Abstract: Objective To apply sacrogenital fold as an anatomical landmark to avoid injury during laparoscopic total mesenteric excision (TME) and evaluate its clinical effect. Methods From June 2015 to October 2017, 109 rectal cancer patients received laparoscopic surgery in our hospital were randomly divided into the control group (n=52) and the observation group (n=57). Both two groups received routine laparoscopic TME, and the observation group received laparoscopic TME with the sacrogenital fold as an anatomical landmark, and all operations were performed behind the sacrogenital fold. The intraoperative blood loss, operating time, postoperative gastrointestinal function recovery, urinary function, postoperative complications and hospital stay were recorded. Results There was no death and conversion to laparotomy during hospitalization in two groups. The operating time, intraoperative blood loss, postoperative gastrointestinal function recovery time, urine output, maximal bladder detrusor systolic blood pressure and IPSS showed no significant difference between the two groups (P＞ 0.05). Compared with the control group, the postoperative catheter indwelling time (1.3±0.5) d vs (1.5±0.5) d, residual urine volume (18.5±7.5) ml vs (21.4±6.9) ml, the incidence rate of urine retention 0(0) vs 4(7.8%) and the length of hospital stay (8.9±2.8) d vs (10.1±3.4) d decreased significantly in the observation group (all P＜ 0.05), while the maximum urinary flow rate increased (P＜ 0.05). Ureteral injury was not found in the observation group. There was no significant difference in rectal injury, presacral venous plexus injury and vaginal/vasovasic/seminal vesicle injury between two groups (all P＞ 0.05). No significant complications occurred after surgery in two groups. Conclusion As an anatomical landmark, sacrogenital fold can be used for laparoscopic TME to avoid ureteral injury and improve urinary function, which is worthy of clinical promotion.