Effect of senescence marker protein 30 on the proliferation and apoptosis of human lens epithelial cells SRA01/04

AIM： To study the effect of senescence marker protein 30（SMP30） on the proliferation and apoptosis of human lens epithelial cell（HLEC） SRA01/04.METHODS： SMP30 overexpression（OE） and knock down（KD） type cell lines were cultivated by using two groups regucalcin（RGN; SMP30） lentiviral vectors（LVRGN, LV-RGN-RNAi） and the respective negative control virus infect SRA01/04 cells. Western blot and real-time quantitative polymerase chain reaction（q-PCR） analysis were used to determine RGN overexpression and knock down efficiency. We use cell counting kit-8（CCK8） assay to measure cell viability and 5-bromodeoxyuridine（Brd U） assay to test cell proliferation. Cell cycle was measured by PI FACS assay and cell apoptosis was tested by Annexin V-APC assay through flow cytometry. We use Western blot to measure the content of caspase-3 in SRA01/04.RESULTS： We used PCR and Western blot techniques to determine the successful transfection of SMP30 OE and KD SRA01/04 cell lines. By CCK8, Brdu and PI FACS cell cycle assay, it was found that the SMP30 OE group promoted cell proliferation（P〈0.05） compared with the control group, and the KD group inhibited cell proliferation（P〈0.05）. The results of Annexin V-APC signal staining detection indicated that compared with respective control group, the cell apoptosis rate was higher in KD group（P〈0.05） but lower in OE group（P〈0.01）. The expression of caspase-3 was down-regulated in OE group through Western blot assay and up-regulated in KD group compared with respective control group. CONCLUSION： Proliferation of SRA01/04 was promoted by SMP30 OE and apoptosis was suppressed. Increasing the expression of SMP30 may protect HLEC SRA01/04 against apoptosis in cataract.

EDIL3 depletion suppress epithelial-mesenchymal transition of lens epithelial cells via transforming growth factor β pathway

AIM： To study the effect of discoidin I-like domaincontaining protein 3（EDIL3） depletion on the proliferation and epithelial-mesenchymal transition（EMT） in human lens epithelial cells（LECs）. METHODS： RNA interference was used to inhibit the expression of EDIL3 in human LECs in vitro. The morphology of cells was observed using an inverted microscope. Cell proliferation was assessed using Ed U kit. Cell migration was investigated using Transwell chamber and EMT of LECs was assessed using confocal microscope and Western blotting. The transforming growth factor β（TGFβ） pathway was investigated using Western blotting. RESULTS： The data showed that silencing EDIL3 expression changed LECs morphology and suppressed LECs proliferation（P〈0.05） and migration（P〈0.01）. Furthermore, the result of Western blotting showed that EDIL3 depletion reduced the expression of α-smooth muscle actin（α-SMA）（P〈0.001） and vimentin（P〈0.01）, while increased the expression of E-cadherin（P〈0.001）. EDIL3 depletion could suppress the phosphorylation of Smad2（P〈0.01） and Smad3（P〈0.01） and the activation of exracellular signal regulated kinase（ERK）（P〈0.05）. CONCLUSION： The findings indicate that EDIL3 might participate in the proliferation and EMT in LECs via TGFβ pathway and may be a potential therapeutic target for the treatment of posterior capsule opacification.

免疫印迹法检测老年白内障晶状体热休克蛋白27

目的探讨热休克蛋白27(H eat Shock P rotein-27,HSP-27)在老年性白内障晶状体中的表达及意义。方法采用免疫印迹法(W estern B lotting)检测17例老年性白内障和7例正常人水溶性(W SF)、水不溶性(W IF)及SDS溶解的晶状体中的HSP-27含量。结果HSP-27在老年性白内障晶状体中的表达低于健康人透明晶状体,老年性白内障皮质型、核型、后囊下型HSP-27表达各不相同。结论HSP-27可能与老年性白内障的形成、发展具有重要关系;HSP-27在不同类型的白内障中所起的作用可能不完全相同。

激光拉曼光谱在白内障研究中的应用

激光拉曼光谱技术通过对物质的分子振动-转光谱的检测来识别和区分不同的物质结构,从而可以在分子水平上了解晶状体蛋白的构成及其变化规律。国内外学者利用这一技术检测了多种白内障晶状体的拉曼光谱,通过光谱中氨基酸侧链、巯基、二硫键和酰胺键的谱峰强度和频移变化行为了解白内障形成过程,为防治白内障提供新思路和理论依据。

Inhibitory Effects of Tetandrine on Experimental Anterior Uveitis Rabbits

Anterior uveitis was induced by injection of lens protein into anterior chamber of rabbit eye.Tetrandrine (Tet) (50mg/kg ip) showed marked inhibition on this ocular inflammation, the total content of prostaglandin E in irises, and leukocyte chemotaxis.The pathological examination revealed that inflammation of irises in Tet group was lighter than that in control group. The antiinflammatory mechanism of Tet may be related to reduction of intracellular free calcium and inhibition of synthesis of inflmmatory factor.

