Organ fibrosis is an important pathological basis for the progression of many chronic diseases to the end stage. Its development is closely associated with chronic injury, inflammatory and immune responses, fibroblast activation, extracellular matrix (ECM) deposition, and tissue structural remodeling. Current antifibrotic therapies have limited efficacy, and advanced fibrosis remains difficult to reverse. Traditional Chinese medicine (TCM), characterized by multiple components and multiple targets, has been investigated as a potential approach for the prevention and treatment of organ fibrosis. This review summarizes recent progress in the use of TCM for organ fibrosis, including the pathological basis of fibrosis and TCM concepts of “root deficiency with branch excess” and “chronic illness entering the collaterals.” Particular attention is given to mechanisms involving transforming growth factor-β/Smad, nuclear factor-κB, and nuclear factor E2 related factor 2/heme oxygenase-1 signaling and to processes including inflammation, oxidative stress, ECM metabolism, cell phenotypic transition, autophagy, ferroptosis, cellular senescence, and metabolic reprogramming. The current research on representative TCM formulas, proprietary Chinese medicines, and active compounds for liver, pulmonary, renal, and myocardial fibrosis is also reviewed. Current studies on TCM antifibrosis still have limitations such as insufficiently clarified mechanisms of action, unclear pharmacodynamic material basis, and inadequate evidence of long-term efficacy and safety. Future studies should identify the major active components and molecular targets of TCM by integrating multi-omics, single-cell sequencing and other technologies, and evaluate long-term efficacy and safety in well-designed clinical trials, so as to provide evidence for the clinical application of TCM in the prevention and treatment of organ fibrosis.

Liver fibrosis is a chronic wound-healing response of the liver, essentially characterized by diffuse excessive deposition and abnormal distribution of the extracellular matrix (ECM). Although etiological therapy can slow fibrosis progression, drug development directly targeting the fibrotic process still faces substantial challenges, and clinical options remain extremely limited. Traditional Chinese medicine (TCM) has attracted extensive attention in the field of liver fibrosis research due to its multi-component, multi-target, and multi-pathway properties. However, current studies are still predominantly based on preclinical experiments, and systematic integration of mechanisms and sorting of clinical evidence are still inadequate. This review outlines the pathogenesis of liver fibrosis, with emphasis on the polarization of Kupffer cells, the capillarization of liver sinusoidal endothelial cells, the activation of hepatic stellate cells, and ECM metabolic imbalance. Antifibrotic effects of active components from herbs are summarized by chemical categories. Furthermore, the interactive network of signaling pathways is integrated from three hierarchical levels—the “inflammation initiation, metabolic regulation, and fibrotic progression”—to explore the systemic mechanisms underlying synergistic modulation by multiple components of TCM. Finally, the limitations of current research are identified, and future directions driven by cutting-edge technologies, including multi-omics integration, single-cell sequencing, artificial intelligence-based screening, and etiology-stratified precision therapy, are discussed. This review provides insights into mechanism interpretation and clinical translation of TCM against liver fibrosis.

Pulmonary fibrosis (PF) is the terminal manifestation of various interstitial lung diseases. It is pathologically characterized by progressive destruction of alveolar structure and functional alveolar units, as well as persistent fibrosis of lung tissue. The prognosis is extremely poor, seriously affecting the quality of life of patients. In the pathological process of PF, there are typical “senescence-associated features” such as telomere shortening, resistance to apoptosis, and senescence-associated secretory phenotype. Cellular senescence, as one of the pathogenic mechanisms, plays an important role in the occurrence and development of PF. Studies have shown that the active ingredients of traditional Chinese medicine (TCM), single medicinal herbs and compound preparations of TCM can achieve the therapeutic effect of delaying the progression of PF by directly or indirectly inhibiting cellular senescence. Therefore, this article elaborates on the connection between cellular senescence and PF, summarizes the current research status of TCM in treating PF by inhibiting cellular senescence in recent years, and provides a reference for the clinical treatment of PF.

Hepatic fibrosis is the pathological consequence of abnormal extracellular matrix deposition in the liver during the self-repair process following chronic injury. Persistent hepatic fibrosis can progress to malignant conditions such as cirrhosis and liver cancer. Currently, no specific therapeutic drugs exist for hepatic fibrosis, therefore, early prevention and intervention are of great significance for delaying disease progression and reducing the incidence of severe liver diseases. In recent years, terpenoids have emerged as a research hotspot due to their significant multi-target and multi-pathway anti-hepatic fibrosis effects. Their mechanisms are associated with anti-inflammation, antioxidative stress, anti-apoptosis, and the regulation of extracellular matrix deposition and degradation. This article systematically reviews natural terpenoids with anti-hepatic fibrosis activity reported in domestic and international literature in recent years, and summarizes their chemical structures and molecular mechanisms, providing a theoretical basis and research perspectives for the research and development of innovative anti-hepatic fibrosis drugs derived from terpenoids.

Objective To explore the mechanisms by which by which senkyunolide A (SenA) ameliorated liver fibrosis via inducing hepatic stellate cells (HSCs) senescence in vitro and in vivo. Methods SenA targets and liver fibrosis related targets were obtained using the SwissTargetPrediction database and GeneCards database. The intersection targets were used for gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. Metascape was used to analyze protein-protein interaction (PPI) network, core targets were screened and confirmed, functional enrichment analysis on core targets was performed, and molecular docking with SenA was performed. A mouse model of liver fibrosis was established using common bile duct ligation. Human hepatic stellate LX-2 cells and rat hepatic stellate HSC-T6 cells were induced using transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor-BB (PDGF-BB), respectively. After SenA intervention, senescence indicators were characterized by senescence-associated-β-galactosidase (SA-β-Gal) staining, EdU staining, qRT-PCR, etc. Results The results of network pharmacology revealed that SenA and liver fibrosis shared 39 common targets, the common target genes were enriched in cellular senescence pathways. A total of 19 genes obtained based on PPI network may be the core targets of SenA on regulation of liver fibrosis progression, and it was found that core targets related to cellular aging are highly expressed in HSCs. Combining molecular docking experiments, it was found that SenA had the strongest binding ability to Aurora kinase A (AURKA), suggesting that AURKA may be a key molecular target for SenA in HSCs. The results of in vivo and in vitro experiments showed that SenA significantly induced HSCs senescence (P < 0.05, 0.01, 0.001), significantly downregulated the expressions of proliferation related genes Cyclin D1 (CCND1) and cyclin dependent kinase 2 (CDK2) (P < 0.05, 0.01), and upregulated the expressions of senescence related genes p21, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) (P < 0.05, 0.01), thereby significantly inhibiting HSC activation and collagen deposition (P < 0.05, 0.01, 0.001). Conclusion SenA induces HSC senescence to inhibit HSC activation, thereby improving liver fibrosis.

Objective To explore the mechanism of Sedum sarmentosum extract in improving liver fibrosis based on adenosine monophosphate-activated protein kinase (AMPK)/silent information regulator 1 (SIRT1)/liver X receptors (LXRs) signaling pathway. Methods A rat model of liver fibrosis was established using ip dimethylnitrosamine (DMN), and after intervention with S. sarmentosum extract or Fuzheng Huayu Capsules (扶正化瘀胶囊) for five weeks, serum and liver tissue were collected. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were determined. Hematoxylin-eosin (HE) and Masson staining were used to detect pathological changes in liver tissue. Immunohistochemistry, Western blotting and qRT-PCR were used to detect fibrosis, apoptosis, inflammation, AMPK/SIRT1/LXRs signaling pathway related proteins and gene expressions in liver tissue. Transforming growth factor-β (TGF-β) was used to stimulate hepatic stellate cells (HSCs) for 2 h, followed by treatment with S. sarmentosum extract or AMPK agonist AICAR or LXRβ agonist GW3965 for 6 h. Immunofluorescence staining and Western blotting were used to detect the expressions of fibrosis and AMPK-SIRT1-LXRs signaling pathway related proteins. After transfecting LX-2 cells with AMPK or LXRβ siRNA, TGF-β stimulation was given for 2 h, followed by treatment with S. sarmentosum extract for 6 h. Western blotting was used to detect the expressions of AMPK/SIRT1/LXRs signaling pathway related proteins. Results The results of in vivo experiments showed that S. sarmentosum extract significantly reduced the activities of ALT and AST in serum of rats induced by DMN (P < 0.001), improved pathological changes and extracellular matrix deposition in liver tissue, inhibited cell apoptosis and inflammatory factor expressions in liver tissue (P < 0.001), and upregulated the expressions of AMPK/SIRT1/LXRs signaling pathway related proteins (P < 0.05, 0.01, 0.001). The in vitro experimental results showed that S. sarmentosum extract significantly inhibited the expressions of extracellular matrix related proteins in activated HSCs (P < 0.001), and upregulated the expressions of AMPK-SIRT1-LXRs signaling pathway related proteins (P < 0.001), with similar effects as AMPK agonists or LXRβ agonists. In addition, the absence of AMPK or LXRβ significantly weakened the inhibition of α-smooth muscle actin (α-SMA) in activated LX-2 cells and the activation of AMPK/SIRT1/LXRs signaling pathway by S. sarmentosum extract (P < 0.001). Conclusion S. sarmentosum extract effectively improves the progression of liver fibrosis by mediating AMPK/SIRT1/LXRs signaling pathway, to regulate extracellular matrix homeostasis and inflammatory responses.

