Objective To establish the HPLC fingerprint of Heleocharis dulcis and simultaneously determine the contents of chlorogenic acid, caffeic acid, ferulic acid, luteolin and jaceosidin, and to evaluate the quality of H. dulcis from different regions by chemical pattern recognition method, so that a reference can be provided for quality control of H. dulcis. Methods The fingerprint of H. dulcis was established on a ZORBAX SB C₁₈ (250 mm × 4.6 mm, 5 μm) column using acetonitrile-0.1% phosphoric acid solution as the mobile phase with gradient elution. the “Chinese medicine chromatographic fingerprint pattern similarity evaluation system”(2012.130723 edition) was used for similarity evaluation, SPSS27.0 and SIMCA14.1software was performed for hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were utilized to evaluate the quality of H. dulcis from diverse origins. The five chemical constituents were identified and content determined by comparison with the reference substance, and the results were synthesized using chemical pattern recognition and entropy weighted TOPSIS method. Results A total of 15 common peaks were matched to 26 batches of H. dulcis HPLC fingerprints, and five components including chlorogenic acid, caffeic acid, ferulic acid, luteolin and jaceosidin were identified. The similarity of fingerprints was between 0.865 and 0.995. The 26 batches of H. dulcis were clustered into two classes by HCA. PCA analysis yielded a cumulative variance contribution of 83.283% for the three primary constituents. Four components, including luteolin and chlorogenic acid, were screened by OPLS-DA as the indicator components for the quality difference of H. dulcis from different regions. The mass fractions of chlorogenic acid, caffeic acid, ferulic acid, luteolin and jaceosidin in 26 batches of H.dulcis were 5.43—60.65 , 0.44—2.27, 1.65—6.46, 2.58—59.94 and 0.31—24.40 μg/g, successively. Both PCA and entropy weighted TOPSIS results indicated that samples from Hubei Province exhibited the best quality. Conclusion The established HPLC fingerprinting method is characterised by its stability, reliability, and reproducibility. which can be used for the quality evaluation of H. dulcis in combination with chemical pattern recognition, and provides a reference for the quality control and resource development of H. dulcis.

Objective To systematically investigate the chemical constituents of 70% acetone extract of Polygonatum cyrtonema processed by nine cycles of steaming and sun-drying. Method Compounds were isolated and purified by MCI gel, silica gel, Toyopearl HW-40C, ODS and semi-preparative HPLC. Their chemical structures were comprehensively elucidated by spectroscopic methods, such as HRESIMS, NMR, IR, UV and ECD. Results A total of 14 nitrogen-containing compounds were finally isolated and identified as 6-[(8R,9S)-8,9,10-trihydroxybutyl]-3,4-dimethylpyridazine (1), 6-ethyl-3-(hydroxymethyl)-1H-pyridin-2-one (2), (+)-(1R,3R)-1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (3), 1H-indole-3-carboxaldehyde (4), cyclo-(L-Leu-L-Phe) (5), cyclo-(L-Phe-L-Ile) (6), cyclo-(L-Asn-L-Phe) (7), (3S,6S)-3-benzyl-6-(1-hydroxyethyl)-2,5-piperazinedione (8), cyclo-(D-Phe-Gly) (9), cyclo-(L-Tyr-L-Tyr) (10), cyclo-(L-Val-L-Tyr) (11), pedatisectine L (12), cyclo-(Ala-Tyr) (13), and (2S,5S)-5-[(1S)-1-methylpropyl]-3,6-dioxo-2-piperazinepropanoic acid (14). Conclusion Compounds 1 and 2 were characterized as two new compounds named cyrtoneine J and cyrtoneine K, while compounds 3 and 5—14 were isolated from the genus Polygonatum for the first time.

