Objective: This study aims to assess the mental well-being and factors that influence medical personnel engaged in safeguarding tasks at medical observation and isolation sites amid COVID-19 prevention efforts in Qingpu District, Shanghai. The findings will provide evidence to inform the development of effective preventive strategies. Methods: A survey employing the Self-Rating Anxiety Scale (SAS), Pittsburgh Sleep Quality Index (PSQI), and a custom basic information questionnaire was conducted among medical personnel stationed at medical observation isolation sites in Qingpu District, Shanghai, during COVID-19 pandemic. Results: Out of the 379 medical personnel surveyed, 132 individuals (34.8%) exhibited symptoms of anxiety, while 252 individuals (66.5%) reported sleep disorders. The multivariate logistic regression analysis revealed that sex (OR=2.44, 95%CI: 1.28-4.65) and living with elderly (OR=1.64, 95%CI: 1.03-2.62) significantly affected the anxiety levels among healthcare workers. Although five variables including age, marital status, professional title, living with children, and living with elderly were examined for their impact on sleep quality, none of these factors showed statistically significant differences. Conclusions: During the COVID-19 pandemic, anxiety symptoms and sleep disorders among medical personnel stationed at medical observation isolation sites varied based on sex, age, marital status, education level, professional title, and living arrangements. These findings underscore the importance of addressing these concerns and implementing measures to enhance and alleviate anxiety and sleep disorders among medical personnel in isolation settings.