BackgroundMen who have sex with men (MSM) are a significant population contributing to human immunodeficiency virus (HIV) transmission. Nevertheless, few studies have focused on the heterogeneity in MSM. The characteristics of men who have sex with men and women (MSMW) may differ from men who have sex with men only (MSMO). MethodsFrom October 2022 to March 2023, a cross-sectional study for MSM was conducted in Eastern China. An electronic questionnaire was used to collect the demographic characteristics and sexual behaviors of participants. A multivariable logistic regression model was used to assess the differences in HIV-related risky sexual behaviors between MSMO and MSMW. ResultsA total of 1993 MSM were enrolled in the study, with 1221 MSMO and 772 MSMW. Compared to MSMW, MSMO reported younger age at first anal sex, higher coerced homosexual experiences, and observed different characteristics of sexual behavior in the past six months. Multivariable analysis indicated that MSMW were more likely to seek casual partners and use stimulants online and offline. MSMW showed inconsistent condom use with online partners (aOR = 0.51, 95 %CI: 0.38–0.68) and more frequent anal sex after drinking (aOR = 2.27, 95 %CI: 1.54–3.35). They also had higher risks of commercial (aOR = 2.47, 95 %CI: 1.71–3.57) and group sexual behaviors (aOR = 1.88, 95 %CI: 1.24–2.83) compared to MSMO. ConclusionOur findings suggest that the differential factors may lead MSMW to become a subgroup at high risk of HIV infection. The gender identity and sexual orientation within MSM should be considered when formulating acquired immune deficiency syndrome (AIDS) prevention and control strategies.