|This study investigate the relationship of working hours, stress studies and workplace fatigue in nursing staff, as the formulation of evidence-based workplace health promotion nursing staff plan to extend the life of nursing staff career. This study used to cross-sectional of quantitative research design, the research objects are nurses who work in the regional hospitals of Kinmen County, by conducting questionnaires of 155 copies. 150 questionnaires were recovered, Deducting 2 copies of invalid questionnaires, and totally we collected 148 copies of valid questionnaires. By using descriptive statistics, multivariate analysis of variance, MANOVA and Pearson correlation, the between variables was significant differences, we continue to analysis the Scheffe's, the significance level set at α = .05. The effects of working time on the workplace fatigue, such as fatigue index scores were significantly higher than moderate height working time working time (HT＞ MT) and low working hours (HT＞ LT) in showing four dimensions. The effects stress studies of fatigue in the workplace, showing personal fatigue, work fatigue and fatigue, such as fatigue index scores were significantly higher than in those no studies (p＜ .05). The relationship of working hours, stress studies and workplace fatigue in between variables, the display has a learning pressure and fatigue scores (r = .35, p ＜ .05) and the height of working time (r = .31, p ＜ .05) was significant between the positive related. The significantly relationship of working hours, stress studies and workplace fatigue in nursing staff, The variables of personal fatigue and workplace fatigue in the nurses, there may by a high degree of stress studies and working hours was significant impact. If you can effectively improve these two factors in order to reduce workplace fatigue of nurses, and thus enhance the domestic care quality.