Objective To investigate the mechanism of total triterpenes from fruits of Chaenomeles speciosa (CST) in alleviating N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and (NIG)-induced human gastric mucosal epithelial cells damage based on microRNA-210 (miR-210)/nuclear factor-κB (NF-κB)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome signaling pathway. Methods MNNG and NIG induced GES-1 cells model were constructed, and then GES-1 cells were treated with CST and/or miR-210 mimic, miR-210 inhibitor, NF-κB siRNA, NLRP3 siRNA. MTT assay was used to detect cell viability, immunofluorescence was utilized to measure reactive oxygen species (ROS) level, flow cytometry was used to test pyroptosis, colorimetric and ELISA methods were utilized to measure the levels of interleukin-4 (IL-4), IL-1β, IL-6, IL-10, IL-18, lactate dehydrogenase (LDH), and tumor necrosis factor-α (TNF-α) in cell supernatant. The colorimetric method was used to determine the levels of catalase (CAT), glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in cells. The dual luciferase reporter assay was employed to verify the targeted intervention between miR-210 and NF-κB. Immunofluorescence was utilized to detect the NF-κB p65 nuclear translocation, the co-localization of NLRP3, apoptosis related spot like protein (ASC) and pro-cystein-asparate protease-1 (pro-Caspase-1). qRT-PCR was used to detect the mRNA expression levels of miR-210, NF-κB, NLRP3. Western blotting was utilized to determine the protein expression levels of cytoplasmic NF-κB, nuclear NF-κB, NIMA-related kinase 7 (NEK7), thioredoxin-interacting protein (TXNIP), NLRP3, ASC, Caspase-1, pro-Caspase-1, gasdermin D (GSDMD), N-terminal domain of GSDMD (GSDMD-N), pro-IL-18 and pro-IL-1β. Results miR-210 was a pro-inflammatory/pyroptosis factor, the dual luciferase reporter assay confirmed that it could bind to 3’-UTR in mRNA of NF-κB. Compared with model group, CST significantly inhibited pyroptosis and LDH release in MNNG and NIG induced GES-1 cells (P < 0.01), reduced levels of IL-1β, IL-6, IL-18, TNF-α in supernatant and ROS, MPO, MDA in cells (P < 0.01), elevated levels of IL-4, IL-10 in supernatant and activities of CAT, GSH, SOD, T-AOC in cells (P < 0.01), suppressed the NF-κB p65 nuclear translocation and the co-localization of NLRP3, ASC and pro-Caspase-1 (P < 0.01), downregulated the mRNA expressions of miR-210, NF-κB, NLRP3 and the protein expressions of nuclear NF-κB p65, TXNIP, NEK7, NLRP3, ASC, pro-Caspase-1, Caspase-1, GSDMD, GSDMD-N, pro-IL-1β, pro-IL-18 (P < 0.01), upregulated the cytoplasmic NF-κB p65 protein expression (P < 0.01). The combination of CST with miR-210 inhibitor, NF-κB siRNA and NLRP3 siRNA transfection could further improve the above indicators (P < 0.01). Conclusion CST has a good preventive and therapeutic effect on MNNG and NIG-induced GES-1 cell injury, and its mechanism is closely related to inhibiting the abnormal elevation of miR-210 and activation of NF-κB/NLRP3 inflammasome signaling pathway, alleviating oxidative stress and subsequent suppressing pyroptosis.

Objective To investigate the effect of andrographolide on head and neck squamous cell carcinoma, and reveal its potential anti-tumor mechanism from the perspective of ferroptophagy through lipidomics. Methods CAL27 cells were cultured in vitro. CCK-8 assay and colony formation assay were used to detect the inhibitory effect of andrographolide on cell proliferation, and flow cytometry was used to detect the cell apoptosis. A patient-derived xenograft (PDX) mouse model of human head and neck squamous cell carcinoma was established. Control group and andrographolide (10 mg/kg) group were set up, and after intervention with administration, the tumor growth was observed dynamically, and the expression of proliferation-related index Ki67 in tumor tissues was detected. Lipidomics technology was used to analyze the metabolic profile changes of CAL27 cells after andrographolide treatment, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. Western blotting was used to detect the expressions of ferroptophagy-related proteins in cells. Results The in vitro experimental results showed that andrographolide could significantly inhibit the proliferation of CAL27 cells and induce cell apoptosis (P < 0.01, 0.001), and showed a dose-dependent effect. The results of in vivo PDX model experiment showed that andrographolide could significantly inhibit the growth of tumors in vivo (P < 0.001), and the positive expression of Ki67 in tumor tissues was significantly reduced compared to the control group (P < 0.001). The lipidomics analysis and KEGG pathway enrichment results showed that the differential metabolic pathways of cells treated with andrographolide were enriched in ferroptosis and autophagy-related pathways. Western blotting results confirmed that andrographolide could significantly regulate the expressions of ferroptophagy-related proteins (P < 0.05, 0.01, 0.001), indicating that andrographolide could effectively induce ferroptophagy in CAL27 cells. Further validation with the nuclear receptor coactivator 4 (NCOA4) targeted degrader and ferroptosis inhibitor demonstrated that both agents markedly reversed the effects of andrographolide (P < 0.05, 0.01, 0.001), verifying that its antitumor effect depends on the ferroptophagy signaling pathway. Conclusion Andrographolide could significantly inhibit the proliferation and induce apoptosis of head and neck squamous cell carcinoma in vitro and in vivo. Its anti-tumor mechanism is related to inducing ferroptophagy in tumor cells.

