青光眼滤过性手术中联合应用抗代谢药物后低眼压性黄斑病变

青光眼滤过性手术中联合应用抗代谢药物后较常见且严重的共发症之一为低眼压性黄斑病变（HM），可导致视功能损害。其发生率与抗代谢药物的浓度、时间、患者年龄和屈光状态有关；抗代谢药物影响滤过泡组织结构致滤过功能过强，加之药物对睫状体的毒性作用使房水生成减少，而发生HM；目前对HM的治疗多在矫正低眼压，常用的方法有滤过泡烧灼法、滤过泡注射自家血疗法、激光、冷冻及手术治疗等。     

PRK后房水中微量元素的变化

目的通过测定准分子角膜切削(PRK)术后1、2、3月兔眼房水中铜、锌、钙、镁、铁含量的变化,探讨其在PRK术后角膜创伤愈合中的作用.方法健康新西兰大白兔12只(24眼)平均分成4组,每组3只(6眼),1组对照,另外3组行PRK术,切削深度为100μm,消融直径5.6mm.分别于术后1、2、3月抽取房水,测定铜、锌、钙、镁、铁含量.结果PRK术后1个月时,钙和铁的含量降低,铁持续到术后3月时仍较低.术后1月锌的含量升高而钙的含量降低.镁及铜的含量没有变化.结论锌的含量增加有利于创伤修复;铁含量减少提示铁参与角膜创伤修复过程且在角膜内再分布;钙在房水中降低体现了角膜能量代谢及能量利用加强.     

关于眼科学教学团队建设的思考

建设高绩效的眼科教学团队,对于眼科教师的专业发展和人才培养质量的提高具有十分重要的作用。文章就眼科教学团队建设现状及存在的问题,眼科教学团队建设的意义及建设策略进行了思考。     

对“光明的偷盗者”说不

青光眼是世界范围内第一位不可逆性致盲性眼病。由于大多数青光眼具有隐匿性,症状不明显,往往不易被患者察觉,等到发现时,视功能已经严重损害,甚至失明,因此青光眼被喻为“光明的偷盗者”。人们往往认为只有老年人才会患青光眼,其实青光眼并非老年人的专利,所有年龄段人群（从出生到老年）都可以患青光眼。通常40岁以上的人比较容易患青光眼,女性较男性常见。     

色素上皮衍生因子对糖尿病大鼠视网膜谷氨酰胺合成酶表达的影响

目的　观察色素上皮衍生因子（PEDF）对糖尿病大鼠视网膜Müller细胞谷氨酰胺合成酶（GS）表达的影响。方法　将Sprague-Dawley大鼠分为模型组、模型对照组、PEDF干预组（干预组）、干预对照组,每组均为8只大鼠。模型组、干预组、干预对照组大鼠链脲佐菌素诱导糖尿病大鼠模型。模型组大鼠不作任何干预,模型对照组为相同月龄的正常大鼠,干预组大鼠左眼玻璃体腔注射0.1 μg/μl的PEDF 10 μl,干预对照组大鼠左眼玻璃体腔注射相同容积的磷酸盐缓冲液。采用免疫组织化学法和实时荧光聚合酶链反应（PCR）法检测视网膜GS和白细胞介素-1β（IL-1β）的表达变化。将视网膜Müller细胞置于高糖环境下培养,实验干预组中加入100 ng/ml PEDF,空白对照组加入相同容积的培养液,24 h后通过蛋白质免疫印迹（Western blot）法和实时荧光PCR法检测PEDF对Müller细胞GS和IL-1β表达的改变。流式细胞仪锚定蛋白-异硫氰酸荧光素和碘化丙啶（Annexin V-FITC-PI）双染色法检测100ng/mlPEDF对高糖状态下Müller细胞凋亡的影响。结果　实时荧光PCR法从基因水平和免疫组织化学法从蛋白质水平检测均显示,相对于模型对照组大鼠,模型组大鼠视网膜GS表达降低,而IL-1β的表达升高,实时荧光PCR法：GS: t=4.23, P＜0.01;IL-1β: t=16.73,P＜0.01;免疫组织化学法：GS:t=5.13,P＜0.01;IL-1β: t=9.32, P＜0.01;干预组大鼠玻璃体腔注射PEDF 48 h后,IL-1β的表达下降,GS的表达升高,与干预对照组比较,实时荧光PCR法：GS: t=3.87,P＜0.01;IL-1β: t=3.61,P＜0.05;免疫组织化学法：GS:t=3.32, P＜0.05;IL-1β: t=2.63, P＜0.05。在高糖环境下,通过实时荧光PCR法和Western bot 法检测均显示PEDF可以下调IL-1β的表达,而上调GS的表达,与空白对照组比较,实时荧光PCR法：GS: t=2.89, P＜0.05;IL-1β: t=3.37, P＜0.05;Western blot：GS: t=2.66, P＜0.05;IL-1β: t=3.23, P＜0.05。流式细胞仪检测结果显示,PEDF可以抑制高糖环境下Müller细胞的凋亡,实验组凋亡率与空白对照组凋亡率比较,差异有统计学意义(t=3.21,P＜0.05)。结论　对于糖尿病大鼠,PEDF可能通过下调视网膜Müller细胞中IL-1β的表达来上调GS的表达,从而改善谷氨酸循环,抑制神经节细胞的死亡      