Objective To evaluate the quality difference evaluation of Hedychium flavum from different origins based on multi-index content determination, using principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (PLS-DA) and CRITIC-TOPSIS methods, so as to provide a basis for further development and utilization. Methods HPLC was used with BetaMax Neutral C₁₈ column, acetonitrile -0.4%phosphoric acid solution as mobile phase with gradient elution, detection wavelength of 320, 230, 360 and 210 nm. A total of 17 batches of H. flavum from different origins were collected. The contents of 13 chemical constituents including protocatechuic acid, trans-4-hydroxycinnamic acid, coniferaldehyde, isocoronarin D, coronarin D, coronarin A, coronarin E, quercetin, kaempferol, isorhamnetin, daucosterol, stigmasterol, β-sitosterol were detected, while extractives and total ash were examined. The 17 batches of H. flavum were grouped using PCA and PLS-DA methods, and quality differential markers were screened out. The weights of each index was calculated using the CRITIC method, and a comprehensive quality evaluation model for H. flavum was constructed using the TOPSIS method. Results The contents of the above 13 indexes among 17 batches of H. flavum were 0.121—0.295, 0.044—0.091, 0.036—0.104, 0.400—1.071, 0.300—0.515, 0.902—1.677, 0.089—0.153, 0.473—0.916, 0.177—0.378, 0.136—0.249, 0.068—0.117, 0.223—0.453, 0.240—0.863 mg/g, while the ranges of extractives and total ash were 11.4%—21.5% and 3.1%—9.2 %, respectively. PCA results showed that the cumulative variance contribution rate of two principal components was 87.564%, and 17 batches of H. flavum were divided into three categories. Kaempferol, coronarin A, protocatechuic acid, extractives, β-sitosterol, isocoronarin D and quercetin were selected as quality differential markers by PLS-DA methods. The results of CRITIC-TOPSIS method showed that the comprehensive relative proximities of 17 batches of H. flavum samples was 0.421 6—0.618 1. The quality of samples from Guizhou and Sichuan provinces exhibited superior comprehensive quality. Conclusion The established multi-index quantitative combined with PCA, PLS-DA and CRITIC-TOPSIS method can comprehensively evaluate the quality difference of H. flavum from different habitats. The method is scientific, intuitive, accurate and reliable, which can provide a theoretical and technical reference for the quality difference evaluation of H. flavum.

Objective CiteSpace and VOSviewer softwares were used to conduct a visualized analysis of the current status of research on Gentiana worldwide, so as to predict its future research hotspots and developmental trends based on bibliometric methods. Methods Relevant literatures on Gentiana from January 1, 2005 to December 31, 2025 were retrieved from the China National Knowledge Infrastructure (CNKI) and Web of Science (WOS) Core Collection databases. After manually excluding duplicate and ineligible literatures and standardizing keywords, Excel, VOSviewer and CiteSpace were used to perform visualized analyses on the annual publication volume, publication journals, institutions, authors, countries and keywords of Chinese and English literatures in the field of Gentiana, respectively. Results A total of 7 449 eligible literature were retrieved, including 6 121 Chinese literature and 1 328 English literature. The annual publication volume of Chinese literature showed a fluctuating trend, while that of English literature presented a rapid upward trend. The Chinese and English journals publishing the most Gentiana-related studies were Chinese Traditional and Herbal Drugs (114 publications) and Journal of Ethnopharmacology (44 publications), respectively. The authors with the highest publication outputs were Wang Yuanzhong (61 publications) and Zhang Ji (27 publications). The institutions with the largest number of Chinese and English publications were Heilongjiang University of Chinese Medicine (118 publications) and Chinese Academy of Sciences (110 publications), respectively. The top three countries in terms of the number of articles published were China (709 publications), India (111 publications), and Japan (105 publications). The keyword analysis revealed that the current research hotspots mainly focused on the analysis of compound formulations, clinical applications, species identification, chemical composition, pharmacological effects, and mechanisms of action related to the Gentiana genus. In addition, traditional Chinese medicine preparations containing Gentiana have also been used to treat symptoms such as coughing and wheezing in patients during the COVID-19 pandemic. Conclusion Research on the Gentiana genus both domestically and internationally is currently showing an overall upward trend. In recent years, studies are gradually shifting from the single analysis of chemical components and pharmacological effects to the exploration of “multi-components and multi-targets” synergistic mechanisms. This research is deeply integrating modern technological methods such as genomics, molecular docking, and network pharmacology, providing important directions for the analysis of the efficacy mechanisms and innovative applications of Gentiana-related species. Future research in this field can also promote the precision and multi-layered development of Gentiana studies by strengthening international collaboration and deepening the integration of modern technologies.

