同轴微切口超声乳化白内障吸出术后角膜生物力学变化

目的　比较同轴微切口超声乳化白内障吸出术（ｐｈａｃｏｅｍｕｌｓｉｆｉｃａｔｉｏｎ,Ｐｈａｃｏ）及标准切口Ｐｈａｃｏ术后角膜生物力学的变化。方法　年龄相关性白内障患者３１２例（３１２眼）随机分成两组。其中研究组（２．２ｍｍ同轴微切口组）１５９例,对照组（３．０ｍｍ标准切口组）１５３例。记录两组术前数据,包括年龄、性别、裸眼视力（ｕｎｃｏｒｒｅｃｔｅｄｖｉｓｕａｌａｃｕｉｔｙ,ＵＣ-ＶＡ）、最佳矫正视力（ｂｅｓｔｃｏｒｒｅｃｔｅｄｖｉｓｕａｌａｃｕｉｔｙ,ＢＣＶＡ）、角膜滞后性（ｃｏｒｎｅａｌｈｙｓｔｅｒｅｔｉｅ,ＣＨ）、角膜阻力因数（ｃｏｒｎｅａｌｒｅｓｉｓｔａｎｃｅｆａｃｔｏｒ,ＣＲＦ）、Ｇｏｌｄｍａｎｎ相关眼压（ｇｏｌｄｍａｎｎｃｏｒｒｅｌａｔｅｄｉｎｔｒａｏｃｕｌａｒｐｒｅｓｓｕｒｅ,ＩＯＰｇ）、角膜补偿眼压（ｃｏｒｎｅａｌｃｏｍｐｅｎｓａｔｅｄＩＯＰ,ＩＯＰｃｃ）、中央角膜厚度（ｃｅｎｔｒａｌｃｏｒｎｅａｌｔｈｉｃｋｎｅｓｓ,ＣＣＴ）、角膜内皮细胞计数（ｅｎｄｏｔｈｅｌｉａｌｃｅｌｌｃｏｕｎｔ,ＥＣＣ）;术中数据包括累积能量复合参数（ｃｕｍｕｌａｔｉｖｅｄｉｓｓｉｐａｔｅｄｅｎｅｒｇｙ,ＣＤＥ）、手术时间。术后１ｄ、１周、２周、１个月复查,比较两组ＵＣＶＡ、ＢＣＶＡ、ＥＣＣ、ＣＣＴ、ＣＨ、ＩＯＰｇ、ＣＲＦ和ＩＯＰｃｃ。结果　术后１ｄ两组ＣＨ、ＣＲＦ均较术前明显降低,差异均有统计学意义（均为Ｐ＜０．０５）。术后１周,研究组ＣＨ、ＣＲＦ与术前差异均无统计学意义（均为Ｐ＞０．０５）;对照组ＣＨ、ＣＲＦ较术前降低,差异均有统计学意义（均为Ｐ＜０．０５）。术后２周,两组ＣＨ、ＣＲＦ均恢复至术前水平（均为Ｐ＞０．０５）;两组ＩＯＰｇ、ＩＯＰｃｃ仍高于术前（均为Ｐ＜０．０５）,而低于术后１周（均为Ｐ＜０．０５）;两组ＣＣＴ恢复至术前水平（均为Ｐ＞０．０５）。术后４周,两组ＣＨ、ＣＲＦ、ＩＯＰｇ、ＩＯＰｃｃ、ＣＣＴ均恢复至术前水平（均为Ｐ＞０．０５）。术前,两组ＣＨ、ＣＲＦ与ＣＣＴ存在正相关性（研究组:ｒ１＝０．４３,ｒ２＝０．５２,对照组:ｒ１＝０．５６,ｒ２＝０．５３;均为Ｐ＜０．０５）。术后１ｄ,两组ＣＨ与ＣＣＴ均无相关性（ｒ１＝０．１３,ｒ２＝０．１０,均为Ｐ＞０．０５）。两组的ＣＲＦ值与ＣＣＴ在不同时相始终存在相关性（均为Ｐ＜０．０５）。两组间不同时相的ＣＨ与ＣＲＦ均存在正相关性（均为Ｐ＜０．０５）。结论　同轴微切口Ｐｈａｃｏ和标准切口Ｐｈａｃｏ均会改变角膜生物力学特征。同轴微切口Ｐｈａｃｏ比标准切口Ｐｈａｃｏ术后角膜生物力学特征恢复更快。     

Effects of mitogen-activated protein kinase signal pathway on heat shock protein 27 expression in human lens epithelial cells exposed to sodium salicylate in vitro

The roles of mitogen-activated protein kinase （MAPK） signal pathway in sodium salieylate-induced expression of heat shock protein 27 （HSP27） in human lens epithelial cells （HLECs-B3） in vitro were investigated. HLECs-B3 were incubated in the fresh media containing sodium salicylate at different concentrations for different durations, and allowed to be recovered in fresh medium without sodium salicylate for different durations with or without pretreatment with p38MAPK inhibitor （SB203580）, ERK1/2 inhibitor （PD98059） and JNK/SAPK inhibitor （SP600125）. The expression of P38MAPK, ERK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 was detected by Western blot. The expression of HSP27 mRNA and protein was detected by RT-PCR and immunohistochemistry respectively. It was found there was only weak expression of HSP27 in normal HLECs. The expression of HSP27 was not detectable in HLECs-B3 that were exposed to sodium salicylate （55 retool/L） for 1-5 h. It was indicated that recovery from sodium salicylate （〉35 mmol/L） significantly increased the synthesis of HSP27. The expression of HSP27 was up-regulated in HLECs-B3 under sodium salicylate recovery for 3 h, reached the peak level for 6 h, and returned to the level of control cells by 24 h. Activation of P38MAPK from sodium salicylate stimulation occurred at 30th rain, and increased significantly at 1st h, then declined and renamed to baseline level at 3rd h under sodium salicylate recovery. Activation of ERK1/2 occurred at 1st h and reached the peak level at 6th h under sodium salicylate recovery. However, JNK/SAPK was inactivated by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059 jointly. It is concluded that sodium salicylate can induce the expression of HSP27 in HLECs-B3. The effects are mediated, at least in part, through the activation of P38MAPK and ERK1/2 signaling pathway.     

Effect of p27Kip1 Inhibition on Proliferation of Bovine Corneal Endothelial Cells by RNA Interference

Three plasmids （pGenesil-P1, pGenesil-P2, pGenesil-P3） with different p27Kip1-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endothelial cells （bCEC） were investigated. Plasmid expressing irrelevant shRNA with a random combination was used as negative control （pGenesil-HK）. The recombination of four plamids was confirmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kip1 mRNA and p27Kip1 protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group （non-transfected group）, pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The proliferation rates of the pGenesil-P3 group, the pGenesil-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was detected by flow cytometry （FCM）. Compared with the control groups, the proliferation rate of the pGenesil-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kip1 could down-regulate the expression of p27Kip1 effectively and increase the proliferation of bCEC. RNA interference （RNAi） may be an effective means to promote the proliferation of CEC.     

水杨酸钠对体外培养的人晶状体上皮细胞中HSP27表达的影响

本实验观察水杨酸钠对H：O：诱导体外培养的人晶状体上皮细胞（lens epithelial cell．LECs）中热休克蛋白27（heat shock protein 27，HSP27）表达的影响．探讨水杨酸钠在延缓白内障发病中的作用机制．