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Amide proton transfer–weighted imaging (APTWI) is a novel molecular MR imaging technique developed to detect and quantitatively visualize endogenous proteins and peptides. APTWI is usually reported in terms of asymmetry in the magnetization transfer ratio at 3.5 ppm. This study included 42 patients with low-grade (n = 28) or high-grade (n = 14) glioma, all of whom underwent conventional MR imaging, proton MR spectroscopy imaging, and amide proton transfer–weighted imaging on the same 3T scanner within 2 weeks before surgery. The asymmetric magnetization transfer ratio at 3.5 ppm values measured by different readers showed good concordance and were significantly higher in high-grade gliomas than in low-grade gliomas, with sensitivity and specificity values of 92.9% and 71.4%, respectively, at a cutoff value of 2.93%. The asymmetric magnetization transfer ratio at 3.5 ppm may serve as a potential biomarker not only for assessing proliferation, but also for predicting histopathologic grades in gliomas.