低度近视组,并将77 例 （154 眼）无近视的志愿者作为正常对照组。首先利用“DNA元件百科全书”计划（ENCODE）和基因 型组织表达数据库（GTEx）确定眼组织细胞相关功能性调控元件中的5个单核苷酸多态性（SNP）,并 利用多重连接酶检测反应技术进行候选SNP的基因分型。在不同遗传模式下,采用卡方检验、非 条件Logistic回归分析近视患者和正常对照人群的基因型频率分布差异。结果：rs8033417 T/C在显 性模式下含等位基因C的个体近视的发病风险明显降低（P=0.035）;rs8033417 与甘肃地区中低度近 视发病风险无相关性,但在显性模式和加性模式下能明显降低高度近视患者的患病风险（P=0.043、 0.032）。进一步的生物信息学分析发现,rs8033417 T/C与RASGRF1基因的反义长非编码RNA基因 RP11-16K12.1的表达相关。结论：rs8033417 可能是甘肃地区近视发生相关的遗传变异位点,推测其 可能通过影响反义长非编码RNA基因RP11-16K12.1的表达,继而调控RASGRF1基因表达从而影响 近视尤其是高度近视的患病风险。     

An Investigation of the Disc Hyperfluorescence in Fuchs Uveitis Syndrome Using Optical Coherence Tomography Imaging

Abstract

Purpose: To identify the frequency of disc hyperfluorescence, and to use optical coherence tomography to look for vitreopapillary traction as a possible underlying cause.

Methods: Eight patients with presumed Fuchs uveitis syndrome were included. A complete ocular examination, fundus fluorescein angiography, and spectral-domain optical coherence tomography for optic nerve head were performed.

Results: There were 4 males and 4 female patients, and the mean age at diagnosis was 41.7 years. The most common ocular symptom was floaters (5/9). The range of initial visual acuity was 6/5–6/12. The most frequent clinical sign was inflammatory cells in the anterior chamber (9/9). Fundus fluorescein angiography showed disc hyperfluorescence in all but 1 patient. Optical coherence tomography did not show evidence of vitreopapillary traction in all eyes but one eye.

Conclusion: We think that the high frequency of disc hyperfluorescence on fundus fluorescein angiography is an indication of an inflammatory process rather than a mechanical one.     

慢病毒载体介导TLR2基因干扰大鼠角膜上皮细胞和基质细胞的实验研究

目的　观察慢病毒载体（ｌｅｎｔｉｖｉｒａｌｖｅｃｔｏｒ,ＬＶ）介导ＴＬＲ２基因干扰大鼠角膜上皮细胞（ｃｏｒｎｅａｌｅｐｉｔｈｅｌｉａｌｃｅｌｌ,ＣＥＣ）和基质细胞（ｃｏｒｎｅａｌｓｔｒｏｍａｌｃｅｌｌ,ＣＳＣ）的有效性和安全性。方法　分别培养大鼠ＣＥＣ和ＣＳＣ,转染组用携带绿色荧光蛋白（ｅｎｈａｎｃｅｄｇｒｅｅｎｆｌｕｏｒｅｓｃｅｎｔｐｒｏｔｅｉｎ,ｅＧＦＰ）和ＴＬＲ２小干扰ＲＮＡ（ｓｍａｌｌｉｎｔｅｒｆｅｒｅｎｃｅＲＮＡ,ｓｉＲＮＡ）的ＬＶ转染,空白对照组加入空白培养液,阴性对照组加入不携带目的基因的ＬＶ。转染组按最佳感染复数（ｍｕｌｔｉｐｌｉｃｉｔｙｏｆｉｎｆｅｃｔｉｏｎ,ＭＯＩ）分别为１０、５０、１００、２００加入ＬＶ-ＴＬＲ２-ｓｉＲＮＡ-ｅＧＦＰ,选择ｅＧＦＰ表达最强的ＭＯＩ进行后续实验,观察细胞形态,ＣＣＫ８检测细胞增殖情况。流式细胞仪检测两种角膜细胞的转染效率,ＲＴ-ＰＣＲ检测转染后ＴＬＲ２ｍＲＮＡ的表达情况。结果　ＭＯＩ＝２００时转染ＣＥＣ和ＣＳＣ荧光表达最高,和空白对照组相比细胞形态未发生明显改变;ＣＣＫ８结果显示转染组与空白对照组、阴性对照组的ＩＯＤ值差异均无统计学意义（均为Ｐ＞０．０５）;流式细胞仪检测ＣＥＣ和ＣＳＣ转染效率分别为７７．６００％ ±１．１００％和７６．３００％ ±１．３８７％,和阴性对照组相比差异均有显著统计学意义（均为Ｐ＜０．００１）。ＲＴ-ＰＣＲ检测转染后ＣＥＣ和ＣＳＣＴＬＲ２ｍＲＮＡ相对表达量均较对照组明显下降,差异均有显著统计学意义（均为Ｐ＜０．００１）。结论　ＬＶ-ＴＬＲ２-ｓｉＲＮＡ-ｅＧＦＰ可在体外稳定有效转染ＣＥＣ和ＣＳＣ,且对细胞安全性无影响,可有效下调ＴＬＲ２ｍＲＮＡ的表达。     