Objective To investigate the antifibrotic effect of combined treatment with sodium arsenite (SA, biotransformation product of arsenic trioxide) and N-acetylcysteine (NAC) on pulmonary fibrosis. Methods L929 fibroblasts were treated with SA or NAC for 24 h before measuring cell viability. The combination drug ratio was determined by examining the effects of different ratios of SA and NAC on viability of L929 cells, and the cell migration ability was detected using scratch assay and Transwell assay. Immunofluorescence assay was used to detect the effects of combined administration of SA and NAC on the expressions of α-smooth muscle actin (α-SMA), collagen type I α1 (COL1A1), and reactive oxygen species (ROS) level in transforming growth factor-β1 (TGF-β1)-induced L929 cells. A mouse model of idiopathic pulmonary fibrosis was established induced by bleomycin, after intervention with SA or NAC for 28 d, the survival and body weight changes of mice were observed. Hematoxylin-eosin (HE) and Masson staining were used to observe pathological changes in lung tissue. The total protein content, TGF-β1 level in bronchoalveolar lavage fluid (BALF), and hydroxyproline (Hyp) level in lung tissue were detected. Immunofluorescence assay was used to detect the expressions of α-SMA, COL1A1, CD86 and CD206 in lung tissue. Results The synergistic effect of SA and NAC combination therapy was related to their ratio. When SA∶NAC = 2∶1, the combination could significantly inhibit viability and migration of L929 cells (P < 0.001), and was superior to the monotherapy group (P < 0.01, 0.001). The combined administration of SA and NAC (2∶1) significantly down-regulated the expressions of α-SMA and COL1A1 in L929 cells induced by TGF-β1 (P < 0.001), and reduced ROS level (P < 0.01). The in vivo experimental results showed that compared with model group, the combined administration of SA and NAC (2∶1) could prolong the survival cycle of mice, increase body weight, reduce lung tissue damage and collagen deposition, decrease the total protein content, TGF-β1 level in BALF and Hyp level in liver tissue (P < 0.01, 0.001), reduce the expression of α-SMA and COL1A1 in lung tissue (P < 0.001), inhibit M2 polarization of lung tissue macrophages (P < 0.001), and had no significant toxic effects on major organs. Conclusion The combination of SA and NAC has a synergistic anti-pulmonary fibrosis effect, providing experimental evidence for the treatment of idiopathic pulmonary fibrosis with arsenic agents combined with thiol donors.

Objective To investigate the mechanism by which Huangqi (Astragali Radix)-Danshen (Salviae Miltiorrhizae Radix et Rhizoma) herb pair (HQ-DS) in treating hepatic fibrosis (HF) via regulating the endogenous retroviruses (ERVs)/retinoic acid-inducible gene I-like receptors (RLRs)/type I interferon (IFN-I) signaling pathway. Methods The components of HQ-DS extract and drug-containing serum were analyzed and identified using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). A total of 48 male C57BL/6J mice were randomly divided into control group, model group, HQ-DS low-, medium-, high-dose (4.55, 9.10, 18.20 g/kg) groups and silymarin (100 mg/kg) group, with eight mice in each group. A mouse model of HF was established using carbon tetrachloride (CCl₄), and after drug intervention, liver function related indicators in serum were detected. The pathological changes of liver tissue through hematoxylin eosin (HE) staining, Masson and Sirius red staining were observed. Flow cytometry was used to detect macrophage infiltration in liver tissue. Western blotting was used to detect the expressions of fibrosis related proteins in liver tissue. High-throughput transcriptome sequencing was used to analyze changes in gene transcription levels in liver tissue. The expressions of factors related to the ERVs/RLRs/IFN-I signaling pathway in liver tissue were detected using qRT-PCR, Western blotting and ELISA. A HF model was established in Mavs^-/- mice and intervened with HQ-DS to verify the mechanism of HQ-DS’s anti-HF effect. In vitro experiments were conducted using CCK-8 assay to evaluate the effect of HQ-DS on the activity of iBMDM cells stimulated by lipopolysaccharides (LPS) and nigericin. Immunofluorescence detection of dsRNA expression was performed, qRT-PCR and Western blotting were used to further verify the inhibitory effect of HQ-DS on the activation of ERVs. Results Compared with control group, the body weight of mice in model group was increased slowly, with vacuolar degeneration and inflammatory cell infiltration in liver tissue, liver function related indicators in serum were significantly increased (P < 0.001), and the expressions of factors related to the ERVs/RLRs/IFN-I signaling pathway in liver tissue were significantly increased (P < 0.001). Compared with model group, HQ-DS and silymarin intervention significantly improved the slow growth of body weight in mice (P < 0.01, 0.001), significantly reduced the degree of vacuolar degeneration and inflammatory infiltration in liver tissue, significantly reduced liver function related indicators in serum (P < 0.01, 0.001), effectively alleviated the process of HF, and significantly reduced the expressions of factors related to the ERVs/RLRs/IFN-I signaling pathway in liver tissue (P < 0.05, 0.01, 0.001). Subsequent verification of the underlying mechanisms revealed that in Mavs^-/- mice, the elevation of pro-inflammatory factors induced by CCl₄ was markedly suppressed (P < 0.001), the mRNA expression levels of factors associated with the ERVs/RLRs/IFN-I signaling pathway were significantly downregulated (P < 0.001). However, following the administration of HQ-DS, there was no significant change in the mRNA expression levels of inflammatory factors and factors related to the ERVs/RLRs/IFN-I pathway. In cell experiments, compared with model group, HQ-DS significantly increased iBMDM cell viability (P < 0.001), reduced dsRNA accumulation, and downregulated the expressions of core genes in ERVs and IFN-I pathways (P < 0.05, 0.01, 0.001), while inhibited the expressions of key proteins [phosphorylated TANK binding kinase 1 (p-TBK1), phosphorylated interferon regulatory factor 3 (p-IRF3)] in RLRs pathway (P < 0.05, 0.01, 0.001). Conclusion HQ-DS alleviates inflammatory injury and ameliorates the progression of HF in mice by inhibiting ERVs/RLRs/IFN-I signaling pathway.