色素上皮衍生因子对糖尿病大鼠视网膜谷氨酸代谢的影响

目的 观察色素上皮衍生因子(PEDF)对糖尿病大鼠视网膜谷氨酸代谢的影响.方法 Sprague_Dawley大鼠78只,分为模型组、模型对照组、PEDF干预组(干预组)、干预对照组,实验结束时去除血糖恢复鼠和实验期间死亡鼠,每组以12只大鼠作为统计样本.模型组、干预组、干预对照组大鼠采用链脲佐菌素诱导糖尿病大鼠模型.模型组大鼠不作任何干预,模型对照组为相同月龄的正常大鼠.干预组大鼠左眼玻璃体腔注射0.1 μg/μl的PEDF 5.0μl,干预对照组大鼠左眼玻璃体腔注射相同容积的磷酸盐缓冲液.采用蛋白免疫印迹法(Western bolt)和实时荧光聚合酶链式反应(PCR)法检测视网膜L-谷氨酸-L-门冬氨酸转运体(GLAST)表达的变化,高压液相色谱法(HPLC)观察视网膜谷氨酸的含量变化.将体外培养的大鼠视网膜Müller细胞随机分为对照组、实验组、PEDF干预组(干预组)和干预对照组,荧光免疫法和实时荧光PCR法检测Müller细胞GLAST表达的改变,根据[3H]标记的D,L-谷氨酸摄入量判断Müller细胞的摄取功能.结果 实时荧光PCR法和Western bolt检测结果显示,相对于模型对照组大鼠,模型组大鼠视网膜GLAST表达降低(实时荧光PCR法:t=8.86,P＜0.01;Western blot:t=3.42,P＜0.05),视网膜谷氦酸含量升高(t=4.01,P＜0.05);干预组大鼠视网膜GLAST表达与干预对照组视网膜GLAST表达比较,干预组大鼠视网膜GLAST的表达升高(实时荧光PCR法:t=3.56,P＜0.05;Western blot:t=3.52,P＜O.05);视网膜谷氨酸含量下调(t=4.36,P＜0.05).实时荧光PCR法和荧光免疫法检测结果显示,高糖可以降低视网膜Müller细胞GLAST的表达(实时荧光PCR法:t=3.48,P＜O.05;荧光免疫法:t=4.72,P＜O.05);[3H]标记的D,L-谷氨酸摄人量结果显示,高糖可以下调视网膜Mü1ler细胞GIAST的功能(t=3.81,P＜0.05);经PEDF处理后,可以明显改善高糖状态下视网膜Müller细胞GLAST的表达(实时荧光PCR法:t=6.82,P＜O.01;荧光免疫法:t=3.72,P＜0.05)和对谷氨酸的摄取功能(t=4.14,P＜0.05).结论 PEDF可通过改善糖尿病大鼠视网膜Müller细胞中GLAST功能从而改善谷氨酸循环,抑制神经节细胞的死亡.

