寒、热、虚、实是中医最基本的证候类型，是辨证论治所必须的诊断要素。证候的生物学基础是中医证候分类研究的关键科学问题。我们前期研究通过基因组、代谢组及生物信息学方法，分别找到了类风湿关节炎（RA）寒、热证的关键分子、生物学通路及生物网络基础。然而RA虚、实证分类的生物学基础尚不清楚。本研究拟通过蛋白质双向凝胶电泳（2-DE）和质谱（MS）分析技术，分别鉴定临床典型RA虚证、实证患者与健康人外周血的差异蛋白质；借助生物信息学方法，分别构建RA虚、实证候生物网络，分析网络生物学特征；并采用分子生物学技术Western-blot蛋白印记实验对网络关键蛋白质、生物学通路及调控模块实验验证，得到验证的关键蛋白质、生物通路及调控模块即为RA虚、实证的生物基础。本研究旨在揭示RA虚、实证分类的生物学基础的同时，结合前期研究结果，归纳RA寒、热、虚、实证候分类的生物学特征及关联关系，为中医证候分类研究提供实验依据及方法学参考。

Cold, hot, deficiency, and excess patterns are the most basic pattern types in Traditional Chinese Medicine (TCM), which are the necessary diagnosis elements in process of diagnosis and treatment in TCM. The biological basis of TCM patterns is the key point in research of TCM patterns. In our previous work, we discovered the characteristic molecular basis of cold and heat patterns of RA by microarray and metabonomics experiments, including the key molecules, biological pathways and networks. However, the biological basis of deficiency and excess patterns of RA was unknown. Therefore, we are going to collect blood samples from clinical RA patients with classic deficiency and excess patterns, using two-dimensional gel electrophoresis of protein (2-DE) and mass spectrometry (MS) methods to explore the differentially expressed proteins in blood samples, using bioinformatics techniques to build and analysis the molecular networks, in order to find out key proteins, pathways and regulators in deficiency and excess patterns patients compared with the healthy respectively. Then the Western-blot experiments are going to be used to verify the key proteins, pathways and regulators in molecular networks of deficiency and excess patterns patients, which may be considered as the potential protein biomarkers in TCM deficiency and excess pattern of RA patients. The purpose of this study is to explore the biological basis of TCM deficiency and excess pattern of RA patients, and compare the biological characters of cold, hot, deficiency, and excess patterns of RA further, in order to provide the experiment evidences and research methods on classification study of TCM patterns.

寒、热、虚、实是中医最基本的证候类型，是辨证论治所必须的诊断要素。证候的生物学基础是中医证候分类研究的关键科学问题。我们前期研究通过基因组、代谢组及生物信息学方法，分别找到了类风湿关节炎（RA）寒、热证的关键分子、生物学通路及生物网络基础。然而RA虚、实证分类的生物学基础尚不清楚。本研究通过蛋白质双向凝胶电泳（2-DE）和质谱（MS）分析技术，分别鉴定临床典型RA虚证、实证患者与健康人外周血的差异蛋白质，得到了虚证差异蛋白：ST8SIA2蛋白等蛋白质，实证差异蛋白：AMBP蛋白前体等蛋白质；借助生物信息学方法，分别构建RA虚、实证候生物网络，分析网络生物学特征，得到了虚证特有的儿茶酚胺生物合成信号通路等7个信号通路和实证特有的凝血系统信号通路等8个信号通路；并采用分子生物学技术Western-blot蛋白印记实验对网络关键蛋白质、生物学通路及调控模块实验进行验证，为中医证候分类研究提供了实验依据及方法学参考。