目的：本研究通过观察经地塞米松处理前后培养的人小梁细胞的变化,探讨激素性青光眼房水排出阻力增加的机制。方法：将人类小梁细胞培养至接近体内的高分化状态，用地塞米松处理后,观察细胞形态的变化，并检测以下蛋白的分布和表达：(1)myocilin/TIGR蛋白；(2)纤连蛋白；(3)肌动蛋白交联网的形成；(4)血清淀粉样物质A蛋白。结果：人小梁细胞经地塞米松处理后(1)胞体变大，排列不规则，边界模糊，呈“融合”状；(2)细胞内外myocilin/TIGR蛋白表达均明显增加，其胞外表达与纤连蛋白位置一致,提示相互作用；(3)纤连蛋白表达增加；(4)CLANs形成，细胞间连接增强；(5)血清淀粉样物质A蛋白表达增加。结论：人类小梁细胞经地塞米松处理后形态发生变化，可能与应力纤维变化及myocilin蛋白在细胞内积聚有关。细胞间边界不清，与细胞外基质蛋白过度表达、沉积有关。小梁细胞外基质沉积以及异常的细胞间连接等改变与房水排出阻力增加的病理过程有关。
Objective To study the expression of the fibronectin (FN) and beta;1 integrin (beta;1) in epiretinal membranes(ERM) of eyes with proliferative vitreoretinopathy(PVR). Methods wenty epiretinal membranes were obtained from eyes undergone vitrectomy for retinal detachment complicated with PVR and observed by immunohistochemical methods. Results Overexpression of FN and beta;1 were observed in 18 and 16 membranes respectively． Conclusion The synergism of FN and beta;1 in their action mignt be one of the important roles in the development of PVR. (Chin J Ocul Fundus Dis, 2001,17:119-121)
ObjectiveTo construct a prokaryotic expression strain of Staphylococcus aureus fibronectin binding protein A (FnBPA) r10-11 truncated fusion protein, and explore the immunogenicity of FnBPAr10-11. MethodsPloymerase chain reaction (PCR) amplification was carried out from the whole genome sequence of Staphylococcus aureus Newman strain by recombinant PCR technique. The amplified product was purified and transformed into Escherichia coli DH5α for cloning. The recombinant plasmid was extracted and identified by double enzyme digestion. The recovered fragment was ligated into the pET-32a plasmid and transformed into Escherichia coli BL21 (DE3) for prokaryotic expression. The FnBPAr10-11 was purified by HIS protein purification column, identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and used to immunize mice, and the mice were divided into phosphate buffered saline (PBS) group, FnBPA group, and FnBPAr10-11 group. The serum levels of immunoglobulin G (IgG) and cytokines, and the immune protection rate of the mice were detected. ResultsSDS-PAGE result showed that the relative molecular mass of the protein was about 33.1×103. The titers of IgG antibody in FnBPAr10-11 group and FnBPA group reached 1∶128 000, and were significantly different compared with PBS group (P<0.05). The cytokine level in FnBPAr10-11 group was not significantly different compared with that in FnBPA group, and they were extremely significant (P<0.01) compared with that in PBS group. The immuno-protective effect of the FnBPAr10-11 group was over 50%. ConclusionsThe prokaryotic expression strain of Staphylococcus aureu FnBPAr10-11 truncated fusion protein was successfully constructed. The truncated protein has good immunogenicity.