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To investigate the association of CYP3A5*3 with acute lymphoblastic leukemia susceptiveness in Chinese han children. Methods: CYP3A5 genotype was analyzed from 172 Chinese han children, including 52 acute lymphoblastic leukemia patients and 120 healthy. PCR-RFLP was used to identify the polymorphism. Results: The allelic frequency of CYP3A5*3 was 0.721 (95% CI: 0.549, 0.893) in patients while 0.758 (95% CI: 0.674, 0.842) in healthy controls. No significant difference existed in CYP3A5*3 frequencies between patients and controls (P=0.275), males and females (P=0.432) in the total population. Conclusion: The CYP3A5*3 variant represents a common polymorphism in Chinesehealthy children and ALL, and this polymorphism is poor association with childhood acute lymphoblastic leukemia susceptiveness.