Gene expression of cell diferenciation related factors of respiratory epithelium and its roll in the mucosal pathology of rhino sinusitis

遗传学研究：用差异显示法提取漂浮后胶原蛋白凝胶上出现的几条带。通过北方印迹法测定了25条测序条带中5条在漂浮前后的蛋白质体积表达。蛋白质表达在两个测量期之间没有差异。这种差异的缺乏归因于使用北方印迹的高频率假阳性结果。细胞角蛋白的表达通过免疫组织化学技术使用抗细胞角蛋白8、13和14单时抗体按时间顺序检查培养的细胞，尽管培养的细胞在漂浮之前没有显示阳性发现，该分析显示，在以下模式中漂浮后阳性染色逐渐增加;抗细胞角蛋白8从细胞中层到细胞顶层呈阳性反应，抗细胞角蛋白13仅在细胞中层呈阳性反应，抗细胞角蛋白13仅在细胞中层呈阳性反应。纤毛发生特异性物质的检测：制备了几种单时性抗体，其中RPCS 3a是纤毛发生特异性的最佳候选抗体。根据细胞ELISA，从漂浮后的第3天起，RPCS 3a在培养的细胞中表达，在第10天，RPCS 3a表达最强，并且许多培养的细胞在其顶端部位含有分泌囊泡以及许多中心粒，表明是前纤毛细胞。ELISA结果表明，RPCS 3a对一种岩藻糖基化碳水化合物H 3型具有特异性。RPCS 3a识别的H 3型的表达量似乎反映了纤毛发生的阶段。因此，我们认为H-3是纤毛发生的相对特异性标记，为阐明纤毛发生的机制提供了有用的工具。

Genetic investigation : Several bands that appeared on the collagen gel following floating were extracted by the differential display method. The protein volume expression of 5 of the 25 sequenced bands was determined before and after floating by Northern blotting. Protein expression did not differ between the two measurement period. This lack of difference was attributed to the high frequency of false positive results using Northern blotting.Morphological confirmation of cell differentiation : Expression of cytokeratin the cultured cells was examined chronologically by an immunohistochemical technique using anti-cytokeratin 8, 13 and 14 monochronal antibodies, although the cultured cells showed no positive findings before floating, this analysis showed a gradual increase in positive staining after floating in the following pattern ; anti-cytokeratin 8 was positively stained from the middle to apical layer of the cultured cells and anti-cytokeratin 13 was positively stained only in the middle layer and anti-cytokeratin 14 was positively stained in the basal layer.Detection of ciliogenesis-specific substance : Several monochronal antibodies were produced, of which RPCS3a, the most eligible candidate specific to ciliogenesis, was characterized. RPCS3a was expressed in the cultured cell from the 3rd day after floating according to cellular ELISA.On the 10th day, RPCS3a was expressed most strongly, and many cultured cells contained secretory vesicles at their apical site as well as numerous centrioles indicating to be preciliated cells. ELISA revealed that RPCS3a is specific to the a sort of fucosylated carbohydrate H type 3. The expressed volume of H type 3 recognized by RPCS3a appears to reflect the stage of ciliogenesis. Based on the present data, we consider H type 3 to be a relativity specific markers of ciliogenesis and thus may provide a useful tool to clarify the mechanisms of ciliogenesis.