Notch信号对高糖诱导人视网膜血管内皮细胞凋亡的保护作用

目的　探讨Ｎｏｔｃｈ１信号对高糖情况下多聚二磷酸腺苷核糖聚合酶-１［Ｐｏｌｙ（ＡＤＰ-ｒｉｂｏｓｅ）ｐｏｌｙｍｅｒａｓ-１,ＰＡＲＰ-１］和核因子-κＢ（ｎｕｃｌｅａｒｆａｃｔｏｒｋａｐｐａＢ,ＮＦ-κＢ）的ｐ５０亚单位（ＮＦ-κＢ５０）介导的人视网膜血管内皮细胞（ｒｅｔｉｎａｌｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｃｅｌｌｓ,ＲＶＥＣｓ）凋亡的抑制作用。方法　体外培养人ＲＶＥＣｓ及ＨＥＫ２９３Ｔ细胞,并进行传代,同时构建高糖（葡萄糖浓度为３０ｍｍｏｌ?Ｌ－１）人ＲＶＥＣｓ细胞模型。构建ｐＣＭＶ-Ｓｃｒｉｐｔ-Ｎｏｔｃｈ１和ｐＣＭＶ-Ｓｃｒｉｐｔ-ＤＬＬ４质粒,酶切鉴定后ＷｅｓｔｅｒｎＢｌｏｔ法检测重组质粒表达目的基因的效果,同时合成ｓｉＮｏｔｃｈ１并转染高糖培养的人ＲＶＥＣｓ,ＷｅｓｔｅｒｎＢｌｏｔ法检测Ｎｏｔｃｈ基因敲除效果。应用免疫共沉淀及ＷｅｓｔｅｒｎＢｌｏｔ法检测ｓｉＮｏｔｃｈ１对高糖下ＰＡＲＰ-１和ＮＦ-κＢ的相互作用的影响;应用ＷｅｓｔｅｒｎＢｌｏｔ法检测高糖条件下Ｎｏｔｃｈ１／ＤＬＬ４上调和下调后人ＲＶＥＣｓ中ＰＡＲＰ-１、ｐ-ＡＫＴ、ＮＦ-κＢ５０及ｃａｓｐａｓｅ-３的表达变化。结果　人ＲＶＥＣｓ复苏后２４ｈ贴壁,细胞呈扁平梭形,３ｄ左右细胞开始融合呈铺路石样,单层生长,铺满瓶底,并可见接触抑制现象。免疫共沉淀结果显示高糖组培养的人ＲＶＥＣｓ其ＰＡＲＰ-１结合的ＮＦ-κＢ５０的量较正常对照组增加;将ｓｉＮｏｔｃｈ１转染高糖组培养的人ＲＶＥＣｓ后或同时加入ＤＬＬ４,则人ＲＶＥＣｓ中ＰＡＲＰ-１结合的ＮＦ-κＢ５０的量较前增加显著,提示Ｎｏｔｃｈ能抑制高糖下ＰＡＲＰ-１和ＮＦ-κＢ５０的相互作用。ＷｅｓｔｅｒｎＢｌｏｔ检测结果显示高糖条件下,Ｎｏｔｃｈ１和ＤＬＬ４上调后其ＰＡＲＰ-１及ｃａｓｐａｓｅ-３表达量较前显著降低;Ｎｏｔｃｈ１下调后其ＰＡＲＰ-１及ｃａｓｐａｓｅ-３表达量较前增加;ｐ-ＡＫＴ表达量则相反;提示Ｎｏｔｃｈ信号能抑制高糖诱导的ＰＡＲＰ-１及ｃａｓｐａｓｅ-３的激活及表达。结论　高糖情况下Ｎｏｔｃｈ信号可调控ＰＡＲＰ-１和ＮＦ-κＢ的相互作用,并抑制ＰＡＲＰ-１激活引起的人ＲＶＥＣｓ的凋亡。     

糖尿病视网膜病变患者红细胞膜Na^＋—K^＋—ATP酶活性的变化及相 …

目的 探讨糖尿病视网膜病变（ｄｉａｂｅｔｉｃ ｒｅｔｉｎｏｐａｔｈｙ,ＤＲ）患者红细胞膜Ｎａ＾＋－Ｋ＾＋－ＡＴＰ酶活性进行检测,并将其结果与红细胞Ｎａ＾＋,Ｋ＾＋浓度、红细胞变形指数（ｅｒｙｔｈｒｏｃｙｔｅ ｄｅｆｏｒｍａｂｉｌｉｔｙ ｉｎｄｅｘ,ＥＤＩ）、空腹血糖（ｆａｓｔｉｎｇ ｂｌｏｏｄ－ｇｌｕｃｏｓｅ,ＦＢＧ）、糖基化血红蛋白（ｇｌｙｃｏｓｙｌａｔｅｄ ｈｅｍｏｇｌｏｂｉｎ,ＨｂＡ１ｃ）及     

