Night ventilation has great energy saving potentials for public buildings in summer. However, night mechanical ventilation inevitably causes more fan energy consumptions, even though it can reduce the cooling loads for the next day and save the air conditioning energy consumption correspondingly. Thus the optimization of night mechanical ventilation strategy is highly significant for building energy efficiency. In this paper, the simplified two-plate room model is used and the optimization model of the mechanical ventilation and air conditioning system is established. To minimize the total energy consumption, the optimal night ventilation strategy is determined through inverse problem method. Taking an office room in Beijing as the illustrative example, the results show that the optimal air exchange rate for night mechanical ventilation is much higher than the traditional value (ACH ¼ 0.5 h À1) and it highly depends on the indoor-outdoor air temperature difference and cooling load demands. Moreover, the total electricity usage amount can decline from 210 kWh to 111 kWh after optimization, reduced by about 47% in the whole summer season. It also indicates that the energy saving effect of night ventilation mainly derives from the cooling storage and load shifting of building internal walls. Compared to traditional ventilation strategy, the energy saving ratio increases with increasing thermal conductivity and specific heat of building internal walls. This work can provide guidance for practical ventilation system optimization design and operation strategy determination.