目的探讨血清新型循环免疫炎症相关蛋白复合物(IIRPCs)水平在炎症性肠病(inflammatory bowel disease, IBD)评估中的价值。方法纳入2012年11月至2016年5月于北京协和医院门诊或住院部就诊的69例UC患者和67例CD患者,收集其完整的临床资料。收集血清,通过Native-PAGE对IIRPCs进行分离,后进行定量分析。结果对于UC患者,活动期较缓解期白蛋白水平下降,血小板、血沉和hsCRP水平升高,差异均有统计学意义( P 均<0.05)。活动期IIRPCs水平较缓解期升高,其中中度活动组b型的b2、b4、b5水平与缓解期相比,差异有统计学意义( P <0.05),重度活动组a型的a3及b型的b2、b5水平与缓解期相比,差异有统计学意义( P <0.05),b2、b5对UC疾病活动度评估的ROC曲线下面积分别为0.826、0.859。对于CD患者,活动期血红蛋白、白蛋白水平下降,血小板、血沉水平升高,差异有统计学意义( P 均<0.05),活动期IIRPCs水平较缓解期呈升高趋势,但差异无统计学意义( P >0.05)。UC患者中,IIRPCs与ESR、hsCRP呈正相关。结论 IIRPCs与IBD疾病活动度相关,活动期IBD患者血清IIRPCs水平较缓解期升高,且IIRPCs含量与传统炎症指标ESR、hsCRP呈正相关。
Objective To screen the transient and stable cell lines with high production of Nectin-like 4(Necl-4)*** First,c DNA sequences encoding the extracellular domain of Necls were cloned into the modified vector p APtag at...
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Objective To screen the transient and stable cell lines with high production of Nectin-like 4(Necl-4)*** First,c DNA sequences encoding the extracellular domain of Necls were cloned into the modified vector p APtag at the N terminus of alkaline phosphatase(AP)for fusion ***,293ET cells stably expressed Necls-AP fusion protein and secreted it into the culture medium which were detected by the AP activity assay and Western blot ***,by adding N-glycosylation processing inhibitor kifunensine into the medium,complex glycan was inhibited to *** residual glycan of purified protein was removed by endoglycosidase ***,AP protein was removed by using human rhinovirus protease and size exclusion *** concentration of purified Necl-4 protein was monitored by measuring the absorbance at280 nm and analyzed by *** The transient and stable cell lines with high production of Necl-4 protein were screened by the color reaction with the AP-tag in the recombinant *** soluble and active form of purified Necl-4 protein was obtained after deglycosylation of native N-glycan protein with an expression level of 4 mg/L culture and purity of 95%.Conclusions By using modified AP mammalian protein expression system,we can easily screen the high productive stable cell lines by using AP activity *** adding mannosidase inhibitor kifunensine into the medium and cutting purified protein by using endoglycosidase H,we can obtain deglycosylated Necl-4 protein in milligram *** method might throw a light on the expression and purification of glycoprotein for structural and functional studies.
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