Objective To investigate the molecular mechanism by which Linggui Zhugan Decoction (苓桂术甘汤, LGZGD) ameliorates heart failure (HF) after myocardial infarction (MI) via regulating mitophagy mediated by mitochondrial calcium uniporter (MCU)-mitochondrial calcium (mtCa²⁺) signaling pathway. Methods Rat models of post-MI HF and hydrogen peroxide (H₂O₂)-induced H9c2 cardiomyocyte injury models were established. For in vivo experiments, rats were assigned to sham group, model group, LGZGD group and captopril group. For in vitro experiments, H9c2 cells were divided into corresponding groups with additional interventions of MCU knockdown or overexpression. Echocardiography, Masson staining and transmission electron microscopy were performed to evaluate rat cardiac function, myocardial fibrosis and mitochondrial ultrastructure, respectively. The mRNA and protein expression levels of genes related to mitophagy and mitochondrial function were detected by qRT-PCR and Western blotting. The levels of reactive oxygen species (ROS) and mtCa²⁺ were measured, myocardial injury markers were quantitatively analyzed, and changes in cellular autophagic flux were observed using confocal laser scanning microscopy. Results Compared with sham group, rats with post-MI HF exhibited significantly impaired cardiac function, aggravated myocardial fibrosis and mitochondrial damage, dysregulated transcription of mitophagy-related genes, and markedly upregulated MCU transcription level (P < 0.001). In H₂O₂-stimulated H9c2 cells, ROS and mtCa²⁺ levels were significantly increased, autophagic flux was blocked, and myocardial injury marker levels were notably elevated (P < 0.01).Compared with model group, LGZGD treatment significantly improved cardiac function, alleviated myocardial fibrosis and mitochondrial damage, downregulated MCU expression (P < 0.01), regulated the mRNA expressions of autophagy and mitophagy-related genes in post-MI HF rats (P < 0.05, 0.01). At the cellular level, LGZGD reduced ROS and mtCa²⁺ levels in H₂O₂-injured H9c2 cells, restored mitochondrial morphology and autophagic flux, and decreased myocardial injury marker levels (P < 0.01). MCU knockdown enhanced the cardioprotective effect of LGZGD, whereas MCU overexpression exacerbated cellular injury. LGZGD effectively reversed MCU overexpression-induced cellular damage and ameliorated the dysregulation of oxidative stress-related enzymes (P < 0.05, 0.01). Conclusions LGZGD alleviates oxidative stress injury and improves post-MI heart failure by restoring MCU-mtCa²⁺ homeostasis and promoting mitochondrial autophagic flux.

