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With the characteristics of numerous constraints, large density and high speed of train, High-speed train operation adjustment problem is a typical large-scale combinatorial optimization problem and also a kind of difficult problem of NP. In combination with the characteristic of China’s high-speed railway, this paper builds the mathematical model of high-speed railway operation adjustment, and the model is solved and optimized with differential evolution algorithm. The result of the experiment shows that the Differential Evolution has very high rationality and feasibility in solving the high-speed train operation adjustment problem.