Abstract: Based on the basic geographic information data, street population data and hospital beds data of Nanjing, the Two-step floating catchment area and Gini coefficient are used to study the spatial distribution equilibrium and accessibility of medical resources in Nanjing. The results show that the spatial distribution of medical resources in Nanjing is unbalanced, which is characterized by a low level of new urban areas in the north and south and a high level of old urban areas in the central part. Moreover, the existing medical resources cannot fully meet the needs of the permanent residents, but can basically meet the needs of the elderly population. The accessibility level of walking is low, while that of driving is high. The comprehensive medical accessibility of two modes shows the characteristics of stratification decreasing from the old city center to the outer circle. The research suggests that the radiation effect of high-quality medical resources in old urban areas and some new urban areas should be increased to the surrounding areas, solve the problem of unbalanced distribution of medical resources byincreasing the supply of medical resources to the northern municipal districtand build a regional medical complex to solve the problem of inaccessible medical treatment by walking through hierarchical diagnosis and treatment.