组织激肽释放酶在糖尿病视网膜病变患者血清中的变化及其调节血管新生机制

目的　检测组织激肽释放酶（ｔｉｓｓｕｅｋａｌｌｉｋｒｅｉｎ,ＴＫＬＫ）、血管内皮细胞生长因子（ｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｇｒｏｗｔｈｆａｃｔｏｒ,ＶＥＧＦ）和可溶性细胞间黏附分子-１（ｓｏｌｕｂｌｅｉｎｔｒａｃｅｌｌｕｌａｒａｄｈｅｎｓｉｏｎｍｏｌｅｃｕｌ-１,ｓＩＣＡＭ-１）在糖尿病视网膜病变（ｄｉａｂｅｔｉｃｒｅｔｉｎｏｐａｔｈｙ,ＤＲ）患者血清中的变化,观察ＴＫＬＫ在低氧条件下对人视网膜微血管内皮细胞（ｈｕｍａｎｒｅｔｉｎａｌｍｉｃｒｏｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｃｅｌｌｓ,ＨＲＭＥＣｓ）中ＶＥＧＦ和ＩＣＡＭ-１表达的影响。方法　收集２型糖尿病患者６０例,按照ＤＲ分期标准将患者分为糖尿病无ＤＲ组（ＤＭ组）、非增生性ＤＲ组（ＮＰＤＲ组）和增生性ＤＲ组（ＰＤＲ组）,收集同期在本院体检的志愿者作为对照组,每组２０例。ＥＬＩＳＡ法检测血清中ＴＫＬＫ、ＶＥＧＦ和ｓＩＣＡＭ-１的水平。体外ＨＲＭＥＣｓ分别进行常氧和低氧培养,不同浓度重组ＴＫＬＫ处理后,检测细胞增殖、凋亡以及ＶＥＧＦ和ＩＣＡＭ-１的表达。结果　ＤＭ、ＮＰＤＲ和ＰＤＲ组患者血清中ＴＫＬＫ、ＶＥＧＦ和ｓＩＣＡＭ-１水平明显高于对照组,４组总体差异有统计学意义（Ｆ＝２８．８０５,Ｐ＝０．００２;Ｆ＝３２．０４１,Ｐ＝０．００２;Ｆ＝２６．１６９,Ｐ＝０．００１）;ＰＤＲ患者血清中ＴＫＬＫ、ＶＥＧＦ和ｓＩＣＡＭ-１水平显著高于ＤＭ和ＮＰＤＲ组（均为Ｐ＜０．００１）。ＴＫＬＫ与ＶＥＧＦ（ｒ＝０．６２３,Ｐ＜０．０１）和ｓＩＣＡＭ-１水平（ｒ＝０．５９８,Ｐ＜０．０１）均呈正相关。１０μｇ?ｍＬ－１ｒｈＴＫＬＫ可显著抑制低氧诱导的ＨＲＭＥＣｓ增殖以及ＶＥＧＦ和ＩＣＡＭ-１的表达,并可促进细胞凋亡（Ｐ＜０．０５）。结论　ＴＫＬＫ通过与ＶＥＧＦ和ＩＣＡＭ-１相互作用而影响ＤＲ的进展。     

内皮素_(-1)及其激动剂、拮抗剂对兔眼睫状体组织环磷酸腺苷的影响

为测定内皮素－１（Ｅｎｄｏｔｈｅｌｉｎ－１，ＥＴ－１）及其激动剂Ｓ６ｂ（ｓａｒａｆｏｔｏｘｉｎ）、拮抗剂ＢＱ１２３对兔眼睫状体组织环磷酸腺苷（ｃｙｃｌｉｃａ－ｄｅｎｏｓｉｎｅ－ｍｏｎｏｐｈｏｓｈａｔｅ，ｃＡＭＰ）的影响，运用内源性蛋白结合法测定兔睫状体组织ｃＡＭＰ的含量。结果：单独使用ＥＴ－１、Ｓ６ｂ以及两者联合应用均使睫状体ｃＡＭＰ升高，且呈剂量效应关系；单独应用ＢＱ１２３未引起ｃＡＭＰ改变，但ＥＴ－１与ＢＱ１２３共同作用，可有效地拮抗ＥＴ－１的升高ｃＡＭＰ效应。结论：ＥＴ－１可使兔睫状体组织ｃＡＭＰ升高；兔眼睫状体上有ＥＴＡ亚型受体。     

