BackgroundGout is a prevalent metabolic disorder characterized by hyperuricemia and inflammation. Verbenalin, an iridoid glycoside from Verbena officinalis, possesses anti-inflammatory properties; however, its therapeutic potential and underlying mechanisms in gout remain underexplored.ObjectiveThis study aimed to evaluate the pharmacological effects and elucidate the molecular mechanisms of verbenalin in a rat model of gout.MethodsHyperuricemia and acute gouty arthritis were induced in rats using potassium oxonate/hypoxanthine and monosodium urate, respectively. Verbenalin was administered orally for 7 days. Therapeutic efficacy was assessed via physical symptom scores (inflammation, gait, swelling), renal/hepatic function indices, and histopathology. Furthermore, a multi-omics strategy integrating transcriptomics, metagenomics, and metabolomics, combined with Western blotting, was employed to investigate the pharmacological mechanisms.ResultsVerbenalin treatment significantly alleviated joint inflammation and swelling while improving gait scores. It effectively lowered serum uric acid (UA), creatinine, and BUN levels, inhibited hepatic xanthine oxidase (XOD) activity, and promoted urinary UA excretion. Histopathological damage in the joints, kidneys, and liver was markedly mitigated. Mechanistically, verbenalin downregulated the expression of urate transporters (URAT1, GLUT9) and inflammatory mediators (NLRP3, IL-1β) by inhibiting the PI3K-AKT and MAPK signaling pathways. Multi-omics analysis further revealed that verbenalin restored gut microbiota diversity and modulated purine metabolism, correlating with reduced UA levels.ConclusionThese findings demonstrate that verbenalin may exert anti-gout effects through the potential synergy of modulating purine metabolism, shifting gut microbiota composition, and suppressing PI3K-AKT and MAPK inflammatory signaling pathways. This study provides a preliminary scientific basis for further investigation into verbenalin as a prospective multi-target therapeutic candidate.

Automated quantification of immunohistochemistry (IHC) supports reproducible biomarker assessment, but routine use remains limited by cellular crowding and staining variability. We present an ImageJ/Fiji-native workflow for batch quantification of nuclear and cytoplasmic H-scores without task-specific model training. It integrates StarDist nuclear detection, a Voronoi-based Cell Distance (VCD) topology-guided segmentation strategy and user-guided adaptive calibration (Mode 2). VCD constructs Skeleton by Influence Zones (SKIZ)-derived boundaries from detected nuclei, reducing boundary fusion and spatial dilution during morphological dilation. Mode 2 uses reference regions from a representative positive-control image to derive batch-specific intensity thresholds and reduce staining-related bias. In stress tests, 99.7% of detected cell boundaries were within 2 pixels of manual ground-truth annotations. Across 752 IHC images representing diverse staining conditions, Mode 2 closely agreed with clinician-assigned H-scores (CCC = 0.997), with a mean Bland–Altman bias of −2.35 H-score units. Across two Human Protein Atlas pan-cancer cohorts, automated H-scores correlated with expert annotations for nuclear Timeless (n = 330; Spearman’s ρ = 0.760) and cytoplasmic CDK1 (n = 290; ρ = 0.660). This open-source ImageJ/Fiji plugin provides an accessible, reproducible workflow for batch IHC quantification.

The development of simple, sensitive, and reliable methods for detecting prostate-specific antigen (PSA) holds significant importance for the early screening and diagnosis of prostate cancer. In this work, a novel signal "off-on" electrochemical aptamer (Apt) sensing platform was constructed for the first time to detect PSA, utilizing a synthesized nanoflower-shaped three-dimensional cobalt-metal-organic framework (Co-MOF) as an oxidase-mimicking nanozyme and employing 1,2-diaminobenzene as the catalytic substrate. In this design, the Co-MOF nanozyme can directly catalyze the oxidation of 1,2-diaminobenzene to generate diaminophenazine (DAP), an electroactive substance, without the need for H2O2. However, when the prepared Co-MOF nanozyme binds to PSA-specific aptamers, its enzyme-like activity becomes inhibited due to the blockage of active sites by the aptamer, leading to the disappearance of the DAP current that corresponds to the signal "off" state. Notably, in the presence of PSA, the catalytic activity of the Co-MOF is restored as the specific binding between Apt and PSA causes Apt to detach from the Co-MOF surface, resulting in the recovery of the DAP current and switching the signal to the "on" state. After the optimization of key experimental parameters, the proposed "off-on" nanozyme-based electrochemical aptasensor demonstrates excellent PSA detection performance. Moreover, this platform may provide a novel, simple, and reliable strategy for detecting a wide range of biomarkers by simply replacing the corresponding aptamer.