Objective To explore the protective effect of herb pair Kuxingren (Armeniacae Semen Amarum)-Kushen (Sophorae Flavescentis Radix) on bleomycin-induced idiopathic pulmonary fibrosis (IPF) in rats and investigate its mechanism. Methods SD rats were randomly divided into control group, model group, pirfenidone (200 mg/kg) group, Armeniacae Semen Amarum-Sophorae Flavescentis Radix low-, medium-and high-dose (1, 2, 4 g/kg) groups. The IPF rat model was established by one-time tracheal instillation of bleomycin sulfate (5 mg/kg). After drug intervention, the pathological changes in lung tissue were observed using hematoxylin-eosin (HE) and Masson staining. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6 and hydroxyproline (Hyp) in serum and bronchoalveolar lavage fluid. TUNEL staining was used to detect cell apoptosis in lung tissue. The expressions of α-smooth muscle actin (α-SMA), collagen type I (collagen-I) and fibronectin-1 (FN-1) in lung tissue were detected by immunofluorescence. Western blotting was used to detect the expressions of p-p38, phosphorylated extracellular signal regulated kinase (p-ERK), phosphorylated c-Jun N-terminal kinase (p-JNK), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in lung tissue. Transforming growth factor-β1 (TGF-β1) was used to induce human lung fibroblasts (MRC-5) in vitro model. After intervention with Armeniacae Semen Amarum-Sophorae Flavescentis Radix containing serum, cell proliferation activity was detected using CCK-8 method. TUNEL staining was used to detect cell apoptosis. The expressions of α-SMA, collagen-I and FN-1 were detected by immunofluorescence. Western blotting was used to detect the expressions of p-RAF, phosphorylated mitogen activated protein kinase (p-MEK), p-ERK, Cyclin D1, proliferating cell nuclear antigen (PCNA), Bax and Bcl-2 proteins. Results In the in vivo experiment, compared with control group, the body weight of rats in model group was decreased (P < 0.01), the dry wet mass ratio and lung index of lung tissue were increased (P < 0.01), the lung tissue showed significant inflammatory infiltration, thickening and edema of alveolar space, and the cell apoptosis rate of lung tissue was decreased (P < 0.01), the levels of inflammatory factors in serum and bronchoalveolar lavage fluid were increased (P < 0.01), the protein expressions of α-SMA, collagen-I, FN-1, p-p38, p-ERK, p-JNK and Bcl-2 in lung tissue were significantly up-regulated (P < 0.01), while the protein expression of Bax was significantly down-regulated (P < 0.01). Compared with model group, the body weight of rats in each treatment group were increased (P < 0.01), the dry wet mass ratio and lung index of lung tissue were decreased (P < 0.01), the pathological changes of lung tissue were significantly improved, the cell apoptosis rate of lung tissue was increased (P < 0.01), the levels of inflammatory factors in serum and bronchoalveolar lavage fluid were decreased (P < 0.05, 0.01), the expressions of α-SMA, collagen-I, FN-1, p-p38, p-ERK, p-JNK and Bcl-2 proteins in lung tissue were significantly down-regulated (P < 0.01), and the expression of Bax protein was significantly up-regulated (P < 0.01). In the in vitro experiments, compared with control group, cell proliferation activity in model group was increased (P < 0.01), cell apoptosis rate was decreased (P < 0.01), α-SMA, collagen-Ⅰ, FN-1, p-RAF, p-MEK, p-ERK, Cyclin D1, PCNA, Bcl-2 protein expressions were increased (P < 0.01), and Bax protein expression was decreased (P < 0.01). Compared with model group, Armeniacae Semen Amarum-Sophorae Flavescentis Radix containing serum and RAF inhibitor GW5074 intervention resulted in a decrease in cell proliferation activity (P < 0.05, 0.01), an increase in cell apoptosis rate (P < 0.01), a decrease in the expressions of α-SMA, collagen-I, FN-1, p-RAF, p-MEK, p-ERK, Cyclin D1, PCNA, Bcl-2 proteins (P < 0.05, 0.01), and an increase in Bax protein expression (P < 0.05, 0.01). Conclusion Armeniacae Semen Amarum-Sophorae Flavescentis Radix exerts an anti-IPF effect by regulating RAF/MEK/ERK signaling pathway, inhibiting inflammatory response, suppressing cell proliferation and promoting cell apoptosis.

Objective To explore the effect and mechanism of Yanggan Ruanjian Wan (养肝软坚丸, YGRJW) in prevention and treatment of hepatic fibrosis (HF) based on in vivo and in vivo experiments and transcriptome sequencing technology. Methods LX-2 cell activation model was induced by transforming growth factor-β1 (TGF-β1), CCK-8 method, cell scratch assay, qRT-PCR and Western blotting were used to detect the effects of YGRJW-medicated serum on cell viability, proliferation, expressions of inflammatory cytokines [interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α)] and fibrosis markers [α-smooth muscle actin (α-SMA), collagen type I alpha 1 chain (COL1A1), collagen type III alpha 1 chain (COL3A1)]. Transcriptome sequencing was employed to identify differentially expressed genes and key pathways in activated LX-2 cells treated with YGRJW-containing serum, followed by validation through qRT-PCR and immunofluorescence experiments. A HF model was induced in mice via intraperitoneal injection of a 10% carbon tetrachloride (CCl₄)-containing corn oil solution. After drug intervention, the therapeutic effects of YGRJW on HF mice were comprehensively evaluated by measuring body weight, organ indices, liver and kidney function, pathological changes in liver tissue, expressions of inflammatory factors and fibrosis markers in liver tissue, as well as levels of inflammatory factors and four liver fibrosis indicators in serum, while also verifying the effect of YGRJW on potential pharmacodynamic targets and key pathway indicators. Results In vitro experiments demonstrated that 10% YGRJW-containing serum significantly inhibited LX-2 cell activation after 24 h (P < 0.01) and downregulated mRNA expression levels of inflammatory factors and fibrosis markers (P < 0.05, 0.01, 0.001), with superior inhibitory effects compared to 10% silybinin-containing serum. Transcriptomic analysis and validation results showed that YGRJW may exert anti-HF effects through the colony stimulating factor 3 receptor (CSF3R)/Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway. Compared with model group, YGRJW drug containing serum intervention could significantly downregulate the mRNA and protein expressions of CSF3R, JAK2 and STAT3 (P < 0.05, 0.01). The in vivo experimental results showed that YGRJW could significantly improve the slow growth of body weight in HF mice (P < 0.05), reduce liver index, spleen index, liver and kidney function indicators (P < 0.05, 0.01, 0.001), alleviate liver tissue inflammation infiltration and collagen deposition, reduce the mRNA expression levels of TGF-β, TNF-α, IL-6, α-SMA, COL1A1, COL3A1 and tissue inhibitor of metalloproteinases-1 (TIMP-1) in liver tissue (P < 0.05, 0.01, 0.001), increase the mRNA expression level of fibroblast growth factor 7 (FGF7) (P < 0.05, 0.01), and reduce levels of inflammatory factors and four indicators of liver fibrosis in serum (P < 0.05, 0.01, 0.001), and downregulated mRNA and protein expressions of CSF3R, JAK2, STAT3 in liver tissue (P < 0.05, 0.01). Conclusion YGRJW exerts anti-HF effects by regulating CSF3R/JAK2/STAT3 signaling pathway, to inhibit hepatic stellate cell (HSC) activation and reducing extracellular matrix (ECM) deposition.

Objective To evaluate the causal associations of gut microbiota, immune cells and serum metabolites with idiopathic pulmonary fibrosis (IPF) based on the multi-omics Mendelian randomization (MR) framework, screen core causal genes, and predict intervening traditional Chinese medicine (TCM) and compounds, so as to provide genetic evidence for the early diagnosis and TCM intervention of IPF. Methods Genetic instrumental variables (IVs) of gut microbiota, serum metabolites and immune cells were extracted, and their causal associations with IPF were analyzed by two-sample MR. Adjacent genes of IVs were annotated to obtain the "microbiota-immunity-metabolism" causal gene set. Integrated with IPF transcriptome data, candidate causal genes were screened by differentially expressed genes (DEGs) analysis and weighted gene co-expression network analysis (WGCNA). Diagnostic models were constructed by machine learning to screen core genes. Immune cell infiltration and single-cell RNA sequencing profiles were analyzed. Potential intervening TCM and compounds were predicted via Comparative Toxicogenomics Database (CTD) and Integrated Traditional Chinese Medicine Database (ITCM), followed by molecular docking verification. Results Two-sample MR identified 2 gut microbiota taxa (Lactobacillaceae as protective factor, Proteobacteria as risk factor), 19 immune cell phenotypes (nine protective, ten risk) and 26 serum metabolites (12 protective, 14 risk) that were significantly causally associated with IPF (P < 0.05). A total of 21 candidate causal genes were obtained by integrated analysis, and 5 core genes were screened by machine learning: adaptor protein complex 5 subunit mu 1 (AP5M1), SRY-box transcription factor 11 (SOX11), zinc finger and MIZ-type containing 2 (ZMIZ2), fragile histidine triad (FHIT) and cyclin dependent kinase 6 (CDK6), all with potential diagnostic value. Immune infiltration showed increased activated mast cells and monocytes, and decreased M0/M2 macrophages and activated natural killer cells in IPF tissues. Single-cell analysis revealed the highest enrichment of FHIT in multiple cell types. A total of 150 kinds of potential intervention Chinese medicine were screened. The high frequency Chinese medicine included Sangbaipi (Mori Cortex), Yuejihua (Rosae Chinensis Flos), Yanhusuo (Corydalis Rhizoma), etc. The active ingredients included kaempferol, naringenin, wogonin, sanguinarine, etc. Molecular docking confirmed that the core compound could stably bind to the target. Conclusions This study reveals the causal associations of gut microbiota, immune cells and serum metabolites with IPF, and identifies AP5M1, SOX11, ZMIZ2, FHIT and CDK6 as potential candidate diagnostic biomarkers for IPF. It also predicts the potential intervention value of active compounds such as kaempferol, naringenin and palmatine, as well as TCM including Morus alba and Rosa chinensis, providing genetic evidence and theoretical support for the diagnosis and treatment of IPF.