Circulating S100A8/A9 Levels Reflect Intraocular Inflammation in Uveitis Patients

ABSTRACT

Purpose: To investigate whether there is an association between circulating S100A8/A9 levels and uveitis activity.

Methods: A total of 549 plasma samples were collected from uveitis patients and non-uveitic controls.

Results: S100A8/A9 plasma levels were elevated in uveitis patients compared to non-uveitic controls (P < 0.001). S100A8/A9 plasma levels in patients with active acute anterior uveitis (AAU) were significantly elevated and remarkably decreased in parallel with the severity of intraocular inflammation after corticosteroid treatment (P < 0.001). S100A8/A9 plasma levels were also higher in AAU patients with ankylosing spondylitis (AS) than in patients without AS (P = 0.02). S100A8/A9 plasma levels were significantly increased in uveitis patients with elevated C-reactive protein (CRP, P = 0.004) or erythrocyte sedimentation rates (ESR, P = 0.049) levels compared to uveitis patients with normal CRP or ESR values.

Conclusion: Circulating S100A8/A9 might be a useful biomarker for the measurement of intraocular inflammation.     

Association of Lysyl oxidase (LOX) Polymorphisms with the Risk of Keratoconus in an Iranian Population

Background: Keratoconus is a connective tissue-related eye disease with unknown etiology that causes the loss of visual acuity. Lysyl oxidase (LOX) is an amine oxidase that catalyzes the covalent cross-link of collagens and elastin in the extracellular environment, thus determining the mechanical properties of connective tissue. The current study aimed to investigate the possible associations between two LOX polymorphisms, rs1800449 and rs2288393, and susceptibility to keratoconus.

Methods: A total of 262 Iranian subjects including 112 patients with keratoconus and 150 healthy individuals as controls were recruited. Genotyping for the LOX variants was performed using allele-specific PCR.

Results: A significant difference was found between two groups regarding allelic and genotyping distribution of LOX polymorphism at position rs1800449 G>A. The frequency of AA and GA?+?AA genotypes were increased in patients compared to controls (17% versus 8% and 62.5% versus 50%, respectively), showing a statistically significant difference (OR?=?2.827, 95% CI: 1.251–6.391, p?=?0.012). The A allele was associated with an increased risk for keratoconus, with the frequency of 39.9% and 29% in patients and controls, respectively (OR?=?1.614, 95% CI: 1.119–2.326, p?=?0.011). Furthermore, the haplotype analysis revealed that the rs1800449G/rs2288393C is a protective factor against keratoconus (OR?=?0.425, 95% CI?=?0.296–0.609, p?=?0.001). Conversely, the +473A/rs2288393C (OR?=?3.703, 95% CI?=?2.230–6.149, p?=?0.001) and +473G/rs2288393G (OR?=?15.48, 95% CI?=?3.805–63.03, p?=?0.001) haplotypes were identified as risk factors for keratoconus.

Conclusion: Our study demonstrated that the LOX rs1800449 genotypes (AA and GA?+?AA) and allele (A) appears to confer risk for susceptibility to keratoconus.     

