Objective To construct a humanistic care training curriculum for nurses working in comprehensive hospitals, and to explore its effect. Methods The curriculum of humanistic care training for nursing staff was established through literature review and expert consultation. Then the curriculum was applied to a humanistic care training program, which provided 1-year training to nurses working in a comprehensive hospital. The effects were evaluated by assessing nurses′ caring ability and patient satisfaction. Results The finalized humanistic care training curriculum contains 4 training modules (humanistic care cognition and knowledge, humanistic care technique and practice, humanistic teaching and training, and research on humanistic care) and 15 knowledge key points. After training, the scores of caring ability among nurses and patient satisfaction scores were significantly higher than those before the training （P＜0.01 for both）. Conclusion The humanistic care training curriculum for nurses working in comprehensive hospitals is scientific and reliable, and can improve nurse′s humanistic care ability and patient satisfaction after training.