Astragali Radix (HQ) and Curcumae Rhizoma (EZ), collectively termed AC, are commonly used as an herb pair in Traditional Chinese Medicine (TCM) for colorectal cancer (CRC). Although AC has been used as an adjuvant to chemotherapy, the molecular basis by which it enhances the efficacy of 5-fluorouracil (5-FU) remains incompletely defined. The antitumor effect of AC alone and in combination with 5-FU was evaluated in a HCT116 xenograft mouse model and in CRC cell lines with different genetic backgrounds. Potential systemic toxicity was assessed using serum biochemical markers. Transcriptomic changes induced by AC were analyzed by RNA sequencing and integrated with network pharmacology and The Gene Expression Omnibus (GEO) (GSE17536, GSE17537, GSE41258, GSE20916). Key targets were validated by Western blotting and immunofluorescence. Cell viability, drug interaction, apoptosis, cell-cycle distribution and DNA damage were assessed using CCK-8 assays, Synergy Finder analysis, flow cytometry, and γ-H2AX staining. Bioavailable AC-derived constituents were characterized by UPLC-QTOF-MS, and candidate active compounds were further evaluated by molecular docking, molecular dynamics simulation, cellular thermal shift assay, and functional validation. AC potentiated the antitumor efficacy of 5-FU in vivo and did not exacerbate 5-FU-associated hepatic or renal biochemical toxicity under the tested conditions. In vitro, AC-containing serum extracts inhibited CRC cell viability, induced reversible G2/M-phase arrest, and enhanced 5-FU sensitivity in HCT116, CT26, and HCT15 cells. AC treatment caused limited early apoptosis, whereas delayed apoptosis was observed after prolonged exposure, suggesting that cell-cycle arrest represents the primary early response. RNA-seq and integrative analyses showed that AC suppressed pathways associated with DNA replication, mismatch repair, ribosome biogenesis, and cell-cycle progression. AC downregulated key regulators involved in cell-cycle control, DNA repair, and stress-survival signaling, including PLK1, CDK1, CCNB1, CDC25A, c-JUN, MSH2, BRCA1, and p-AKT. Combination treatment further increased γ-H2AX accumulation and reduced cell-cycle regulatory proteins. Functional validation indicated that c-JUN overexpression partially attenuated AC-mediated 5-FU sensitization, whereas pharmacological AKT inhibition partially mimicked the sensitizing effect of AC. Among the serum-exposed constituents, Demethoxycurcumin and Kumatakenin were identified as bioavailable active compounds that partially recapitulated the effects of AC by inducing G2/M-phase arrest and enhancing the antiproliferative activity of 5-FU. Molecular docking, molecular dynamics simulation, and CETSA supported their target engagement with cell-cycle regulators, particularly CCNB1 and CDK1. AC enhances the therapeutic efficacy of 5-FU in CRC by weakening DNA repair-cell-cycle coordination and stress-survival signaling, thereby increasing 5-FU-induced DNA damage and promoting cell-cycle arrest. Demethoxycurcumin and Kumatakenin may represent important bioavailable contributors to this chemosensitizing effect, although the complete AC extract likely acts through multi-component and multi-target mechanisms. These findings provide mechanistic support for the use of AC as an adjunctive strategy to improve 5-FU-based chemotherapy in CRC.
The traditional habitat of the Chinese medicinal plant Cistanches Herba (CH) is the deserts of northwestern China. Recently, CH cultivation has been introduced to saline-alkali land (SAL) in the Yellow River delta region. However, the effects of this SAL environment on the accumulation of secondary metabolites in CH remain unclear. This research aims to systematically characterize the chemical profiles of CH grown in SAL and to identify key chemical markers associated with the SAL environment. CH samples were collected from both SAL and traditional sandy land (SL). Untargeted metabolomics was performed using ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) to characterize the global metabolic profiles of CH from two habitats. Chemical markers associated with SAL environments were screened via weighted gene co-expression network analysis (WGCNA), targeted quantification by ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-MS), and compound loading contributions of principal component analysis (PCA). Subsequently, desorption electrospray ionization mass spectrometry imaging (DESI-MSI) was employed to visualize the in situ spatial distribution of partial screened constituents in mouse organs. Untargeted UPLC-QTOF-MS metabolomics identified 74 chemical constituents from CH samples, including 40 phenylethanoid glycosides, 13 iridoid glycosides, 9 lignans, and other minor constituents. Orthogonal projections to latent structures-discriminant analysis (OPLS-DA) clearly separated the SAL and SL sample groups, with 30 constituents showing prominent contributions to the discrimination. WGCNA revealed 23 metabolites correlated with SAL. By integrating these two screening methods, 12 constituents were preliminarily selected as candidate markers associated with SAL. Subsequently, combining PCA loading coefficients (with the PCA model validated by Kaiser–Meyer–Olkin and Bartlett’s sphericity tests) and targeted UPLC-TQ-MS quantification, 4 constituents-verbascoside, echinacoside, isoacteoside, and pinoresinol- were ultimately identified as characteristic chemical markers, all exhibiting significantly higher levels in SAL samples. DESI-MSI results further demonstrated that echinacoside predominantly accumulated in brain tissue, whereas verbascoside or isoacteoside displayed the strongest ion signal in the lung. This study developed an integrated strategy that combines hierarchical filtering of mass spectrometry data to successfully identify 4 characteristic chemical markers of CH associated with SAL. This strategy provides a scientific basis for future quality control and resource development.
