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Objective: To report 4 cases of biphenotypic acute leukemia (BAL) with t(8;21)(q22;q22), and analyze the characteristics of morphology, immune phenotype, chromosome karyotype (MIC) and clinical manifestations. Methods: The BAL patients with t(8;21)(q22;q22) (group A) were compared with the randomly selected BAL patients with other clonical chromosomal changes (group B) and acute myeloid leukemia M2 cases with t(8;21)(q22;q22) (group C)in MIC and clinical features.Results: BAL with t(8;21)(q22;q22) showed acute myeloid leukemia with high percentages of blast cells morphologically;revealed co-positive to B-lymphoid and myeloid lineages, frequent and high expressions of CD34 and CD33; were responsive to chemotherapy for myeloid and lymphocytic leukemia simultaneously well. Conclusion: A new subset of BAL with t(8;21)(q22;q22) was reported, and this suggests that the leukemia colony with t(8;21)(q22;q22) might originate from early phase of hematopoiesis.