Abstract：

Objective To observe the effect of pigment epithelium-derived factor(PEDF)on glutamate metabolism in diabetic rat retina.Methods 78 Sprague-Dawley rats were randomly divided into the model group,model control group,PEDF intervention group and intervention control group.There were some dead and euglycemia rats at the end of experiment,so only 12 rats in each group were included in the statistical analysis.The diabetic retinopathy rat model of the model,PEDF intervention and intervention control group were induced with streptozotocin injection.The rats in the model group were not intervened.The monthly-age matched normal rats of model group were in the model control group.The left eyes of rats were received intravitreal injection with 5μl(0.1μg/μl)PEDF(PEDF intervention group)or 5μlphosphate buffer solution(intervention control group).The expressions of L-glutamate/L-aspartate transporter in retina were analyzed by western blot and real time RT-PCR techniques and glutamate content in retina was analyzed by high-pressure liquid chromatography(HPLC).Cultured rat Mailer cells were divided into the control,experimental,PEDF intervention and intervention control group,GLAST expressions were detected by fluorescence immunofluorescence and real-time RT-PCR techniques.The glutamate up-take activity of Müller cells was determined by intracellular[3H] labeled D,L-glutamate concentration with scintillation counting.Results Western blot and real-time RT-PCR showed that GLAST expression decreased(real-time RT-PCR:t=8.86,P＜0.01;Western blot:t=3.42,P＜0.05).glutamate content increased(t=4.01,P＜0.05)in model group compared with the model control group:GLAST expression increased(real-time RT-PCR:t=3.56,P＜0.05;Western blot:t=3.52,P＜0.05),glutamate content decreased(y=4.36,P＜0.05)in the PEDF intervention group compared with the intervention control group.Real-time RT-PCR and fluorescence immunofluorescenee showed that high glucose down-regulate GLAST expressions in Müller cells(real-time RT-PCR:t=3.48,P＜0.05;fluorescence immunofluoreseence:t=4.72,P＜0.05)and impair glutamate up-take activity of Müller cells(t=3.81,P＜0.05).Under high glucose conditions,PEDF up-regulated GLAST expression significantly(real-time RT-PCR:t=6.82,P＜0.01;fluorescence immunofluorescence:t=3.72,P＜0.05)and ameliorated the glutamate up-take activitv of Mailer cells(t=4.14,P＜0.05).Conclusions In diabetic rats,PEDF may improve the activitv of GLAST in Müller cells,thus ameliorate retinal glutamate metabolism and inhibit death of retinal ganglion cells.     