Objective To investigate the effect and underlying mechanism of Prunella vulgaris-derived exosomes loaded with orlistat (PVENs-Orl) on lipid metabolism and malignant biological behaviors of hepatoma cells. Methods PVENs were extracted by ultracentrifugation. After characterization by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA), a PVENs-Orl drug delivery system was constructed. The inhibitory effects of PVENs-Orl on the proliferation, migration and invasion of HepG2 cells were evaluated through CCK-8, cloning, scratch and Transwell assays. The free fatty acids (FFA) uptake, fatty acid synthase (FASN) activity, triglyceride (TG) content, lipid peroxidation indicators and mitochondrial membrane potential and morphology were detected. The expressions of genes related to lipid metabolism and ferroptosis [acyl-CoA synthetase long-chain family member 4 (ACSL4), glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), B-cell lymphoma-2 interacting protein 3 (BNIP3L), lysophosphatidylcholine acyltransferase 3 (LPCAT3), transferrin receptor (TFRC)] was detected by qRT-PCR. Results The PVENs were successfully extracted and characterized, and the constructed PVENs-Orl could be effectively internalized by HepG2 cells. PVENs and PVENs-Orl exhibited dose-dependent inhibition of the proliferation, migration and invasion of HepG2 cells (P < 0.001). In terms of lipid metabolism phenotype, PVENs-Orl could inhibit FASN activity (P < 0.01), and simultaneously cause intracellular accumulation of FFA and TG (P < 0.01). PVENs-Orl significantly increased the levels of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) (P < 0.01, 0.001), and increased the release of lactate dehydrogenase (LDH) (P < 0.001). PVENs-Orl significantly induced a decrease in mitochondrial membrane potential (P < 0.01) and triggered mitochondrial dysfunction (P < 0.001). At the molecular mechanism level, PVENs-Orl upregulated the expressions of genes related to lipid peroxidation and ferroptosis promotion (P < 0.001), and downregulated the expressions of genes related to antioxidant and ferroptosis inhibition (P < 0.001). Conclusion PVENs could effectively deliver orlistat, disrupt lipid homeostasis by inhibiting FASN activity, induce mitochondrial dysfunction and lipid peroxidation, and synergistically regulate the expressions of key genes involved in ferroptosis, thereby inhibiting the growth of liver cancer cells. This provides experimental evidence for targeted lipid metabolism therapy based on natural exosomes.

Objective To explore the therapeutic effect and related mechanism of citronellol on methicillin-resistant Staphylococcus aureus (MRSA)-induced skin infections in mice via NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome pathway. Methods The minimum inhibitory concentration (MIC) of citronellol against standard MRSA strains was determined using the microbroth dilution method. A full-thickness skin MRSA infection model was established in female C57BL/6 mice, control group, model group, positive control (2% mupirocin) group, citronellol low-, medium- and high-dose (0.75%, 1.50%, 3.00% citronellol gel) groups were set up. Local administration was performed for 7 d. Wound healing rates were dynamically monitored, hematoxylin-eosin (HE) staining was used to observe histopathological changes in skin tissues, and ELISA was used to detect levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD) and malondialdehyde (MDA) in skin tissues. Masson staining was used to observe collagen fiber deposition in skin tissues. RNA-seq transcriptome sequencing, qRT-PCR and Western blotting were employed to analyze the expressions of NLRP3 inflammasome pathway-related genes and proteins in skin tissues, while molecular docking was used to predict the binding affinity of citronellol to relevant inflammatory targets. The lactate dehydrogenase (LDH) release assay was used to evaluate the effect of citronellol on pyroptosis in MRSA-infected immortalized keratinocytes. Results Citronellol exhibited significant antibacterial activity against MRSA, with an MIC ranging from 0.312 5—0.625 0 mg/mL. In vivo experiments revealed that compared with model group, citronellol significantly accelerated wound healing, reduced inflammatory cell infiltration and tissue edema, decreased IL-1β and TNF-α levels in skin tissues (P < 0.05, 0.01, 0.001), markedly increased collagen fiber deposition at wound sites (P < 0.05, 0.01), and inhibited the activation of NLRP3 inflammasome-cell pyroptosis pathway (P < 0.05, 0.01, 0.001). Molecular docking results indicated that citronellol could bind to target proteins such as IL-1β. In vitro experiments demonstrated that citronellol significantly suppressed LDH release in MRSA-infected HaCaT cells (P < 0.05, 0.01). Conclusion Citronellol exerts significant antibacterial, anti-inflammatory and wound repair-promoting effects on MRSA-induced skin infections in mice. Its mechanism may be related to inhibiting NLRP3 inflammasome-cell pyroptosis pathway, reducing the release of pro-inflammatory cytokines, improving the local immune microenvironment, and promoting collagen deposition.