Objective To investigate the chemical constituents from the whole plant of Physalis angulata and their anti-inflammatory activities. Methods Various chromatographic techniques were employed for the isolation and purification of compounds, and their structures were elucidated by spectroscopic methods including NMR, MS, and UV. The cytotoxicity against mouse macrophage RAW264.7 cells was evaluated using the SRB assay, while the in vitro anti-inflammatory activity was determined via the Griess assay. Results Five compounds were isolated from the methanol extract of the whole plant of P. angulata and identified as 6- 5β,6β:14α,17:14β,26-triepoxy-13α,15β-dihydroxy-1-keto-11,15-cyclo-13,14-secoergostan-2-en-18,26-olide (1), physalin B (2), physalin F (3), physalin XI (4), and aspergillus triazolate A (5). Compounds 2—5 exhibited inhibitory effects on NO production in RAW264.7 cells, with IC₅₀ values ranging from (1.97 ± 0.12) to (6.73 ± 0.71) μmol/L. Conclusion Compound 1 is a new compound, named 5,6-epoxyphysalin IX. Compounds 4 and 5 showed significant anti-inflammatory activities.

Objective Novel Diels-Alder-type adducts from the stems and branches of Morus nigra were investigated, and their peroxisome proliferator-activated receptor gamma (PPARγ) binding activity was preliminarily evaluated. Methods Systematic isolation and purification were carried out by multiple chromatographic methods including silica gel, AB-8 macroporous resin, polyamide, Sephadex LH-20 gel and ODS reversed-phase column chromatography. Structures of the isolated compounds were identified by modern spectroscopic techniques including high-resolution electrospray ionization mass spectrometry（HR-ESI-MS）and nuclear magnetic resonance spectroscopy (¹H-NMR, ¹³C-NMR). The in vitro activities of some compounds were evaluated by PPARγ ligand-binding assay. Results Six Diels-Alder adducts were isolated from the ethanol extract of M. nigra twigs and identified as (5aR,10aR,14S)-2-((1S,2R,3S)-2-(2,4-dihydroxybenzoyl)-2',4'-dihydroxy-5-methyl-1,2,3,6-tetrahydro-[1,1'-biphenyl]-3-yl)-1,3,8, 14-tetrahydroxy-13,13- dimethyl-11H-10a,5a-(epoxypropano)benzofuro[3,2-b]chromen-11-one (1), sanggenon O (2), sanggenon C (3), mulberrofuran F (4), cathayanin B (5), and mulberrofuran G (6). Compound 1 exhibited weak PPARγ ligand-binding activity at 10 μmol/L. Compound 3 exhibited moderate ligand-binding activity toward PPARγ at 10 μmol/L, which was obviously weaker than that of its biosynthetic precursor sanggenol F (SGF). Conclusion Compound 1 is a new compound, named nigragenon Q, it is formed by an intermolecular [4+2] cycloaddition between a sanggenone-type dihydroflavone and a chalcone. Oxidation of the C-2 isoprenyl side chain enables its connection to C-3 via an oxygen bridge, furnishing a six-membered ring and constructing a unique [4,4,3] bridged ring system. Compound 3 exhibited moderate PPARγ ligand-binding activity.

Objective The in vitro co-incubation method was employed to investigate the metabolic transformation regularity of Semen Cuscutae-Fructus Lycii (SC-FL) mediated by gut microbiota in premature ovarian insufficiency (POI) rats. Methods SC-FL and its thirteen monomer components were co-incubated with gut microbial communities from POI rats in vitro, followed by UPLC-Q-TOF-MS for qualitative analysis. Results A total of 85 compounds were characterized from the SC-FL co-incubated samples, including 61 prototypical components (e.g., rutin), 13 metabolites (e.g., methyl hyperoside), and 11 kinds of components that exist in the prototype at the same time and can be metabolized by other components, (e.g., isorhamnetin). Hyperoside was characterized by five metabolites, rutin by seven, quercitrin by six, astragalin by seven, isoquercitrin by five, quercetin by seven, isorhamnetin by four, kaempferol by five, apigenin by four, chlorogenic acid by nine, ferulic acid by three, scopoline by four, and scopoletin lactone by three. Conclusion The metabolic transformation reactions mediated by the intestinal microbiota in POI rats mainly include Phase I metabolic reactions such as hydrolysis, reduction-hydrogenation, and hydroxylation reactions, and Phase II metabolic reactions such as methylation reactions. Reverse traceability analysis of the metabolic conditions of each component revealed that isorhamnetin, quercetin, kaempferol and apigenin were mutually metabolically transformed in the intestinal flora of POI rats These findings provide important evidence for elucidating the pharmacodynamic material basis and mechanisms of SC-FL in POI, and lay a foundation for subsequent research on optimizing SC-FL efficacy through gut microbiota modulation.

Objective On the basis of the requirements of ancient prescriptions, combined with modern technical means, the ancient decoction and modern decoction methods of Gualou (Trichosanthis Fructus) were compared. Methods Trichosanthis Fructus was soaked in rice wine (ancient method) and purified water (modern method) for 30 min, then decocted for 180 min. Samples were analyzed by UHPLC-Q-Exactive Orbitrap MS on a Hypersil GOLD aQ column (100 mm × 2.1 mm, 1.9 μm), with 0.1% formic acid acetonitrile–0.1% formic acid water as mobile phase by gradient elution (0.3 mL/min, 35 °C, 1 μL). ESI data were acquired in positive and negative modes (m/z 100—1 500). Data were processed using Thermo Scientific Xcalibur. Chemical constituents in different Trichosanthis Fructus decoctions were identified and analyzed based on MS data, reference standards, literature, and fragmentation patterns. Results Compared with the conventional modern decoction method, the ancient decoction method of Trichosanthis Fructus yielded more compound types. A total of 44 compounds were identified in ancient decoction samples versus 39 in modern decoction samples. The contents of major components such as arginine, lysine, guanine, adenine,and hypoxanthine were significantly higher in the ancinet decoction than in the modern decoction. Histidine, citrulline, 5-hydroxymethylfurfural, medicarpin, vanillic acid, chrysoeriol, diosmetin, kaempferide, and paeonol were identified exclusively in the ancient decoction samples of Trichosanthis Fructus. Conclusion This study confirmed that the ancient decoction method can more effectively extract or transform the chemical components in Trichosanthis Fructus, which may be the material basis for its unique clinical efficacy. It is worthy of attention and further research in clinical application.

