AIM:To investigate the effect of vision therapy system 4D combined with stereoscopic 3D technology training for the treatment of amblyopia.METHODS: Prospective study. A total of 102 children with amblyopia who attended the clinic from January 2018 to January 2022 were selected, and they were randomly assigned into two groups by computer, with 51 cases in each group. Control group received stereoscopic 3D technology training, while observation group participated in vision therapy system 4D on the basis of control group. Then the overall effective rate, binocular visual function, spherical equivalent(SE), axial length(AL), mean corneal curvature(Km), best corrected visual acuity(BCVA)and visual evoked potential were compared between two groups.RESULTS: The overall efficacy rate was 94.1% in observation group, which was obviously higher than control group(74.5%; P<0.05). The improvement in binocular vision parameters simultaneous perception, total fusion, and stereoacuity were all more remarkable in observation group than in control group(P<0.05). The △SE, △AL and △Km yielded no statistical difference between two groups(P>0.05). The latency of two spatial frequencies(1°grid and 15'grid)showed a decline in both groups, and the decline was more notable in observation group than in control group(P<0.05). In both groups, BCVA improved, and the improvement was more significant in observation group compared with control group(P<0.05).CONCLUSION: Application of vision therapy system 4D combined with stereoscopic 3D technology training for amblyopia can effectively ameliorate the visual acuity, promote the reconstruction of simultaneous perception, total fusion, and stereoacuity without additional risk of myopic shift, and improve visual pathway function in children.