病理性近视与HLA—DPB1基因相关性的初步探讨

目的研究病理性近视与ＨＬＡ－ＤＰＢ１基因的相关性,以便得到其易感或抵抗基因。方法用多聚酶链反应（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ,ＰＣＲ）扩增４０例患者ＨＬＡ－ＤＰＢ１基因的第二个外显子后,用Ｂｓｐ１２８６Ⅰ,ＦｏｋⅠ,ＤｄｅⅠ,ＢｓａＪⅠ,ＢｓＨⅡ,ＲｓａⅠ,ＡｖａⅡ和ＥｃｏＮⅠ共８种等位基因特异的限制性内切酶酶切,依据限制性片段长度多态性（ｒｅｓｔｒｉｃｔｉｏｎｆｒａｇｍｅｎｔｌｅｎｇｔｈｐｏｌｙｍｏｒｐｈｉｓｍ,ＲＦＬＰ）格局对每例患者进行基因型确定,然后计算各等位基因频率并与正常人进行比较。结果ＨＬＡ－ＤＰＢ１＊０３０１等位基因的频率显著低于正常对照组（Ｙａｔｅｓ校正χ２＝４．３２,Ｆｉｓｈｅｒ确切Ｐ＜０．０５）,ＦｉｓｈｅｒＰ值经所比较的等位基因数修正后,Ｐｃｏｒ＞０．０５;ＨＬＡ－ＤＰＢ１＊０５０１／０５０１纯合子的比率与正常人差异有显著性（ｕ＝２．２７,Ｐ＜０．０５）。结论病理性近视在ＨＬＡ－ＤＰＢ１基因座位不存在易感或抵抗基因。ＨＬＡ－ＤＰＢ１＊０５０１／０５０１纯合子的频率在患者中显著升高可能为一种连锁信号。     

准分子激光角膜屈光性手术治疗近视的疗效分析

对准分子激光屈光性角膜切削术（ｐｈｏｔｏｒｅｆｒａｃｔｉｖｅｋｅｒａｔｃｅｃｔｏｍｙ，ＰＲＫ）和准分子激光原位角膜磨镶术（ｌａｓｅｒｉｎｓｉｔｕｋｅｒａｔｏｍｉｌｅｕｓｉｓ，ＬＡＳＩＫ）治疗近视的疗效进行对比研究。方法，对术前近视度为－１．５０Ｄ－２０．００Ｄ的患者的６００只眼，接近视度分为三组：中低度近视组（－１．５０Ｄ－－６．００Ｄ）２３４只眼，１９４眼行ＰＲＫ手术，４０眼行ＬＡＳＩＫ手术，高     

2.0mm小切口飞秒激光角膜基质透镜取出术与飞秒激光辅助的LASIK矫正近视的疗效比较

目的　比较２．０ｍｍ微切口飞秒激光角膜基质透镜取出术（２．０ｍｍｓｍａｌｌｉｎｃｉｓｉｏｎｌｅｎｔｉｃｕｌｅｅｘｔｒａｃｔｉｏｎ,２．０ｍｍＳＭＩＬＥ）与飞秒激光辅助的ＬＡＳＩＫ（ｌａｓｅｒｉｎｓｉｔｕｋｅｒａｔｏｍｉｌｅｕｓｉｓｗｉｔｈｆｅｍｔｏｓｅｃｏｎｄｌａｓｅｒ,ＦＳ-ＬＡＳＩＫ）矫正近视的疗效。方法　研究纳入行２．０ｍｍＳＭＩＬＥ的近视患者４８例（９６眼）,同期行ＦＳ-ＬＡＳＫ者５０例（１００眼）,记录并比较两组术前及术后１ｄ、１周、１个月、３个月及６个月的裸眼视力（ｕｎｃｏｒｒｅｃｔｅｄｖｉｓｕａｌａｃｕｉｔｙ,ＵＣＶＡ）、最佳矫正视力（ｂｅｓｔｃｏｒｒｅｃｔｅｄｖｉｓｕａｌａｃｕｉｔｙ,ＢＣＶＡ）、屈光度、生活质量量表（ｑｕａｌｉｔｙｏｆｌｉｆｅ,ＱＯＬ）得分及手术满意度量表评分。结果　在术后１周以后ＳＭＩＬＥ组视力高于ＦＳ-ＬＡＳＩＫ组,且状态较ＦＳ-ＬＡＳＩＫ组更为稳定。术后１个月、３个月、６个月ＳＭＩＬＥ组ＵＣＶＡ≥术前ＢＣＶＡ的比例高于ＦＳ-ＬＡＳＩＫ组,差异均有统计学意义（均为Ｐ＜０．０５）。术后早期（术后１周）ＳＭＩＬＥ组存在１例过矫,而ＦＳ-ＬＡＳＩＫ组存在１例欠矫,但在术后１个月、３个月、６个月两组手术患者均在±１．００Ｄ范围内。术后平均角膜前表面形态变异指数及垂直不对称指数在各时间点均为ＦＳ-ＬＡＳＩＫ组大于ＳＭＩＬＥ组,差异均有统计学意义（均为Ｐ＜０．０５）。两组术前及术后６个月ＱＯＬ得分组间比较,差异均无统计学意义（均为Ｐ＞０．０５）,术后６个月ＱＯＬ得分均较术前有所增加,且差异均有统计学意义（ＳＭＩＬＥ:ｔ＝－１３．８５,Ｐ＝０．００;ＦＳ-ＬＡＳＩＫ:ｔ＝－１３．２１,Ｐ＝０．００）,而两组术后６个月ＱＯＬ得分比较,差异无统计学意义（Ｐ＞０．０５）。两组均有较高的再次手术选择率及手术推荐率,并且未发生严重的并发症。结论　２．０ｍｍＳＭＩＬＥ与ＦＳ-ＬＡＳＩＫ均具有良好的有效性、稳定性以及可预测性,术后都可获得良好的视力及良好的生活质量,前者更好地保持了角膜前表面的形态。     

