Objective To observe the effectiveness of introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride. Method A total of 80 rabbits of both genders for operation were randomly divided into A, B and C groups. The A group was injected with Sumianxin intramuscularly (0.3 mL/kg by weight). The B group was injected with Sumianxin and ketamine hydrochloride intramuscularly (0.3 mL/kg by weight). The C group was injected with Diazepam intravenously (1.5 mL/kg by weight) combined with Sumianxin and ketamine hydrochloride injected intramuscularly (0.3 mL/kg by weight). The aesthetic effects, induction time, anesthesia maintaining time, total anaesthetic dose and operation time were observed, recorded and compared. Result The induction time of the C group was significantly shorter than A and B groups (P<0.01). The initial anesthesia maintaining time of the C group was the longest among the three (P<0.01) with least total anaesthetic dose (P<0.01). The operation time of the C group was the least with best aesthetic effects (P<0.01). Conclusion Introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride can improve the aesthetic effects. Therefore, this is an optional aesthetic method for time-consuming animal operation or sensitive surgical sites of rabbits.