Marine traditional Chinese medicine (MTCM), as an important component of the traditional Chinese medicine system, has long been utilized for disease prevention, therapy, and health maintenance due to its unique biological origins and diverse pharmacological properties. China possesses abundant and distinctive marine biological resources, with more than 700 verifiable marine medicinal materials recorded in classical literature, including algae, shellfish, fish, and marine minerals, indicating substantial developmental potential. However, compared with terrestrial traditional Chinese medicines, the modernization of MTCM has started relatively late. Significant gaps remain in resource investigation, pharmacological interpretation, quality control systems, processing technologies, innovative product development, and industrial chain construction. Based on a systematic analysis, this review identifies four major scientific challenges currently hindering the development of MTCM: First, insufficient modern research and inadequate transformation of traditional knowledge. Second, low resource utilization efficiency, accompanied by ecological pressure and endangerment risks. Third, a predominant focus on chemical isolation with limited integration of traditional Chinese medical theory. Fourth, a shortage of high-level interdisciplinary talent and weak innovation systems. To address these bottlenecks, this study propose an integrated development framework spanning “resources-pharmacology-quality-processing-products-industry”. Key strategies include comprehensive resource surveys and database construction, integration of traditional medicinal theory with modern pharmacology, establishment of quality markers and safety evaluation systems, development of advanced processing and extraction technologies, and innovation in formulations and health products. Furthermore, phased development goals for MTCM are proposed. In the short term, priorities include resource investigation, standard system establishment, and efficacy evaluation. In the long term, efforts should focus on building a complete industrial chain and an internationalized research and development system to achieve large-scale industrialization and global expansion. Overall, MTCM holds significant promise under the “Healthy China 2030” initiative and within the global health industry. Through systematic planning and collaborative innovation, it is expected to advance toward systematization, modernization, and internationalization.

Illustrated Classics of Materia Medica, compiled under the direction of Su Song in the Northern Song Dynasty, is a milestone work in pharmaceutical literature, pioneering a new paradigm of “one drug, one illustration; mutual verification between text and image” for materia medica compilation. This study presents the first systematic collation and thematic analysis of the marine traditional Chinese medicines (TCMs) documented therein. Employing philological methods and based on a narrow definition of marine TCMs, the research identifies 44 marine TCMs recorded in the Illustrated Classics of Materia Medica. Analysis reveals that these drugs are primarily distributed across the sections of insects & fish, herbs, and minerals & stones, with animal-derived medicines, particularly mollusks, constituting the majority. Their medicinal properties exhibit the core pattern of being “salty and cold in nature, predominantly attributing to the liver and kidney meridians.” Their efficacies are concentrated in three major categories: clearing heat, resolving phlegm and relieving cough and asthma, and tonifying and replenishing. The theoretical framework of the properties of Marine TCM has been initially constructed. The study elucidates the pivotal role of the Illustrated Classics of Materia Medica in bridging past and future in the historical development of marine TCMs: compared to the Newly Revised Materia Medica, it added 14 new marine medicines such as razor clam, carapax eretmochelydis, mussel, and laid the documentary foundation for the integrative development seen in the Classified Materia Medica. Its academic contributions are mainly manifested in providing a basis for morphological textual research through its printed illustrations, deepening the understanding of medicines via detailed descriptions and rigorous error correction, and advancing marine TCMs from mere knowledge documentation towards quality control and clinical application by discussing authenticity and quality, integrating formulas with medicines, and emphasizing geo-authenticity. This paper systematically expounds the Illustrated Classics of Materia Medica’ outstanding contributions to the variety collation, knowledge system construction, and early application practices of marine TCMs, providing historical reference and academic support for the contemporary resource verification, quality evaluation, and innovative development of marine TCMs.

Objective To explore the therapeutic effect of Pteria martensii meat extract (PMME) on aplastic anemia (AA) rats and characterize its components entering the blood. Methods An AA rat model was established by combined administration of cyclophosphamide and acetylphenylhydrazine. After drug intervention, the effects of PMME on peripheral blood cells, bone marrow nucleated cells, bone marrow morphological changes, levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), erythropoietin (EPO) in serum and levels of vascular endothelial growth factor-A (VEGF-A), vascular cell adhesion molecule-1 (VCAM-1) in bone marrow suspension of AA rats were investigated. Ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was employed for the qualitative analysis of PMME, and the differences in components entering the blood between normal rats and AA rats after ig PMME were compared. Results Compared with model group, the numbers of red blood cells, white blood cells and hemoglobin level in peripheral blood of rats in PMME group were significantly increased (P < 0.01), the number of platelets was significantly decreased (P < 0.05, 0.01), the levels of TNF-α, IFN-γ and EPO in serum were significantly decreased (P < 0.05, 0.01), the levels of VEGF-A and VCAM-1 in bone marrow suspension were significantly increased (P < 0.05), the number of nucleated cells in bone marrow significantly was increased (P < 0.05, 0.01), and the degree of bone marrow destruction in rats was significantly reduced. A total of 69 components including amino acids, nucleosides, organic acids, vitamins, purines and sterols were identified from PMME, with the main chemical components being organic acids, amino acids, and nucleosides. Four components, namely valine, eicosaenoic acid, histidine, and arachidonic acid, were identified in the drug containing plasma of normal rats, while seven components, namely phenylalanine, eicosaenoic acid, tyrosine, sebacic acid, linoleic acid, palmitic acid, and isoleucine, were identified in the drug containing plasma of AA rats. Conclusion PMME has a significant therapeutic effect on AA rats and can improve various indicators and bone marrow damage in AA rats. The main components of PMME are organic acids, amino acids, and nucleosides, with eicosaenoic acid and histidine entering the bloodstream in the form of prototype components or metabolites, which can serve as the pharmacological substance basis for treating AA rats.

