The major features of different road sections were identified. Methods quantitatively determining the ecological impact of highway construction were discussed, and a demonstration was presented. The Dabao highway (from Dali to Baoshan) and the Sixiao highway (from Simao to Xiaomengyang) passing through the Longitudinal Range-Gorge Region were used as examples in this paper. For the normal road sections having no important species to protect, a method of ecosystem health assessment was introduced to calculate the overall range of the ecological impact of highway construction. For the road sections having plants or soils of interest, indexes were selected and the range of impact was determined using mathematical methods such as regression analysis and variance analysis. The range of impact on animal populations was also discussed in terms of the minimum living areas required by animal populations and the fragmentation caused by highway construction. The results indicate that the zones impacted by highway construction were composed of both regular and anomalous figures, the range of impacts for different landforms of the two sample highways were substantially different; highway construction had dramatic effects on roadside ecosystem health; the impact on soil factors of farmland was greater than 200 m, and the distance may exceed 1000 m when important large animals were considered.