JNK细胞通路在高压氧和过氧化氢诱导人晶状体上皮细胞凋亡中的作用

目的　研究ＪＮＫ细胞信号通路在高压氧（ｈｙｐｅｒｂａｒｉｃｏｘｙｇｅｎ,ＨＢＯ）和过氧化氢（ｈｙｄｒｏｇｅｎｐｅｒｏｘｉｄｅ,Ｈ２Ｏ２）影响晶状体上皮细胞活力和凋亡中的作用。方法　培养人晶状体上皮细胞系ＳＲＡ０１／０４,实验分为对照组、ＳＰ６００１２５组（２０μｍｏｌ?Ｌ－１）、ＨＢＯ组（体积分数９５％ Ｏ２＋体积分数５％ＣＯ２,６００Ｐａ）、ＨＢＯ＋ＳＰ６００１２５组、Ｈ２Ｏ２组（２００μｍｏｌ?Ｌ－１）和Ｈ２Ｏ２＋ＳＰ６００１２５组,处理６ｈ后收集细胞,应用ＭＴＴ法检测各组细胞活力,ＡｎｎｅｘｉｎＶ-ＦＩＴＣ凋亡试剂盒处理细胞后用流式细胞仪检测各组细胞的凋亡率,通过Ｗｅｓｔｅｒｎ-ｂｌｏｔ法检测各组细胞ＪＮＫ、ｐ-ＪＮＫ、ｃ-Ｊｕｎ、ｐ-ｃ-Ｊｕｎ、ｃａｓｐａｓｅ３、ｃａｓｐａｓｅ９的表达情况。结果　作用６ｈ后,ＨＢＯ和Ｈ２Ｏ２处理的晶状体上皮细胞Ａ５７０分别为０．４２３±０．０４７和０．４０６±０．０６９,较对照组的０．８２４±０．０６５明显下降（Ｐ＜０．０５）,细胞凋亡率为（１９．７７３±１．１４７）％和（２９．２０６±１．６７８）％,较对照组的（２．１８４±１．０１５）％明显上升（Ｐ＜０．０５）,ＳＰ６００１２５可部分抑制ＨＢＯ和Ｈ２Ｏ２对晶状体上皮细胞活力和凋亡的影响,ＨＢＯ和Ｈ２Ｏ２处理的晶状体上皮细胞ｐ-ＪＮＫ、ｐ-ｃ-Ｊｕｎ蛋白表达量及ｃａｓｐａｓｅ３、ｃａｓｐａｓｅ９蛋白表达量升高,Ｓｐ６００１２５可部分抑制高压氧和Ｈ２Ｏ２ 诱导的晶状体上皮细胞ｐ-ＪＮＫ、ｐ-ｃ-Ｊｕｎ、ｃａｓｐａｓｅ３、ｃａｓｐａｓｅ９蛋白高表达。结论　ＨＢＯ和Ｈ２Ｏ２可激活晶状体上皮细胞中ＪＮＫ细胞信号通路,促使细胞凋亡,提示ＪＮＫ细胞信号通路在ＨＢＯ和Ｈ２Ｏ２诱导晶状体上皮细胞凋亡的发生发展中起重要作用。     

去整合素echistatin对糖尿病兔后发性白内障形成中信号通路PI3-K/Akt和ERK1/2的影响

目的　观察去整合素ｅｃｈｉｓｔａｔｉｎ对糖尿病兔后发性白内障形成中信号通路ＰＩ３-Ｋ／Ａｋｔ和ＥＲＫ１／２的影响,从分子水平上探讨ｅｃｈｉｓｔａｔｉｎ的作用。方法　建立糖尿病兔模型（ｎ＝２４）,随机分组并行透明晶状体囊外摘出术,术毕前房分别注入０．２ｍＬ灭菌蒸馏水（对照组,ｎ＝１２）或０．２ｍＬ１０．０ｍｇ·Ｌ－１ｅｃｈｉｓｔａｔｉｎ溶液（ｅｃｈｉｓｔａｔｉｎ干预组,ｎ＝１２）。术后１０ｄ及６周（每个时间点６眼）,裂隙灯显微镜观察两组术眼ＰＣＯ分级情况,同时摘取术眼应用ＲＴ-ＰＣＲ法检测后囊膜上信号因子Ａｋｔ和ＥＲＫ１／２ｍＲＮＡ表达情况。结果　术后１０ｄ对照组和ｅｃｈｉｓｔａｔｉｎ干预组ＰＣＯ分级比较差异无统计学意义（Ｐ＝０．０９３）,但ｅｃｈｉｓｔａｔｉｎ干预组ＰＣＯ１级眼数少于对照组;术后６周ｅｃｈｉｓｔａｔｉｎ干预组ＰＣＯ级别明显低于对照组（Ｐ＝０．００６）。对照组和ｅｃｈｉｓｔａｔｉｎ干预组ＡｋｔｍＲＮＡ相对表达量术后１０ｄ分别为０．９８１７±０．３８０４、０．４８１７±０．１６６５,术后６周分别为０．６５１７±０．２０４７、０．４０１７±０．１５１３,ｅｃｈｉｓｔａｔｉｎ干预组均明显低于对照组（Ｐ＝０．０１５,０．０３７）。对照组和ｅｃｈｉｓｔａｔｉｎ干预组ＥＲＫ１／２ｍＲＮＡ相对表达量术后１０ｄ分别为０．９４８３±０．２２７５、０．６１００±０．２８０６,术后６周分别为０．９２１７±０．２９９４、０．４６５０±０．１８００,ｅｃｈｉｓｔａｔｉｎ干预组均明显低于对照组（Ｐ＝０．０４５,０．００９）。结论　去整合素ｅｃｈｉｓｔａｔｉｎ对糖尿病兔后发性白内障的发生和发展有一定的抑制作用,其发生的机制可能与抑制信号因子Ａｋｔ和ＥＲＫ１／２表达,进而阻断信号通路ＰＩ３-Ｋ／Ａｋｔ和ＥＲＫ１／２的转导有关。     