Metastasis remains the leading cause of mortality in breast cancer, and effective prevention strategies are urgently needed. Sodium ferulate (SF), a bioactive compound derived from ferulic acid, has demonstrated diverse pharmacological activities, yet its role in suppressing cancer metastasis remains unclear. In this study, we demonstrate that SF significantly inhibits breast cancer metastasis through comprehensive modulation of the bloodstream microenvironment. At non-toxic concentrations, SF inhibited the migration and invasion of MCF-7 and MDA-MB-231 cells by suppressing the BMP4/Smad1/5/9 signaling pathway, reversing epithelial-mesenchymal transition (EMT), and disrupting cell cycle progression. SF also enhanced the sensitivity of breast cancer cells to paclitaxel. In addition, SF reduced tumor cell adhesion to endothelial cells by blocking TNF-α-induced expression of ICAM-1 and VCAM-1 through inhibition of NF-κB signaling cascade. Furthermore, SF attenuated platelet activation and angiogenesis, two key processes involved in metastatic dissemination and colonization. In 4T1 mouse metastasis model, SF treatment significantly decreased pulmonary metastatic nodules without causing organ toxicity. Immunohistochemical analysis further confirmed decreased expression of BMP4, PI3K, and N-cadherin in lung tissues following SF treatment. Additionally, SF enhanced systemic immunity by increasing the proportion of cytotoxic T cells and natural killer cells in peripheral blood. Collectively, these findings reveal that SF exerts potent anti-metastatic effects by modulating tumor-bloodstream-immune interactions rather than direct cytotoxicity. By remodeling the bloodstream microenvironment and enhancing immune defense, SF may represent a promising and safe candidate for the prevention of breast cancer metastasis.

Ginsenoside Rd (Rd) is a bioactive compound predominantly found in Panax ginseng C.A. Meyer and Panax notoginseng (Burkill) F.H. Chen ex C.H. Chow, both species belonging to genus Panax in the Araliaceae family. However, its hepatic protective effect against acute liver injury and related mechanistic action remain unexplored. To investigate the protective effect of Rd against thioacetamide (TAA)-induced acute liver injury and assess its underlying regulatory mechanisms related to autophagy and inflammation. Forty-eight 8 weeks old C57BL/6 mice were treated with saline (control or model group), Rd (12.5 mg/kg, 25 mg/kg or 50 mg/kg), and diammonium glycyrrhizinate (DG, 30 mg/kg) for three days. Then the mice were stimulated with TAA to establish acute liver injury model, excluding the control group. HSC-T6 cells were treated with Rd at concentrations of 2.5, 5, or 10 µM, for 12 h with or without Lipopolysaccharide (LPS) stimulation at 100 ng/mL. Immunofluorescence staining, qPCR and Western blot were employed to analyze the expressions of genes and proteins associated with inflammation and autophagy. To validate the role of Rd in regulating autophagy and inflammation, the autophagy inducers, rapamycin and GSK621, were utilised in reverse validation experiments in cells. Rd exhibited significant hepatic protective effects in mice by reducing the serum levels of Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Glutathione S-transferase (GST) and Lactate dehydrogenase (LDH) with acute liver injury. It exhibited strong anti-inflammatory effect by reducing inflammation associated protein, such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), nod-like receptor protein 3 (NLRP3), associated speck-like protein containing a CARD (ASC), interleukin-18 (IL-18) and interleukin-1β(IL-1β) proteins and the mRNA expression levels of COX-2, Tumor Necrosis Factor α (TNF α), interleukin-6 (IL-6) and iNOS were decreased in liver tissue. And Rd inhibited LPS-induced inflammation by reducing the expression of COX-2 and NLRP3 in HSC-T6 cells. Moreover, not only in vivo but also in vitro, Rd downregulated the expression of LC3II, Beclin1, phosphorylation-AMP-activated protein kinase (p-AMPK), phosphorylation-ULK1 (p-ULK1) and upregulated the expression of p62 and phosphorylation-mechanistic target of rapamycin (p-mTOR) to suppress autophagy via the AMPK/mTOR/ULK1 pathway. Finally, the inhibitory effects of Rd on autophagy and inflammation in HSC-T6 cells were partially blocked by rapamycin and GSK621. Rd is a promising therapeutic agent to protect liver against TAA-induced acute liver injury by regulating the autophagy-NLRP3 inflammasome pathway.

