碱性磷酸酶快速染色法的实验研究

目的 建立一种简便快速中性粒细胞碱性磷酸酶染色方法。方法 在改良 Gomori钙钴法的基础上 ,利用 2 -氨基 - 2 -甲基 - 1 -丙醇为缓冲体系和磷酸基受体 ,加快转磷酸作用的酶促反应速度。结果 快速法的孵育时间从改良 Gomori钙钴法的 4h缩短为 1 5min。整个实验可在 30 min内完成。与改良 Gomori钙钴法比较 ,1 2 0例标本配对试验 ,快速法 NAP积分为 97.7± 8.4,改良 Gomori法 NAP积分为 96.5±1 0 .6(x± s) ,两者结果无显著性差异 (P>0 .0 5)。批内和批间 CV分别为 2 .9%和 5.2 %。结论 该法设计合理 ,具有快速、简便。

中性粒细胞碱性磷酸酶的快速染色法

中性粒细胞的碱性磷酸酶(neutrophil alkalinephosphatase,简称NAP)是一种胞内水解酶。测定其活性对于某些血液病的诊断、鉴别诊断和对疗效的观察有重要价值。目前国内推荐的改良Gomori钙钴法(简称改良法),其孵育时间长达4h,而且试剂配制繁琐,笔者将生物化学中有关磷酸基受体缓冲浪的原理应用于NAP染色,即在改良的基础上,以2-氨基-2-甲基-1-丙醇(AMP)作为缓冲体系,建立了一种简便快速的染色法(简称快速法),孵育时间仅15min,现报告如下:1 材料和方法1.1 主要试剂1.1.1 固定液 10％甲醛甲醇溶液。

EFFECTS OF INTEGRIN ALPHAⅡb^(R995A) MUTATION ON RECEPTOR AFFINITY AND pp125 (FAK) PHOSPHORYLATION

Objective To investigate the role of cytoplasmic domain of integrin alphaⅡb in platelet signal transduction. Methods Binding capacity of integrin alphaⅡb R995A to antibody platelet activation complex-1 (PAC-1) and pp125 focal adhesion kinase (FAK) phosphorylation of cells were detected by flow cytometry, immune precipitation, and Western blotting. Results Without activation, wild-type alphaⅡbbeta3 Chinese hamster ovary (CHO) cells failed to bind to PAC-1, but mutant chimera alphaⅡb R995A beta3 CHO cells were able to bind with PAC-1. Furthermore, phosphorylation of pp125 (FAK) in wild-type alphaⅡbbeta3 CHO cells occured only when cells were adhered to fibrinogen, but could not be detected in bovine serum albumin suspension. However in the mutant chimera group, it could be detected in both conditions. Conclusion The mutation in integrin alphaⅡb R995A alters its affinity state as a receptor, thus also mediating cytoplasmic signal transduction leading to the phosphorylation of pp125 (FAK) without ligand binding.