Introduction: Jian-Pi-Yi-Shen pill (JPYSP) is a Chinese medicine formula developed for the treatment of anaemic patients with chronic kidney disease (CKD). Objective: To investigate the chemical profile of JPYSP in the treatment of renal anaemia. Methods: A method coupling ultra-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was established to characterise the chemical constituents present in JPYSP. Subsequently, a high-performance liquid chromatography method coupled with triple-quadrupole tandem mass spectrometry (HPLC-QQQ-MS/MS) was developed to quantify the major constituents from the identified compounds related to the treatment of CKD and anaemia. Results: A total of 71 compounds were tentatively identified from JPYSP, including saponins, flavonoids, sesquiterpenoids, coumarins, phenylpropanoids, anthranones, anthraquinones, tannins, phenolic acids and others. Amongst them, 12 compounds (i.e. astragaloside IV, calycosin, calycosin 7-O-glucoside, salvianolic acid A, rosmarinic acid, rhein, liquiritin, formononetin, atractylenolide I, dioscin, tanshinone IIA, and acteoside) were further quantified simultaneously by HPLC-QQQ-MS/MS. Conclusion: The newly developed approach is suitable for the chemical profiling analysis and quality control of JPYSP, and could lead to additional pharmacodynamic studies involving the components of JPYSP.

Huo-Xue-Jiang-Tang Yin (HXJTY) is a Chinese medicine formulation, which has been widely used for the treatment of various lipometabolism- and glycometabolism-related diseases in clinics. Currently, HXJTY is mainly prescribed to treat patients with type 2 diabetes mellitus (T2DM), yet its chemical and pharmacologic profiles remain to be elucidated. Here, the potential bioactive compound and action mechanism were investigated using chemical and network pharmacology analysis. A rapid HPLC-MS was employed to identify and quantify the component of HXJTY. On the basis of the identified chemical markers from HXJTY, a network pharmacology study, including target gene prediction and functional enrichment, was applied to screen out the main quality markers of HXJTY and explore its potential mechanism for the treatment of T2DM. The results showed that a total of 22 components were identified and quantified from HXJTY by HPLC-MS. Furthermore, 12 active components such as astragaloside IV, calycosin-7-O-β-D-glucoside, hydroxysafflor yellow A, and others were proposed as quality markers of HXJTY for treating T2DM based on network pharmacology analysis. In addition, 125 corresponding possible therapeutic target genes of T2DM were obtained. These target genes are mainly related to peptidase activity, hydrolase activity, phosphatase activity, and cofactor binding, suggesting the involvement of PI3K-Akt, MAPK, AGE-RAGE, and Rap1 signaling pathways in HXJTY-treated T2DM. Our results may provide a useful approach to identify potential quality markers and molecular mechanism of HXJTY for treating T2DM.

Melanoma is the most lethal type of skin cancer. Despite the breakthroughs in the clinical treatment of melanoma using tumor immunotherapy, many patients do not benefit from these immunotherapies because of multiple immunosuppressive mechanisms. Therefore, there is an urgent need to determine the mechanisms of tumor-immune system interactions and their molecular determinants to improve cancer immunotherapy. In this study, combined analysis of microarray data and single-cell RNA sequencing data revealed the key interactions between immune cells in the melanoma microenvironment. First, differentially expressed genes (DEGs) between normal and malignant tissues were obtained using GEO2R. The DEGs were then subjected to downstream analyses, including enrichment analysis and protein–protein interaction analysis, indicating that these genes were associated with the immune response of melanoma. Then, the GEPIA and TIMER databases were used to verify the differential expression and prognostic significance of hub genes, and the relationship between the hub genes and immune infiltration. In addition, we combined single cell analysis from GSE123139 to identify immune cell types, and validated the expression of the hub genes in these immune cells. Finally, cell-to-cell communication analysis of the proteins encoded by the hub genes and their interactions was performed using CellChat. We found that the CCL5-CCR1, SELPLG-SELL, CXCL10-CXCR3, and CXCL9-CXCR3 pathways might play important roles in the communication between the immune cells in tumor microenvironment. This discovery may reveal the communication basis of immune cells in the tumor microenvironment and provide a new idea for melanoma immunotherapy.

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