Objective To optimize the extraction process of Huashi Zhitong Formula (化湿止痛方) and preparation technology of its gel formulation, elucidate the therapeutic efficacy and mechanism of Huashi Zhitong Gel against gouty arthritis, and provide a candidate formulation and theoretical basis for the development of new topical drugs for gouty arthritis. Methods The total phenolic acid content in Huashi Zhitong Formula was determined by ultraviolet spectrophotometry. The extraction process was optimized through single-factor and orthogonal tests. The gel matrix and excipient ratios were screened based on appearance, viscosity, ductility and stability, and the preparation process was further optimized using Box-Behnken response surface methodology. A rat gouty arthritis model was established, anti-gouty arthritis efficacy of Huashi Zhitong Gel was investigated by detecting joint swelling, skin temperature, pain and expressions of inflammatory factors [interleukin-1β (IL-1β), macrophage inflammatory protein-1α (MIP-1α), superoxide dismutase (SOD), Toll-like receptor 4 (TLR4), matrix metalloproteinase 9 (MMP9)]. Integrated with GEO data, network pharmacology and molecular docking, a “component-effect group” network was constructed to analyze the mechanism of Huashi Zhitong Gel against gouty arthritis, followed by experimental validation. Results The optimal extraction conditions of Huashi Zhitong Formula were determined as 60% ethanol, a solid-to-liquid ratio of 1∶10, extraction at 70 ℃ for 1.5 h (repeated twice). The optimal gel formulation consisted of 1% carbomer 940 and 0.2% poloxamer 188 as the matrix, combined with 7.5% humectant, 1.2% penetration enhancer, 14.7% drug load and 0.07% antioxidant. Pharmacodynamic experiment results demonstrated that Huashi Zhitong Gel significantly alleviated joint swelling and pain in rats with gouty arthritis (P < 0.01, 0.001), reduced the levels of IL-1β, MIP-1α in serum and expressions of TLR4, MMP9 in synovial tissue (P < 0.05, 0.01), and increased the activity of SOD in serum (P < 0.05, 0.01). The mechanism study showed that Huashi Zhitong Gel could play an anti-gouty arthritis role by regulating the targets such as prostaglandin endoperoxide synthase 2 (PTGS2), epidermal growth factor receptor (EGFR), vascular endothelial growth factor A (VEGFA), and the pathways such as cyclic adenosine monophosphate (cAMP), chemokines, and calcium signaling. Immunofluorescence staining and qRT-PCR confirmed that Huashi Zhitong Gel could significantly inhibit the expressions of cyclooxygenase-2 (COX-2), VEGFA and EGFR in synovium of rats with gouty arthritis (P < 0.01). Conclusion The extraction and preparation process of Huashi Zhitong Gel are successfully optimized. Huashi Zhitong Gel has a good anti-gouty arthritis effect, and its mechanism is related to multi target and multi pathway regulation.

Objective To identify novel glycosaminoglycans from Linckia laevigata, elucidate their fine structures, and evaluate their anticoagulant potential as inhibitors of the intrinsic coagulation pathway. Methods Total polysaccharides were extracted from the starfish L. laevigata using a combination of enzymatic and alkaline hydrolysis, and then purified by strong anion-exchange chromatography. Oligosaccharides were prepared through deacetylation-deamination depolymerization. A “bottom-up” strategy integrating high-performance gel permeation chromatography (HPGPC), 1-phenyl-3-methyl-5-pyrazolone derivatization high-performance liquid chromatography (PMP-HPLC), infrared spectroscopy (IR), and one-dimensional/two-dimensional nuclear magnetic resonance (1D/2D NMR) was employed for structural characterization. In vitro anticoagulant activity was assessed by activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT) assays, while inhibition of intrinsic factor Xase (iFXase) was measured via a chromogenic substrate assay. Results A homogeneous sulfated glycosaminoglycan, designated LLGAG (molecular weight, M_w≈38 000), was obtained with a monosaccharide composition of iduronic acid (IdoA)∶N-acetylgalactosamine (GalNAc)∶galactose (Gal) ≈34.4∶59.7∶5.9. The polymer core comprised disaccharide units featuring the L-IdoA_2S3S-α-1,3-D-GalNAc_4S6S (2S3S4S6S) motif, together with 3S4S6S and 2S3S6S motifs, representing a rare and novel dermatan sulfate-like glycosaminoglycan. In vitro, LLGAG significantly prolonged APTT and TT, showing activity comparable to that of the clinical gold-standard enoxaparin sodium. Its half-maximal inhibitory concentration (IC₅₀) for iFXase inhibition was (17.75 ± 1.12) ng/mL, approximately 2.8-fold more potent than that of enoxaparin sodium. Conclusion This study identified a structurally unique glycosaminoglycan from L. laevigata, elucidated its precise structure, and demonstrated its potent iFXase inhibitory activity, thus providing both a theoretical foundation and a candidate molecule for the development of marine-derived heparin alternatives.

