Objective: To evaluate the feasibility and safety of single-port laparoscopic hysterectomy comparing with multi-port laparoscopic hysterectomy in treatment of benign uterine diseases. Methods: Data were collected retrospectively by review of the medical records of 252 patients who underwent multi-port or single-port laparoscopic surgery for treatment of benign gynecologic diseases. Laparoscopy assisted vaginal hysterectomy (LAVH) was performed for single-port surgery and LAVH and total laparoscopic hysterectomy (TLH) were performed for multi-port surgery. Demographic variables were collected and analyzed by independent t-test and Pearson Chi-Square test. The primary outcome was analyzed by independent t-test and Fisher’s Exact test. Results: A longer operative time was observed in the multi-port surgery group compared with that of the single-port group (p < 0.05). No difference with respect to change of Hemoglobin between the preoperative level and that of the postoperative first day, the number of days from the operation to discharge, uterine weight, and the rate of laparotomy conversion and complications were observed between the two groups. Conclusion: Single-port laparoscopic hysterectomy for treatment of benign uterine diseases is a safe and feasible method.