紫外光A/核黄素角膜交联后兔角膜基质内基质金属蛋白酶-1含量的变化

目的　检测紫外光Ａ／核黄素角膜交联（ｕｌｔｒａｖｉｏｌｅｔＡ／ｒｉｂｏｆｌａｖｉｎｃｏｒｎｅａｌｃｒｏｓｓ-ｌｉｎｋ-ｉｎｇ,ＵＶＡＲＣＸＬ）后兔角膜基质内基质金属蛋白酶（ｍａｔｒｉｘｍｅｔａｌｌｏｐｒｏｔｅｉｎａｓｅ,ＭＭＰ）-１的短期含量变化。方法　１５只新西兰白兔,随机分为３组,每组５只,Ａ组为空白对照组,Ｂ组为角膜交联后３ｄ,Ｃ组为角膜交联后７ｄ。Ｂ、Ｃ两组行ＵＶＡＲＣＸＬ。各组取角膜后去除角膜上皮和内皮细胞,应用ＥＬＩＳＡ法检测角膜基质内ＭＭＰ-１的含量。结果　Ａ组角膜基质内ＭＭＰ-１／总蛋白含量为（０．１４０±０．０３６）×１０－６,Ｂ组为（０．２４２±０．０５９）×１０－６,Ｃ组为（０．３７２±０．０６１）×１０－６,３组之间差异有统计学意义（Ｆ＝２４．０５１,Ｐ＝０．０００）。ＵＶＡＲＣＸＬ后３ｄ,兔角膜基质内ＭＭＰ-１含量显著升高,与Ａ组相比差异有统计学意义（Ｐ＜０．０５）,术后７ｄ其含量进一步升高,与Ｂ组相比差异有统计学意义（Ｐ＜０．０１）。结论　ＵＶＡＲＣＸＬ后７ｄ内,兔角膜基质内ＭＭＰ-１含量持续增加,推测ＭＭＰ-１可能对ＵＶＡＲＣＸＬ后的角膜基质重塑起一定作用。     

Association of HLA with Vogt-Koyanagi-Harada syndrome in Koreans

PURPOSE: To study the distribution of human leukocyte antigen HLA-A/B antigens and HLA-DR/-DQ/-DP alleles and to investigate the immunogenetic background of Korean patients with Vogt-Koyanagi-Harada (VKH) syndrome and clinical course with different types of HLA. METHODS: Human leukocyte antigen typings were performed in 18 Korean patients with VKH syndrome and in 128 healthy control subjects. HLA-A/B loci serologic typing was performed according to the standard microlymphocytotoxicity technique. DNA was extracted through the salting out method, and HLA-DR phenotyping and HLA-DR4, HLA-DQ, and HLA-DP subtyping were performed with the polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) method. RESULTS: Among HLA-A/B antigens typed by the standard microlymphocytotoxicity method, the frequencies of HLA-A31 (RR = 6.1, P<1x10(-2)) and HLA-B55 (RR = 15.8, P<.05) were significantly increased in the patient group compared with the control group. Among HLA-DR/-DQ/-DP alleles subtyped by DNA methods, the frequencies of HLA-DRB1*04 (RR = 45.1, P<1x10(-7)) and HLA-DRB1*07 (RR = 3.2, P<.05) were significantly increased. However, significant decreases in HLA-DRB1*08 (RR = .1, P<.05), HLA-DRB1*13 (RR = .1, P<.05), and HLA-DRB1*14 (RR = .1, P<.05) frequencies were observed. The result of HLA-DR, HLA-DQ, and HLA-DP subtyping showed the significant increase in DRB1*0405 (RR = 45.1, P<1x10(-7)), DQA1*0302 (RR = 12.0, P<1x10(-4)), DQB1*0303 (RR = 5.0, P<1x10(-2)), DQB1*0401 (RR = 18.9, P<1x 10-6), and DPB1*0501 (RR = 3.8, P<.05). However, significant decreases in DQA1*0101 (RR = .1, P< .05), DQA10102 (RR = .1, P<1x10(-2)), DQA1*0103 (RR = .1, P<.05), DQA1*0501 (RR = .1, P<1x10(-2)), DQB1*0301 (RR = .1, P<.05), DQB1*0601 (RR = .1, P<.05), DPB1*0201 (RR = .3, P<.05), and DPB1*0401 (RR = .1, P<.05) frequencies were also observed. In patients with DRB1*0405 itself or HLA-DRB1*0405-DQA1*0302-DQB1*0401 haplotype, a reduction in visual acuity and ocular complications was common. CONCLUSIONS: These results suggest that HLA-DRB1*0405 itself or HLA-DRB1*0405-DQA1*0302-DQB1*0401 haplotype is greatly increased and may play the most important role in the development and the clinical course of VKH syndrome in Korean patients.     

