Asthma III

Bronchial Epithelial Cells were isolated processed as described (Chu et al., 2002 and Zhao et al., 2011). The objective of the study was to identify differentially expressed genes between normal control (NC), mild-moderate asmathic (notSA) and severe asthmatic (SA) patients. For demographics data, contact Dr. Sally Wenzel ([e-mail]) Bronchoscopy with endobronchial epithelial brushing was performed as previously described (Chu et al., 2002; Zhao et al., 2011). Bronchial alveolar lavage fluids were spun down on 4000 g for 10 minutes. 0.5- 1X10^6 cells were stored in Trizol for RNA extraction. RNA in Trizol solution was extracted using the QIACube system (Qiagen, Valencia, CA). RNA quality was determined using the Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA), and only samples with an RIN higher than 7 used for microarray experiments.

本研究的支气管上皮细胞（Bronchial Epithelial Cells）分离与处理参照已发表方法（Chu等，2002；Zhao等，2011）完成。本研究旨在筛选正常对照（normal control, NC）、轻中度哮喘患者（mild-moderate asthmatic, notSA）与重度哮喘患者（severe asthmatic, SA）之间的差异表达基因。若需获取人口统计学数据，请联系Sally Wenzel博士（邮箱：[e-mail]）。经支气管镜行支气管上皮刷检的操作参照既往方法（Chu等，2002；Zhao等，2011）进行。支气管肺泡灌洗液以4000 g离心10分钟，收集0.5×10^6至1×10^6个细胞并保存于Trizol试剂中用于后续RNA提取。采用QIACube系统（QIAGEN公司，美国加利福尼亚州瓦伦西亚）提取Trizol试剂中的RNA。使用Agilent Bioanalyzer 2100（安捷伦科技公司，美国加利福尼亚州圣克拉拉）检测RNA质量，仅选取RNA完整性数（RNA Integrity Number, RIN）大于7的样本用于基因芯片实验。