Objective: to explore associations and gender differences between OHI-seeking (online health information seeking) behaviors and eHealth (electronic health) literacy among Chinese university students. Methods: Online questionnaires of eHealth literacy scale and OHI-seeking behaviors created in software Wenjunxing were used in this survey. Chi-squared tests, t-test, and Pearson correlation analysis were performed using SPSS for data analysis. Results: Among 5,383 participants, 72.4% were girls, 77.5% lived in rural areas, 51.2% majored in liberal arts, 76.6% with low education parents. The average C-eHEALS scores of boys and girls were 26.53 ± 5.861 and 26.84 ± 5.816, respectively, with no significant difference (P = 0.084). The top three OHI-seeking behaviors for boys and girls, as well as for the C-eHEALS low and high groups, were “finding information about physical exercises” “reading or sharing health information via social media” “finding information about nutrition and diet”, all of which had significant gender and eHealth literacy differences. Conclusions: Gender and eHealth literacy differences should be focused for intervention when developing and implementing eHealth intervention training for parents and adolescents in schools and communities.