Objective To explore the effect of usnic acid on inflammatory injury in mice with intracerebral hemorrhage (ICH) based on high mobility group box 1 (HMGB1)/receptor for advanced glycation end products (RAGE) pathway. Methods C57BL/6 mice were randomly divided into sham group, model group, HMGB1 inhibitor ethyl pyruvate (40 mg/kg) group, usnic acid low-, high-dose (20, 30 mg/kg) groups and usnic acid (30 mg/kg) + rHMGB1 (8 μg/kg) group, with 18 mice in each group. The ICH model was constructed by intracranial injection of 0.5 μL collagenase. After continuous administration for 3 d, the modified neurological severity score (mNSS) and left turn rate of mice in each group were evaluated. The water rate of brain tissue in mice was determined. ELISA was used to measure the levels of interleukin-6 (IL-6), IL-1β and IL-18 in brain tissue. Hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue. Immunofluorescence was used to observe microglia activation in brain tissue. TUNEL staining was used to detect cell apoptosis of brain tissue. Western blotting was used to detect the expressions of HMGB1/RAGE pathway related proteins in brain tissue. Results Compared with sham group, the striatum tissue of mice in model group was severely damaged, mNSS, left cerebral hemisphere water rate, ionized calcium-binding adapter molecule 1 (Iba1) positive cells number, inflammatory factors levels, cell apoptosis rate, HMGB1, RAGE, phosphorylated nuclear factor-κB p65 (p-NF-κB p65)/NF-κB p65 and TNF-α protein expression levels were significantly increased (P < 0.05), and the left turn rate was significantly decreased (P < 0.05). Compared with model group, the damage degree of striatum tissue in usnic acid low-, high-dose group and ethyl pyruvate group was alleviated (P < 0.05), mNSS, left cerebral hemisphere water rate, Iba1 positive cells number, inflammatory factors levels, cell apoptosis rate and HMGB1/RAGE pathway related proteins expression levels were significantly decreased (P < 0.05), and the left turn rate was significantly increased (P < 0.05). rHMGB1 significantly reversed the inhibitory effect of usnic acid on inflammatory injury in ICH mice (P < 0.05). Conclusion Usnic acid may alleviate inflammatory injury in ICH mice by inhibiting HMGB1/RAGE pathway.