Objective To clarify whether the combined use of sodium dodecyl sulfate (SDS) and hydroxypropyl methylcellulose (HPMC) can improve the dissolution behavior of baicalin-imperatorin co-amorphous system (Bai-Imp-CAS), and explore the mechanism of combination benefit. Methods A series of Bai-Imp CAS with different ratios of SDS and HPMC were prepared by the spray-drying method. Powder X-ray diffraction (PXRD) and differential scanning calorimetry (DSC) were adopted to characterize the solid-state properties of samples. Dispersion performance was evaluated by determining caking rate and dispersion time, and wettability was analyzed via contact angle and surface free energy measurements. The optimal ratio of SDS to HPMC was determined based on dispersion performance. According to the requirements of the 2025 Edition of Chinese Pharmacopoeia, the in vitro dissolution of the optimal SDS/HPMC (2∶1)-CAS was investigated and compared with those of SDS-CAS and HPMC-CAS. UV-visible spectrophotometry was used to detect the nucleation time and crystal growth rate of the two drugs. Results All spray-dried products were identified as co-amorphous systems. The caking rate of SDS/HPMC (2∶1)-CAS was (15.30 ± 1.55)%, which was significantly lower than that of single excipient groups. There was no significant difference in dispersion time between SDS/HPMC (2∶1)-CAS and SDS-CAS. The water contact angle of SDS/HPMC (2∶1)-CAS was 24.23°, with a surface free energy of 57.54 mJ/m², indicating favorable wettability. The in vitro dissolution results showed that the area under curve (AUC) values of baicalin and imperatorin in SDS/HPMC (2∶1)-CAS were (38 661.00 ± 375.32) and (4 565.00 ± 434.65) μg·h/mL, respectively, both significantly higher than those of SDS-CAS and HPMC-CAS. The early dissolution profile of SDS/HPMC (2∶1)-CAS was similar to that of SDS-CAS, while its dissolution decline rate in the middle and late stages was consistent with HPMC-CAS. Crystallization kinetics demonstrated that HPMC alone or combined with SDS could inhibit the nucleation of the two drugs and the crystal growth of baicalin, but exert no obvious effect on the crystal growth of imperatorin. SDS had no significant influence on the nucleation and crystal growth of both drugs. Conclusion SDS/HPMC (2∶1)-CAS shows the best dispersibility, and SDS and HPMC can jointly boost the dissolution of both drugs. The mechanism works by sequentially complementing each other's functions to improve the dissolution of Bai-Imp-CAS: SDS mainly drives fast initial dispersion and prevents aggregation, while also indirectly inhibiting crystallization by lowering local supersaturation; HPMC directly inhibits crystal growth during the mid to late stages of dissolution. Together, they create a combined “indirect and direct” effect on crystal inhibition, helping to maintain a supersaturated state. This strategy offers a fresh approach for designing co-amorphous systems for poorly soluble drug combinations, and it can reduce the amount of SDS needed, giving a “less is more” advantage for clinical translation.

Objective To prepare prostate specific membrane antigen (PSMA)-targeting peptide-modified lipid-polymer nanoparticles co-loaded with homoharringtonine and triptolide (HHT/TPL-PSMA-Lips), optimize its preparation process, and evaluate its in vitro anti-prostate cancer activity. Methods The solubility and oil-water partition coefficient (lgP) of HHT and TPL were determined, and PSMA-targeting peptide-modified PEGylated phospholipids were synthesized. HHT/TPL-PSMA-Lips was prepared by the emulsification-solvent evaporation method, and the formulation was optimized using central composite design-response surface methodology (CCD-RSM). Indicators including particle size, ζ potential, and encapsulation efficiency were investigated. The in vitro release behavior of the formulation in different media was evaluated. MTT assay and flow cytometry were used to assess the in vitro inhibitory effect and apoptosis-inducing activity of the nanoparticles on prostate cancer cells, while cell uptake assay was performed to verify the tumor cell targeting efficiency of the nanoparticles. Results HHT and TPL showed low solubility in various media and strong lipophilicity. The molecular weight of the synthesized PSMA-targeting peptide-modified PEGylated phospholipids was 4 643.8. HHT/TPL-PSMA-Lips prepared with the optimal formulation exhibited uniform particle size, with an average particle size of (209.12 ± 3.24) nm, a ζ potential of (-34.4 ± 2.5) mV, and a comprehensive encapsulation efficiency of (81.20 ± 0.40)% [HHT (80.73 ± 0.86)%, TPL (82.00 ± 0.40)%]. In vitro release experiments demonstrated that HHT/TPL-PSMA-Lips could effectively improve the solubility of HHT and TPL, and exhibite a certain pH-responsive release property. Cell experiments showed that the HHT/TPL combined administration group displayed stronger inhibitory effect on prostate cancer cells and better apoptosis-inducing activity than the single drug groups. HHT/TPL-Lips enhanced the anti-tumor activity, and the targeted modified nanoparticles (HHT/TPL-PSMA-Lips) showed the strongest inhibitory effect on prostate cancer cells. Cell uptake assay indicated that modification with PSMA-targeting peptide could effectively enhance the cellular uptake of the nanoformulation in tumor cells. Conclusion The prepared HHT/TPL-PSMA-Lips not only solves the solubility problem of the drugs but also significantly enhances tumor targeting and anti-tumor efficacy, providing a new approach for the development of new traditional Chinese medicine formulations for the treatment of prostate cancer.

Objective Given that the liquid formulation of the medicine-food homology formula Wuzhiyin is susceptible to non-enzymatic browning during storage, this study systematically investigates the underlying browning mechanism and evaluates the gastroprotective effect of Wuzhiyin prepared using an anti-browning processing technology against alcoholic gastric injury. Methods The differential metabolites and browning intermediates before and after browning of Wuzhiyin were analyzed by non-targeted ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and Spe-Pak microcolumn separation technology. The degree of browning was characterized by color changes, browning degree, and alterations in nutritional components. An anhydrous ethanol-induced acute gastric injury mouse model was established to evaluate the gastroprotective function of anti-browning prepared Wuzhiyin. Results Total sugars, total polyphenols, and total amino acids were significantly decreased by browning (P < 0.05). Nineteen key differential compounds were screened, among which fructose, sucrose, valine, isoleucine, tyrosine, and p-coumaric acid were identified as major driving factors. 2-Hydroxy-3-methyl-2-cyclopenten-1-one was identified as the core browning intermediate, and the browning mechanism was elucidated as a synergistic process involving the Maillard reaction, acid-promoted sugar degradation, and oxidative cross-linking of phenolic compounds. High-dose anti-browning prepared Wuzhiyin (43.72 g/kg) significantly reduced the gastric ulcer index (inhibition rate 72.42%) and alleviated pathological damage to the gastric mucosa. The underlying mechanisms include inhibiting nuclear factor-κB (NF-κB) pathway, reducing inflammatory cytokines [interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6], regulating oxidative stress [elevation of superoxide dismutase (SOD)/malondialdehyde ratio], and protecting mucosal barrier [modulation of NO, prostaglandin E₂ (PGE₂), trefoil factor 2 (TFF2)]. Conclusion Fructose, sucrose, valine, isoleucine, tyrosine, and p-coumaric acid are the key drivers of browning in Wuzhiyin; the Maillard reaction, acid-catalyzed sugar degradation, and phenolic oxidative cross-linking constitute the major browning pathways; and Wuzhiyin prepared via the anti-browning processing technology exhibits a significant gastroprotective effect against alcoholic gastric injury.

Objective To investigate the phase changes and component distribution patterns during the decoction process by performing phase separation and characterization of Baihe Dihuang Decoction (BDD, 百合地黄汤) at different decoction times, and provide a reference for elucidating the material basis of synergistic effects of components in traditional Chinese medicine formulas. Methods BDD was decocted for 20, 40, 60, 80, 100, and 120 min to obtain the decoctions. The sediment phase and nanophase solutions were separated using the “high-speed centrifugation-dialysis” method. The two phases were characterized by scanning electron microscope, laser particle size analyzer, ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy. The fingerprint spectra of BDD and its different phases were constructed using HPLC method, and chemometric methods were applied to screen key differential components among different phases, followed by content determination of the main differential components. Results The nanophase of BDD exhibited the smallest particle size, while the original decoction showed the largest particle size. The particle size of the nanophase initially increased and then decreased with prolonged decoction time, eventually stabilizing, and the stability of nanophase gradually enhanced. The contents of polysaccharides and proteins increased progressively with prolonged decoction time, and both the nano-sized fraction and the precipitated fraction were found to be rich in polysaccharides. Spectral analysis revealed that the chemical structures of components in the original decoction and each phase at different decoction times were generally similar, but differences in functional group contents were observed. Fingerprint analysis combined with hierarchical cluster analysis (HCA) and partial least squares-discriminant analysis (PLS-DA) showed that the types of chemical components in different phases of BDD were similar, but their contents differed significantly. The effect of decoction time on components was smaller than that of phase state. A total of 12 key differential components among different phases were screened. The average contents of five indicator components catalpol, regaloside C, regaloside A, regaloside B, isoverbascoside in the nanophase were higher than those in the sediment phase and the original decoction. Conclusion During the decoction process, BDD interacts with each other, resulting in significant differences in the content of active ingredients among various phases. The nanophase achieves enrichment of active components, suggesting that it may be the main effective phase of BDD.