Background:Ginsenoside Re (G-Re), a unique ginsenoside almost exclusively found in Araliaceae plants, is a promising therapeutic agent for attenuating liver injury. This study aims to investigate the liver-protective effects of G-Re and the underlying mechanisms in acute liver injury models. Methods:Male C57BL/6 mice were intraperitoneally injected with various agents induce the acute liver injury model after pre-treatment with G-Re (5-20 mg/kg, oral gavage). Additionally, the phosphoinositide 3-kinases (PI3K) inhibitor LY294002 and the mammalian target of rapamycin (mTOR) inhibitor RAPA were co-administered with G-Re in the thioacetamide (TAA)-induced rat hepatic stellate cell line (HSC-T6) to explore the mechanisms associated with G-Re. Results:G-Re at (20 mg/kg) protected liver against thioacetamide (TAA), ethanol, acetaminophen, and D-Galactosamine-induced liver injury in C57BL/6 mice. G-Re reduced serum levels of aspartate aminotransferase (AST) from 151.98 to 40.24 U/L and alanine aminotransferase (ALT) from 392.04 to 49.43 U/L. Both in vivo and in vitro studies consistently showed that G-Re decreased mRNA expression levels of key pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Additionally, G-Re dose-dependently downregulated the protein expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase -1 (caspase-1), interleukin-18 (IL-18), and IL-1β. In addition, our results suggested that the suppression of autophagy by G-Re may play a crucial role in its ability to inhibit the NLRP3 inflammasome. Notably, this regulatory effect on autophagy appears to be mediated through the phosphatidylinositide 3-kinases/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR signaling pathway). G-Re inhibits autophagy in both cellular and animal models by downregulating the expression of light chain 3-II (LC3-II), Beclin-1, and sequestosome-1 (p62) through this pathway. Furthermore, the PI3K inhibitor LY294002 and the mTOR inhibitor rapamycin (RAPA) were shown to partially reverse the inhibitory effects of G-Re on autophagy and inflammation in HSC-T6 cells. These results further support the notion that reactivation of autophagy can counteract G-Re-mediated suppression of NLRP3 and caspase-1 expression. Conclusion:This study highlights G-Re as a promising therapeutic candidate for liver injury, acting through inhibition of autophagy and inflammation via the PI3K/AKT/mTOR signaling pathway.

Metastasis is the leading cause of cancer-related deaths and poses a treatment challenge. Although studies have shown the importance of epithelial-mesenchymal transition (EMT) and metabolic reprogramming during cancer metastasis, the link between EMT and metabolic reprogramming, as well as the underlying molecular mechanisms by which both mediate cancer cell invasion and metastasis have not been elucidated. Here, we observed that interactions between platelets and cancer cells promote the secretion of TGF-β, thereby initiating EMT, promoting the invasion, and altering the metastatic and metabolic potential of colon cancer cells. TGF-β activates the AKT signaling pathway to enhance HK1 and HK2 expression in cancer cells, leading to increased glucose consumption, ATP production, and precise modulation of cell cycle distribution. In an energy-deficient model induced by oxidative phosphorylation (OXPHOS) inhibition with oligomycin A, TGF-β-induced highly metastatic HCT116 (H-HCT116) cells adapt by upregulating HK expression and glycolytic metabolism, while concurrently decreasing cell proliferation to conserve energy for survival. Mechanistically, H-HCT116 cells regulate cell division rates by downregulating CDK2, CDK4, and Cyclin D1 protein expression and upregulating p21 expression. Furthermore, H-HCT116 cells display enhanced motility, which is linked to increased mitochondrial metabolic activity. These findings indicated that cancer cells-platelets interaction secreted TGF-β activates cancer metastasis potential by inducing metabolic reprogramming and bioenergetic adaptation. The present study provides new insights into the adaptive strategies of highly metastatic cancer cells under adverse conditions and indicates that targeting glycolysis and metabolic reprogramming could serve as a viable approach to prevent cancer metastasis.