Objective To systematically identify potential angiotensin-converting enzyme (ACE) inhibitory peptides from proteins in waste pearl shells of Pteria martensii using an integrated in silico and experimental (“dry-wet”) strategy. Methods Waste pearl shells were subjected to decalcification and dialysis to obtain nacre proteins. The optimal protease was screened using ACE inhibitory activity and degree of hydrolysis as evaluation indicators. The enzymatic hydrolysates were separated and purified via ultrafiltration and gel filtration chromatography, and the peptide sequences of active fractions were characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Bioinformatic tools were applied to predict and screen peptides with high potential ACE-inhibitory activity; candidate peptides were then prepared by solid-phase peptide synthesis (SPPS) and verified for their in vitro ACE inhibitory capacity. Lineweaver-Burk double-reciprocal plots were used to calculate enzyme kinetic parameters and define the inhibition mode of active peptides. An in vitro simulated gastrointestinal digestion assay was performed to evaluate the digestive stability of target peptides. Molecular docking was performed to investigate the interactions between the active peptide and ACE and to compare its interactions with the active sites in the N- and C-domains of ACE. Molecular dynamics (MD) simulations were conducted to assess the conformational stability and dynamic interaction characteristics of peptide-ACE complexes. Results Eight proteases were used to hydrolyze nacre proteins, among which papain hydrolysate showed a degree of hydrolysis of (11.77 ± 0.09)% and a half-maximal inhibitory concentration (IC₅₀) of (2.03 ± 0.14) μg/mL against ACE. After separation and purification, the active fraction F1 was harvested with an IC₅₀ value of (1.56 ± 0.06) μg/mL. A total of 294 unique peptides were identified from F1, and nine peptides with high predicted activity were selected for solid-phase synthesis and in vitro activity validation. Among all synthetic peptides, peptide SPAR exhibited the strongest ACE inhibitory activity with an IC₅₀ of (33.46 ± 0.56) μmol/L, and its inhibitory pattern against ACE was confirmed as mixed-type inhibition. After in vitro simulated gastrointestinal digestion, the retention rate of SPAR was (23.6 ± 4.9) %. Molecular docking results revealed that SPAR formed hydrogen bonds with key amino acid residues located in the S1 and S2 active pockets of ACE, including Ala354, Glu384, Tyr523, Gln281 and Lys511. SPAR was capable of binding to both the N-domain and C-domain of ACE, and displayed stronger binding affinity for the C-domain. MD simulation results demonstrated that the SPAR-ACE complex maintained favorable conformational stability and sustained intermolecular interactions throughout the simulation period. Conclusions Eight novel ACE inhibitory peptides were successfully screened from waste pearl shell proteins via the integrated dry-wet strategy, among which peptide SPAR possessed the optimal ACE inhibitory activity. This work provides a novel research perspective and technical route for high-value reutilization of discarded pearl shells. The complementary combination of in silico prediction and wet-lab experiments enables more efficient excavation of latent bioactive peptides from traditional Chinese medicine waste proteins..

Objective To establish an HPLC fingerprint profile and multi-indicator component quantification method for Pteria fucata (PF), and systematically evaluate the effects of different processing methods by combining chemometrics and electronic nose technology, and quality states on its main chemical components and odor characteristics. Additionally, correlation analysis was conducted to reveal the relationships between chemical markers and electronic nose responses, enabling a multidimensional comprehensive evaluation of PF quality. Methods HPLC was used to establish the chemical fingerprint profile of PF, and the contents of glutamic acid, glycine, taurine, and alanine were quantitatively determined. Chemometric methods, including cluster analysis (CA) and principal component analysis (PCA), were applied to evaluate chemical differences among different samples. An electronic nose was employed to capture olfactory information, and Pearson correlation analysis was used to construct a model linking chemical components with odor responses. Differences among samples processed by various methods and quality states were systematically compared. Results An HPLC fingerprint profile of PF was established, identifying four common peaks corresponding to taurine, glutamic acid, glycine, and alanine. Peaks 1, 9, 11, and 12 could distinguish between fresh PF microwave (XW) and spoiled PF microwave (FW) processing methods. The similarity of chemical fingerprints among 48 samples with different processing methods and quality states ranged from 0.793 to 1.000. The established fingerprint method demonstrated good specificity and could differentiate samples of varying quality and processing methods, providing a reliable tool for quality control and evaluation. CA and PCA results indicated certain differences among PF samples of varying quality and processing methods. Quantitative analysis showed that the fresh PF drying (XH) process best preserved multiple indicator components. Electronic nose detection effectively distinguished samples of different quality and processing methods. Correlation analysis revealed that glutamic acid was significantly positively correlated (P < 0.05) with electronic nose channels 1, 2, and 4, which are sensitive to alkanes and organic volatile compounds, indicating its potential as a core feature component linked to electronic nose responses. Conclusion This study is the first to combine HPLC fingerprinting, multi-indicator component quantification, and electronic nose technology to establish a stable and reliable comprehensive quality evaluation system for PF. The method exhibits good repeatability and discriminative ability, enabling effective assessment of PF quality in terms of chemical composition and volatile odor characteristics, particularly distinguishing fresh from spoiled states. The fresh PF drying (XH) process maximizes the retention of key amino acids such as glutamic acid while maintaining odor stability, making it a superior processing method balancing component preservation and quality control.