Objective To explore the characteristics, medication patterns, and the mechanisms of action of traditional Chinese medicines for treating angle-closure glaucoma. Methods Literature and prescriptions related to the treatment of angle-closure glaucoma with traditional Chinese medicine were retrieved from databases including CNKI, Wanfang, VIP, China Biology Medicine (CBM), and the Ancient and Modern Medical Case Cloud Platform. Through data mining and analysis, core prescriptions were identified. Subsequently, a network pharmacology approach was employed. Active ingredients with oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18 were screened using the TCMSP and BATMAN-TCM 2.0 databases. Targets were uniformly annotated via the UniProt database and intersected with disease targets obtained by integrating and deduplicating data from four major databases (GeneCards, TTD, OMIM, and CTD) to identify pharmacodynamic targets. A “core prescription-active ingredient-pharmacodynamic target” interaction network was constructed using Cytoscape 3.10.2. The pharmacodynamic targets were then imported into the Metascape platform (restricted to human species, with a threshold of P < 0.05) for gene ontology (GO) (biological process, molecular function, cellular component) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis, with the top 20 significant pathways and functions visualized. Additionally, a protein-protein interaction (PPI) network was constructed using the String database, and core targets were screened using the Cytoscape MCODE plugin (cut-off = 0.05) to elucidate the potential molecular mechanisms underlying the core prescriptions in treating angle-closure glaucoma. The binding energy between the active ingredient and the core target was investigated through molecular docking. Sodium dithionite was used to induce hypoxic injury in mouse retinal ganglion cells (RGCs) to establish an in vitro model. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was employed to detect the effects of these active components on mRNA expression of core target genes. ELISA was used to measure the impact of these active components on tumor necrosis factor-α (TNF-α) levels. Immunofluorescence staining was applied to evaluate the effects of these active components on phosphorylated protein kinase B (p-AKT) and nuclear factor-κB (NF-κB) protein expression. Results The top three most frequently used traditional Chinese medicines for angle-closure glaucoma were Fuling (Poria), Danggui (Angelicae Sinensis Radix), and Chaihu (Bupleuri Radix). The core syndromes were mainly liver qi stagnation and liver-kidney deficiency. A core prescription consisted of Mudanpi (Moutan Cortex)-Angelicae Sinensis Radix-Zhizi (Gardeniae Fructus)-Bupleuri Radix-Juhua (Chrysanthemi Flos)-Baizhu (Atractylodis Macrocephalae Rhizoma)-Gancao (Glycyrrhizae Radix et Rhizoma)-Baishao (Paeoniae Radix Alba)-Poria. Cluster analysis yielded three clusters: modified Chaihu Shugan San (柴胡疏肝散), modified Yijing Buyang Huanwu Decoction (益精补阳还五汤), modified Qiju Dihuang Pills (杞菊地黄丸). Network pharmacology analysis showed that the core prescription regulated key targets such as AKT1, TNF-α, NF-κB by modulating signalling pathways such as TNF, phosphatidylinositol-3-hydroxykinase (PI3K)-Akt, and MAPK (mitogen-activated protein kinase), thereby participating in biological processes such as inflammatory response, and oxidative stress, and thus intervening in the pathological process of glaucoma. Quercetin and luteolin were identified as the core components of the core prescription for the intervention of angle-closure glaucoma through molecular docking, and in vitro experiments were conducted for verification. The results showed that all luteolin dose groups (5, 10, 20 μmol/L) upregulated AKT1 mRNA expression (P < 0.01), while the medium and high dose groups downregulated NF-κB and TNF-α mRNA expression (P < 0.05), significantly increased p-AKT protein expression, and decreased NF-κB protein and TNF-α levels (P < 0.05). The medium and high dose groups of quercetin (40, 80 μmol/L) also significantly upregulated AKT1 mRNA expression and downregulated NF-κB and TNF-α mRNA expression (P < 0.05). All low (20 μmol/L), medium, and high dose groups of quercetin increased p-AKT protein expression and decreased NF-κB protein expression (P < 0.05), while the medium and high dose groups significantly reduced TNF-α expression (P < 0.05). Conclusion Data mining techniques were systematically employed to analyze ancient and modern literature and medical records concerning the treatment of angle-closure glaucoma. From multiple dimensions including medication frequency, nature-flavor-meridian tropism, efficacy categories, association rules, and core combinations, the “high-frequency-core-differential” three-dimensional characteristics were progressively analyzed to extract core prescriptions and reveal their compatibility patterns. Network pharmacology analysis further confirmed that the above high-frequency prescriptions synergistically regulate key biological processes such as oxidative stress, inflammatory response, and apoptosis through a “multi-component-multi-target-multi-pathway” mechanism, reflecting the holistic therapeutic advantages of traditional Chinese medicine compound formulas, thereby providing scientific evidence for clinical syndrome differentiation and medication, with certain guiding significance. In vitro experiments simultaneously validated that, in the hypoxic injury model, both luteolin and quercetin significantly upregulated AKT1 mRNA and p-AKT protein expression, inhibited NF-κB and TNF-α mRNA and protein expression, effectively reduced levels of inflammation and apoptosis-related factors, and exerted protective effects on RGCs through a bidirectional mechanism of “anti-inflammatory and pro-survival” effects.

Objective To conduct a bibliometric analysis of Chinese and English literature on the prevention and treatment of knee osteoarthritis (KOA) by traditional Chinese medicine (TCM) from 1995 to 2025, and visualize its research status, development context, cooperation network, hot topics and frontier trends. Methods Relevant Chinese literature from January 1995 to December 2025 were retrieved from CNKI, Wanfang and VIP, and relevant English literatures from January 1998 to December 2025 were retrieved from Web of Science (WOS). CiteSpace and VOSviewer software were used to visually analyze the trend of publication, countries, institutions, authors, keywords, etc., and draw a knowledge map. Results A total of 6 093 literature were included, including 5 242 Chinese literature and 851 English literature. The overall number of publications showed a fluctuating upward trend, and Chinese research output and cooperation in this field were dominant. The core research institutions are represented by Guangzhou University of Chinese Medicine, Fujian University of Traditional Chinese Medicine and Beijing University of Chinese Medicine. Keyword analysis showed that the current research hotspots focused on traditional therapies such as clinical efficacy, mechanism of action, traditional Chinese medicine compound, acupuncture and massage. Emergent words and cluster analysis showed that the frontier trend was turning to the mechanism exploration and application research of interdisciplinary and new drug delivery systems such as network pharmacology, metabolomics, programmed cell death, intestinal flora-articular axis, self-assembled nanocarriers and metal-organic frameworks. Conclusion Research on TCM for KOA prevention and treatment has entered a phase of rapid development and deep integration. While continuing to validate clinical efficacy, it is increasingly converging with modern technologies, shifting the research paradigm toward systems biology, precision medicine, and intelligent drug delivery. Through the integration of multi-omics, molecular subtyping, and the development of innovative formulations, TCM holds promise for playing a more pivotal role in the prevention, early intervention, and disease-modifying treatment of KOA.

