Background: Esophageal squamous cell carcinoma (ESCC) is a common digestive tract malignant tumor with high incidence and dismal prognosis worldwide. However, the reliable biomarkers for clinical diagnosis and the underlying signaling pathways insights of ESCC are not unequivocally understood yet. The serum proteome may provide valuable clues for the early diagnosis of ESCC and the discovery of novel molecular insights. Methods: In the current study, an optimized proteomics approach was employed to discover novel serum-based biomarkers for ESCC, and unveil abnormal signal pathways. Gene ontology (GO) enrichment analysis was done by Gene Set Enrichment Analysis (GSEA) and Metascape database, respectively. Pathway analysis was accomplished by GeneCards database. The correlation coefficient was assessed using Pearson and distance correlation analyses. Prioritized candidates were further verified in two independent validation sets by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) staining. Results: A total of 633 non-redundant proteins were identified in the serum of patients with ESCC, of which 59 and 10 proteins displayed a more than 1.5-fold increase or decrease compared with healthy controls. Verification was performed for six candidate biomarkers, including S100A8/A9, SAA1, ENO1, TPI1 and PGAM1. Receiver operating characteristics (ROC) curve plotting showed the high diagnostic sensitivity and specificity of these six protein molecules as a biomarker panel: the area under characteristic curve (AUC) is up to 0.945. Differentially expressed proteins were subjected to functional enrichment analysis, which revealed the dysregulation of signaling pathways mainly involved in glycolysis, TLR4, HIF-1α, Cori cycle, TCA cycle, folate metabolism, and platelet degranulation. The latter finding was all the more noteworthy as a strong positive correlation was discovered between activated glycolysis and TLR4 pathways and unfavorable clinicopathological TNM stages in ESCC. Conclusions: Our findings propose a potential serum biomarker panel for the early detection and diagnosis of ESCC, which could potentially broaden insights into the characteristics of ESCC from the proteomic perspective.

Ligustici Rhizoma et Radix (LReR) is the dried rhizomes and roots of Ligusticum sinese Oliv. (LS) or Ligusticum jeholense Nakai et Kitag. (LJ). However, in the market, LS and LJ are frequently confused with each other. Since the volatile oils are both the main active components and quality control indicators of LReR, a strategy combining gas chromatography-mass spectrometry (GC-MS) and chemical pattern recognition (CPR) was used to compare the volatile components of LJ and LS. Total ion chromatography (TIC) revealed that phthalides (i.e., neocnidilide) and phenylpropanoids (i.e., myristicin) could be thought of as the most critical components in the volatile oils of LJ and LS, respectively. In addition, the chemical components of the volatile oils in LJ and LS were successfully distinguished by hierarchical cluster analysis (HCA) and principal component analysis (PCA). Moreover, two quality markers, including myristicin and neocnidilide, with a very high discriminative value for the classification of LJ and LS, were found by orthogonal partial least squares discriminant analysis (OPLS-DA). The relative contents of myristicin and neocnidilide were 10.86 ± 6.18% and 26.43 ± 19.63% for LJ, and 47.43 ± 12.66% and 2.87 ± 2.31% for LS. In conclusion, this research has developed an effective approach to discriminating LJ and LS based on volatile oils by combining GC-MS with chemical pattern recognition analysis.

Several monoterpene glycoside compounds were extracted from Paeonia lactiflora Pall. Among them, paeoniflorin, a water-soluble monoterpene glycoside found in the root of Paeonia lactiflora Pall, exhibits excellent antioxidant pharmacological functions. Initially, Sc(CF₃SO₃)₃ was employed as the catalyst for paeoniflorin’s dehydration and rearrangement reactions with alcohols. Subsequently, structural modifications were performed on paeoniflorin through a series of responses, including acetylation, deacetylation, and debenzoylation, ultimately yielding 46 monoterpene glycoside derivatives. The potential inhibitory effects on the pro-inflammatory mediators interleukin-1 beta (IL-1β) and nitric oxide (NO) were assessed in vitro. The results revealed that compounds 29 and 31 demonstrated notable inhibition of NO production, while eight derivatives (3, 8, 18, 20, 21, 29, 34, and 40) displayed substantial inhibitory effects on the secretion of IL-1β. Computational research was also undertaken to investigate the binding affinity of the ligands with the target proteins. Interactions between the proteins and substrates were elucidated, and corresponding binding energies were calculated accordingly. The findings of this study could provide valuable insights into the design and development of novel anti-inflammatory agents with enhanced pharmacological properties.