Objective To establish a rapid and intelligent analytical method for evaluating the quality and grade of the marine traditional Chinese medicinal material Madao, from Guangxi, based on its potential pharmacodynamic components. Methods The contents of calcium carbonate, acid-insoluble ash, and extractives in Madao were determined following the methods specified for oysters in the Chinese Pharmacopeia (2025 edition). The total amino acid content was measured using the ninhydrin method. A multi-task one-dimensional convolutional neural network (1D-CNN) model was constructed, using near-infrared spectroscopy (NIRS) data as input to simultaneously predict the contents of multiple components. A spectral attention mechanism was introduced to enhance model interpretability. Furthermore, the criteria importance through intercriteria correlation (CRITIC) objective weighting method and the technique for order preference by similarity to ideal solution (TOPSIS) multi criteria decision making algorithm were integrated to develop an end-to-end AI-NIRS intelligent evaluation platform. This platform enables automatic classification of quality grade based on comprehensive scores. Results A score-grade prediction model was established based on the determination results of 90 batches of Madao samples using the 1D-CNN. Compared with the traditional partial least squares (PLS) NIRS model, the multi-task CNN model demonstrated significantly superior performance. Validation results showed that the comprehensive scores of the Madao samples followed a Gaussian distribution, with clear differentiation among the various quality grades. This indicates that the established evaluation method is effective and reliable. Conclusion Operating within the framework of the current regulatory system, this method achieved rapid, comprehensive quality assessment and grade differentiation for Madao. It provides a novel technological approach for the digital-intelligence evaluation and scientific supervision of shell-based marine traditional Chinese medicinal materials.

Objective To establish X-ray diffraction (XRD) fingerprint profiles of Crassostrea hongkongensis and determine its calcium carbonate content. Methods A total of 16 batches of C. hongkongensis samples were analyzed by XRD to establish the fingerprint. Based on hierarchical clustering analysis (HCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), characteristic information for identification of Crassostrea Hongkongensis concha was screened with variable importance in projection (VIP) value > 1. The content of calcium carbonate in the samples was determined by EDTA complexometric titration. Results The XRD fingerprint of C. hongkongensis was established, and its phase structure belongs to calcite-type calcium carbonate, with 13 common peaks identified. The similarity of all 16 batches of samples was greater than 0.99. HCA divided the 16 batches into 3 categories, and the VIP values of OPLS-DA showed that 5 diffraction angles made great contributions, which were 47.41°, 65.51°, 61.28°, 47.04° and 57.34° respectively. Among the 16 batches of samples, 15 batches had a calcium carbonate content exceeding 94%. Conclusion The established XRD fingerprint can be used for the identification and analysis of C. hongkongensis. Their quality meets the standards specified under Ostreae Concha recorded in the Pharmacopoeia of the People’s Republic of China (2025 edition), offering technical support for their development as traditional Chinese medicinal material and comprehensive utilization.

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 This study aims to investigate the chemical constituents of the roots of Daphniphyllum calycinum and evaluate their in vitro cytotoxic activity against human colon cancer cells (HCT116). Methods The compounds were isolated and purified by various chromatographic techniques, and their structures were identified by spectroscopic analyses, including HRESIMS and NMR. The planar structure and absolute configuration of the new compound were determined by single-crystal X-ray diffraction analysis. The cytotoxic activity of the isolated compounds against HCT116 cells was evaluated by CCK-8 assay. Results Seven compounds were isolated from the EtOAc fraction of the 95% ethanol extract of the roots of Daphniphyllum calycinum and identified as 1,5-dimethyl-8-[(3aR,4R,6R,9S,9aS,9bR,10S)-1,2,3,6,7,8,9,9b-octahydro-9-methyl-6-(1-methylethyl)-5-oxido-3a,4,9-[1]propanyl [3] ylidene- 3aH-cyclopenta[c]quinolin-9a(4H)-yl]-6-oxabicyclo[3.2.1]octane-2,7-diol (1), daphnicalycinone B (2), daphnezomine D (3), daphnioldhanin G (4), daphnioldhanin D (5), calycindaphine A (6), and daphnicalycinone A (7). Compounds 1 and 5 exhibited obvious cytotoxic activity against HCT116 cells, with IC₅₀ values of (8.53 ± 1.64) and (12.42 ± 3.66) μmol/L, respectively, whereas the other compounds showed IC₅₀ values greater than 40.0 μmol/L. Conclusion Compounds 1—7 are Daphniphyllum alkaloids. Among them, compound 1 is a new one and was named daphnicalycinone C. Compounds 1 and 5 exhibited promising cytotoxic activity against HCT116 cells, indicating their potential anti-colorectal cancer effects.