汉族人群MMP-9基因单核苷酸多态性与PACG的相关性

目的：探讨基质金属蛋白酶9(extracellular matrix metalloprotease 9,MMP-9)基因单核苷酸多态性(single nucleotide polymorphism,SNP)与汉族人群原发性闭角型青光眼(primary angle closure glaucoma,PACG)易感性的关联。

方法：选取我院2014-01/2016-12收治的200例汉族PACG患者作为PACG组,同时招募200例汉族体检健康者作为正常对照组。常规外周静脉采集,提取全血基因组DNA,采用聚合酶链反应-限制性片段长度多态性技术(polymerase chain reaction- restriction fragment length polymorphism,PCR-RFLP)检测两组受检者MMP-9基因rs2250889、rs2274755及rs2664538位点等位基因及基因型,分析等位基因和基因型频率分布及其与PACG易感性之间的关系。

结果：两组受检者年龄、性别、体质量指数、血压等一般资料差异均无统计学意义(P>0.05)。MMP-9基因rs2250889、rs2274755及rs2664538位点基因型频率均符合哈迪-温伯格平衡(Hardy-Weinberg equilibrium)。两组受检者rs2250889和rs2664538位点基因型及等位基因频率分布差异均有统计学意义(P<0.05),而rs2274755位点基因型及等位基因频率分布差异均无统计学意义(P>0.05)。rs2250889位点中携带CC基因型人群为PACG的易感人群。同样,rs2664538位点中携带GG基因型人群为PACG的易感人群。

结论：汉族人群中,MMP-9基因rs2250889和rs2664538位点多态性与 PACG 的发病存在一定关联,而rs2274755位点多态性与PACG的易感性无关。     

Analysis of HLA Class I and Class II Gene Polymorphisms in Japanese Patients with Human T-Cell Lymphotropic Virus Type 1-Associated Uveitis

Purpose: To investigate the immunogenetic background of human T-cell lymphotropic virus type 1 (HTLV-1)-associated uveitis (HAU) that presents immune-mediated reactive changes in the uvea. Methods: HLA class I and class II genes were studied in 51 patients with HAU, 192 asymptomatic HTLV-1 carriers, and 266 HTLV-1-seronegative controls using a high-resolution method of HLA DNA typing. The HLA alleles of HAU were compared with those of HTLV-1 carriers and healthy controls. Results: We identified 62 distinct alleles of HLA-A, HLA-Cw, and HLA-B and 49 distinct alleles of HLA-DRB1 and HLA-DQB1 in patients with HAU, asymptomatic HTLV-1 carriers, and healthy controls. The relative frequencies of these HLA alleles did not differ among the three groups. Conclusion: The results suggest that HLA class I and class II genes do not contribute to susceptibility to HAU.     

基质金属蛋白酶-9在人正常角膜及真菌性角膜溃疡中的基因表达

目的:研究基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)在人正常角膜及真菌性角膜溃疡中的基因表达,探讨MMP-9在真菌性角膜溃疡形成中的作用。方法:搜集24份真菌性角膜溃疡标本(溃疡组)及12份人正常角膜组织标本(对照组),其中12份真菌性角膜溃疡组织和6份正常的角膜组织分别抽提组织总的RNA,以Primer5.0软件设计MMP-9的特异引物,并对标本RNA进行RT-PCR扩增,以MMP-9目的电泳条带与β-actin内参照条带灰度值比值代表相对表达量。分12份角膜溃疡组织和6份正常的角膜组织进行免疫组化检测MMP-9表达,通过免疫组化染色的灰度值进行统计学分析。结果:MMP-9在真菌性角膜溃疡中的表达明显高于正常角膜,且主要位于角膜上皮层、基底膜以及基质层。结论:真菌性角膜溃疡组织中MMP-9的基因表达明显上调。MMP-9在真菌性角膜溃疡的发生、发展及角膜穿孔中起着重要作用。加强MMP-9在真菌性角膜溃疡中的作用机制的研究,有望对真菌性角膜溃疡的治疗提供新的途径。     

CFH (rs1410996), HTRA1 (rs112000638) and ARMS2 (rs10490923) Gene Polymorphisms are Associated with AMD Risk in Spanish Patients

ABSTRACT

Purpose: Age-related macular degeneration (AMD) is the main cause of legal blindness in the western adult population. We investigated the association between SNPs located in CFH, ARMS2 and HTRA1 and AMD in Spanish patients.

