Listening comprehension plays a pivotal role in second language (L2) acquisition, but the process of listening is complex. L2 learners' listening ability is influenced by both cognitive and affective factors. Among the latter, anxiety is one of the most commonly studied variables in the field of education (Horwitz, 2001); however, its impact on learners' listening performance is inconsistent in previous research and there have been contradictory findings regarding whether anxiety levels differed significantly according to gender. To address these limitations, this study investigated the effect of gender and foreign language listening anxiety (FLLA) on listening comprehension among 187 university students in China. In addition, it explored whether there were significant differences in FLLA levels between genders. Data sources included the FLLA scale and the Oxford Online Listening Level Test to measure FLLA and L2 listening comprehension, respectively. Results from between-groups analysis of variance showed that listeners with low anxiety listened significantly better than listeners with moderate and high levels of anxiety. Gender differences were examined in two aspects of listening: listening comprehension and FLLA. Specifically, females performed better than males in listening comprehension, and females had a higher level of FLLA compared to males. These results provide implications for teaching L2 listening considering the role of anxiety and gender.