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Background: HIV pre-exposure prophylaxis (PrEP) is a key component of HIV combination prevention strategies and has the potential to decrease the incidence of HIV in men who have sex with men (MSM). This study aimed to evaluate levels of PrEP acceptability and explore factors associated with willingness to use PrEP among MSM in Liuzhou, China. Methods: Between November 2017 and May 2019, a cross-sectional survey was conducted among 829 MSM. The study participants were recruited through a nonprobability sampling method. The participants' demographics, HIV/AIDS knowledge, sexual behaviors, drug use and HIV test history were collected. Multivariate logistic regression was performed to identify factors associated with willingness to use PrEP. Results: A total of 829 MSM completed the survey, and 30.28% (95% CI 27.3–33.4) were willing to use PrEP. In multivariate logistic regression, factors associated with a higher willingness to use PrEP included Zhuang or Han ethnic origin, recruitment through peer introduction or gay venues, pursuit of a higher education level, previous HIV testing and oral sex with a man. Conversely, having casual sex in the past 6 months was associated with a lower willingness to use PrEP. Conclusions: MSM in Liuzhou reported a relatively low level of willingness to use PrEP. The results indicate the need for effective education, targeted intervention, and implementation strategies to promote PrEP acceptance among MSM. [ABSTRACT FROM AUTHOR]