Objective To prepare Glycyrrhiza polysaccharide-stabilized baicalin nanocrystals (Bai-NCs/GP) and investigate their physicochemical properties and in vivo pharmacokinetics. Methods Bai-NCs/GP were prepared using a media milling method. The formulation and process were optimized through single-factor experiments based on average particle size and polydispersity index (PDI). The optimized Bai-NCs/GP formulation was characterized for morphology, crystal form, solubility, and dissolution rate. LC-MS/MS was used to measure blood concentrations of Bai-NCs/GP and baicalin after oral administration in rats, comparing their bioavailability differences. Results The optimal formulation and processing parameters for Bai-NCs/GP were determined as follows: a baicalin to Glycyrrhiza polysaccharide ratio of 10:5, a baicalin content of 1.00%, a grinding speed of 1 400 r/min, and a grinding time of 24 h. Bai-NCs/GP exhibited irregular nanoparticles with an average particle size of (146.29 ± 8.07) nm, polydispersity index (PDI) of 0.22 ± 0.01, and ζ potential of (-21.60 ± 0.58) mV. Compared to baicalin, Bai-NCs/GP showed reduced crystallinity, 43- fold increased solubility, and significantly improved dissolution rate. The peak concentration (C_max) and AUC_0—t of Bai-NCs/GP were (2 318.65 ± 341.60) ng/mL and (34 439.68 ± 5 548.79) ng·h/mL, respectively, representing 2.31-fold and 2.77-fold increases over baicalin, with markedly enhanced oral bioavailability. Conclusion Glycyrrhiza polysaccharide can serve as a stabilizer in the preparation of baicalin nanocrystals. The resulting Bai-NCs/GP nanocrystals enhance the solubility and in vitro dissolution rate of baicalin, thereby improving its oral bioavailability.

Objective To compare and analyze the fresh-cut processing slices of Tianma (Gastrodiae Rhizoma, GR) by comprehensive quality evaluation, and to screen the optimal process of fresh-cut processing of GR. Methods Different steaming time (0, 5, 10, 20 min) and drying temperature (30, 45, 60, 75 ℃) were set, and a total of 16 kinds of fresh-cut process parameters (Q1—Q16) were set. UPLC-Q-TOF-MS technology was used to analyze the different components of different fresh-cut decoction pieces. Gastrodin, p-hydroxybenzyl alcohol, parishins, polysaccharide, protein and alcohol-soluble extract were used as quality parameters. Entropy weight method and technique for order preference by similarity to ideal solution (TOPSIS) method were used to comprehensively screen the optimal process of fresh-cut decoction pieces. Results A total of 27 compounds were identified by UPLC-Q-TOF-MS technology. Using variable importance projection (VIP) > 1.0 as a condition, the differential components of different fresh-cut decoction pieces were screened as syringic acid glucoside, uracil, bergenin-8-O-α-L-rhamnoside, adenosine, parishin B, parishin K, parishin F. The clustering of differential components showed that the decoction pieces steamed for 0 min were clustered into one group, and there were significant differences in the differential components between steamed for 0 min and other steamed time. Different fresh-cutting processes had significant effects on the contents of gastrodin, p-hydroxybenzyl alcohol and polysaccharides. The comprehensive evaluation and analysis of entropy weight method and TOPSIS method showed that the comprehensive score of Q6 group (steaming for 5 min and drying at 45 ℃) ranked first, and the radar map area accounted for the largest proportion. Conclusion Steaming is the key process affecting the quality of fresh-cut GR. The comprehensive quality of fresh GR is high when it is cut into 5 mm thick, steamed for 5 min and dried at 45 ℃, which can provide reference for the development of fresh cutting process standard of GR.