色素上皮衍生因子对糖尿病大鼠视网膜谷氨酸代谢的影响

Objective To observe the effect of pigment epithelium-derived factor(PEDF)on glutamate metabolism in diabetic rat retina.Methods 78 Sprague-Dawley rats were randomly divided into the model group,model control group,PEDF intervention group and intervention control group.There were some dead and euglycemia rats at the end of experiment,so only 12 rats in each group were included in the statistical analysis.The diabetic retinopathy rat model of the model,PEDF intervention and intervention control group were induced with streptozotocin injection.The rats in the model group were not intervened.The monthly-age matched normal rats of model group were in the model control group.The left eyes of rats were received intravitreal injection with 5μl(0.1μg/μl)PEDF(PEDF intervention group)or 5μlphosphate buffer solution(intervention control group).The expressions of L-glutamate/L-aspartate transporter in retina were analyzed by western blot and real time RT-PCR techniques and glutamate content in retina was analyzed by high-pressure liquid chromatography(HPLC).Cultured rat Mailer cells were divided into the control,experimental,PEDF intervention and intervention control group,GLAST expressions were detected by fluorescence immunofluorescence and real-time RT-PCR techniques.The glutamate up-take activity of Müller cells was determined by intracellular[3H] labeled D,L-glutamate concentration with scintillation counting.Results Western blot and real-time RT-PCR showed that GLAST expression decreased(real-time RT-PCR:t=8.86,P＜0.01;Western blot:t=3.42,P＜0.05).glutamate content increased(t=4.01,P＜0.05)in model group compared with the model control group:GLAST expression increased(real-time RT-PCR:t=3.56,P＜0.05;Western blot:t=3.52,P＜0.05),glutamate content decreased(y=4.36,P＜0.05)in the PEDF intervention group compared with the intervention control group.Real-time RT-PCR and fluorescence immunofluorescenee showed that high glucose down-regulate GLAST expressions in Müller cells(real-time RT-PCR:t=3.48,P＜0.05;fluorescence immunofluoreseence:t=4.72,P＜0.05)and impair glutamate up-take activity of Müller cells(t=3.81,P＜0.05).Under high glucose conditions,PEDF up-regulated GLAST expression significantly(real-time RT-PCR:t=6.82,P＜0.01;fluorescence immunofluorescence:t=3.72,P＜0.05)and ameliorated the glutamate up-take activitv of Mailer cells(t=4.14,P＜0.05).Conclusions In diabetic rats,PEDF may improve the activitv of GLAST in Müller cells,thus ameliorate retinal glutamate metabolism and inhibit death of retinal ganglion cells.     

灯盏细辛对眼压已控制青光眼患者视野的保护作用

-目的:探讨灯盏细辛对眼压已控制青光眼患者视野的保护作用。方法:选择有青光眼视野缺损,眼压控制在18mmHg以内的原发性青光眼患者24例40眼。按随机、双盲法予药物口服,药物分别为灯盏细辛片和安慰剂。患者每日口服3次,每次2片。2mo为一疗程,连续3个疗程,每2mo随访1次。试验结束由药物提供方拆盲并反馈信息。结果:①用药前后各疗程对照组和治疗组收缩压、舒张压、脉搏、眼压、C/D、视力均无显著性统计学差异(P>0.05),且所有患者用药过程中无明显不良反应。②治疗组用药6mo后的平均缺损(MD)、平均敏感度(MS)与用药前的MD、MS相比,差异有显著性意义(P<0.05)。③中晚期治疗组用药2,4,6mo后的MD、MS分别与用药前MD、MS相比差异均有显著性意义(P<0.05)。结论:降低眼压对部分青光眼患者的视功能有保护作用。灯盏细辛对原发性青光眼患者的视功能有一定的保护作用,且应用疗程越长,视野缺损改善越明显;而对于原发性中晚期青光眼患者,灯盏细辛改善视野更显著。灯盏细辛对血压、脉搏、眼压、视力、C/D比均没有影响。     

促红细胞生成素对谷氨酸作用的视网膜神经细胞的保护作用

目的评价促红细胞生成素(Epo)对视网膜神经细胞的生存影响。方法原代新生大鼠视网膜神经细胞体外混合培养;免疫细胞化学法检测培养细胞Epo和EpoR的表达;经与不同浓度Epo共同培养,用MTT比色法检测细胞存活,评价Epo对细胞存活的影响。经不同浓度Epo预培养12 h,再用谷氨酸刺激,MTT法检测细胞存活,观察Epo是否能抵抗谷氨酸兴奋毒性。结果培养的神经样细胞表达Epo和EpoR;Epo不影响正常培养神经细胞的存活,但是能提高谷氨酸刺激后的神经细胞存活。结论Epo能部分抵抗谷氨酸对混合培养视网膜神经细胞的兴奋毒性。