Knowledge-motivation-psychological model was set up and tested through structural equation model to provide evidence on HIV prevention related strategy in Men who have Sex with Men (MSM). Snowball sampling method was used to recruit a total of 550 MSM volunteers from two MSM Non-Governmental Organizations in Urumqi, Xinjiang province. HIV prevention related information on MSM was collected through a questionnaire survey. A total of 477 volunteers showed with complete information. HIV prevention related Knowledge-motivation-psychological model was built under related experience and literature. Relations between knowledge, motivation and psychological was studied, using a ' structural equation model' with data from the fitting questionnaires and modification of the model. Structural equation model presented good fitting results. After revising the fitting index: RMSEA was 0.035, NFI was 0.965 and RFI was 0.920. Thereafter the exogenous latent variables would include knowledge, motivation and psychological effects. The endogenous latent variable appeared as prevention related behaviors. The standardized total effects of motivation, knowledge, psychological on prevention behavior were 0.44, 0.41 and 0.17 respectively. Correlation coefficient of motivation and psychological effects was 0.16. Correlation coefficient on knowledge and psychological effects was -0.17 (P<0.05). Correlation coefficient of knowledge and motivation did not show statistical significance. Knowledge of HIV and motivation of HIV prevention did not show any accordance in MSM population. It was necessary to increase the awareness and to improve the motivation of HIV prevention in MSM population.