Objective To conduct a visual analysis of hotspots and development trends in Curcumae Radix research domestically and internationally using bibliometric software CiteSpace and VOSviewer, and provide a reference for in-depth research and application of Curcumae Radix. Methods The China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Platform, Web of Science (WOS) Core Collection, and PubMed databases were searched as data sources. The search period was set from January 1, 2016 to December 12, 2025. Analyses of publication volume, authors, and keywords were performed on the selected literature using CiteSpace6.4.R1 and VOSviewer1.6.20 softwares. Results A total of 1 354 Chinese literature and 77 English literature were included. Shi Zhenggang was identified as the core author of Chinese publications with 21 papers, while Ji De was the core author of English publications with six papers. Chinese research has formed a bidirectional knowledge network of “clinical experience-computational mechanism” represented by “data mining” and “network pharmacology”. English research focuses on “Curcumae Radix” and “molecular docking”, demonstrating an in-depth path of “computational prediction-disease focus”. The research hotspots are rapidly evolving from early-stage clinical efficacy observation to data-driven and computational pharmacological mechanistic elucidation. Conclusion Research on Curcumae Radix has evolved from simple efficacy observation into a systematic exploration that equally emphasizes “clinical problems” and “mechanistic elucidation”, while deeply integrating computational simulation and systems biology methods. Future efforts should focus on establishing a closed-loop research system of “computational prediction-experimental verification”, promoting high-quality evidence-based transformation, thereby driving the transformation and upgrading of this field toward precision medicine and modernized pharmacology.

Objective To integrate the correlation between the administration routes of traditional Chinese medicine essential oils and diseases, as well as their core biomarkers in disease treatment, analyze their pathways and mechanisms of action on the body, and promote the rational application and innovative development of traditional Chinese medicine essential oils. Methods China National Knowledge Infrastructure(CNKI), VIP Information(VIP), Wanfang Data, Web of Science were used to analyze the distribution of administration routes for traditional Chinese herbal essential oils and the types of diseases treated. The correlation between different administration routes and disease types was examined, along with frequency statistics of biomarkers involved in the therapeutic process. After identifying the most closely associated biomarkers, their mechanisms of action in disease treatment by traditional Chinese herbal essential oils were analyzed. Results Among all administration routes, oral administration accounted for 66.23%, inhalation for 21.37%, topical application for 8.18%, intraperitoneal injection for 3.17%, and intravenous injection for 1.06%. The disease types most strongly associated with oral and inhalation routes were neurological and psychiatric disorders, while those most associated with topical application were skin diseases and immune-related conditions. Intraperitoneal injection showed the strongest association with tumor diseases. Among biomarkers appearing more than 10 times, those related to oxidative stress regulation included malondialdehyde (MDA), nitric oxide (NO), and superoxide dismutase (SOD); those involved in inflammatory response regulation included tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, and IL-8; those associated with apoptosis regulation included B-cell lymphoma-2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), and cysteinasparate protease-3 (Caspase-3); and those involved in neural regulation included 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), norepinephrine (NE), and acetylcholinesterase (AchE). Conclusion Traditional Chinese medicine essential oils enter the human body through three primary routes: inhalation, transdermal absorption and oral administratio to exert their biological effects. They can exert effects via by regulating multiple mechanisms such as oxidative stress, neurotransmitter, inflammatory responses, and apoptosis, demonstrating significant advantages in disease prevention and treatment.

