Few studies have assessed knee range of motion (ROM) and moments of patients with mobile-bearing unicompartmental knee arthroplasty (MB UKA) during active deep flexion activities. We analyze knee kinematic and kinetic parameters during postoperative squatting-standing activity, aiming to evaluate the efficacy of MB UKA and postoperative rehabilitation progress. This was a clinical cohort study. We followed up with 37 patients diagnosed with medial knee osteoarthritis (OA) with primary UKA. After screening 31 patients were recruited to take gait tests. Squatting-standing activities were performed under the test of 10-camera motion analysis system and force plates preoperatively at different stages after UKA (12, 18, and 24 months). The average duration of follow-up was 24.4 months (from 22.8 to 26.7 months). Hip-knee-ankle angle improved significantly compared with pre-UKA as well as scores of American Knee Society Score, numeric rating scale, ORS, and Western Ontario and McMasters. University Osteoarthritis Index. About 83.6% (31/37) of follow-up patients completed squatting-standing activity independently. At 1-year follow-up, peak varus angle (20.6 ± 2.8 degrees), internal rotation angle (13.6 ± 1.8 degrees), extensor moment (1.44 ± 0.04N*m/kg), and internal rotator moment (0.02 ± 0.005N*m/kg) of UKA knees were inferior to contralateral knees. Peak adductor moment (0.76 ± 0.05N*m/kg) was superior to contralateral knees. At 2-year follow-up, peak flexion angle (125.0 ± 2.8 degrees) showed a growing trend meanwhile extensor (1.70 ± 0.03N*m/kg) and adductor (0.68 ± 0.06 N*m/kg) moment closely resembled those of the contralateral knee. MB UKA could alleviate the affected knee mainly in flexion-extension ROM and moment meanwhile did not affect the biomechanical indicators of healthy limbs. OA knees in the early postoperative period showed decreased extensor moment and increased adductor moment during active deep flexion activity. Better ROM and relatively more natural extensor and adductor moment of UKA knee with rehabilitation time increasing may predict ideal rehabilitation outcome in the medium or longer term.