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As an important means of transportation, urban rail transit provides effective mobility, sufficient punctuality, strong security, and environment-friendliness in large cities. However, this transportation mode cannot offer a 24-h service to passengers with the consideration of operation cost and the necessity of maintenance, that is, a final time should be set. Therefore, operators need to design a last train timetable in consideration of the number of successful travel passengers and the total passenger transfer waiting time. This paper proposes a bi-level last train timetable optimization model. Its upper level model aims to maximize the number of passengers who travel by the last train service successful and minimize their transfer waiting time, and its lower level model aims to determine passenger route choice considering the detour routing strategy based on the last train timetable. A genetic algorithm is proposed to solve the upper level model, and the lower level model is solved by a semi-assignment algorithm. The implementation of the proposed model in the Beijing urban rail transit network proves that the model can optimize not only the number of successful transfer directions and successful travel passengers but also the passenger transfer waiting time of successful transfer directions. The optimization results can provide operators detailed information about the stations inaccessible to passengers from all origin stations and uncommon path guides for passengers of all origin–destination pairs. These types of information facilitate the operation of real-world urban rail transit systems. [ABSTRACT FROM AUTHOR]