COVID-19 is a major public health event affecting the people worldwide. Nurses are still under immense psychological pressure. This study aimed to explore the relationship between mental fatigue and negative emotions among frontline medical staff during the COVID-19 pandemic. The study was conducted in August 2020, which included 419 medical staff between 17 to 28 years. The Fatigue Scale, Multidimensional Mental Flexibility Questionnaire, Cognitive Fusion Scale, and Depression-Anxiety-Stress Brief Version Scale were used. During the data collection period, the pandemic was under control in China and continued worldwide. The results indicated that 27.7% of the medical staff experienced depression, and 32.3% of them feel stressed. Specifically, first, correlation analyses showed significant positive pairwise correlations between mental fatigue, psychological inflexibility, cognitive fusion, and negative emotions among nurses. Second, mediation model tests showed statistically significant mediating effects of psychological inflexibility and cognitive fusion between mental fatigue on nurses' negative emotions, and statistically, significant chain mediating effects of psychological inflexibility and cognitive fusion. Mental fatigue indirectly affects nurses' negative effects through the mediating effects of psychological inflexibility, cognitive fusion, and the chain mediating effects of psychological inflexibility and cognitive fusion, respectively. the negative effects of mental fatigue come from impairment of cognitive functioning, and interventions using acceptance and commitment therapy for mental fatigue and negative emotions are more effective since both psychological inflexibility and cognitive fusion are important components of the therapy.