Environmental and social factors influencing resource allocation in rural, developing regions are critical social determinants of health that necessitate cross-sector collaboration to improve health opportunities. Thus, we sought to evaluate the spatial distribution and accessibility of medical resources to assess existing disparities, identify best practices for resource allocation, and inform regional health planning policies. In this study, inequality in the frequency distribution of medical resources in Wenzhou, China, was measured using the Gini coefficient and agglomeration degree. We evaluated the spatial accessibility of medical institutions throughout the city using the modified hierarchical two-step floating catchment area (H2SFCA) method. Using the Spearman correlation analysis, we investigated the factors influencing accessibility differences. The results indicate that Wenzhou’s spatial distribution of medical resources is unbalanced and unequal. According to the population and geographic distribution, the distribution of medical resources in Wenzhou is unequal. Wenzhou’s overall spatial accessibility is poor. The east region is more accessible than the west region, and the accessibility of medical institutions at different levels varies greatly. The correlation between accessibility and the number of institutions, doctors, population density, road density, and GDP is positive. There is a need for policies and initiatives to enhance the geographical distribution of resources, construct interconnected road networks, and improve residents’ access to medical resources.