Mutations in Gap Junction Beta 2 (GJB2) have been reported to be a major cause of non-syndromic heating loss in many populations worldwide.The spectrums and frequencies of GJB2 variants vary substantially among different ethnic groups, and the genotypes of GJB2 in these populations are poorly understood.In the present study, we carried out a systematic and extended mutational screening of GJB2 gene in a cohort of 1067 Han Chinese non-syndromic hearing loss subjects.Molecular analysis revealed 25 (23 known and 2 novel) GJB2 variants in these affected subjects, including 11 known hearing loss-associated mutations.