Objective To evaluate the clinical efficacy and safety ofKushen Gel combined with oral fluconazole in the treatment of vulvovaginal candidiasis (VVC). Methods A multicenter, randomized, double-blind, placebo-controlled trial was conducted. Patients with VVC who visited four hospitals between June 2024 and October 2025 were recruited and randomly assigned in a 1∶1 ratio to the placebo group or the Kushen Gel group. Both groups received a single oral dose of fluconazole 150 mg. Additionally, the Kushen Gel group received intravaginal Kushen Gel, while the placebo group received a placebo gel with identical appearance and properties, both for 14 consecutive days. The primary efficacy endpoint was the clinical response rate at 7—10 days after drug discontinuation. Secondary efficacy endpoints included the proportion of patients with normalization of at least four vaginal microecological indicators, time to complete resolution of vulvar pruritus, pain, and abnormal discharge, vaginal cleanliness, and mycological recurrence rate at 25—31 days after drug discontinuation. Adverse events were monitored to evaluate safety. Results A total of 93 patients were randomized. At the 7—10 days post-treatment follow-up visit, the clinical response rate in the Kushen Gel group was significantly higher than that in the placebo group (87.50% vs 65.00%, P = 0.033 9). The proportion of patients achieving normalization of at least four vaginal microecological indicators was significantly higher in the Kushen Gel group compared with the placebo group [97.50% vs 77.50%, P = 0.0143]. The times to resolution of vulvar pruritus, pain, and abnormal discharge in the Kushen Gel group were all shorter than those in the placebo group, but the differences were not statistically significant (P > 0.05). At the 25—31 days post-treatment follow-up, the mycological recurrence rate was 10.53% (2/19) in the Kushen Gel group, which was lower than that in the placebo group (23.81%, 5/21). The incidence of adverse events was low in both groups, with no statistically significant difference between groups (P > 0.05), and no serious adverse events occurred. Conclusion The combination of Kushen Gel with oral fluconazole can alleviate clinical symptoms, enhance short-term clinical efficacy, and may help reduce the recurrence rate, as well as, the recovery of vaginal microecology in patients with VVC. The treatment is safe and possesses significant clinical value.

Objective To investigate the medication regularity of traditional Chinese medicine (TCM) for preventing and treating alcohol-related liver and brain injuries, and to explore the intervention mechanisms of Jiejiu Formula (JJF) with homology of medicine and food on alcoholic liver disease (ALD) and alcohol-related brain injury (ARBI) through network pharmacology and experimental validation. Methods Herbal formulas targeting ALD and ARBI were collected from TCM classics of the Tang, Song, Yuan, and Ming dynasties. Frequency analysis was performed on the constituent herbs to identify high-frequency ingredients. Their properties, flavors, meridian affinities, and dosages were analyzed. Association rule mining and cluster analysis were then applied to identify a core herbal combination for the JJF. Network pharmacology was used to screen the active components and potential targets of the JJF against ALD and ARBI. Core targets were subjected to gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses. Molecular docking and molecular dynamics simulations were employed to validate the predicted mechanisms and binding stability. Subsequently, an alcohol-induced liver and brain injury mouse model was established to verify the findings in vivo. Results A total of 301 formulas containing 263 distinct herbs were identified. Herbs with a warm nature, pungent flavor, stomach meridian affinity, and heat-clearing functions were the most frequently used. The combination of “Gancao (Glycyrrhizae Radix et Rhizoma)-Chenpi (Citri Reticulatae Pericarpium)-Ganjiang (Zingiberis Rhizoma)” demonstrated the highest frequency and strongest association, and all three herbs are classified as medicinal food ingredients. Therefore, this combination was designated as the JJF. Network pharmacology identified 289 active components and 254 potential targets for the JJF against ALD/ARBI. Protein-protein interaction (PPI) network analysis revealed five core targets. KEGG enrichment analysis highlighted pathways related to endocrine resistance. Molecular docking suggested that 2-[(3R)-8,8-dimethyl-3,4-dihydro-2H-pyrano[6,5-f]chromen-3-yl]-5-methoxyphenol and phaseolinisoflavan have good binding affinity and stability with protein kinase B1 (AKT1), which was further confirmed by stable binding in MD simulations. In vivo experiments showed that the JJF alleviated alcohol-induced damage in liver and brain tissues to varying degrees. Conclusion JJF exerts preventive and therapeutic effects against ALD and ARBI. It potentially acts on key targets such as Src proto-oncogene tyrosine-protein kinase (SRC), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), and AKT1, and modulates pathways like endocrine resistance. Furthermore, the JJF may accelerate alcohol metabolism, enhance anti-oxidative stress capacity, upregulate mRNA expression within the phosphatidylinositol-3-hydroxykinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) signaling pathway, and inhibit mitogen-activated protein kinase 3 (MAPK3) expression, thereby regulating cell proliferation, apoptosis, and inflammatory responses to achieve its protective effects.

