Abstract: Rural human settlements refer to the organic combination of material and non-material that is required for the production and living of residents in rural areas. Moreover, it is also considered a dynamic and complex giant system. However, the rough and disorderly development mode has led to a series of prominent “negative effects” of rural human settlements in recent years, including rural ecological pollution, land abandonment, low infrastructure, and hollow villages. It is a practical demand for the better development of rural human settlements in rural revitalization. This study aims to quantitatively evaluate the quality of rural human settlements, and then diagnose its obstacle factors. An important basis was also offered to promote the scientific and orderly development of villages in transition. A comprehensive evaluation index system was constructed for the rural human settlements quality from three dimensions of production, living and ecological space. The multi-source data was collected from the Dongting Lake area as a typical case area at the county scale from 2010 to 2020. Afterwards, the dynamic evaluation of the rural human settlements quality was carried out to combine with the entropy value method, comprehensive index and GIS spatial analysis. Furthermore, the obstacle degree model was used to diagnose the main obstacle factors constraining the improvement of rural human settlements quality. The results showed that: 1) The comprehensive index of rural human settlements quality continued to rise, indicating the remarkable effectiveness of human settlements construction. But the differences among regions were still relatively outstanding. Additionally, the quality measurement showed an evolutionary trend of relative decline in the southwest, while the gradual rise in the northeast in space. 2) The production space quality index increased steadily from the internal perspective of the rural human settlements system. The distribution of counties with higher and above quality grades was shifted from the southern to the eastern, indicating the great variation in the spatial pattern. The living space quality index increased significantly with the removal of low-quality grade areas, and the high-quality grade areas gradually expanded outward along the urban areas of the city centers. There were spatial evolution features from the “scattered” pattern to “high in the west and low in the east” pattern. The ecological space quality index remained basically unchanged, and the spatial distribution of quality grade areas shared a decreasing trend in the western, eastern and central regions. 3) Production and living space were the main subsystems to restrict the improvement of rural human settlements quality. However, the tightening of resource constraints had led to an increasing degree of obstacles in the ecological space subsystem, with the rapid development of the economy and society. Among them, the main obstacle factors of rural human settlements quality were the factors of rural economic activity intensity, biological richness index, density of leisure facilities, level of agricultural mechanization, level of rural electric facilities, medical condition and hydrology index. The primary regulating objects were determined in the future. In conclusion, the finding can provide empirical guidance and decision-making to promote the rural human settlements quality in rural revitalization.