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Objectives: To comprehensively analyze the copper metabolism in Breast cancer, we established a prognostic signature for breast cancer (BC) related to copper metabolism. Methods: Copper metabolism-related genes were sourced from previous literatures and were selected by the Univariate Cox regression. Cu-enrichment scores were calculated via ssGSEA. Differentially expressed genes were identified with limma between high and low Cu-enrichment scores group, then we used the Random Survival Forest and LASSO to build the CuScore for BC. Kaplan-Meier analysis, ROC curves, and Cox regression were used to evaluate CuScore. Genomic mutations were analyzed with GISTIC. Immune cells were examined using ESTIMATE, ssGSEA and TIMER. Enrichment analysis used clusterProfiler and GSVA. The GDSC database and oncoPredict package analyzed chemotherapeutic sensitivity. MMP13 was selected for in vitro assays. Results: Four copper metabolism-related genes (UBE2D2, SLC31A1, ATP7A, and MAPK1) with prognostic value were identified. Higher expression levels of these genes were associated with higher Cu-enrichment scores, a factor of malignancy in breast cancer. Among 115 differentially expressed genes, 19 prognostic genes were identified, with three (CEACAM5, MMP13, and CRISP3) highlighted by Random Survival Forest and LASSO. Higher CuScores correlated with worse prognoses and were effective in predicting breast cancer outcomes. CuScore and metastasis were independent prognostic factors. Tumor-infiltrating immune cells were associated with lower CuScores. GO-GSEA analysis indicated six immune-related pathways might be regulated by CuScore. Patients with higher CuScores had lower TMB and were more sensitive to Sapitinib and LCL161, while those with lower CuScores might respond better to anti-PD1 therapy. High MMP13 expression in breast cancer was linked to malignancy, affecting cell proliferation and migration. Conclusion: The identified copper metabolism-related gene signature has the potential to predict prognosis and guide clinical treatment for BC. Among these genes, MMP13 may act as a malignant factor in BC.

Hot water extract of Chlorella vulgaris (CVE) is a biologically substance that enhances organism’s immune function and antioxidative capacity. This study evaluated the effect of supplementation with various concentrations of CVE on muscle nutritional components, non-specific immunity, antioxidation, and resistance to non-ionic ammonia (NH3-N) stress in Litopenaeus vannamei over 45 days using diets supplemented with CVE at five different concentrations (0%, 0.5%, 1%, 5%, 10%, and 15%). Specifically, fresh and sweet amino acids (Asp, Glu) significantly increased (P < 0.05) in shrimp fed the 1% CVE diet, reaching 18.12 g/kg and 33.08 g/kg, respectively. Bitter amino acids (Leu) and Hypoxanthine (Hx) significantly decreased (P < 0.05) in shrimp fed the 1% CVE diet, at 10.56 g/kg and 10.56 ug/g. CVE supplementary enhanced the activities of acid phosphatase, alkaline phosphatase, and nitric oxide synthase while decreasing malondialdehyde levels. Shrimp fed with a 1% CVE diet exhibited significantly higher enzyme activity than the control group under NH3-N conditions (P < 0.05). Overall, this study demonstrated that 1% CVE as a feed additive significantly improved the muscle mass, boosted immunity and reduced the stress response to NH3-N in L. vannamei. This research provides a valuable reference for the application of CVE as a feed additive in crustacean aquaculture.