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Traditional Chinese medicine (TCM) formulas have attracted increasing attention worldwide in the past few years for treating complex disease including rheumatoid arthritis.However,their mechanisms are complex and remain unclear.Guan-Jie-Kang (GJK),a prescription modified from "Wu Tou Decoction," was found to significantly relieve arthritis symptoms in rats with adjuvant-induced arthritis after 30-day treatment,especially in the 24 g/kg/day group.By analyzing 1749 targets related to 358 compounds in the five herbs of GJK,we identified the possible anti-arthritis pathways of GJK,including the calcium signaling and metabolic pathways.Bone damage levels were assessed by micro-computed tomography,and greater bone protective effect was observed with GJK treatment than with methotrexate.Receptor activator of nuclear factor κB ligand (RANKL)-RANK signaling,which is related to calcium signaling,was significantly regulated by GJK.Moreover,a target metabolomics assay of serum was conducted;17 metabolic biomarkers showed significant correlations with treatment.An integrated pathway analysis revealed that pyruvate metabolism,purine metabolism,and glycolysis metabolism were significantly associated with the effects of GJK in arthritis treatment.Thus,this study establishes a new omics analytical method integrated with bioinformatics analysis for elucidating the multi-pathway mechanisms of TCM.