Objective To understand the current status and influencing factors of healthy eating behavior among young and middle-aged people, and to analyze its relationship with the risk of gallstone disease, so as to provide guidance for developing dietary intervention programs for this population. Methods A total of 1 038 young and middle-aged people undergoing health examination were selected, and they were investigated utilizing the basic information questionnaire and the Adult Healthy Eating Behavior Evaluation Scale. Results The total score of healthy eating behavior in the participants was 73.93±12.85. Multiple linear regression results showed that, gender, educational background, living status, past medical history of gallstone disease and family history of gallstone disease were the influencing factors of healthy eating behavior in young and middle-aged people (P<0.05, P<0.01). During the 1-year follow-up, the prevalence of gallstone disease in the young and middle-aged people with unhealthy diet was significantly higher than their counterparts with healthy diet (P<0.01). The results of multivariate Cox regression analysis showed that, age, past medical history of gallstone disease, family history of gallstone disease, awareness of healthy eating, regular eating attitude, and healthy eating habits were the influencing factors of gallstone disease in young and middle-aged people (HR:0.091-0.299, P<0.01 for all). Conclusion The healthy eating behavior among young and middle-aged people is at a medium level, which is affected by several factors. Medical staff should strengthen health education, so as to guide them establish healthy eating behavior to prevent gallstone disease.