Abstract: Objective To investigate the awareness, usage and requirement of Massive Open Online Courses (MOOC) among preventive medicine students in a medical university in Anhui Province, and provide the scientific basis for promoting applications of MOOC in public health. Methods Preventive medicine undergraduates and some graduate students were recruited from the university. The related data was collected though a self-designed questionnaire. Results A total of 317 students participated in this investigation. In the aspect of MOOC awareness, only 2.5% students were well aware of the MOOC, and 37.5% were completely unaware. It should be noted that 57.1% students (113/198) knew about MOOC though recommendation of their teachers. For the usage of the MOOC, 45.1% students said that they had taken the course, and 29.7% students had completed at least one MOOC course. In the aspect of MOOC requirement, 72.9% students considered it is necessary to be offered MOOC courses. In addition, sex and grade may have an influence on awareness, usage and requirement of MOOC among these students. Conclusions The study show a low level of the awareness, usage of MOOC but a relatively high level of requirement in these preventive medicine students. Enhanced MOOC propaganda among students by high schools and teachers may be beneficial to improve the utilization of MOOC.