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AIM:To explore the effectiveness of a new non-inverted pedicle internal limiting membrane(ILM)flap transposition technique in the treatment of large macular holes(MH).<p>METHODS: This was a prospective pilot study which included 9 patients with 10 eyes in Jiangsu Province People's Hospital from December 2016 to February 2017. All patients was diagnosed with large MH(size >400μm)by the spectra-domain optical coherence tomography(SD-OCT)and underwent the non-inverted pedicle ILM flap transposition surgery. Best-corrected visual acuity(BCVA), SD-OCT images, and MP-1 microperimetry tests were performed pre-operation, 1d, 1wk, 1, 3, and 6mo post-operation. <p>RESULTS: The macular hole closure rate after 6mo was 100%. The averaged BCVA improved from 1.19±0.54(LogMAR)pre-operation to 0.70±0.50(LogMAR)post-operation(<i>P</i>=0.005). The mean retinal sensitivity within 8° and 2° improved from 3.14±4.52dB to 8.91±5.53dB(<i>P</i>=0.008), and 1.46±2.94dB to 6.33±4.90dB(<i>P</i>=0.008)respectively. Preoperative unstable fixation in seven eyes resolved at the last postoperative follow-up. <p>CONCLUSION: Our non-inverted pedicle internal ILM flap transposition technique shows effectiveness in the treatment of large macular holes with high MH closure rate and improving visual function.