Objective: To investigate the clinical effect of using pelvic autonomic nerves as anatomical landmarks to plot a safe surgical plane for performing laparoscopic total mesorectal excision (TME). Methods: The clinical data and operation videos of 157 male patients who underwent laparoscopic low rectal resection between January 2010 and December 2015 were retrospectively analyzed. Of the patients, 81 cases treated after 2012 were categorized as observation group, who underwent TME using pelvic autonomic nerves as landmarks which were dissected and freed through medial approach, and digestive tract reconstruction by double-stapling technique; 76 cases treated before 2012 served as control group who underwent surgery without using the above anatomical landmarks. The relevant surgical variables as well as postoperative urinary and sexual function between the two groups were compared. Results: In observation group compared with control group, the intraoperative blood loss was significantly reduced (14.9 mL vs. 26.5 mL), the rate of level-3 operation quality was significantly elevated (89.2% vs. 59.6%), the number of resected lymph nodes was significantly increased (19 vs. 15), and the incidence of postoperative erectile dysfunction (2.3% vs. 4.5%) and urinary retention (6.2% vs. 10.5%) were significantly decreased, all the differences had statistical significance (all P<0.05). Conclusion: Using pelvic autonomic nerves as anatomical landmarks for TME can achieve maximally complete mesorectal resection and meanwhile reduce pelvic splanchnic nerves injuries. In addition, it may help to standardize and improve the technique of laparoscopic TME in low rectal cancer surgery.