Abstract: Objective: To evaluate the prevalence of myopia among preschool children in Wuhan, and to explore its influencing factors and compare the urban-rural differences. Methods: Using a cluster sampling method, all preschool children in kindergartens affiliated with 13 primary-level myopia prevention and control stations in Wuhan City, Hubei Province, were selected as study subjects. The refraction was tested and questionnaires were administered to collect information on children's basic characteristics, daily living habits, screen exposure and dietary habits. Univariate analysis, and multivariate logistic regression analysis were used to explore influencing factors for myopia in preschool children and compare urban-rural differences. The Shapley value analysis was further employed to calculate the contribution degree of different influencing factors. Results: A total of 7,352 preschool children were recruited in this study, among whom 420 were identified with myopia, resulting in an overall myopia prevalence of 5.71%. The numbers of myopic children were 185 in urban areas and 235 in rural areas, with myopia prevalence rates of 7.61% and 4.78%, respectively. In the urban area, parental myopia (OR=5.58, P＜0.01） and parental astigmatism (OR=1.81, P=0.02） were risk factors for myopia in children, whereas correct pen-holding and reading/writing posture (OR=0.68, P=0.02） was a protective factor; while in the rural area, older age (OR=1.55, P＜0.01）, parental myopia (OR=3.05, P＜0.01）, and prolonged eye use at a close distance (OR=1.80, P＜0.01）, and frequent television viewing (OR=1.90, P=0.02） were risk factors for children's myopia, whereas matching of desk and chair to height (OR=0.27, P＜0.01）, and correct pen-holding and reading/writing posture (OR=0.68, P=0.01） were protective factors for children's myopia. Shapley value analysis indicated that the primary contributing factor to myopia among preschool children in urban areas was parental myopia (77.25%）, whereas in rural areas, lifestyle behaviors accounted for the largest proportion (43.62%）. Conclusion: The prevalence rate of myopia in preschool children in Wuhan in this study is generally moderate, with a higher rate in children in urban areas than in rural areas. The influencing factors affecting myopia in children differs between urban and rural areas, and interventions should be targeted separately.