To compare the clinical effects of uterine myomectomy by vaginal orifice transluminal endoscopic surgery (V-NOTES) and by singleport laparoscopy. Methods: A total of 96 patients with single myoma of the anterior uterine wall were selected , and were randomly divided into two groups (48 cases in each group) according to the random number table from May 2018 to November 2020. The patients in the control group were treated with single-hole laparoscopic uterine myomectomy, and the patients in the observation group were treated with V-NOTES uterine myomectomy. The operation indexes, the situations of complication and postoperative pain of the patients were compared between the two groups. The levels of hemoglobin (Hb) and stress response factors of the patients were compared between before and after the operation. Results: The postoperative exhaust time, the time of postoperative getting out of bed, and the hospital stay time of the patients in the observation group were significantly lower than those of the patients in the control group (P<0.05), but there were no significant differences in the operative time and intraoperative blood loss of the patients between the two groups (P>0.05). There were no significant differences in the incidences of the intraoperative and postoperative adjacent organ injury, the postoperative infection, and the poor wound healing of the patients between the two groups (P>0.05). The VAS scores of the patients in both groups at postoperative 4h, 12h and 24h had decreased gradually, and which of the patients in the observation group was significantly lower than that of the patients in the control group (P<0.05). The levels of serum Hb and malondialdehyde (MDA) of the patients in the two groups after operation were significantly higher than those before operation, but the levels of superoxide dismutase (SOD) of the patients in the two groups after operation was significantly lower than those before operation (P<0.05). There was no significant difference in the Hb level of the patients between the two groups (P>0.05), the serum MDA level of the patients in the observation group was significantly lower than that of the patients in the control group, the SOD level of the patients in the observation group was significantly higher (P<0.05). Conclusion: V-NOTES and single-port laparoscopic myomectomy are both safe and effective. However, compared with those of single-port laparoscopic myomectomy, V-NOTES myomectomy has more advantages in improving postoperative recovery, relieving postoperative pain and stress reaction.