Objective By conducting a whole-genome identification and functional analysis of auxin response factors (ARFs) family members in Gentiana rigescens, this study investigate their potential roles in the thermosensitive corolla opening-closing movement of its corolla. Methods Based on whole-genome data and transcriptome data, bioinformatics methods were used to analyze the physicochemical properties, conserved motifs, gene structure, cis-acting elements, and evolutionary relationships of the G. rigescens ARFs (GrARFs) gene family. Real-time quantitative reverse transcription PCR (RT-qPCR) technology was employed to detect the expression patterns of GrARFs genes under different opening and closing states. Additionally, subcellular localization experiments were conducted to verify the protein localization of GrARF9, so as to preliminarily analyze the biological functions of the GrARFs gene family. Results A total of 21 GrARFs were identified in the G. rigescens genome, encoding 612—1 065 amino acids. Their relative molecular weights ranged from 68 082.29 to 118 117.97, and their isoelectric points spanned 5.52—8.43. The secondary structure was dominated by random coils. Phylogenetic tree analysis classified the ARFs in G. rigescens into four subgroups, namely Group A, Group B, Group C, and Group D. Most GrARFs contained Auxin-resp and B3 DNA-binding domains. The promoter cis-acting elements of GrARFs were rich in hormone-responsive and light-responsive elements. The 21 GrARFs were unevenly distributed across eight chromosomes. Collinearity analysis revealed that the GrARFs in G. rigescens were relatively conserved among dicotyledonous plants and shared a considerable number of collinear pairs with the functionally characterized ARFs from Rosa hybrida. All 21 GrARFs genes exhibited differential expression during the thermosensitive process of G. rigescens corollas. Results of subcellular localization experiments showed that the GrARF9 protein was localized in the nucleus, displaying the nuclear localization characteristic of transcription factors. Conclusion The results of this study preliminarily clarify the expression pattern of GrARFs in the thermosensitive movement of G. rigescens, and lay a foundation for further research on the function of GrARFs in the corolla movement of G. rigescens.

Objective To identify members of the cycloartenol synthase (CAS) gene family (PgCAS) using the Panax ginseng transcriptome database, and analyze the expression patterns of PgCAS04-5 in overexpressed and RNAi-positive hair root materials), as well as explore the function of this gene in ginsenoside biosynthesis. Methods Bioinformatics approaches were adopted to characterize the chromosomal localization, conserved motifs, phylogenetic relationships and expression profiles, and the correlations with the gene expression of key saponin synthesis enzymes and the content of monomer saponin of PgCASs. The protein structure and subcellular localization of PgCAS04-5 were predicted. Meanwhile, the expression levels of key enzyme genes and variations in ginsenoside contents were detected in overexpression and RNAi-positive hairy root lines. Results A total of 20 PgCAS transcripts were identified from the P. ginseng transcriptome database. Chromosomal mapping showed that PgCAS genes were unevenly distributed on five chromosomes of P. ginseng. Phylogenetic tree analysis divided the PgCAS family into three subfamilies, and most PgCAS members belonged to subfamily Ⅱ and subfamily Ⅲ. The PgCAS genes displayed obvious spatiotemporal specificity in expression across different cultivars, growth years and tissues. Additionally, the PgCAS family formed an interaction network with key genes involved in ginsenoside biosynthesis, and their content was significantly correlated with the accumulation of monomeric ginsenosides. Subcellular localization prediction demonstrated that PgCAS04-5 was localized to chloroplasts. Moreover, the overexpression of this gene was inversely regulated with the expression of four genes including PgCAS04-5, PgDS-1, PgSE2-4 and PgCYP137 in RNAi positive materials, with significant differences. Conclusion A total of 20 members of the PgCAS gene family were identified in this study, and functional differentiation exists among PgCAS family members. PgCAS04-5 positively regulates the biosynthesis of ginsenosides Rg₂and Rf, and negatively modulates the synthesis of ginsenosides Rb₂and Rd. The results provide a valuable reference for further research on the PgCAS gene fami

Objective To clone the full-length cDNA sequence of the CoMYB62 gene from Cornus officinalis, and analyze the structural characteristics, phylogenetic relationships, subcellular localization, and tissue expression patterns of its encoded protein, thereby providing a molecular basis for elucidating the potential roles of MYB transcription factors in the growth, development, and environmental adaptation of C. officinalis. Methods Based on transcriptome data of C. officinalis, the CoMYB62 gene sequence was screened and its full-length cDNA sequence was cloned using RT-PCR. Bioinformatics analyses were conducted to predict the physicochemical properties, conserved domains, secondary and tertiary structures, and phylogenetic relationships of the CoMYB62-encoded protein. A pCAMBIA1300-CoMYB62-GFP fusion expression vector was constructed and transiently expressed in tobacco leaves, and the subcellular localization of the CoMYB62 protein was observed using a laser confocal inverted microscope. In addition, quantitative real-time PCR (qRT-PCR) was performed to analyze the expression patterns of CoMYB62 in four tissues, including root, stem, leaf, and fruit. Results The full-length cDNA of the CoMYB62 gene was successfully cloned. Sequence analysis indicated that the encoded protein belonged to a typical R2R3-MYB transcription factor and contained a highly conserved DNA-binding domain at the N-terminus, which was predicted to be mainly localized in the nucleus. Phylogenetic analysis revealed that CoMYB62 shared high evolutionary conservation with MYB homologous proteins from various plant species. Laser confocal microscopy further confirmed that the CoMYB62 protein was localized in the nucleus. The qRT-PCR results showed that CoMYB62 was expressed in all examined tissues of C. officinalis, although the expression levels varied significantly among different tissues. Conclusion CoMYB62 is a conserved MYB transcription factor in C. officinalis with tissue-specific expression characteristics, which may participate in the regulation of plant growth, development, and stress responses to provide a foundation for further biological functional analysis of CoMYB62 in C. officinalis.