Abstract Background Increasing studies suggest that supplementing flavonoids from Eucommia ulmoides Oliv. Leaves (EUL) exerts liver protective effects via reshaping gut microbiota and regulating bile acids (BAs) metabolism. However, the relationship between EUL, gut microbiota and BAs metabolism remains unclear and the potential mechanisms of action has not been expounded clearly. Purpose To demonstrate the important role of gut microbiota and microbial BAs metabolism in alleviation acetaminophen-induced liver injury (AILI) by EUL. Methods AILI mice model was employed to evaluate the therapeutic efficacy of EUL. The hepatic function was assessed, and biological samples were collected for histopathological examination, bacterial 16S rRNA sequencing analysis, BAs-targeted metabolomics, and the evaluation of the intestinal barrier function. Additionally, antibiotic cocktail-triggered gut microbiota deficiency and fecal microbiome transplantation were investigated to the relationship of molecular mechanistic between the protective efficiency of EUL in AILI, gut microbiota, and the metabolism of microbial BAs. Results EUL alleviate liver damage, involving reduction of liver function index, down-regulation of inflammatory cytokines and oxidative stress index, and restoration of pathological changes of liver tissue. EUL also improved intestinal barrier dysfunction and reshaped gut microflora dysbiosis in AILI mice. Moreover, antibiotic cocktail-triggered intestinal microbiota deficiency and fecal microbiome transplantation (FMT) evidenced that the liver protection of EUL was tightly related with gut microbiota. BAs-targeted metabolomics assay demonstrated that EUL mitigated the altered metabolism of microbial BAs during the development of AILI. EUL promoted the abundance of Firmicutes phylum, and elevating the beneficial microbes in Ruminococcaceae and Lachnospiraceae family, which were associated with up-regulated proportion of secondary bile acids (including DCA, NorDCA and 7-ketoLCA), leading to the reduction intestinal inflammation and preservation intestinal integrity. In particular, EUL enriched the beneficial microbes in Lachnoclostridium, Lachnospiraceae_UCG-006, [Eubacterium]_xylanophilum_group, Oscillibacter, Ruminiclostridium, and Desulfovibrio genera. Furthermore, EUL administration increased levels of CDCA, which could activate FXR-FGF15, enabling to decreased synthesis of BAs in the liver and promoting excretion. Conclusions EUL supplementation alleviate AILI by reshaping the dysfunctional gut microbiota and attenuating disordered BAs metabolism, resulting to improvements in BAs enterohepatic circulation and gut barrier function. The mechanism may be closely related to the gut microbiome-BAs-FXR signaling in gut-liver axis.
Abstract Background Traditional Chinese medicine (TCM) is a complex medical system characterized by multi-source data, implicit knowledge representation, and syndrome-based diagnostic and therapeutic reasoning. With the increasing digitization of classical texts and clinical records, large language models (LLMs) have attracted growing interest as computational tools for organizing and utilizing knowledge related to TCM. However, existing studies remain scattered, and a comprehensive overview of data resources, modeling strategies, evaluation practices, and application scenarios is still lacking. Overview In this review, we summarize current research on the application of LLMs in TCM based on published literature. We focus on commonly used data sources, including classical texts, clinical records, and related structured resources, and review representative modeling approaches such as knowledge-enhanced and multimodal methods. Domain-specific training and fine-tuning strategies, as well as reported evaluation practices, are also summarized. Furthermore, we review representative application scenarios described in the literature, including medical consultation support, syndrome differentiation assistance, prescription-related support, education, and research assistance. Key limitations, such as limited interpretability, safety concerns, and the lack of standardized evaluation frameworks, are discussed. Conclusion By organizing existing studies along the workflow from data construction to model development, evaluation, and application, this review aims to clarify the current research landscape and highlight methodological challenges that should be addressed to support cautious and appropriate use of LLMs in TCM research and practice.
Abstract Background Scalp acupuncture (SA) is used as adjunctive rehabilitation for post-stroke aphasia, but its effect on naming recovery and structural language-network changes remains unclear. We evaluated SA coupled with speech-language therapy (SLT) in patients with post-stroke anomia and explored diffusion tensor imaging markers of ventral-dorsal structural language networks. Methods This randomized controlled trial recruited 85 patients with post-stroke anomia and randomly assigned them to SA or control. Both groups received conventional stroke treatment and SLT; the SA group additionally received scalp acupuncture synchronously with SLT during needle retention. Treatment was delivered once daily, 5 days/week for 4 weeks. The primary outcome was WAB naming total score. Diffusion tensor imaging assessed white matter integrity and topological properties before and after treatment. Results Compared to the control group, the SA group showed greater improvements in the WAB naming total score ( P = 0.009), object naming, sentence completion, responsive speech, BNT spontaneous naming, and SAQOL-39 communication. In the ventral-dorsal structural network, the SA group showed smaller changes in the area under the curve (AUC) of small-worldness properties ( P = 0.008) and normalized clustering coefficient ( P = 0.017), and greater changes in clustering coefficient and local efficiency. In the SA group, the change in small-worldness was negatively correlated with the change in the WAB naming total score (r = −0.451, P = 0.027). The SA group also showed smaller axial diffusivity (AD) changes in the left uncinate fasciculus ( P = 0.003, Bonferroni corrected), which correlated negatively with the WAB naming total score change (r = −0.440, P = 0.031). Exploratory adjusted regression showed that SA-group assignment was associated with greater naming improvement after adjustment for baseline naming and clinical covariates. Conclusions SA coupled with SLT was associated with greater naming recovery in patients with post-stroke anomia. The benefit was accompanied by changes in structural language-network topology and white matter integrity, particularly involving the left UF. These findings support further evaluation of scalp acupuncture as an adjunct to language rehabilitation and suggest structural network plasticity as a relevant imaging correlate.
