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As the impacts by climate changes and human activities are intensified, variability may occur in river's annual runoff as well as flood and low water characteristics. In order to understand the characteristics of variability in hydrological series, diagnosis and identification must be conducted specific to the variability of hydrological series, i.e., whether there was variability and where the variability began to occur. In this paper, the mainstream of Yangtze River was taken as the object of study. A model was established to simulate the impounding and operation of upstream cascade reservoirs so as to obtain the runoff of downstream hydrological control stations after the regulation by upstream reservoirs in different level years. The Range of Variability Approach was utilized to analyze the impact of the operation of upstream reservoirs on the variability of downstream. The results indicated that the overall hydrologic alterations of Yichang hydrological station in 2010 level year, 2015 level year and the forward level year were 68.4, 72.5 and 74.3% respectively, belonging to high alteration in all three level years. The runoff series of mainstream hydrological stations presented variability in different degrees, where the runoff series of the four hydrological stations including Xiangjiaba, Gaochang and Wulong belonged to high alteration in the three level years; and the runoff series of Beibei hydrological station in 2010 level year belonged to medium alteration, and high alteration in 2015 level year and the forward level year. The study on the impact of the operation of cascade reservoirs in Upper Yangtze River on hydrological variability of the mainstream had important practical significance on the sustainable utilization of water resources, disaster prevention and mitigation, safe and efficient operation and management of water conservancy projects and stable development of the economic society. [ABSTRACT FROM AUTHOR]