Objective To summarize the imaging features and differential diagnosis of diffuse midline glioma(H3 K27-altered)in order to improve the accuracy of preoperative diagnosis.Methods The clinical,imaging and pathological in-formation of 26 pathologically confirmed diffuse midline glioma(H3 K27-altered)cases were retrospectively analyzed,and MRI manifestations and the corresponding pathological features were summarized.Results Of the 26 cases,7 cases were located in the spinal cord,6 in the thalamus,5 in the pons,and 3 in the medulla oblongata.Diffuse midline gliomas(H3 K27-altered)located supratentorially had a significant occupying effect and multiple morphologies,pontine lesions were swollen and tend to be non-enhancing,medullary lesions showed markedly enhancing nodules or masses,and spinal cord le-sions showed multiple long segments and patches of marked enhancement.There were more females than males in children's group,and most of the lesions were found in the pontine brain,while in the adult group there were slightly more males than females,and most of the lesions were found in the thalamus and spinal cord.More lesions were ill-defined in the adult group than in the children's group(P=0.028);a higher proportion of lesions in the children's group showed necrosis than in the adult group(P=0.036).The percentage of Ki67 expression ≥30%was greater in children's group than in the adult group(77.8%>41.2%).Conclusion Diffuse midline gliomas(H3 K27-altered)at different sites and ages have relative specificity of images.The combination of clinical symptoms,MRI plain scan and contrast-enhanced scan,DWI and PWI helps to improve the diagnosis accuracy of diffuse midline gliomas(H3 K27-altered).