Human leukocyte antigen serologic and DNA typing of Beh?et's disease and its primary association with B51.

Ninety Japanese patients with Beh?et's disease (BD) were typed for human leukocyte antigen (HLA)-DRB1, -DQA1-, -DQB1, and -DPB1 alleles by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and for HLA-A, -B, -C, -DR, and -DQ antigens by conventional serologic typing. Serologic HLA typing showed a remarkably significant increase of HLA-B51 and a significant decrease of HLA-DQw1 in the patients with BD, especially those with ocular lesions including complete type, as compared with the control group (for B51, chi-squared = 46.75, P corrected < 0.001, relative risk [RR] = 7.9; for DQw1, chi-squared = 12.10, P corrected < 0.01, RR = 0.4). By PCR-RFLP genotyping, no significant difference was revealed in any class II alleles between the patient and the control groups in the corrected P value test, but P value analysis showed the significantly high frequency of DRB1*0802 and the significantly low frequencies of DQA1*0103, DQB1*0601, and DQB1*0501. No significant difference was observed in any DPB1 alleles by either P value analysis. These results indicated that the primary and primordial gene(s) responsible for the susceptibility to BD, especially related to ocular lesions, were not located in the HLA class II gene region but were in or very close to the HLA-B locus in the class I region. They also suggested the possibility that BD was a symptom complex associated with some independent diseases.     

我国汉族Vogt-Koyanagi-Harada综合征患者人类白细胞抗原DQB1等位基因多态性研究

目的探讨人类白细胞抗原（HLA）-DQB1等位基因多态性与沃格特-小柳-原田综合征（VKH）遗传易感相关性及与临床表现的关系。方法应用聚合酶链式反应-序列特异性引物（PCR—SSP）,对我国汉族VKH患者和非VKH正常对照者HLA-DQB1等位基因进行分型,分析HLA-DQB1等位基因与患者临床表现的关系。结果收集我国汉族VKH患者88例,非VKH正常对照者88例。VKH患者中男性41例（46．6％）,女性47例（53．4％）;发病年龄15—67岁,平均36岁。VKH患者HLA—DQB1各等位基因频率：HLA-DQB1＊0401为31．8％,DQB1＊0201为17．6％,DQB1＊0301／＊0304为17．1％,DQB1＊0602为12．5％,DQB1＊0303为6．8％,DQB1＊0302为6．3％,DQB1＊0402为1．7％,DQB1＊0502为1.7％,DQB1＊0601为1．7％,DQB1＊0501为1．1％,DQB1＊050为31．1％,DQB1＊0604为0．6％;DQB1＊0603未检出。VKH患者中HLA—DQB1＊0401（VKH组31．8％与对照组4．5％比较,r=44．00,P=0．000,OR=9．8,95％口为4．51—21．31）和HLA—DQB1＊0303（VKH组6．82％与对照组0．57％比较,〈=9．67,P=0．002,伽=12．81,95％CI为1.65—99．58）等位基因频率高于正常人对照组,差异有统计学意义。而VKH患者HLA—DQB1＊0601（VKH组1.7％与对照组9．7％比较,r=10．39,P=0．001,OR=0．16,95％CI为0．05—0．56）和HLA—DQB1＊0302（VKH组6．3％与对照组19．3％比较,r=13．48,P=0．000,OR=0．28,95％CI为0．14—0．57）等位基因频率显著低于正常人对照组,差异有统计学意义。HLA-DQB1＊0401阴性患者与阳性患者之间的临床表现差异无统计学意义（P〈0．01）。结论（I）HLA—DQB1＊0401和DQB1＊0303是VKH的易感等位基因,而HLA-DQB1＊0601和DQB1＊0302是抗性等位基因。HLA-DQB1＊0401基因与临床表现无明显相关性。（2）PCR-SSP可用于快速检测HLA—DQB1等位基因型别。     