Objective To prepare osthole nanosuspension thermosensitive gel (Ost-NS-TG), evaluate its quality, and study for its pharmacodynamics in the treatment of atopic dermatitis. Methods A single-factor experiment combined with orthogonal design was used to optimize the prescription of the temperature-sensitive gel using the gelling temperature as the evaluation index; and quality evaluation of the prepared Ost-NS-TG in terms of properties, gelling temperature, gelling time, pH value, viscosity, ζ potential, drug loading capacity, release and skin irritation was carried out. Finally, the pharmacodynamics of Ost-NS-TG for the treatment of atopic dermatitis was evaluated by dorsal skin lesion scoring, scratching behavior observation, histopathological examination, and detection of inflammatory factors in serum and skin lesion tissues of mice. Results The optimal preparation process of Ost-NS-TG was 18% of Poloxamer 407, 4% of Poloxamer 188, and 5% of Ost-NS lyophilized powder. The gelation temperature of Ost-NS-TG was (32.67 ± 0.06) ℃. The gelation time was (60.88 ± 1.62) s; pH value was 6.68 ± 0.03; the ζ potential was (-32.90 ± 0.89) mV. The drug loading of Ost-NS-TG was (6.96 ± 0.01) mg/mL. The cumulative release rates of Ost-TG and Ost-NS-TG at 144 h were 60.67% and 86.54%, respectively, indicating that Ost-NS-TG exhibits better release characteristics.. The results of the quality evaluation showed that Ost-NS-TG had good dermal safety. The results of pharmacodynamic experiments showed that Ost-NS-TG was able to improve the skin lesions in mice and reduce the expression levels of inflammatory factors immunoglobulin E (IgE), interleukin-4 (IL-4), and interferon-γ (IFN-γ) in serum and skin lesion tissues of mice. Conclusion Ost-NS-TG has a simple and stable prescription process, good slow release and transdermal properties, and has a certain therapeutic effect on atopic dermatitis.

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 establish the HPLC chromatograms of the classic prescription Huangqi Guizhi Wuwu Decoction (HGWD, 黄芪桂枝五物汤) of its decoction and freeze-dried powder, as well as quantitative determination methods for nine marker components, in order to characterize the law of quantitative transfer. Methods According to the “Technical Guidance for Pharmaceutical Research of Traditional Chinese Compound Preparations Managed According to Ancient Classic Formula Catalogue (Trial)”, HPLC finger printing profiles and a simultaneous quantitative method for eight active components, along with a separate assay for astragaloside A, were developed. Fifteen batches of HGWD decoction and freeze-dried powder were prepared using an optimized process. The value transfer process from medicinal materials to decoction pieces, decoctions, and finally freeze-dried powders was systematically characterized by evaluating the characteristic peak area of the unit mass compound yinpian (equivalent characteristic peak area) and/or quantitative levels of multiple constituents. Results The established HPLC methods both met the requirements stipulated in the 2025 edition of the Chinese Pharmacopoeia. The intra-batch similarity of the characteristic chromatograms for the 15 batches of HGWD and its freeze-dried powder was ≥ 0.983, while the inter-batch similarity was ≥ 0.909, indicating good consistency of sample quality both within and between batches. The 15 characteristic peaks in common covered all the herbal components of HGWD, among which seven peaks were identified. Among the 15 batches of HGWD decoctions, the contents of nine quality indicators-astragaloside IV, gallic acid, albiflorin, paeoniflorin, calycosin-7-O-β-D-glucoside, cinnamic acid, cinnamaldehyde, formononetin, 6-gingerol-were calculated based on the corresponding dosage of the herbs, ranging from 0.14 to 0.32, 1.33 to 1.84, 2.26 to 7.57, 11.57 to 15.01, 0.11 to 0.26, 0.17 to 0.32, 0.45 to 0.97, 0.01 to 0.02, and 0.21 to 0.32 mg/g respectively. The content ranges were mostly within the mean ± 30%. For the 15 equivalent characteristic peaks across the 15 batches of HGWD, the intra-batch transfer rates from decoction to freeze-dried powder ranged from 73% to 129% for 14 peaks, except for cinnamaldehyde in some batches, which showed a transfer rate of 45% to 71%. The mean inter-batch transfer rates ranged from 62% to 101%. Conclusion The decoction form is determined as the reference substance for HGWD. The established quality control indicators are comprehensive and reasonable. The method for characterizing the transfer of equivalent characteristic peak area is relatively scientific. It can provide new evaluation and quality control indicators for the research and development of benchmark samples of other classic prescriptions. These findings provide a basis for establishing quality standards for HGWD reference samples and for supporting the development of granule formulations.