Objective To comprehensively sort out and visually analyze the research status, hotspots and trends of knowledge graph technology in the field of traditional Chinese medicine (TCM) from the perspectives of bibliometrics and global patents, provide references for in-depth research and development in this field. Methods The relevant Chinese and English literature published between January 1, 2012, and December 31, 2025, was systematically retrieved from the CNKI, Wanfang, VIP, and Web of Science databases. Using tools such as CiteSpace, VOSviewer, and Excel, we performed bibliometric and visual analyses of publication trends, institutional and author collaboration networks, journal sources, and keyword co-occurrence and evolution. Additionally, we analyzed global patent applications from the Incopat database, focusing on application trends, geographical distribution, applicants, and technical themes. Results A total of 745 valid publications were included (618 in Chinese, 127 in English) and 432 patent families. Research on knowledge graph technology in TCM is in a phase of rapid development. Current hotspots focus on refining knowledge extraction techniques, developing KG-assisted diagnosis and treatment systems, intelligent question-answering, and synergistic applications with large language models. The field is moving toward deeper interdisciplinary integration and clinical application . China occupies a dominant position in both research output and patent applications (96% share), with established collaboration networks. However, cross-regional and interdisciplinary cooperation requires further strengthening. Conclusion Research on knowledge graph technology in the field of traditional Chinese medicine is deeply evolving from fundamental construction to intelligent applications. Despite China’s quantitative advantage in patents, challenges persist, including a relative lack of high-value patents, technical homogenization, and insufficient international competitiveness.

Objective To analyze the knowledge evolution context, patent genealogy characteristics and prospective innovation directions of Euphorbia plant research, and clarify the efficiency of achievement transformation and the global competitive pattern, this study adopts bibliometric and knowledge graph analysis methods, combined with patent information lineage analysis technology to carry out systematic research. Methods The relevant literature in the CNKI, Wanfang, VIP and Web of Science Core Collection were systematically searched with Euphorbia pekinensis and Euphorbia as the subject words, imported into NoteExpress literature management software for deduplication, screening and format conversion, and CiteSpace and VOSviewer software were used to visualize the annual publication trends, the publishing country, institution, author, journal distribution, discipline distribution and other dimensions of Chinese and English literature. At the same time, based on the Incopat patent database, the application and distribution status of patents related to Euphorbia plants were systematically analyzed from aspects such as the number of patent applications and geographical distribution patterns. Results After the screening procedure, a total of 1 682 Chinese literature and 1 339 English literature were incorporated into the study, indicating that the research of Euphorbia plants was at the international stage. A total of 98 countries had issued documents, among which China had published the most achievements. The author with the largest number of publications in both Chinese and English was Zhang Li. The core publishing institutions of the Chinese and English literature were Nanjing University of Chinese Medicine and the United States Department of Agriculture. The cooperation connections between these institutions were still relatively scattered, but some authors had formed stable research teams. The Chinese and English journals with the highest number of articles published were China Journal of Chinese Materia Medica and Journal of Ethnopharmacology. The English literature mainly involved plant science and extended to pharmacology, medicinal chemistry and other fields. The Incopat patent database retrieved 8 350 patents related to Euphorbia plants, and 5 603 domestic patents, accounting for 67.1% of the total, which was in a dominant position. Conclusion Research on Euphorbia plants shows a pattern dominated by China, coexisting with global participation. The fundamental research system in this field has become increasingly well-established, with remarkable progress in the modernization of traditional medicinal applications and broad prospects for industrial transformation, but there are still problems such as regional development imbalance. In the future, it should deepen the basic theory, promote interdisciplinary research, optimize the patent layout, and realize the efficient transformation and sustainable development of resource value.

Objective To clone the cycloartenol synthase (CAS) gene PcCAS from Polygonatum cyrtonema, perform bioinformatic and recombinant protein expression analyses, and determine its subcellular localization etc, thereby providing essential resources for further elucidating the pivotal role of PcCAS in saponin biosynthesis. Methods Based on the transcriptome data of P. cyrtonema, the PcCAS gene was screened and cloned, and bioinformatics approaches were employed to analyze the physicochemical properties, secondary structure, and tertiary structure of its encoded protein. qRT-PCR was used to analyze the tissue-specific expression of PcCAS. A prokaryotic expression vector was constructed and transferred into Escherichia coli for prokaryotic expression analysis. Yeast expression vectors were constructed and transformed into Pichia pastoris for subsequent yeast expression analysis. A subcellular-localization vector was built and infiltrated into Nicotiana benthamiana leaves for the analysis of heterologous expression in tobacco. Results The open reading frame (ORF) of PcCAS is 2 286 bp in length and encodes 759 amino acids. The protein is hydrophilic and lacks a signal peptide. The total saponin content and PcCAS expression level in the rhizome are significantly higher than those in the stem, leaf, and flower, indicating the expression level of PcCAS is positively related with the total saponin content. Yeast-expressed recombinant protein matched the expected molecular weight of 86 400. Subcellular-localization assays in tobacco leaves demonstrated that the PcCAS protein is predominantly localized to the endoplasmic reticulum. Conclusion PcCAS protein belongs to the OSC superfamily of enzymes and is localized in the cytoplasm, positively regulating the total saponin biosynthesis.