我国汉族Vogt-Koyanagi-Harada综合征患者人类白细胞抗原DQB1基因多态性研究

目的 探讨人类白细胞抗原(HLA)DQB1启动子和编码子在Vogt-Koyanagi-Harada(VKH)综合征发病中的作用.方法 88例汉族VKH综合征患者和88名正常对照者纳入本研究.VKH综合征患者中,男性41例,女性47例,发病年龄15～67岁,平均年龄(36±11)岁.正常对照者中,男性42例,女性46例,年龄24～68岁,平均年龄(41±19)岁.采用聚合酶链式反应-序列特异性引物(PCR-SSP)方法进行HLA-DQB1等位基因分型,聚合酶链式反应-单链构象多态性(PCR-SSCP)-克隆-测序法检测HLA-DQB1基因启动子区(HLA-QBP)等位基因.应用x2检验或Fisher精确概率法进行统计分析.结果 VKH综合征患者中HLA-DQB1*0401(0.318比0.045,x2=44.00,P=0.000,OR=9.8)和DQB1*0303(0.068比0.006,x2=9.67,P=0.002,OR=12.81)频率明显高于正常对照者,差异有统计学意义.而HLA-DQB1*0601(0.017比0.096,x2=10.39,P=0.001,OR=0.16)和HLA-DQB1*0302(0.063比0.193,x2=13.48,P=0.000,OR=0.28)在VKH综合征患者中出现的频率显著低于正常对照者,差异有统计学意义.VKH综合征患者HLA-DQB1基因启动子区-189C/A的C等位基因频率显著高于正常对照者(0.324比0.074,x2=45.92,P=0.000),而-227G/A等位基因频率低于正常对照者(0.011比0.108,x2=15.63,P=0.000).VKH综合征患者中易感等位基因的组合(-189C和HLA-DQB1*0401)频率明显高于正常对照者,正常对照者中抗性等位基因的组合(-227G和HLA-DQB1*0601)频率显著高于VKH综合征患者.结论 VKH综合征发病可能与HLA-DQB1启动子和编码区相互作用有关.     

Immunogenetics and clinical phenotype of sympathetic ophthalmia in British and Irish patients

BACKGROUND/AIMS: Sympathetic ophthalmia (SO) is a classic example of autoimmune disease where human leucocyte antigen (HLA) genomic associations could provide further understanding of mechanisms of disease. This study sought to assess HLA genetic polymorphism in British and Irish patients with SO, and to assess whether HLA gene variants are associated with clinical phenotype or disease severity. METHODS: High resolution DNA based HLA typing using polymerase chain reaction sequence specific primers was performed in 27 patients with SO and 51 matched healthy controls. Clinical phenotype and markers of disease severity were determined prospectively in 17 newly diagnosed patients and from medical record review and repeat clinical examination in 10 previously diagnosed patients. RESULTS: HLA-Cw*03 (p=0.008), DRB1*04 (p=0.017), and DQA1*03 (p=0.014) were significantly associated with SO. For class II alleles at higher resolution, only HLA-DRB1*0404 (relative risk (RR) = 5.6, p = 0.045) was significantly associated with SO. The highest relative risk for any of the associated haplotypes was with HLA-DRB1*0404-DQA1*0301 (RR=10.9, p=0.019). Patients with the DRB1*04-DQA1*03 associated haplotype were significantly more likely to develop SO earlier, with fewer inciting ocular trauma events, and to require more systemic steroid therapy to control inflammatory activity. CONCLUSIONS: Sympathetic ophthalmia is associated with HLA-DRB1*04 and DQA1*03 genotypes in white patients, similar to Japanese patients. Differences in DRB1*04 gene variant associations (-0404 in Britain and Ireland and -0405 in Japan) may have implications for HLA peptide binding in disease initiation. The DRB1*04-DQA1*03 haplotype is a marker of increased SO susceptibility and severity, as in Vogt-Koyanagi-Harada disease, which also has similar clinicopathological and HLA associations.     

病理性近视与人类白细胞抗原-DQB1基因的关联性研究

目的 研究病理性近视（pathologic myopia,PM)与人类白细胞抗原（human leucocyte antigen HLA)-DQB1 基因的相关性,以探讨PM的发病机制。方法 抽提66例PM患者的基因组DNA,用聚合酶链反应－限制性片段长度多态性方法扩增HLAⅡ类基因DQB1的第2个外显子,扩增产物用HaeⅢ、BasHⅡ,ApaⅠ,BsaHⅠ,HaeⅡ,HpaⅡ,RsaⅠ,Bsp1286Ⅰ特异性的限制性内切酶酶切分型,检测HLA－DQB1的16个等位基因的分布频率,并与正常人进行比较。结果 PM患者HLA－DQB1的＊0301和＊0303等位基因分布频率明显高于正常人（P＜0．0001）;PM患者HLA－DQB1的＊0601和＊0602等位基因分布频率明显低于正常人（P＜0．0001）。结论 HLA－DQB1的＊0301和＊0303等位基因为PM的易感基因,可能与其发病有关;HLA－DQB1的＊0601和＊0602等位基因为抵抗基因,可能具有保护作用。     

人类白细胞抗原—DR4基因亚型与Vogt—Koyanagi—Harada综合征相…

研究人类白细胞抗原－ＤＲ４基因亚型与Ｖｏｇｔ－Ｋｏｙａｗｎａｇｉ－Ｈａｒａｄａ综合征的关系，从免疫遗传学角度阐明ＶＫＨ的易感性及抵抗性。方法 采用聚合酶链反应－限制性片段长度多态性分析方法，检测４例ＶＨＫ综合征患者和１０６例健康对照者的ＥＲ４－ＤＲＢ１基因亚型。结果 ５４例ＶＫＨ综合征患者中，３８例扩增出２６３ｂｐ的特异性ＤＲ－４－ＤＲＢ１基因片段，阳性检出率为７０％，对照组仅为１９％，相对危险度     

HLA-DRB and HLA-DQB loci in the genetic susceptibility to develop glaucoma in Mexicans.