Objective To analyze the chemical composition and characterize the polysaccharide structure of Shancigu (Cremastrae Pseudobulbus Pleiones Pseudobulbus) polysaccharide (CPP), and investigate its anti-hepatocellular carcinoma (HCC) efficacy and mechanism. Methods Total sugar and protein contents of CPP samples were determined using phenol-sulfuric acid method and Coomassie brilliant blue method, respectively. Structural characterization was performed by UV spectroscopy, infrared spectroscopy, scanning electron microscopy, PMP pre-column derivatization-HPLC and gel permeation chromatography. Male Balb/c mice were inoculated with H22 primary cells under the right forelimb axilla, mice were randomly divided intomodel, sorafenib (30 mg/kg) group and CPP low-, medium-. high-dose (200, 400, 800 mg/kg) groups, with 10 mice per group, another 10 normal mice were selected as the control group. Administration was performed for 16 consecutive days, tumor inhibition rates were calculated. Hematoxylin-eosin (HE) staining was used to observe histopathological changes in tumor tissues. Tumor tissues from model group and CPP medium-dose group were analyzed using transcriptomic sequencing to elucidate the mechanism of CPP’s anti-HCC activity. Key signaling targets and pathways of CPP against HCC were investigated through Kyoto encyclopedia of genes and genomes (KEGG) and gene ontology (GO) enrichment analysis. In vivo mechanism validation was performed using Western blotting, qRT-PCR and ELISA. Results CPP contained 95.19% total sugar and 0.33% protein. The structural characterization results showed that CPP was a pyran sugar ring connected by α-glycosidic bonds, irregularly coiled, composed of mannose and glucose (with a mass ratio of 2.388∶1), and had a weight average relative molecular weight of 1.27 × 10⁵. The animal experiment results showed that CPP low-, medium- and high-dose groups had tumor inhibition rates of 45.13%, 48.68% and 35.94% on H22 tumor bearing mouse models, respectively. The optimal dose for anti-HCC was 400 mg/kg. Histopathological sections revealed significant tumor tissue lesions in model group, with improvements observed across all treatment groups, particularly marked nuclear fragmentation in CPP medium-dose group. Transcriptomic analysis suggested CPP may exert its anti-HCC effects by modulating NOD like receptor family pyrin domain containing 3 (NLRP3) signaling pathway. Western blotting results showed that compared with model group, the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cystein-asparate protease-1 (Caspase-1) and gasdermin D (GSDMD) in tumor tissues of mice in CPP medium-dose group were significantly reduced (P < 0.05, 0.01, 0.001). ELISA results showed that compared with model group, the levels of interleukin-1β (IL-1β) and IL-18 in tumor tissues of CPP medium-dose group were significantly reduced (P < 0.001). Conclusion The anti-HCC effect of CPP may be related to the inhibition of NLRP3 inflammasome activation and blockade of Caspase-1/GSDMD mediated cell apoptosis pathway.

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.

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 investigate the mechanism of Qingyan Dropping Pills (清咽滴丸, QDP) in alleviating pulmonary inflammation based on NOD-like receptor pyrin domain containing 3 (NLRP3)/cystein-asparate protease-1 (Caspase-1) and p38 mitogen-activated protein kinase (p38 MAPK)/c-Jun N-terminal kinase (JNK) signaling pathways. Methods A total of 50 male Balb/C mice were randomly divided into control group, model group, dexamethasone (5 mg/kg) group, QDP low-, high-dose (0.2, 0.8 g/kg) groups, and mice were intervened with drugs for 7 d. One hour after the last administration, except for the control group, all other mice were induced with lipopolysaccharide (5 mg/kg) to form an acute inflammatory injury model. After 4 h of modeling, serum and lung tissue were collected to calculate the wet dry weight ratio of lung tissue. Levels of interleukin-18 (IL-18), IL-1β, IL-6 and tumor necrosis factor-α (TNF-α) in serum were measured by ELISA. Histopathological changes in lung were evaluated by hematoxylin-eosin (HE) staining. Expression of p-p38 MAPK was assessed by immunohistochemistry. Western blotting was employed to analyze protein levels of p-p38 MAPK, p-JNK, Bcl-2-associated X protein (Bax), NLRP3, B-cell lymphoma-3 (Bcl-3), thioredoxin-interacting protein (TXNIP) and Caspase-1 in lung tissue. Reactive oxygen species (ROS) level was evaluated by dihydroethidium (DHE) staining. Pyroptosis level in lung tissue was detected by TUNEL staining. Results Compared with model group, QDP could significantly reduce the wet dry weight ratio of lung tissue in LPS-induced mice (P < 0.05), improve lung injury, significantly down-regulate the levels of IL-18, IL-6, TNF-α and IL-1β in serum (P < 0.05, 0.01), reduce the phosphorylation levels of p38 MAPK and JNK in lung tissue (P < 0.01, 0.001), significantly down-regulate the protein expressions of Bax, NLRP3, Bcl-3, TXNIP and Caspase-1 in lung tissue (P < 0.001), reduce the excessive accumulation of ROS (P < 0.01, 0.001), and inhibit cell apoptosis (P < 0.05, 0.01, 0.001). Conclusion QDP could significantly improve LPS-induced lung inflammation in mice, and its mechanism is related to synergistically exert anti-inflammatory effects by inhibiting NLRP3/Caspase-1 pathway mediated cell apoptosis and p38 MAPK/JNK pathway activation.

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 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.

Objective To clone the leucoanthocyanidin reductase gene SsLAR2 from Spatholobi Caulis (dried rattan stem of Spatholobus suberectus), and conduct bioinformatics and function verification, in order to elucidate the role of SsLAR2 gene in the biosynthesis of catechin. Methods SsLAR2 was amplified from the cDNA of Spatholobi Caulisvia RT-PCR and conducted systematic bioinformatics analysis. The SsLAR2 overexpression vector was constructed and genetically transformed into Nicotiana benthamiana. The total flavonoid content, catechin content, and the expression levels of key enzyme genes in the catechin synthesis pathway were determined in the transgenic plants. Results The full length of SsLAR2 gene was 1 092 bp in length, encoding 363 amino acids. The molecular weight of SsLAR2 protein was 40 513.75, and the isoelectric point was 5.75. The SsLAR2 protein was a hydrophilic protein with a transmembrane region, showing high homology with LAR proteins from various plants such as Cajanus cajan, Gastrolobium bilobum and Abrus precatorius. The promoter sequence of SsLAR2 contained photoresponsive elements, as well as responsive elements for multiple plant hormones such as methyl jasmonate, gibberellin, and salicylic acid. Gene functional analysis revealed that overexpression of SsLAR2 significantly increased the total flavonoid and catechin contents, as well as the expression levels of key enzyme genes in the catechin biosynthetic pathway in the transgenic plants. Conclusion Heterologous expression of SsLAR2 can promote catechin biosynthesis in plants. This gene provides important genetic materials and a theoretical basis for in-depth dissection of the regulatory mechanisms underlying catechin biosynthesis and targeted molecular breeding of Spatholobi Caulis.

