Objective To explore the correlation between presenteeism behavior of nurses and their knowledge of patient safety culture. Methods A total of 435 nurses from 5 top-three hospitals in Henan Province were selected and asked to fill out the general information questionnaire, the Presenteeism Behavior Scale and the Patient Safety Culture Scale. Results The nurses scored (2.62±0.93) points in presenteeism behavior, with the incidence rate of presenteeism behavior being 87.82%. The nurses scored (3.73±0.38) points in patient safety culture.Nurses′ presenteeism behavior was negatively correlated with their perception of patient safety culture（P＜0.01）. Conclusion The incidence rate of presenteeism behavior is high among nurses.Nurses′ presenteeism behavior undermines their perception of patient safety culture.It is suggested to reduce nurse presenteeism behavior as far as possible, in a bid to improve patient safety culture and ensure patient safety.