Background: The outbreak of COVID-19 has laid unprecedented threats and challenges to health workers (HWs) in Wuhan, China. We aimed to assess the sociodemographic characteristics and hospital support measures associated with the immediate psychological impact on HWs at Tongji Hospital in Wuhan during COVID-19 outbreak. Methods: We conducted a single-center, cross-sectional survey of HWs via online questionnaires between February 8th and 10th, 2020. We evaluated stress, depression and anxiety by IES-R, PHQ-9, and GAD-7, respectively. We also designed a questionnaire to assess the perceptions of threat of COVID-19, and the satisfactions of the hospital's support measures. Multivariate logistic regressions were used to identify associated variables of acute stress, depression, and anxiety. Findings: We received 5062 completed questionnaires (response rate, 77.1%). 29.8%, 13.5% and 24.1% HWs reported stress, depression and anxiety symptoms. Women (odds ratio [OR], 1.31; 95% CI, 0.47-0.97; p = 0.032), years of working >10 years (OR, 2.02; 95% CI, 1.47-2.79; p<0.001), concomitant chronic diseases (OR, 1.51; 95% CI, 1.27-1.80; p<0.001), history of mental disorders (OR, 3.27; 95% CI, 1.77-6.05; p<0.001), family members or relatives confirmed or suspected (OR, 1.23; 95% CI, 1.02-1.48; p = 0.03), hospital-based and department-based care (OR, 0.76; 95% CI, 0.60-0.97; p = 0.024) and full coverage of all departments for avoiding nosocomial infection (OR, 0.69; 95% CI, 0.53-0.89; p = 0.004) were associated with stress. Interpretation: Women and those who have more than 10 years of working, concomitant chronic diseases, history of mental disorders, and family members or relatives confirmed or suspected are susceptible to stress, depression and anxiety among HWs during the pandemic. In addition, since HWs often have a greater stigma against mental problems than the general public, it is worthwhile to address the needs of the HWs during this pandemic and to provide appropriate psychological supports for those people at high risk of mental problems. (C) 2020 The Author( s). Published by Elsevier Ltd.