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Establishment of physiological reference interval (RI) is important for health status evaluation, disease screening and diagnosis, and the effect assessment of clinical therapy. Currently, there is no review on methodology of RI establishment in China. Therefore, by reviewing of literatures on methodology of RI establishment, this article comprehensively described the purpose and significance of RI establishment,its statistical methods of calculation and its application. Three methods are commonly used for RI calculation: the nonparametric, parametric and robust estimate. Researchers may use a specific method based on research purpose and the characteristics of the data. This review provides methodological reference for RI establishment in China and also some guidelines to better understand the meaning of physiological constant RI in health practices.