Interferon gamma (IFNγ) can amplify immune cell-mediated anti-tumor immunity, as well as directly kill cancer cells. Ginsenoside Rh2 (Rh2), a bioactive compound in traditional Chinese medicine, exhibits anti-cancer effects such as inhibiting proliferation and metastasis. Our earlier research found that Rh2 combined with IFNγ enhanced CXCL10 secretion in cancer cells. Here, we explored whether Rh2 and IFNγ exerted more potent anti-cancer activity in vitro and in vivo, along with its mechanisms and clinical value. Our data showed that Rh2 in combination with IFNγ resulted in a remarkably increased cytotoxicity in colorectal cancer cells including HT29, LoVo and T84 cell lines. Consistently, intratumoral injection with Rh2 plus IFNγ further restricted the HT29 tumor growth in vivo, and importantly, it was demonstrated to be safe for mice. Meanwhile, the combo treatment activated the stimulator of interferon genes (STING) pathway in cancer cells, promoting the transcription of downstream type I interferon. RNA sequencing revealed a dramatically transcriptional alteration in cancer cells with combo treatment and indicated that Rh2 further augmented the activation of interferon signaling pathway, compared with the IFNγ alone. Inhibition of janus kinase (JAK) by ruxolitinib could significantly rescue the cell death-triggered by the combo treatment. Then, a gene set named Rh2+IFNγ signature genes (RISG) was defined, which contained top 20 significantly upregulated genes from the combo treatment. Patients who exhibited a favorable response to the immunotherapy had a higher expression of RISG in tumor compared with those who did not respond. And the high expression of RISG was correlated with better clinical outcome in patients with colorectal cancer (CRC) and skin cutaneous melanoma (SKCM). Herein, the combination of Rh2 with IFNγ served as a promising strategy for cancer treatment, and its-derived RISG gene set also exhibited potential value in predicting clinical outcome.

Amyloid-beta (Aβ) aggregation, phosphorylated tau accumulation and neuroinflammation are considered as three hallmarks of Alzheimer’s disease (AD). Rhynchophylline (RN), the major alkaloid of a Chinese medicinal plant Uncaria rhynchophylla, has been shown to possess potent anti-AD effects. This study explored the effects of RN on Aβ pathology, tauopathy, and neuroinflammation using three AD mouse models, including TgCRND8, 3×Tg-AD, and 5×FAD, with RN treatment lasting for 4, 6, and 6 months, respectively, followed by behavioral tests and biological assays. In addition, BV2 cells were employed to further evaluate the biological effects of RN. RN treatment improved cognitive functions by reducing anxiety-like behaviors, enhancing recognition ability, and ameliorating learning impairments. It modulated Aβ processing through reducing the Aβ-producing enzyme activities and enhancing degradation enzyme activities, thereby diminishing Aβ accumulation. RN also decreased hyperphosphorylated tau proteins at Thr181, Thr205, Ser396, and Ser404 sites. Moreover, RN diminished neuroinflammation by reducing microglia and astrocyte activation and lowering the release of inflammatory cytokines. Furthermore, RN treatment could restore gut microbiota dysbiosis in 5×FAD mice. In BV2 cells, knockdown of p53, HDAC2, and Galectin-3 markedly enhanced the anti-inflammatory effects of RN. Overall, the anti-AD properties of RN were attributed to its regulation of multiple biological pathways, including regulation of the p53/PINK1 signaling pathway, inhibition of the HDAC2/AMPK signaling pathway, suppression of the Galectin-3/C/EBPβ/AEP signaling pathway, and modulation of gut microflora dysbiosis. This pioneering study unambiguously revealed the effects of RN on cognitive impairments, APP processing, tauopathy, and neuroinflammation in different transgenic mouse models with differing AD burdens, highlighting its potential as an anti-AD therapeutic agent and enhancing the scientific basis for its clinical use in treating AD.