PURPOSE: Glaucoma is a clinically heterogeneous disease with a pathophysiology that may include genetic susceptibility, possibly associated with an immunologic disorder. The aim of this study was to determine whether the DNA polymorphisms located in the HLA-DRB1 and HLA-DQB1 genes show a specific association pattern in Mexican mestizo patients with primary open-angle glaucoma. METHODS: This was a cross-sectional, case-control, multicenter study. We analyzed the HLA-DRB1 and DQB1 loci of 81 Mexican mestizo nonrelated patients with primary open-angle glaucoma and 98 healthy ethnic matched control subjects. Patients were diagnosed clinically and by visual fields examination. HLA typing was performed by PCR-SSO reverse dot blot. RESULTS: We documented increased frequencies of HLA-DRB1*0301, DRB1*1101, DRB1*0701, DRB1*1402, DQB1*0302, and DQB1*0301; however, none of them were significantly different from normal control subjects. Haplotype analysis showed that the HLA-DRB1*0407-DQB1*0302 haplotype is significantly increased in patients compared with control subjects (P = .0001). CONCLUSIONS: The haplotype HLA-DRB1*0407-DQB1*0302 is common among Mexican mestizo (haplotype frequency = 0.102), and it was increased in our patients (haplotype frequency = 0.259, P = .0001). This may reflect an independent association of this haplotype with the disease as the result of linkage disequilibrium or the influence of a neighboring gene. The pathophysiology of this illness is uncertain, and further studies are needed regarding the genetic susceptibility to develop primary open-angle glaucoma.     

Strong associations between specific HLA-DQ and HLA-DR alleles and the tubulointerstitial nephritis and uveitis syndrome

PURPOSE: To identify genetic markers for the tubulointerstitial nephritis and uveitis (TINU) syndrome by using human leukocyte antigen (HLA) genotyping. METHODS: Eighteen patients who had TINU syndrome were evaluated at three institutions. Typing of class I and II genes was performed by using DNA-based techniques. RESULTS: Significant associations were found with HLA-B14 (6/18 patients, 33.3%; control subjects, 5.5%; P = 0.0003; relative risk [RR] = 8.5), HLA-DQA1*01 (17/18 patients, 94.4%; control subjects, 46.6%, P = 0.0001; RR = 19.5), HLA-DQA1*0101 (14/18 patients, 77.8%; control subjects 22.2%; P < 0.0001; RR = 12.2), HLA-DQB1*05 (14/18 patients, 77.8%; control subjects 17.7%; P < 0.0001; RR = 16.3), HLA-DQB1*0501 (13/18 patients, 72.2%; control subjects 12.9%; P < 0.0001; RR = 17.6), HLA-DRB1*01 (14/18 patients, 77.8%; control subjects, 12.1%; P < 0.0001; RR = 25.5), and HLA-DRB1*0102 (13/18 patients, 72.2%; control subjects, 1.6%; P < 0.0001, RR = 167.1). The HLA haplotype most frequently identified in the study patients was HLA-DQA1*01/DQB1*05/DRB1*01 (13/18 patients, 72.2%). CONCLUSIONS: TINU syndrome is strongly associated with HLA-DQA1*01, HLA-DQB1*05, and HLA-DRB1*01. The association with HLA-DRB1*0102 is one of the highest reported for any disease. Because these genes are in linkage disequilibrium, the role of the individual alleles is difficult to assess. Based on the results of the present study and on previously reported HLA associations in patients with TINU syndrome, the alphabeta dimer encoded by HLA-DQA1*01/DQB1*05 may be particularly important in conferring risk for development of this disease.     

HLA-DRB1 typing of Vogt-Koyanagi-Harada's disease by PCR-RFLP and the strong association with DRB1*0405 and DRB1*0410.

Vogt-Koyanagi-Harada''s (VKH) disease is reported to be closely associated with the HLA class II antigen, HLA-DR4. Serologically defined DR4 is further divided into 11 alleles by molecular HLA genotyping. However, no study of HLA-DNA typing of VKH patients has been reported. To clarify molecular genetic mechanism underlying the susceptibility/resistance to VKH disease, HLA-DNA typing of DR antigens (DRB1 genotyping) by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was performed. It was found that DRB1*0405 showed a significant association with VKH disease compared with the healthy controls (corrected p value < 1 x 10(-5)) and that all the patients had DRB1*0405 and/or DRB1*0410. The specific amino acid residue shared only by these two alleles is Ser at position 57 which is located in the antigen binding groove and may influence the immunological function as an antigen-presenting molecule, suggesting that Ser at position 57 plays an important role in the susceptibility to VKH disease, although the possibility that the involvement of the HLA-DQ molecule, DQ4, in strong linkage disequilibrium with DRB1*0405 and DRB1*0410, cannot be excluded.