Objective To explore the chemical profile variations of Glycyrrhiza uralensis from different origins and screen origin-specific characteristic markers by using UPLC fingerprinting integrated with chemometrics. This approach provides a scientific basis and reference for the quality evaluation of G. uralensis with unified attributes. Methods This study established UPLC fingerprints for 30 batches of G. uralensis collected from five origins and validated the analytical method. The quality consistency of the samples was preliminarily analyzed through similarity evaluation. Subsequently, chemometric methods such as principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to identify the patterns and differences of G. uralensis samples from different origins by SIMCA 14.0 software. Further, the key discriminant markers were identified based on the variable importance projection (VIP) values. Finally, the contents of the five characteristic markers were determined. Results UPLC fingerprints of 30 batches of G. uralensis samples were established. The similarities between each sample batch and the reference fingerprint varied but all exceeded 0.849. This indicates that while the overall chemical profiles of the samples were generally consistent, quantitative differences existed. Chemometric analysis showed clear clustering trends for samples from different origins, and the consistent results of PCA and HCA confirmed the presence of chemical differences among the origins. The OPLS-DA model effectively distinguished different origins, and the permutation test confirmed the reliability of the model. Five key characteristic markers were selected based on VIP > 1. The content determination results indicated that the contents of the five characteristic markers in G. uralensis from different origins varied significantly, further confirming the differences in chemical components among the origins. Conclusion The UPLC fingerprint combined with chemometrics (PCA, HCA, OPLS-DA) analysis method established in this study can effectively distinguish G. uralensis from different origins, and glycyrrhizin, glycyrrhizic acid, liquiritin apioside, glycyrrhizin B, and violanthin can be used as key characteristic markers for origin discrimination. This analytical system provides a reliable strategy and data support for objectively evaluating the quality differences of G. uralensis from different origins, identifying their origin sources, and improving future quality standards.

Objective To investigate the composition and differential characteristics of volatile organic compounds (VOCs) in Fritillariae Cirrhosae Bulbus (FCB) from six legally authorized origins, and provide a reference for origin identification and quality evaluation of FCB. Methods Headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) was employed to analyze the volatile components in dried bulbs of Fritillaria cirrhosa, F. unibracteata, F. przewalskii, F. delavayi, F. taipaiensis and F. unibracteata var. Wabuensis. Qualitative matching was performed using the built-in NIST and IMS database of GC-IMS Library Search. Principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and hierarchical cluster analysis (HCA) were applied to evaluate the differences in volatile components among samples from different origins. Results A total of 90 volatile organic compounds were preliminarily identified, mainly including aldehydes, alcohols, ketones, esters, carboxylic acids, pyrazines, and sulfur-containing compounds. Among them, aldehydes, alcohols, and ketones were the major categories. PCA and OPLS-DA results indicated that FCB samples from the six origins could be well distinguished. Based on variable importance in projection (VIP) values, 17 potential differential markers were screened out, including ethyl acetate, 2-propanone, α-phellandrene, 3-hydroxy-2-butanone, methyl acetate, 2-propanethiol, 2-isobutyl-3-methoxypyrazine, propyl acetate, 3-methylbutanal, 1-butanol, 2-methyl-2-propenal, hexanal, 3-methyl-1-pentanol, 2-methylpropanoic acid, 3-butenenitrile, benzaldehyde, and 2-octanone. Cluster heatmap analysis further demonstrated that the above differential compounds could reflect the volatile metabolic characteristics of FCB from different origins. Conclusion HS-GC-IMS combined with chemometrics can effectively characterize the differences in volatile organic compounds among FCB from different origins, providing technical reference for origin identification, quality evaluation, and characteristic component screening of FCB.