Objective To address the potential uneven distribution of active ingredients indicated by uneven color within batches of soft material fine powder during the production process of Guizhi Fuling Capsules (GFC, 桂枝茯苓胶囊), a “cascade fault-tolerant model” was constructed based on color difference characteristics to explore a new low-cost approach for intermediate quality control. Methods A total of 177 samples of soft material fine powder from five batches were collected. The paeoniflorin content, determined by HPLC, was divided into low, medium, and high grades at a ratio of 2:6:2. Based on a traditional three-class model, a cascade fault-tolerant quality control model was constructed by comparing 126 algorithm combinations, which involved a primary algorithm for early warning and a secondary physical direction for correction. External validation was introduced to assess the generalization ability, and quality control experiments were conducted based on the model results. Finally, near-infrared spectroscopy (NIRS) technology was used for benchmarking against the upper limit of efficacy. Results Correlation analysis established paeoniflorin as the core grading indicator, with its content showing a significant positive correlation with red-green chroma a^* (r = 0.64, P < 0.001). The analysis indicated that the accuracy of the benchmark model was only 70.34% (RSD was 12.70%), which was difficult to meet quality control requirements. However, the cascade fault-tolerant model achieved an overall industrial accuracy of 78.62%, with significantly enhanced robustness (RSD was 6.79%), and its external validation accuracy was 80.56%. Quality control experiments showed that the RSD of paeoniflorin content in mixed samples decreased from 7.74% in random mixing to 2.34%, indicating a significant increase in material uniformity, and a simultaneous reduction in total mixing process time and load. Benchmarking results with NIRS (accuracy of 90.68%, RSD was 6.22%) indicated that the colorimeter had a high cost-performance ratio and could be used for batch screening; NIRS was more accurate and suitable for high-precision grading. Conclusion The cascade fault-tolerant model constructed in this study achieved the synergistic empowerment of algorithm early warning and physical correction, effectively enhancing the discriminative robustness of low-cost optical sensors. This provides a new strategy for quality control of active ingredients in batches of intermediates during the production process of traditional Chinese medicine in the future.

Objective To prepare galangin-whey protein-nanoparticles (Gal-WP-NPs), and study its oral pharmacokinetic behavior and protective effect on the hepatic ischemia reperfusion injury in vivo. Methods Single-factor experiments were conducted to investigate the effects of various formulation processes on Gal-WP-NPs. Box-Behnken design-response surface methodology (BBD-RSM) was used to obtain its optimal prescriptions. Entrapment efficiency, drug loading, particle size and ζ potential were determined. Gal-WP-NPs powder was prepared by freeze-drying method. Transmission electron microscopy (TEM) was used to observe the microstructure of Gal-WP-NPs. Galangin crystal in Gal-WP-NPs powder was analyzed by X-ray powder diffraction (XRPD). Solubility of galangin and Gal-WP-NPs powder were compared, and drug release behavior of Gal-WP-NPs in gastrointestinal fluid was determined. SD rats were administered galangin and Gal-WP-NPs via ig at a dose of 150 mg/kg (calculated as galangin), the main pharmacokinetic parameters and relative bioavailability of Gal-WP-NPs were calculated. The model of hepatic ischemia reperfusion injury was established, and then the protective effects of galangin and Gal-WP-NPs were compare on the hepatic ischemia reperfusion injury. Results Optimal formulation processes of Gal-WP-NPs: the volume ratio of the anti-solvent phase to the solvent phase was 15.40:1, the amounts ratio of whey protein to galangin was 15.05:1, and the ultrasonic power was 82.00 W. Envelopment efficiency, drug loading, particle size and ζ potential were (94.07 ± 0.98)%, (5.76 ± 0.09)%, (197.39 ± 4.46) nm and (-23.79 ± 0.75) mV, respectively. Gal-WP-NPs are spherical nanoparticles. Galangin existed as an amorphous state in Gal-WP-NPs powder. Solubility of galangin was significantly increased (P < 0.01), and the release process of Gal-WP-NPs in vitro conformed to the Weibull model. The peak time (t_max) of Gal-WP-NPs was delayed to (2.17 ± 0.64) h, half-life period (t_1/2) was increased to (5.11 ± 0.95) h, and oral relative bioavailability was increased to 6.58 times. Gal-WP-NPs significantly enhanced the protective effect of galangin on the hepatic ischemia reperfusion injury. Conclusion Gal-WP-NPs significantly increased the oral relative bioavailability of galangin and enhanced the protective effect on the hepatic ischemia reperfusion injury.

Objective To establish an interpretable deep learning model based on a bidirectional gated recurrent unit (BiGRU) and a temporal attention mechanism to elucidate the dynamic spatiotemporal response mechanisms of the multi-component–multi-target interactions underlying the effects of Buyang Huanwu Decoction (BYHWD) against ischemic stroke (IS). Methods A rat model of middle cerebral artery occlusion (MCAO) was established. Plasma and brain tissue samples were collected at multiple time points after administration. Plasma concentrations of 14 active components of BYHWD were quantified by UPLC–MS/MS, and the expression levels of 15 key targets in brain tissue were determined by ELISA to construct a multi-component–multi-target time series dataset. A BiGRU-based temporal attention network was established using a sliding window strategy, and leave-one-batch-out cross-validation (LOBO-CV) combined with bootstrap resampling was employed to evaluate model performance and stability. The model was further interpreted to identify highly contributive time points, potential key components, and responsive target features. Results The model demonstrated good predictive performance in LOBO-CV, with an R² value greater than 0.80 and a mean squared error (MSE) below 0.20 across all test sets. Temporal attention analysis identified 10 h post-administration as a time point receiving high model attention, with an attention peak value of 0.602 6. Gradient attribution analysis identified paeonol, calycosin, and formononetin as the major driving components, while brain derived neurotrophic factor (BDNF), protein kinase G type II (PKG2), and calmodulin-dependent protein kinase II (CAMK2) were identified as the key responsive targets. Bootstrap resampling showed that the 10 h critical time window was identified with a frequency of 90%, indicating good model stability. Conclusion This study established a BiGRU and temporal attention-based approach for dynamic time series analysis of the multi-component–multi-target actions of BYHWD. The proposed method provides new temporal evidence for elucidating the mechanisms underlying the anti-IS effects of BYHWD and offers a reference for investigating the dynamic pharmacological mechanisms of traditional Chinese medicine formulas.

Objective To systematically evaluate the clinical efficacy of kidney-tonifying and blood-activating traditional Chinese medicines as adjunctive therapies for distal radius fractures using a Meta-analysis, and to preliminarily explore their mechanisms of action through network pharmacology. Methods PubMed, Web of Science, Cochrane Library, Embase, CBM, CNKI, Wanfang, and VIP databases were searched from inception to June 2026 to identify clinical controlled studies comparing traditional Chinese medicine decoctions combined with conventional Western medicine versus conventional Western medicine alone for distal radius fractures. RevMan 5.4 software was used for Meta-analysis, and network pharmacology was employed to screen core active components, predict targets, and conduct pathway enrichment analysis. Results A total of 23 studies involving 3 377 patients were included. The Meta-analysis results showed that both kidney-tonifying and blood-activating traditional Chinese medicines significantly improved the clinical effective rate, shortened fracture healing time, reduced visual analog scale (VAS) pain scores, increased osteocalcin levels, and improved wrist joint function, with no statistically significant difference in overall efficacy between the two categories (all P > 0.05). Blood-activating traditional Chinese medicines showed superior improvements in wrist dorsiflexion range of motion, while kidney-tonifying traditional Chinese medicines exhibited more stable efficacy with lower heterogeneity. Network pharmacology identified core targets such as tumor protein p53 (TP53), proto-oncogene tyrosine-protein kinase Src (SRC), signal transducer and activator of transcription 3 (STAT3), primarily regulating pathways including phosphatidylinositol-3-hydroxykinase (PI3K)-protein kinase B (Akt), mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB). Blood-activating traditional Chinese medicines exerted anti-inflammatory and microcirculation-improving effects mainly through vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF), and Janus kinase (JAK)-STAT pathways, whereas kidney-tonifying traditional Chinese medicines promoted osteogenic differentiation and bone metabolism via the Wnt/β-catenin pathway and cell cycle pathways. Conclusion Kidney-tonifying and blood-activating traditional Chinese medicines are effective and safe as adjunctive therapies for distal radius fractures, both promoting fracture healing through multi-component, multi-target, and multi-pathway mechanisms. Blood-activating traditional Chinese medicines are more suitable for relieving swelling and pain in the early stage, while kidney-tonifying traditional Chinese medicines are better for strengthening bone and promoting healing in the middle and late stages.

