Background: The aim of this study was to investigate the prevalence of the middle mesial canal (MMC) and radix entomolaris (RE) in mandibular first permanent molars in a western Chinese population using cone-beam computed tomography (CBCT). Methods: A total of 1174 CBCT images of the mandibular first molars were collected from West China Hospital of Stomatology, Sichuan University. The following information was recorded and evaluated: the detection rate and location of the MMC and RE, the curvature of the RE, the canal configuration and bilateral symmetry. Results: The detection rates of the MMC and RE were 3.41 and 25.04%, respectively, as calculated by individuals, and 1.79 and 22.15%, respectively, as calculated by total teeth. The average curvature in the buccolingual (BL) orientation (40.63 ± 14.39°) was significantly larger than that in the mesiodistal (MD) orientation (17.64 ± 7.82°) (p < 0.05). Of 587 patients, 71.72% (421/587) had bilateral symmetry according to the root canal morphology. The prevalence of threerooted mandibular first molars was higher in males than in females, while the prevalence of two-rooted mandibular first molars was higher in females than in males. Conclusions: Our results showed that the RE could be detected in almost 1/4 of the western Chinese population; thus, RE detection requires special attention and careful assessment in endodontic treatment.