Background: This research aims to evaluate the clinical outcomes of vaginal natural orifice transluminal endoscopic surgery hysterectomy (vNOTESH) compared with conventional laparoscopic hysterectomy (cLH) on non-prolapsed uteri for presumed benign gynecologic disorders. Methods: We retrospectively analyzed the data of all patients receiving vNOTESH or cLH in Meizhou People’s Hospital, Meizhou, China from January 2018 and December 2020. Relevant patient characteristics and clinical outcomes data were collected after written informed consent. Results: There were no significant differences between cLH and vNOTESH groups regarding age, body mass index (BMI), number of pregnancy, history of abdominal operation, type of surgery, blood loss, changes in hemoglobin levels, postoperative fever, complications, 12th-hour, and 48th-hour Visual Analog Scale (VAS) scores. The vNOTESH group had significantly shorter operative time (67.37 \pm 25.90 vs. 89.71 \pm 36.43 min, p 0.001) and postoperative hospitalization (5.25 \pm 1.67 vs. 5.82 \pm 2.88 d, p = 0.007) than the cLH group. Besides, the 24th-hour VAS scores were noticeably lower (1.50 \pm 0.75 vs. 1.78 \pm 0.77, p = 0.001) in the vNOTESH group. Furthermore, the subgroup analysis showed similar trend in operative time (83.56 \pm 33.37 vs. 95.84 \pm 33.83 min, p = 0.017) and 24th-hour VAS scores (1.38 \pm 0.59 vs. 1.79 \pm 0.75 min, p = 0.001) between the two enlarged uterine subgroups. Conclusions: The vNOTESH can offer a safe and effective alternative to laparoscopy for women with non-prolapsed benign uteri. Even in case with a large uterus, the vNOTESH can be a feasible treatment with promising short-term efficacy and safety as compared with cLH.