Gut microbiota and bile acid metabolism play crucial roles in the progression of nonalcoholic fatty liver disease (NAFLD). Early evidence demonstrates that Ginsenoside Re (Re) possesses pharmacological effects on NAFLD, but its mechanisms of action are not well understood. This study aimed to investigate the hepatic protective effects of Re in NAFLD and elucidate relevant mechanisms. The effects of Re treatments (10, 20, or 40 mg/kg) against high-fat diet-induced NAFLD were initially tested on male C57BL/6 mice. Then, a separate mouse group received Re with or without antibiotics to confirm the regulatory role of microbiota in the effect of Re. Finally, another group of mice received fecal microbiota transplantation (FMT) from the initial experiment of NAFLD mice to further investigate the mechanistic role of gut microbiota. Re significantly improved liver function by reducing hepatic lipid accumulation, injury and hepatocyte steatosis, and inflammation. The liver protection was mediated by the regulation of gut microbiota as evidenced by restored intestinal barrier integrity, normalized Firmicutes/Bacteroidota ratio, enhanced abundances of Adlercreutzia equolifaciens , and reduced Faecalibaculum rodentium. Following that, Re reduced total and primary bile acids and downregulated bile acid synthesis genes and proteins such as farnesoid X receptor and cytochrome P450 family 7 subfamily A member 1. The co-administration of antibiotic cocktail counteracted the effect of Re against NAFLD. Further, the results obtained from the FMT animal study confirmed that Re's liver protective effects were at least partly driven by the regulation of gut microbiota. Re modulated bile salt hydrolase-related microbial genera to alter bile acid synthesis pathways, thereby inhibiting NAFLD progression.

ETHNOPHARMACOLOGICAL RELEVANCE:Curcuma kwangsiensis radix (CKR) is one of the most important herbs in traditional Chinese medicine. It effectively enhances blood circulation and eliminates stasis, which is highly associated with thrombosis. Furthermore, CKR is primarily produced in the Guangxi and Yunnan provinces of China. However, the quality control indicators of CKR in different production regions remain controversial. AIM:To explore the quality marker (Q-Marker) of CKR in different production regions. MATERIALS AND METHODS:First, we determined the UPLC fingerprints of CKR from different production regions. Second, in vitro, antiplatelet aggregation biopotency (AAB) was measured using a parallel-line assay based on the quantitative response method of the bioassay. We identified CKR components and their serum metabolism using UPLC-Q-TOF-MSE. Subsequently, molecular docking technology was used for Q-Marker analysis. Finally, we established a method for the quantitative analysis of Q-Marker. RESULTS:We observed significant differences of CKR between the Guangxi and Yunnan provinces according to the UPLC fingerprint and AAB results. Eight quality control-relevant components were screened using orthogonal partial least squares based on the spectrum-effect relationship. UPLC-Q-TOF-MSE identified 57 CKR components, and 10 prototype components and 11 metabolites, respectively, were detected during serum metabolism. Ultimately, curcumenone was screened as a Q-Marker using the spectrum-effect relationship integrated with serum metabolism, which positively correlated with the quality. The AAB results of the Q-Marker indicated that curcumenone exhibited significant anti-platelet aggregation activity. The results of the Q-Marker molecular docking revealed the strongest binding effect between curcumenone and the GP-IIb/IIIa receptor, whereas that between the P2Y12 receptor and the P2Y1 receptor was the weakest. In addition, quantitative analysis of the Q-Marker indicated that there were significant differences in the contents of the Q-Marker from different production regions. CONCLUSIONS:We identified a Q-Marker for CKR that can provide a foundation for quality evaluation research from different production regions.

The extraction of anticancer agents from medicinal plants represents a highly promising research frontier. Ginkgetin, a natural biflavone, is one of the effective pharmacological components of Ginkgo biloba leaves (GBLs). This natural product exhibits significant anti-cancer efficacy against a variety of cancer cells in vitro and demonstrates a potent inhibitory impact on tumor growth in vivo without severe toxicity. Additionally, ginkgetin synergizes with chemotherapy drugs or adjuvant therapies to potentiate antitumor effects and reduce side effects. These compelling findings underscore Ginkgetin's potential as a promising candidate for novel anti-cancer therapeutics. Therefore, this review systematically summarizes the remarkable anticancer effects of ginkgetin and elucidates its multifaceted anticancer mechanisms, including inducing cell cycle arrest, triggering programmed cell death, and preventing invasion and angiogenesis. From a molecular mechanism perspective, ginkgetin exerts anti-cancer activity by modulating critical signaling pathways (e.g. JAK/STAT, Wnt/β-catenin, AKT/GSK-3β, MAPKs, and estrogen receptor pathways) and regulating microRNA expression levels. Furthermore, target identification, research limitations, future directions, and application prospects are comprehensively outlined, aiming to facilitate the clinical translation of ginkgetin.

