Botiinae was the second largest group of Cobitidae fish. In order to investigate the morphological differences of the four species (Sinibotia superciliaris, S. reevesae, Leptobotia taeniops, L. microphthalrna) of Botiinae fishes, anatomical methods were used in this study. Standard length, body weight and other biological indices of each specimen were measured. Statistical analysis of all data was performed using Microsoft Excel 2007 and SPSS 19.0. The results showed that the morphological structures of the brain in four Botiinae fishes were similar to other teleosts, consisting of five parts: telencephalon, diencephalon, mesencephalon, cerebellum and myelecephalon (Fig. 1 and 2). Among them, there were significant differences in the relative volumes of cerebrum and corpus cerebelli, while there was no significant difference in other parts (Table 3). The more closely related the fish, the more similar the morphological structure of the brain. The L. taeniops and L. microphthalrna prefer to live in the environment with flowing water, suggesting that possession of the larger relative volume of cerebrum and corpus cerebelli might be related to a higher motor coordination ability.