Patients and Methods: We obtained peripheral blood samples from 121 patients with a diagnosis of AMD (84 exudative and 37 atrophic) at the Department of Ophthalmology of the University Hospital of Salamanca. We took 91 subjects as a control group. We studied a single nucleotide polymorphism (SNP) in each patient for each of the genes associated with high susceptibility to developing AMD using Real-time PCR with TaqMan probes for CFH and ARMS2 polymorphisms and PCR-RFLP for HTRA1 polymorphism.

Results: We observed a statistically significant difference between patients and controls in the distribution of CFH rs1410996 genotypes, patients homozygous for the C-allele have twice the risk of developing the disease (p?=?0.010; OR?=?2,176 (1.194–3.964)). The analysis of ARMS2 rs10490923 polymorphism also showed differences in allelic distribution between the case and control groups (p?<?0.001). Carriers of the T-allele appear more frequently in the group of patients (p?<?0.001; OR?=?3.340 (1.848–6.060)). Our results also confirm significant differences in the distribution of HTRA1 rs112000638 polymorphism with an increased representation of the G-allele in the patient’s group (p?<?0.001; OR?=?6.254(3.463–12.280)). Our study also indicates that TTGG ARMS2/HTRA1 (rs10490923/rs112000638) haplotype increases the risk of developing AMD by 9 times.

Conclusions: Our results show that genotypes of ARMS2 (rs10490923), HTRA1 (rs112000638) and CFH (rs1410996) polymorphisms are related to an increased risk of suffering AMD in Spanish patients.     

非球面人工晶状体植入后的视功能观察

目的 观察非球面折叠人工晶状体与传统球面人工晶状体对患者术后视功能的影响.方法 对收治的白内障患者62例62只眼行超声乳化联合人工晶状体植入术.随机分为两组,术中分别选用美国ALCON公司AcrySof R Natural和AcrySof R IQ两种人工晶状体,术后1月行最佳矫正视力、对比敏感度及视觉心理问卷调查,并对各项检查结果行统计分析.结果 术后1月两组患者最佳矫正视.力差异无统计学意义(P>0.05);对比敏感度检查显示在明视有或无眩光状态下,两组患者差异无统计学意义(P>0.05),在3c～18.0c/d 4个空间频率的暗视及暗视有眩光状态下,两组患者差异有统计学意义,非球面人工晶状体组对比敏感度值高于球面人工晶状体组(P<0.05);视觉心理问卷调查结果显示两组差异无统计学意义(P>0.05).结论 与球面人工晶状体相比,非球面人工晶状体能有效提高白内障患者术后的对比敏感度,改善视功能,但对患者术后视觉心理改变不明显,尚需要增大样本量进一步分析.     

Investigation of the Rho-kinase Gene Polymorphism in Primary Open-angle Glaucoma

Purpose: Genetic factors are shown to have a role in the development of primary open-angle glaucoma (POAG). The aim of this study was to determine the effects of genetic polymorphisms of Rho-kinase (ROCK) genes on the risk of POAG in a Turkish population.

Methods: Genomic DNA was extracted from leukocytes of the peripheral blood, and 8 single nucleotide polymorphisms in the ROCK1 and ROCK2 genes were analysed in 179 patients with POAG and in 182 healthy controls of similar age by using BioMark HD dynamic array system.

Results: Neither genotype distributions nor the allele frequencies for the ROCK1 (rs35996865) and ROCK2 [rs2290156, rs965665, rs10178332, rs2230774 (Thr431Asn), rs2230774 (Thr431Ser), rs6755196, and rs726843] gene polymorphisms showed a significant difference between the groups. There were also no marked associations between the haplotype frequencies and POAG.

Conclusions: This is the first study to examine the involvement of ROCK1 and ROCK2 gene variations in the risk of POAG development. This study demonstrated that the polymorphisms studied are not associated with the increased risk of development of POAG in the Turkish population.