Many studies have reported positive contributions of health promotion on the health behavior of nursing staff working in hospitals, including the maintenance of a regular healthy diet, engagement in physical activity, performance of routine screening practices, and participation in a health examination. Despite being considered a role model for healthy lifestyles, little is known about the effect of health-promoting hospital settings on nursing staff. The aim of this study was to perform a nationwide, hospital-based, cross-sectional, survey comparing health practices between full-time nurses of health-promoting hospitals and those of non-health-promoting hospitals in Taiwan. We conducted a nationwide, hospital-based, cross-sectional, survey in 100 hospitals from May to July 2011 using a questionnaire as the measurement tool. Nurses aged between 18 and 65 years from certified health-promoting hospitals (n = 14,769) were compared with nurses in non-health-promoting hospitals (n = 11,242). A multiple logistic regression model was conducted to estimate the effect of certified HPH status on the likelihood of performing health behavior, receiving general physical examination, undergoing cancer screening, and participating in hospital-based health-promoting activities. All nurses of HPH hospitals were more likely to perform physical activity, practice cancer screening, receive at least one general physical examination in the past 3 years, and had a higher chance of participating in at least one hospital-based health-promoting activity in the past year (particularly weight-control groups and sports-related clubs) than those of non-HPH hospitals. This study suggests the effectiveness of implementing health promotion on the health behavior of full-time nursing staff in hospitals.