Abstract: Objective: To investigate and analyze the separation of knowledge and practice among men who have sex with men in a city of Guangxi from 2013 to 2021, and analyze its influencing factors, so as to formulate more effective countermeasures.Methods: Based on the sentinel monitoring data of MSM, a cross-sectional study was conducted to investigate the separation of knowledge and practice among MSM population in a city of Guangxi from 2013 to 2021, and the factors influencing the separation were analyzed with logistic multiple analysis.Results: A total of 931 MSM patients were investigated from 2013 to 2021, and the overall HIV infection rate was 7.196%.Among this group, 415 people had homosexual anal sex with knowledge and practice, accounting for 57.40%, and 161 people had heterosexual sex with knowledge and practice with 51 people, accounting for 33.12%.The main reasons for the separation of knowledge and practice in MSM were high school and technical secondary school and the long time of local residence.Conclusion: In this city, the HIV infection rate of MSM is high, and so is the separation rate of homosexual or heterosexual anal sex.AIDS intervention should be strengthened to reduce the separation of knowledge and practice.