Objective To explore the influence of two-port laparoscopy and four-port laparoscopy on postoperative recovery and stress response in patients with middle and upper rectal cancer. Methods148 patients with middle and upper rectal cancer in the hospital from April 2021 to April 2024 were divided into two-port laparoscopy group (n=76, two-port laparoscopic surgery) and four-port laparoscopy group (n=72, four-port laparoscopic surgery) according to the surgical method. The perioperative indexes, postoperative recovery status, stress response before and after surgery were compared between the two groups, and the complications in both groups were evaluated. Results The surgial time, intraoperative blood loss,The incision length in the two-port laparoscopy group was shorter, and the visual analogue scale (VAS) score at 3 days after surgery was lower than that in the four-port laparoscopy group (P<0.05). The first postoperative ambulation time, first exhaust time, first defecation time and first oral feeding time were shorter in the two-port laparoscopy group than those in the four-port laparoscopy group (P<0.05). At 3 days after surgery, the levels of cortisol and epinephrine were enhanced in both groups than those before surgery, and the above levels in the two-port laparoscopy group were lower compared to the four-port laparoscopy group (P<0.05). No obvious differences were manifested between both groups in terms of postoperative complications (P>0.05). Conclusion Compared with four-port laparoscopic surgery, two-port laparoscopic surgery has more obvious advantages in patients with middle and upper rectal cancer, and the latter one has milder postoperative pain, faster recovery and smaller impact on stress response