Objective To analyze the current application status and hotspots of mass spectrometry imaging (MSI) in traditional Chinese medicine (TCM) research, and use knowledge graphs to display the research process in this field, providing reference for in-depth research and application. Methods Relevant Chinese and English literature from China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform (Wanfang), VIP Data Knowledge Service Platform (VIP) and Web of Science (WOS) databases were retrieved. Literature was screened by NoteExpress literature management software. The CiteSpace software was used to draw a knowledge graph of publishing volume, publishing institutions, authors, keywords, etc. Results A total of 124 Chinese literature and 306 English literature were included. The quantities of related publications significantly increased since the 2021. The Chinese Traditional and Herbal Drugs and Chinese Journal of Chinese Materia Medica are the journals with the highest quantities of publications in Chinese. Journal of Ethnopharmacology is the journal with the highest quantities of publications in English. Sun Bo’s team and Cai Zongwei’s team were the teams with the highest quantities of publications in Chinese and English, respectively. The China Academy of Chinese Medical Sciences and Chinese Academy of Science were the research institutions with the highest quantities of publications in Chinese and English, respectively. Research in Chinese-language literature focused primarily on metabolomics, chemical composition, and tissue distribution, whereas English literature research emphasized component identification, metabolites, and biosynthesis. The main research focus of Chinese and English literature research is the same, both involving the characterization and spatial distribution of chemical components, biosynthesis, and metabolic pathways. Conclusion At present, MSI has been widely used in TCM research, primarily covering the characterization and spatial localization of chemical component, quality evaluation of TCM, biosynthesis of active components, in vivo metabolism and distribution, and elucidation of the TCM mechanism of action. Future research should focus on the following aspects: improvement of MSI-related databases, quantitative MSI technology optimization, the integration of MSI with other analytical technologies, clinical research and application promotion of TCM, the development and application of three-dimensional and in vivo MSI.

Objective To screen stable reference genes in Gastrodia elata and analyze the expression patterns of key enzyme genes in the gastrodin biosynthesis pathway, providing reliable normalization tools for molecular studies of G. elata. Methods Two transcriptome datasets, encompassing various tissues (tubers, stems, and flowers) at the jian-ma stage and four developmental stages post-inoculation, were integrated. Expression matrices were constructed using Transcripts Per Million (TPM) and normalized via the TMM method. Candidate reference genes were screened based on a coefficient of variation (CV) threshold of < 0.05, and specific primers were designed using the PrimerServer2 package. The expression levels of these candidates across different tissues and developmental stages were validated by qRT-PCR. Stability was comprehensively evaluated using delta-Ct, geNorm, NormFinder, BestKeeper, and RefFinder. Furthermore, the identified optimal reference gene was used to normalize the expression of six key enzyme genes in the gastrodin biosynthesis pathway. Results Gelref1 exhibited the highest stability across all samples, serving as a universal reference gene for various tissues and developmental stages of G. elata. Expression analysis revealed distinct patterns for the six key enzyme genes, with most showing high expression levels during the mid-to-late stages of tuber development. Conclusion This study establishes a systematic workflow for reference gene screening in G. elata based on RNA-Seq data, providing a reliable foundation for further research into the molecular mechanisms and functional validation of genes in gastrodin biosynthesis.

Objective To clone the chalcone synthase gene SsCHS1 from Spatholobus suberectus and conduct bioinformatics and functional analyses, in order to elucidate its role in flavonoid biosynthesis. Methods Using cDNA from S. suberectus as a template, SsCHS1 was cloned by RT-PCR. Systematic bioinformatics analyses were performed on the sequence characteristics, protein structure, and function of the gene. An SsCHS1 overexpression vector was constructed and genetically transformed into Nicotiana benthamiana. The total flavonoid content and the expression levels of key enzyme genes in the flavonoid biosynthesis pathway in transgenic plants were determined. Results The full-length SsCHS1 gene was 1 176 bp, encoding a protein consisting of 391 amino acids, with a theoretical relative molecular mass of 42 825.35, an isoelectric point of 6.10, and was characterized as a hydrophilic protein. It showed high homology with CHS proteins from leguminous plants such as Glycyrrhiza uralensis and Glycine max. Promoter analysis revealed that SsCHS1 contains multiple cis-acting elements associated with hormone response, light response, stress response, and transcription factor binding sites. Functional analysis demonstrated that overexpression of SsCHS1 significantly promoted flavonoid accumulation in transgenic tobacco and upregulated the expression levels of key enzyme genes in the flavonoid biosynthesis pathway. Conclusion The SsCHS1 gene from S. suberectus promotes flavonoid biosynthesis. The identification of this gene provides an important theoretical basis and a candidate gene for further elucidation of the biosynthetic mechanisms of active components in S. suberectus.

Objective Root rot has become a bottleneck restricting the industrial development of Monochasma savatieri, however, current understanding regarding its pathogenesis remains poorly understood. Methods This study took three groups of M. savatieri with different root rot progress and their rhizosphere soil as the research objects, and determined the soil physical and chemical properties of rhizosphere soil and the content of main active components of M. savatieri; metagenomic sequencing of the rhizosphere soil of M. savatieri. Results With the development of root rot, the physical and chemical properties of rhizosphere soil changed significantly. Among them, pH, electrical conductivity (EC) and available phosphorus increased significantly with the deepening of disease severity, while the contents of total carbon and total nitrogen showed the opposite trend. And the contents of total flavonoids, luteolin, apigenin and isoverbascoside were significantly reduced, while the content of verbascoside was increased. The microbial diversity increased with the deepening of disease severity, and the abundance of pathogenic bacteria related to soil-borne diseases in the soil, such as Fusarium, Cladosporium and Botrytis, as well as beneficial Trichoderma, increased in the later stage of the disease. The metabolic related pathways under KEGG annotation, such as fatty acid metabolism, glyoxylic acid and dicarboxylic acid metabolism, β-alanine metabolism and pyruvate metabolism, were more enriched than those in the healthy group and the mid-stage disease group. The pH value and EC value were significantly positively correlated with the abundance of potential pathogenic fungi. The genera such as Microbacterium, Drechslerella, Metarhizium and Trichoderma, which show significant positive correlations with total flavonoids of M. savatieri, luteolin, apigenin and verbascoside, may be involved in the improvement of soil microecology and the process of plant disease resistance. Conclusion Through a comprehensive correlation analysis of key indicators in the “M. savatieri -Microorganisms-Soil” system under different progression stages of root rot, this study systematically elucidates the patterns of change and interaction relationships among the three during the disease progression. The findings provide a basis for uncovering the microecological mechanisms underlying the occurrence of root rot in M. savatieri, as well as for developing control strategies through the regulation of soil and rhizosphere microorganisms.

Objective To characterize the chloroplast genomes and phylogenetic relationships of four medicinal species of Disporopsis, namely Disporopsis aspersa, D. fuscopicta, D. longifolia, and D. pernyi, and to identify potential regions for species discrimination, thereby providing a molecular and theoretical basis for species identification, germplasm resource conservation, quality control of medicinal materials, and resource development and utilization of this genus. Methods Young leaves of the four medicinal Disporopsis species were subjected to high-throughput sequencing. The chloroplast genomes were assembled and annotated, followed by bioinformatic analyses of repeat sequences, inverted repeat (IR) boundaries, codon usage bias, nucleotide diversity, selection pressure, and phylogenetic relationships. Results The chloroplast genomes of the four medicinal Disporopsis species exhibited a typical quadripartite circular structure, with genome sizes ranging from 155 975 to 156 109 bp and an overall GC content of 37.68%. A total of 131 genes were annotated in each genome, including 85 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. Simple sequence repeats (SSRs) were dominated by mononucleotide repeats, whereas long sequence repeats (LSRs) were dominated by palindromic repeats. The IR boundaries were generally conserved among the four species. Their codon usage patterns were highly similar, with a preference for codons ending in A or T. Nucleotide diversity analysis showed that rrn23S and trnT-UGU had relatively high Pi values among the genic regions, whereas rrn4.5S–rrn5S, ccsA–ndhD, rps11–rpl36, and rps15–ycf1 were highly variable intergenic regions. Selection pressure analysis showed that most protein-coding genes had Ka/Ks values below 1 and were predominantly constrained by purifying selection. However, ycf1, ycf4, psaI, and cemA had Ka/Ks values above 1 in some species comparisons, suggesting possible signals associated with adaptive evolution. Phylogenetic analysis showed that the four Disporopsis species clustered within the same well-supported branch and formed a sister group with Polygonatum. Conclusion The chloroplast genomes of the four Disporopsis species are generally conserved in genome structure and gene composition, although local sequence variation is present. The candidate highly variable regions identified in this study may provide useful references for species identification and molecular marker development in Disporopsis. The phylogenetic results also provide chloroplast genomic evidence for investigating the classification and relationships of this genus.