Gout is an increasingly prevalent global health issue, characterized by disrupted uric acid (UA) homeostasis, and commonly accompanied by hepatorenal damage and inflammation. This study investigated the effects of limonin on tissue pathology, UA regulation, and inflammation in a gout rat model. Histopathological analysis revealed that limonin significantly alleviated damage in the ankle joints, livers and kidneys. Limonin restored UA balance by enhancing renal UA excretion via transporter modulation and reducing UA production through inhibition of hepatic xanthine oxidase. Urine metabolomic analysis further confirmed that limonin modulates purine and pyrimidine metabolic pathways involved in UA regulation. Transcriptomic analysis of hepatorenal tissues, supported by western blotting, molecular docking, and CETSA, revealed that limonin bound to AMPK and then inhibited NF-κB signaling, thus exhibiting anti-inflammatory and UA-lowering effects. These findings showed limonin's dual role in UA regulation and inflammation inhibition, highlighting its potential as a functional food ingredient for gout management.

ETHNOPHARMACOLOGICAL RELEVANCE:Gouty arthritis (GA), a metabolic disorder, presents as recurrent acute arthritis with hyperuricemia. Huazhuo Sanjie Chubi decoction (HSCD), a traditional Chinese medicine oral decoction, has been extensively utilized for spleen deficiency with damp retention and GA management. However, its mechanisms against GA and associated renal fibrosis remain underexplored. AIM OF THE STUDY:This study aimed to elucidate the mechanisms by which HSCD mitigates GA and hyperuricemia-associated renal fibrosis. METHODS:The chemical components of HSCD were analyzed by UHPLC-Q-Orbitrap-MS. Serum and urine samples were collected to assess uric acid (UA) levels and renal function. H&E, Gomori's methenamine silver, and Masson's trichrome staining were performed to assess pathological changes in the ankles and kidneys. RNA sequencing (RNA-seq), Western blot analyses, and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were conducted to investigate potential mechanisms. RESULTS:Twenty-eight key compounds in HSCD were quantified. HSCD treatment significantly reduced serum and urine UA levels, decreased monosodium urate crystal deposition, inhibited inflammation in the ankle and kidney, and alleviated renal fibrosis. These effects were attributed to the modulation of renal transporters OAT1, OAT2, ABCG2, and Glut9 to promote UA excretion, thereby mitigating GA and renal fibrosis. RNA-seq, Western blot, and RT-qPCR further suggested that the anti-GA and anti-renal fibrosis effects of HSCD were related to the regulation of renal transporters and deactivation of the JAK2/STAT3 signaling pathway. CONCLUSION:HSCD exerts therapeutical effects against GA and renal fibrosis by improving arthritis symptoms, reducing UA levels, regulating renal transporters, and deactivating the JAK2/STAT3 signaling pathway. These effects highlighted its pharmacological potential and enhancing the scientific foundation for its clinical use in treating GA and related renal fibrosis.

Based on ultra-high performance liquid chromatography-quadrupole-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS) technology and network pharmacology, this study explored the pharmacodynamic substances and potential mechanisms of Huazhuo Sanjie Chubi Decoction in the treatment of gouty arthritis(GA). UPLC-Q-Exactive Orbitrap-MS technology was used to identify the components in Huazhuo Sanjie Chubi Decoction, and the qualitative analysis of its active ingredients was carried out, with a total of 184 active ingredients identified. A total of 897 active ingredient targets were screened through the PharmMapper database, and 491 GA-related disease targets were obtained from the OMIM, GeneCards, CTD databases. After Venn analysis, 60 intersecting targets were obtained. The component target-GA target network was constructed through the Cytoscape platform, and the STRING database was used to construct a protein-protein interaction network, with 16 core targets screened. The core targets were subjected to Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses, and the component-target-pathway network was constructed. It was found that the main active ingredients of the formula for the treatment of GA were phenols, flavonoids, alkaloids, and terpenoids, and the key targets were SRC, MMP3, MMP9, REN, ALB, IGF1R, PPARG, MAPK1, HPRT1, and CASP1. Through GO analysis, it was found that the treatment of GA mainly involved biological processes such as lipid response, bacterial response, and biostimulus response. KEGG analysis showed that the pathways related to the treatment of GA included lipids and atherosclerosis, neutrophil extracellular traps(NETs), IL-17, and so on. In summary, phenols, flavonoids, alkaloids, and terpenoids may be the core pharmacodynamic substances of Huazhuo Sanjie Chubi Decoction in the treatment of GA, and the pharmacodynamic mechanism may be related to SRC, MMP3, MMP9, and other targets, as well as lipids and atherosclerosis, NETs, IL-17, and other pathways.

