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Introduction: Traditional Chinese medicines (TCM) contain multi-interactive compounds that have been used for treatment of peri-menopausal syndrome and have become a new phytoestrogens resource.The QiBaoMeiRan formula (QBMR),including Polygoni Multiflori Radix, Angelicae Sinensis Radix, Achyranthis Bidentatae Radix, Semen Cuscutae, Fructus Lycii, Poria, and Fructus Psoraleae, has been used clinically for treating osteoporosis in post-menopausal women by virtue of its kidney-invigorating function.However, no evidence base links QBMR to estrogen replacement therapy.Methods:In this study, we undertook a characterization of estrogenic activity of QBMR using ovariectomized (OVX) rats and immature mice together with in vitro studies for further molecular characterization.OVX rats were treated with QBMR at doses of 0.875, 1.75,and 3.5 grams/kg per day for 8 weeks, and immature mice were treated with QBMR at doses of 1.125, 2.25, and 4.5 g/kg for 7 days, respectively.Results: QBMR treatments demonstrated significant estrogenic activity, as indicated by promoting development of uterus and vagina in immature mice.restoring the estrus cycle and reversal of atrophy of uterus and vagina in OVX mice, up-regulation of estrogen receptor (ER) αand ERβ expressions at protein and mRNA level in the reproductive tissues in immature and OVX mice, where ERα up-regulation was stronger than that of ERβ.Meanwhile, treatment with QBMR significantly increased adrenal gland weight and serum estradiol, clearly decreased follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in circulation of immature and OVX animals.QBMR significantly induced luciferase expression from the ERα/β-estrogen response element (ERE)luciferase reporter and upregulated ERα and ERβ expressions in MCF-7 cells.All activities were significantly inhibited by estrogen antagonist ICI182,780.Conclusion:This study demonstrates that QBMR activity is mediated through estrogenic components and ER-ERE-dependent pathway, and preferentially activates ERβ over ERα.This study also provides an evidence base for QBMR treatment of post-menopausal symptoms.