Fresh traditional Chinese medicine (Fresh herbs) has the unique advantage of being effective due to its freshness, with the core being the complete retention of active ingredients and high physiological activity. However, the clinical promotion of fresh drugs is restricted by the variability of their components during growth and after harvest, the lagging preservation technology and the lack of standards. This article systematically reviews the historical evolution of the application of fresh herbs and explains the material basis for efficacy due to freshness, summarizes the dynamic changes of the active ingredients of fresh herbs, such as spatio-temporal accumulation before harvest, physical escape, enzymatic oxidation and hydrolytic polymerization after harvest, and proposes a classification processing and precise storage strategy centered on retaining dominant active ingredients. It aims to reveal the scientific connotation of efficacy due to freshness of fresh herbs, and provide theoretical basis and ideas for the protection of active ingredients, quality improvement and modern application of fresh herbs.

Objective To study the protective effect and mechanism of taraxasterol on alcoholic liver disease (ALD) from the perspective of gut microbiota and liver metabolites. Methods C57BL/6J mice were randomly divided into control group, model group, silibinin (100 mg/kg) group, taraxasterol high- and low-dose (10, 5 mg/kg) groups, with eight mice in each group. After continuous administration for four weeks and daily administration for 4 h, the ALD model was induced in mice by ig 53° red star erguotou. Liver function, liver tissue oxidative stress and inflammation related indicators in serum were detected. Hematoxylin-eosin (HE) staining was used to observe pathological changes in liver tissue. Western blotting was used to detect the expressions of zonula occludin-1 (ZO-1) and Occludin proteins in small intestine tissue. Metabolomics and 16S rRNA sequencing were used to detect changes in liver metabolites and gut microbiota. Results Compared with model group, taraxasterol significantly reduced the liver index and levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TG) in serum of ALD mice (P < 0.05, 0.01), increased the activity of superoxide dismutase (SOD) in liver tissue (P < 0.05, 0.01), reduced the levels of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in liver tissue (P < 0.05, 0.01), improved liver pathological changes, and upregulated the expressions of ZO-1 and Occludin in small intestine tissue (P < 0.05, 0.01). 16S rRNA sequencing analysis showed that taraxasterol effectively regulated the diversity of gut microbiota in ALD mice, improved the disorder of gut microbiota structure, and increased the abundance of beneficial bacteria such as Bacteroides, Bifidobacterium and Parabacteroides, decreased the abundance of pathogenic bacteria such as Escherichia Schilla and Streptococcus. Metabolomics analysis showed that taraxasterol reversed alcohol induced liver metabolic disorders, particularly in the sphingolipid and linoleic acid metabolic pathways, and increased levels of acetic acid, butyric acid and valeric acid. Conclusion Taraxasterol alleviates liver inflammation and alleviates ALD by regulating changes in gut microbiota and metabolites.

Objective To construct an intelligent agent for the diagnosis and treatment of Alzheimer’s disease (AD) based on knowledge of renowned veteran traditional Chinese medicine (TCM) physicians’ knowledge by adopting artificial intelligence techniques including chain-of-thought reasoning, retrieval-augmented generation (RAG), and prompt engineering, so as to support clinical decision-making with expert TCM experience. Methods Relevant literature and monographs regarding AD treatment by prestigious TCM physicians were retrieved, and a total of 16 renowned veteran TCM physicians, including Tian Jinzhou, Zhou Zhongying, and Shen Baofan, were selected. Their medical records, clinical experience, and academic theories were systematically mined to establish a knowledge base of renowned veteran TCM physicians. To address inconsistencies in syndrome differentiation approaches among different physicians, a syndrome-element classification method was adopted to standardize syndrome differentiation.Based on a large language model and the constructed knowledge base, an intelligent agent for the diagnosis and treatment of AD was developed on the Dify 1.9.2 platform, incorporating both an agent-based individual decision-making scheme and an agent-based group decision-making scheme. Symptom clusters extracted from 108 real-world medical records were used as inputs to the agent, which generated syndrome differentiation results and recommended prescriptions, thereby creating an evaluation dataset. A total of 30 cases were randomly selected for a single-blind expert questionnaire evaluation. Experts evaluated the rationality of syndrome differentiation and prescription recommendations generated by three approaches: the original medical records, the agent-based individual decision-making scheme, and the agent-based group decision-making scheme. Results The agent-based group decision-making scheme achieved the highest estimated marginal mean score (3.93 ± 0.14), outperforming both the agent-based individual decision-making scheme (3.47 ± 0.14) and the original medical records (3.16 ± 0.14). The agent-based individual decision-making scheme also outperformed the original medical records. Conclusion By integrating large language model technology with a syndrome-element classification method, this study incorporated the syndrome differentiation experience of multiple renowned veteran TCM physicians and developed an intelligent agent for the diagnosis and treatment of AD that embodies their collective expertise. The agent-based group decision-making scheme achieved higher scores in expert evaluations, suggesting that this approach can leverage the advantages of collaborative decision-making among multiple renowned physicians. The development of this intelligent agent provides a reusable methodological framework and implementation pathway for applying renowned physicians’ knowledge to support clinical decision-making, and offers a model for developing disease-specific intelligent agents that leverage the advantages of group decision-making, thereby laying the foundation for a subsequent prospective validation platform.