OBJECTIVE:This paper aims to analyse the institutional design and practical challenges of Traditional Chinese Medicine (TCM) in Australia based on its legislative history, current regulatory framework, and educational system development, thereby providing references for TCM internationalisation under China's 'Belt and Road' Initiative. METHODS:This study delineates Australia's TCM legislative trajectory, dissects its registration and regulatory architecture, and examines its TCM education evolution - from private training to university programs - along with curriculum, faculty, and clinical placement features, while identifying existing challenges. RESULTS:Australia established TCM's legal status via state-led pilots through to national legislation, with a registration system centred on public safety. Its education integrates TCM-Western medicine but faces inconsistent curricula and language barriers. TCM lacks Medicare coverage, and some herbal medicines are restricted. CONCLUSIONS:Australia's experience shows a pathway for TCM institutionalisation in Western societies. Refining standards, optimising education through China-Australia cooperation, and enhancing public communication will promote TCM's sustainable development and provide replicable models for its internationalisation.

Ginsenoside Rd (G-Rd), found in Panax species, has shown therapeutic potential against metabolism-associated fatty liver disease (MAFLD), but its mechanism has not been well elucidated. This study investigated the key mechanisms of G-Rd in modulating the gut microbiome and lipid peroxidation-mediated ferroptosis pathway in MAFLD. A high-fat diet-induced MAFLD model was established. Ultrastructural changes in liver tissue were observed using transmission electron microscopy. Metagenomics were employed to detect alterations in gut microbiota and their metabolites. Biochemical analysis and immunohistochemistry were used to examine liver injury, blood lipids, lipid peroxidation-related indicators, and tissue iron content. G-Rd significantly reduced liver injury and steatosis in MAFLD mice and downregulated the elevated relative abundance of Firmicutes and the Firmicutes/Bacteroidetes ratio. It also significantly reduced the abundances of Faecalibaculum rodentium while increasing Muribaculum intestinale, with its functional role being relevant to lipid metabolism regulation. Moreover, G-Rd ameliorated mitochondrial damage and inhibited the ferroptosis pathway in the liver, which was associated with antioxidant-related factors mediated by Nrf2 signaling. The liver protective effect of G-Rd was driven by the regulation of gut microbiota, as demonstrated by antibiotic cocktail treatment and fecal microbiota transplantation. G-Rd attenuated HFD-induced MAFLD by alleviating liver oxidative stress, lipid peroxidation, and ferroptosis through modulation of the gut microbiota. The antioxidant and anti-ferroptotic actions of G-Rd, mediated via the Nrf2 pathway, were found to contribute to the amelioration of liver injury and hepatic steatosis in MAFLD.

ETHNOPHARMACOLOGICAL RELEVANCE:Biyuantong decoction (BYT), a traditional Chinese medicinal formulation, has been used for years to treat chronic rhinosinusitis (CRS) with good clinical results. However, the underlying mechanisms of its treatment for CRS remain to be fully elucidated. STUDY AIM:This research investigates the molecular mechanism by which BYT ameliorates CRS and provide new perspectives for CRS treatment research. MATERIALS AND METHODS:Clinical research is conducted on CRS patients who underwent surgery, and post-operative treatments and observations were performed. The pathological alterations of CRS were inspected by H&E staining and nasal endoscopy. Flow cytometry and ELISA were employed to measure the levels of inflammatory cells and cytokines in the peripheral blood of CRS patients. The cytotoxic impacts of BYT were assessed by cell viability, cell cycle and apoptosis assays. The effects of BYT on the adhesion and invasion of inflammatory cells to endothelial cells were evaluated by hetero-adhesion and transwell assay. Flow cytometry was employed to analyze the expression of cell adhesion molecules (CAMs) on HUVECs. The effects of BYT on NF-κB signaling pathway was analyzed by Western blot and immunofluorescence staining. The chemical components of BYT was determined by UPLC-HRMS, and network pharmacology analysis was adopted to predict potential targets in the NF-κB pathway. RESULTS:Clinical samples demonstrated that BYT treatment could effectively alleviate sinus mucosal edema and significantly decreased the recurrence rate after surgery. H&E staining disclosed obvious inflammatory cell infiltration in the sinus mucosa of CRS patients. Flow cytometry and ELISA results indicated that BYT treatment reduced the levels of eosinophils (median decrease 16.21 %) and cytokines in peripheral blood. Cell adhesion and transwell assays manifested that BYT inhibited the adhesion and invasion of U937 cells to TNF-α-induced HUVECs. Moreover, BYT counteracted the TNF-α-induced upregulation of CAMs on endothelial cells. Western blot and immunofluorescence analyses confirmed that BYT reduced the expression of NF-κB-related proteins and hindered the nuclear translocation of NF-κB. Network pharmacology analysis and component identification of BYT further supported the function of its compounds in synergistically modulating NF-κB signaling. CONCLUSION:BYT enhances the clinical efficacy of CRS by suppressing inflammatory cell adhesion and infiltration into the nasal mucosa via NF-кB pathway regulation. These findings provide a robust foundation for the clinical application of BYT in CRS treatment and suggest interrupting inflammatory cell adhesion as a potential new approach.

