Objective To explore the effect of laughter yoga in chemotherapy patients with non-small cell lung cancer. Methods According to cluster randomization method, 84 patients with non-small cell lung cancer who were treated with chemotherapy in two wards of respiratory department were selected and divided into two groups, 42 cases in each group.The control group received routine nursing care education, while the intervention group additionally received laughter yoga intervention.The effect of intervention was evaluated by the Perceived Stress Scale, Self-rating Anxiety Scale, Self-rating Depression Scale and Psychological Capital Scale. Results The scores of perceived stress, anxiety and depression in the intervention group were significantly lower than those in the control group, and the scores of positive psychological capital were significantly higher than those in the control group (P＜0.05 for all). Conclusion Laughter yoga can effectively relieve patients′ stress perception, as well as anxiety and depression, and improve patients′ positive psychological capital.