Objective An evidence assessment was conducted on five commonly used traditional Chinese patent medicines for the treatment of knee osteoarthritis (KOA), using the integrated evidence chain-based efficacy evaluation of traditional Chinese medicine (Eff-iEC) method and the grading of recommendations assessment,development and evaluation (GRADE) system. Methods The internationally recognized GRADE system and Eff-iEC method, which aligns with the characteristics of TCM, were applied in parallel to evaluate the evidence for five high-frequency recommended CPMs from clinical guidelines and expert consensuses. Simultaneously, a Meta-analysis was performed to evaluate their clinical efficacy and safety, using the clinical effective rate as a core indicator for evidence determination. Results GRADE assessment showed that, due to limitations in randomized controlled trials (RCTs) such as blinding and publication bias, among the five Chinese patent medicines, Jintiange Capsules and Biqi Capsules were rated as Grade C (low-quality evidence), while the rest were rated as Grade B (moderate-quality evidence). In contrast, the Eff-iEC method provided different evidence stratification: Jintiange Capsules (AA+B+), Zhuangguguanjie Capsules (BAA), and Biqi Capsules (AA+B+) were identified as high-level evidence. Wangbi Tablets/Capsules (BA+B+), Xianling Gubao Capsules (BA+B+) were identified as moderatelevel evidence. The Meta-analysis showed that CPMs combined with conventional Western medical therapy significantly improved the overall clinical effective rate (RR range 1.15—1.26, all P < 0.05). Notably, Wangbi Tablets/Capsules (RR = 0.41), Xianling Gubao Capsules (RR = 0.57), significantly reduced the risk of adverse reactions, demonstrating efficacy enhancement and toxicity reduction effects. Conclusion The Eff-iEC system and GRADE each have their own advantages in evidence evaluation. The Eff-iEC evaluation system can more comprehensively assess the efficacy evidence of traditional Chinese medicine compared to the GRADE system, avoiding underestimation of value due to flaws in trial design. However, high-quality RCTs are still needed in the future to further strengthen the certainty of the evidence.