Objective Using cultivar 85-5 as the experimental material, Stereo-seq spatial transcriptomics technology was employed to analyze the in situ gene expression patterns of tuberous roots, thereby revealing the spatial heterogeneity of gene expression in root cells. Methods The optimal permeabilization duration was determined by fluorescence intensity and grayscale quantification, and the best bin resolution was identified by comparing gene detection efficiency and cell clustering performance across multiple parameter groups. Principal component analysis (PCA) and uniform manifold approximation and projection (UMAP) were used for cell clustering. Cluster-specific differentially expressed genes (DEGs) were identified using the Seurat package, and qRT-PCR was performed to validate the reliability of spatial transcriptomic data. Results The optimal permeabilization time for frozen sections of R. glutinosa tuberous roots is 18 min, and a bin size of 200 is more suitable for the analysis of macroscopic cell populations in R. glutinosa tuberous roots. Root cells of R. glutinosa were classified into five distinct populations with unique spatial localization via PCA and UMAP. Specifically, clusters 1 and 2 were predominantly distributed in the phloem, cluster 3 in the periderm, and cluster 4 in the cambium. The qRT-PCR verified that gene DH2020-016897 was specifically expressed in the periderm, whereas DH2020-009767 and DH2020-009768 exhibited predominant expression in the cambium. Conclusion In this study, we established a high-resolution spatial transcriptomic atlas of R. glutinosa tuberous roots, delineating spatial distribution patterns of individual cell clusters and the expression profiles of key genes. These findings provide a fundamental resource for exploring tuberous roots development, secondary metabolite biosynthesis, and spatially resolved molecular regulatory networks underlying stress responses in R. glutinosa.

Fresh medicinal materials refer to unprocessed or non-dried botanical and zoological substances utilized in their fresh state. By circumventing conventional drying and preparation procedures, they preserve a higher concentration of volatile compounds, heat-sensitive constituents, and other bioactive ingredients, thereby offering distinct advantages in treating external-contraction febrile diseases, dermatological conditions, and similar ailments. A systematic comparison of pharmacological activities between fresh and dried medicinal materials reveals that fresh variants of some species generally exhibit elevated levels of volatile oils, alkaloids, and other components, along with more pronounced antibacterial, anti-inflammatory, antioxidant, and antitumor effects. Simultaneously, it provides a comprehensive review of the research progress on modern formulations of fresh medicinal materials. Novel dosage forms, such as juice preparations, capsules, and vesicular carrier systems, have, to some extent, mitigated the challenges associated with the storage and transportation of fresh medicinal materials. Nevertheless, issues persist in practical applications, including inconsistent quality standards and inadequate stability assessment frameworks. This review aims to compare and analyze the chemical constituents and pharmacological activities of fresh and dried Chinese medicinal materials, sort out the existing problems in the application of fresh medicinal materials, prospect the research directions of their preparations, further summarize the current principles for selecting appropriate dosage forms for fresh Chinese medicinal materials, and explore the direction of constructing a suitable quality standard system oriented toward active ingredients. It is expected to provide theoretical basis and technical references for the in-depth development and rational utilization of fresh Chinese medicinal materials, and promote the inheritance, innovative development of traditional Chinese medicine.

Volatile oil in traditional Chinese medicine (TCM) is one of the important material bases for TCM to exert pharmacological effects, and its extraction technology directly determines the efficacy, safety and quality stability of the product. However, traditional extraction methods generally have problems such as low extraction efficiency, susceptible degradation of heat-sensitive constituents, and organic solvent residues, which seriously affect the quality and large-scale application of volatile oils. Therefore, the development of green and efficient new extraction technologies has become a key direction to promote technological progress in this field. This paper systematically summarizes the application status and inherent limitations of conventional extraction technologies such as solvent extraction and steam distillation, and discusses the research strategies to improve the efficiency of traditional methods through process parameter optimization, equipment structure improvement, and combination of ultrasonic, microwave and other physics auxiliary technologies. At the same time, the latest technical progress and research overview of emerging technologies such as supercritical fluid extraction, subcritical extraction, ultrasound-assisted extraction, microwave-assisted extraction, ionic liquid extraction and pulsed electric field-assisted extraction are summarized. Finally, the future development trends of multi-technology collaboration, green solvent substitution and intelligent regulation system construction are prospected, providing references for the innovation and industrial application of TCM volatile oil extraction technologies.

Akkermansia muciniphila (AKK) is a key commensal bacterium colonizing the intestinal mucus layer, playing a crucial role in maintaining intestinal barrier integrity, regulating immune homeostasis and participating in metabolic control. Recent studies indicate that the abundance of AKK is significantly reduced in the intestines of patients with inflammatory bowel disease (IBD), with its dynamic changes closely correlated with disease activity. Animal experiments demonstrate that supplementing this bacterium or its functional proteins can alleviate intestinal inflammation by regulating the “intestinal barrier-immune inflammation-metabolism” triple pathway. However, under specific pathological conditions, AKK may also exacerbate barrier damage by excessively degrading the mucus layer, suggesting that they exhibit significant context-dependent bidirectional regulatory characteristics. Traditional Chinese medicine (TCM) and its active components can bidirectionally regulate the abundance and function of AKK through multiple pathways, including the direct supply of nutritional substrates, promotion of mucus secretion, remodeling of the microbial community structure and regulation of metabolic interactions, thereby maintaining the homeostasis of the gut microbiome and exerting an anti-IBD effect. This article systematically reviews the dual mechanisms of action of AKK in IBD and their roles in intestinal barrier repair, immune regulation and metabolic remodeling. It focuses on summarizing research progress of TCM in improving IBD by targeting the AKK-mediated “microbiota-barrier-immunity” axis, with the aim of providing a theoretical basis and research directions for TCM microbiota intervention strategies based on the regulation of key bacteria.

As a traditional plant resource with both medicinal and aromatic value, Muxiang (Aucklandiae Radix) exhibits distinct regional characteristics and trends of industrial upgrading of resource distribution, development and utilization, and international trade patterns. This paper systematically reviews the evolution of Aucklandiae Radix varieties and their application characteristics in Asian traditional medical systems, summary of the current development status and technical bottlenecks of Aucklandiae Radix-related products, compares quality control standards across countries, and analyzes international trade data and development trends to provide theoretical references and practical ideas for the sustainable utilization of Aucklandiae Radix resources, high-value product development, and international trade layout. The results indicate that there are numerous adulterants of Aucklandiae Radix cannot be used interchangeably; resources are primarily derived from artificial cultivation, with Yunnan, Sichuan, and Chongqing in China serving as core production areas. In terms of medical applications, it is utilized in traditional Chinese medicine, Indian medicine, and Southeast Asian folk medicine, with its core functions of promoting qi circulation to relieve pain and strengthening the spleen to aid digestion being widely applied. Currently, Aucklandiae Radix product development has extended from traditional herbal materials to high-value-added extracts, traditional Chinese patent medicines, health supplements, cosmetics, and veterinary drugs, with the industrial chain continuously improving. There are differences in quality standards across countries regarding detection indicators and limit requirements; pharmacopoeias in Europe, British, China, and South Korea emphasize limits on pesticides and heavy metals, whereas Japan, Vietnam, and India do not specify requirements for the determination of major component content. In terms of internationalrade, Aucklandiae Radix has formed a production and sales network centered on Asia. Meanwhile, the Aucklandiae Radix industry faces challenges such as resource degradation, uneven processing technology, and poor alignment of international standards. This paper provides theoretical references and practical ideas for the sustainable utilization of Aucklandiae Radix resources, high-value product development, and international trade layout.

As one of China’s medicinal herbs with dual food and medicinal properties, Codonopsis Radix sees extensive clinical practice and industrial development, making quality evaluation research critically important. The “quality assessment through morphological analysis” methodology, which synthesizes the practical experience and wisdom of generations of medical experts, holds unique reference value. This paper reviews the herbal textual research of the “quality assessment through morphological analysis” theory for C. pilosula, modern research on the correlation between its morphological features and quality, and the determinants of its quality. This review aims to provide a reference for contemporary research on the quality evaluation of Codonopsis pilosula.