Background: The COVID-19 pandemic brought about great transformation to medical education mode. Although mobile communication devices played a crucial role in online learning among quarantined university students, the potential smartphone addition problems, negative health behaviors, and psychological symptoms need considerable attention. This study examined the relationship of problematic smartphone use (PSU), sleep quality, and daytime fatigue among medical students.Methods: A web-based survey was conducted in six polyclinic hospitals in Beijing between February and May 2020. 1016 participants (26.01 ± 2.46 years, 65.16% female) completed self-report measurements including Short Version Smartphone Addiction Scale (SAS-SV), Athens Insomnia Scale (AIS), and Subjective Fatigue Scale (FS). Spearman correlation coefficients and multiple regression models were used to analyze the association among PSU, sleep quality, and daytime fatigue. We used structural equation modeling to test the mediating effect of sleep quality between PSU and daytime fatigue.Results: 49.70% of the participants had PSU. Significant positive correlations were found among SAS-SV, AIS, and FS scores (r = 0.35–0.61, PS < 0.001). Subjects with PSU were more likely to report sleep disturbance (β = 1.07, P < 0.001, OR = 2.91, 95%CI = 2.17–3.91), physical fatigue (β = 1.16, P < 0.001, OR = 3.18, 95%CI = 2.45–4.15), and mental fatigue (β = 0.88, P < 0.001, OR = 2.42, 95%CI = 1.86–3.14). The indirect effect of PSU on physical fatigue and mental fatigue mediated by sleep quality accounted for 50.03 and 45.43% of the total effect, respectively.Conclusions: PSU was significantly associated with sleep disturbance and fatigue among medical students during the COVID-19 pandemic. Sleep quality mediated the relationship between PSU and daytime fatigue. Our results provide valuable information for maintaining medical students' health status and constructing online education structures.