Objective To explore the differences in characterization, miRNA composition, and target gene enrichment pathways of Astragali Radix-derived extracellular vesicle-like particles (AR-EVLP) obtained through two pretreatment methods: soaking and decoction. Methods Transmission electron microscopy, nanoparticle tracking analysis, nanoscale laser particle sizing, protein quantification, silver staining, real-time quantitative PCR, miRNA sequencing, and bioinformatics analysis were used to compare the morphology, particle concentration, size, ζ potential, protein concentration, protein distribution, miRNA composition, and molecular function differences of AR-EVLP obtained from the two pretreatment methods. Results The quality of AR-EVLP obtained from soaking was better than that obtained from decoction; AR-EVLP from decoction had a higher yield, relatively larger particle size, smaller absolute ζ potential, higher protein concentration, and more concentrated protein distribution. Sequencing results showed 219 differentially expressed miRNAs between the AR-EVLP obtained by the two pretreatment methods. Gene Ontology (GO) analysis indicated that the miRNA target genes from both pretreatment methods were significantly enriched in protein binding, suggesting potential cross-species regulatory effects. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that miRNAs lja-miR1511-3p_R-3_1ss2AC and gma-miR1511_R+1_1ss2AC from AR-EVLP obtained by soaking might have therapeutic potential for fatty liver and neurodegenerative diseases via the glycerophospholipid metabolism pathway. miRNAs mtr-miR164a, stu-miR477a-5p_R+1_1ss21CG, and gma-MIR162c-p5 from AR-EVLP obtained by decoction might treat cancer-related diseases through pathways such as glioma, prostate cancer, p53 signaling, Hedgehog signaling, ErbB signaling, and mitogen-activated protein kinase (MAPK) signaling. Conclusion AR-EVLP obtained by different pretreatment methods exhibited significant differences in characterization, protein composition, miRNA expression profiles, and associated biological functions. AR-EVLP from soaking had better quality and tended to be useful for treating diseases like fatty liver and neurodegenerative disorders, while AR-EVLP from decoction had a higher yield and tended to be geared toward treating cancer-related diseases.

Objective To investigate the mechanism by which Osmanthus fragrans and its active component salidroside improve hyperlipidemia. Methods An oleic acid-induced lipid deposition model was established in HepG2 cells. Lipid-regulating activities of O. fragrans ethanol extract and its components (acteoside, salidroside, isoacteoside, echinacoside) were evaluated using MTT assay and intracellular triglyceride (TG) level, supplemented by oil red O staining to visualize lipid droplet formation. A hyperlipidemic rat model was established via high-fat diet feeding. Control group, model group, O. fragrans ethanol extract group (810 mg/kg), salidroside low-, high-dose (40, 80 mg/kg) groups and simvastatin (2.1 mg/kg) group were set up, with eight rats per group, drugs were continuously administration for five weeks. Serum lipid levels, hepatic index, hepatic total cholesterol (TC) and TG levels in rats of each group were measured. Oil red O staining was used to observe hepatic lipid deposition in each group. Hematoxylin-eosin (HE) staining was used to examine the pathological morphology of epididymal adipose tissue. Transcriptome sequencing of liver tissue was performed to analyze the potential molecular mechanisms by which O. fragrans ethanol extract ameliorates hyperlipidemia. Molecular docking technology was employed to predict the affinity of salidroside, the primary active component, with pyroptosis-related proteins NOD like receptor family pyrin domain containing 3 (NLRP3), cystein-asparate protease-1 (Caspase-1), N-terminal fragment of gasdermin D (GSDMD-N), interleukin-1β (IL-1β), IL-18 and IL-6. Western blotting was used to further validate the effect of salidroside on expressions of pyroptosis-related proteins in liver tissue. Results O. fragrans ethanol extract and salidroside significantly reduced TG levels (P < 0.05, 0.01, 0.001), and diminished lipid droplet accumulation in oleic acid-induced HepG2 cells. Concurrently, they markedly lowered TC, TG and low density lipoprotein cholesterol (LDL-C) levels in serum (P < 0.01, 0.001), increased high density lipoprotein cholesterol (HDL-C) level (P < 0.001), reduced hepatic TC and TG levels (P < 0.01, 0.001), alleviated liver lipid deposition, and improved adipose tissue pathology in model rats. Transcriptomic gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses suggested that the lipid-regulating mechanism of O. fragrans ethanol extract may be related to regulating lipid metabolism and inflammatory responses. Molecular docking results indicated that salidroside exhibits high affinity for proteins involved in regulating inflammatory responses, such as NLRP3, Caspase-1 and IL-1β. Western blotting results showed that compared with model group, except for the cleaved Caspase-1 in liver tissues of salidroside low-dose group, the expression levels of pyroptosis-related proteins were significantly reduced in all other treatment groups (P < 0.05, 0.01, 0.001). Conclusion Salidroside may be one of the primary active components in O. fragrans for improving hyperlipidemia, and its lipid-regulating mechanism is associated with inhibiting NLRP3/Caspase-1/IL-1β signaling pathway to counteract hepatic pyroptosis.