ETHNOPHARMACOLOGICAL RELEVANCE:Huazhuo Sanjie Chubi Decoction (HSCD), a Chinese herbal formula, is traditionally used for the treatment of spleen deficiency with dampness accumulation and is commonly used to treat gouty arthritis (GA). However, the potential active compounds and mechanisms of HSCD remain unclear. AIM OF THE STUDY:To elucidate the key bioactive compounds and pharmacological mechanisms of HSCD in treating GA. MATERIALS AND METHODS:The chemical compounds in HSCD were qualitatively and quantitatively analyzed using ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Network pharmacology and molecular docking were employed to identify key active compounds and associated molecular pathways. Monosodium urate (MSU)-induced RAW264.7 macrophages and GA rat model were used to explore the potential therapeutic effects and mechanisms of HSCD in treating GA. RESULTS:UPLC-MS/MS identified 184 compounds in HSCD, with 28 key compounds quantified. Network pharmacology revealed that verbenalin, limonin, and quercitrin are strongly associated with the molecular mechanisms of HSCD in treating GA via the PI3K-AKT signaling pathway. These compounds exhibited strong binding affinity to PI3K and AKT proteins. In RAW264.7 cells, HSCD and the three identified compounds dose-dependently reduced inflammation by inhibiting nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). They also downregulated both the PI3K-AKT and apoptosis signaling pathways. In rats, HSCD exerted therapeutic effects against acute GA by alleviating swelling and pathological damage to the ankle joints. Moreover, the molecular mechanisms in vivo were confirmed to be associated with the PI3K-AKT and apoptosis signaling pathways. CONCLUSION:This study employed a multivariant approach to demonstrate the main bioactive compounds and molecular mechanisms of HSCD in treating GA, thereby supporting its traditional use.

Distant metastasis is a major cause of treatment failure in cancer patients and a key challenge to improving cancer care today. We hypothesized that enhancing anti-cancer immune response and inhibiting circulating tumor cells (CTCs) adhesion and transendothelial migration through synergistic multi-target approaches may effectively prevent cancer metastasis. “Fuyuan Decoction” (FYD) is a traditional Chinese medicine compound that is widely used to prevent postoperative metastasis in cancer patients, but its underlying mechanism remains unclear. In this work, we systematically elucidated the underlying molecular mechanism by which FYD prevents cancer metastasis through multi-compound and multi-target synergies in vitro and in vivo. FYD significantly prevented cancer metastasis at non-cytotoxic concentrations by suppressing the adhesion of CTCs to endothelial cells and their subsequent transendothelial migration, as well as enhancing anti-cancer immune response. Mechanistically, FYD interrupts adhesion of CTCs to vascular endothelium by inhibiting TNF-α-induced CAMs expression via regulation of the NF-κB signaling pathway in endothelial cells. FYD inhibits invasion and migration of CTCs by suppressing EMT, PI3K/AKT and FAK signaling pathways. Moreover, FYD enhances the anti-cancer immune response by significantly increasing the population of Tc and NK cells in the peripheral immune system. In addition, the chemical composition of FYD was determined by UPLC-HRMS, and the results indicated that multiple compounds in FYD prevents cancer metastasis through multi-target synergistic treatment. This study provides a modern medical basis for the application of FYD in the prevention of cancer metastasis, and suggesting that multi-drug and multi-target synergistic therapy may be one of the most effective ways to prevent cancer metastasis.