评价接触镜除蛋白护理液去蛋白的试验方法

利用人工泪液浸泡镜片．模拟蛋白吸附．采用紫外分光光度计在波长280 nm处测试经护理液洗脱的蛋白质的吸光度, 计算蛋白质含量。测试除蛋白护理液对软性角膜接触镜上的蛋白质沉淀的洗脱效果。结果除蛋白含量的测试结果具有重复性。该试验方法可以对除蛋白护理液的洗脱率效果进行评价,但尚需完善．

软性角膜接触镜片上蛋白质沉淀洗脱的动态观察

目的：研究市售的软镜多功能护理系统对镜片上蛋白质沉淀的清洗效果与动力学。方法：使用58％含水的，EtafilconA材料的角膜接触镜，进行体外人工泪液浸泡和在体戴用实验，AB两种多功能护理波作为洗脱液，用紫外分光光度法测定镜片洗脱液在280nm处的吸光度值，换算为蛋白质量后，进行比较分析。结果：A、B二种洗脱液的实验结果均表明，蛋白质沉淀的洗脱量呈动态变化。体外实验中，镜片浸泡6－8小时已接近最大洗脱量。戴用镜片的数据提示，浸泡1周时，尚未达到最大洗脱量。结论：体外实验的数据表明，存在一个最大洗脱时间点，不同的洗脱法，该时间点不同，不同的多功能护理液在角膜接触镜蛋白沉淀的清除效力上存在差异环使用除蛋白酶片条件下，护理液仅能洗脱掉蛋白质沉淀的一部分。

实验性近视眼视网膜多巴胺转运蛋白研究

目的 研究鸡眼视网膜多巴胺转运蛋白 (dopaminetransporter,DAT)的分布及其在实验性近视眼形成中的变化和可能作用。方法 孵化第 2d的 30只雏鸡随机分为Ⅰ～Ⅳ组 ,Ⅰ、Ⅱ及Ⅲ组小鸡单眼分别戴 - 10 0 0D、- 2 0 0 0D透镜及半透明眼罩 ,对侧眼为正常对照眼 ;Ⅳ组为正常对照组。3周后检测各组小鸡双眼屈光状态及眼轴长度。各组小鸡于一侧大腿肌注射1 2 5碘 2 β 甲氧羰基 3β (4 碘苯基 )托烷 ,2h后将其处死 ,摘出双眼球并解剖出全层视网膜组织 (Ⅰ和Ⅳ组 )或将视网膜神经上皮层与视网膜色素上皮层 (retinalpigmentepithelium ,RPE)分别取下 (Ⅱ和Ⅲ组 ) ,应用γ 计数器检测各组标本的DAT放射性计数 ,并进行比较。结果 Ⅱ组小鸡双眼的RPE组织标本的DAT放射性计数与视网膜神经上皮层比较 ,差异均有非常显著意义 (t=9 173,4 2 95 ;P <0 0 1) ;Ⅲ组小鸡双眼的RPE组织标本的DAT放射性计数与视网膜神经上皮层比较 ,差异均有非常显著意义 (t=13 84 8,9 6 74 ;P <0 0 1)。Ⅰ、Ⅱ及Ⅲ组全层视网膜组织标本或RPE组织标本的DAT放射性计数分别与对侧正常对照眼比较 ,差异均有非常显著意义 (t=5 5 85 ,4 92 3,5 791;P <0 0 1)。结论 鸡眼视网膜DAT主要分布在RPE。视网膜DAT可能参与小鸡形觉剥夺性近视?

扫描电镜法评价护理液对软性接触镜上蛋白的清除效果

目的：通过扫描电镜观察并结合图像计算机统计分析法评价护理液对软性接触镜上吸附蛋白的清除效果。方法：实验研究。将软性接触镜镜片浸泡在人工泪液中65h，模拟蛋白吸附，再分别用隐形眼镜护理液和PBS对镜片进行揉搓、冲洗和浸泡处理（6h）操作；然后镜片经戊二醛水溶液固定后干燥，通过扫描电镜观察镜片表面的蛋白形态及数量，最后通过图像计算机统计分析法对镜片表面的蛋白数量进行定量分析，吸附蛋白的软性接触镜经护理液处理前后蛋白数量减少值占镜片处理前表面蛋白的百分比，即为清除率。采用Tukey多重比较法对不同护理液处理的镜片表面蛋白清除数据进行单因素方差分析。结果：隐形眼镜护理液对镜片表面吸附的蛋白有显著清除效果，100倍放大倍数下，PBS对镜片表面蛋白的清除率为89．58％，2个不同厂家的隐形眼镜护理液对镜片表面蛋白的清除率分别为98．18％和98．38％；1500倍放大倍数下，PBS对镜片表面蛋白的清除率为89．76％，2个不同厂家的隐形眼镜护理液对镜片表面蛋白的清除率分别为98．42％和98．27％。结论：扫描电镜观察并结合图像计算机统计分析法可对隐形眼镜护理液的蛋白清除效果进行评价。隐形眼镜护理液对镜片表面吸附的蛋白有显著清除效果。