Objectives: The authors used the National Aeronautics and Space Administration Task Load Index (NASA-TLX) and general health questionnaire to analyze the factors influencing the mental health status and the workload of support nurses during the COVID-19 epidemic. Material and Methods: The authors conducted a cross-sectional survey of 349 support nurses in April–October 2022. Using QuestionStar, a powerful online survey tool, the authors administered surveys to the participants, collected data on the mental health status and workload of support nurses, and analyzed the influencing factors based on the collected data. Results: A total of 316 questionnaires were successfully collected, with an effective rate of 98.75%. The proportion of support nurses with mental health problems was 25% and the value of the NASA-TLX questionnaire was: M±SD 68.91±7.28 pts. Multi-factor analysis revealed that the number of children, family support, and nursing support location were the influencing factors of mental health status, while the multivariate analysis revealed that the presence of symptoms, nursing support location, support work type, and total 12-item General Health Questionnaire (GHQ-12) score were the influencing factors of the workload of support nurses. Conclusions: Compared to their counterparts in the plains, nurses working in isolated plateau regions who were caring for children and lacked family support, were more likely to have mental health issues. There was a positive correlation between the changes in GHQ-12 and NASA-TLX scores of the study participants. Compared to their counterparts in the plains and the tropical regions, nurses working in plateau regions had a heavier workload. As part of the follow-up measures to prevent and treat patients impacted by the COVID-19 epidemic, it is important to improve the mental health evaluation, consultation, and treatment of the support nurses to guarantee the high quality of the first-line support work. Int J Occup Med Environ Health. 2023;36(6)761–72