In order to investigate the association between masturbation and psychopathological and reproduction relative conditions, an online survey was conducted among campus male students of Zhejiang University, which comprised of basic personal information, and questions for reproduction related health, masturbation frequency, and psychological well-being. Psychological status was evaluated with Middlesex Hospital Questionnaire (MHQ). The psychological and reproduction health related parameters were compared among the groups divided according to masturbation frequency. Finally 143 students were included in analysis. Floating anxiety, somatic, and hysteric scores were significantly associated with masturbation, with higher scores in group with highest masturbation frequency; masturbation is significantly associated with fatigue, soreness and weakness of the lumbar region, memory decline, immunity decline, insomnia dreaminess and gradual increase of frequency of masturbation, and the rates of most of the reproduction related symptoms increased accompanying the increase of the masturbation frequency. MQH scores of obsession, phobic anxiety, and depression were not associated with masturbation. It was concluded that masturbation may adversely affect psychological health as well as reproductive well-being.