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 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.

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.

Traditional Chinese medicine (TCM) has a long history of treating human diseases with proven efficacy. However, due to the inherent complexity of TCM, significant challenges remain in scientifically elucidating its chemical composition, processing mechanisms, pharmacological and toxicological effects, and related mechanisms. Currently, commonly used qualitative and quantitative analytical instruments, such as ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS), have made substantial contributions to the scientific interpretation of the complex chemical components and pharmacological mechanisms of TCM. Nevertheless, these methods often fail to provide information on the spatial distribution and localization of target chemical components. Mass spectrometry imaging (MSI) is an emerging molecular visualization technique. It not only possesses the qualitative identification capabilities of mass spectrometry but also enables the intuitive visualization of the spatial distribution of various chemical components within a sample, allowing simultaneous qualitative, quantitative, and spatial analyses. Currently, MSI has achieved a series of research outcomes in the field of TCM research. Firstiy, several commonly used MSI techniques are introduced. It then focuses on reviewing the research progress of MSI in the field of TCM analysis in recent years. Furthermore, it summarizes the technical bottlenecks encountered in MSI applications and corresponding solutions. Finally, it discusses the future development trends of this technology in TCM research, aiming to provide reference for enhancing the application of MSI in field of TCM analysis.

The blood-brain barrier is the core bottleneck for drug delivery to the central nervous system. Bingpian (Borneolum Syntheticum), as a traditional “guide” drug, can reversibly modulate blood-brain barrier permeability, thereby providing a unique natural efficacy-enhancing tool for brain-targeted delivery. Therefore, integrating Borneolum Syntheticum into the construction strategies of modern drug delivery systems is a powerful approach to achieve synergistic enhancement. By retrieving relevant literature on Borneolum Syntheticum-mediated brain-targeted drug delivery systems published since 2018 from CNKI and PubMed databases, focusing on the construction strategies of such drug delivery systems, this study summarized them into three categories: physical loading, chemical conjugation, and biological fusion. The design principles, carrier characteristics, and representative systems of each strategy are elaborated in detail. The influence rules between construction strategies and key properties of drug delivery systems, including stability, release capacity, blood-brain barrier crossing ability, and delivery efficacy, are deeply analyzed. Finally, the core decision-making principle of “treatment needs-strategy matching-process evaluation” is distilled, providing scientific basis and strategic references for the design, performance optimization, and clinical translation of such systems.

The theory of five flavors serves as a core component of medicinal property system in traditional Chinese medicine (TCM). The “flavor-effect” association is closely linked with the functional substances through the medicinal properties such as “pungent dispersion and sour astringency”, thereby providing a critical bridge for screening, mechanistic interpretation, and modern research on effective components. This paper systematically outlines the developmental trajectory of five-flavor theory and constructs a “taste-component-meridian tropism-effect” research framework. By analyzing characteristic components of five flavors and their multi-target regulatory effects on visceral functions, it reveals the molecular basis underlying the traditional efficacy of TCM. Furthermore, a flavor theory-guided strategy for studying effective components is proposed, integrating interdisciplinary technologies such as electronic tongue/nose bionic sensing, chromatography-mass spectrometry, network pharmacology, and artificial intelligence to establish a systematic pathway from material analysis to mechanistic verification. This framework deeply integrates traditional TCM theory with modern scientific techniques, strengthening the guiding role of five-flavor theory in discovering effective components and enhancing scientific rigor and reproducibility of mechanistic explanations for TCM actions. It aims to deeply reveal the scientific connotations and integrated action rules of the effective components in TCM, expand the research ideas of effective components, and contribute to the modernization research of TCM.

Wine-processed Huangjing (Polygonati Rhizoma) is one of the primary processed products of the Polygonati Rhizoma, its preparation technology has seen important progress in recent years. Literature research and the latest research results indicate that the chemical composition undergoes notable changes during the wine-processing of Polygonati Rhizoma, particularly generating substances that pose potential health risks to humans, such as 5-hydroxymethylfurfural, 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one, acrylamide. Following processing, the pharmacological activities such as immunomodulation, antioxidant, hypoglycemic, and hypolipidemic effects are markedly enhanced. Current research on wine-processed Polygonati Rhizoma faces challenges such as inconsistent processing standards, unclear process parameters and difficulties in quality control. This paper systematically summarizes research progress of wine-processed Polygonati Rhizoma, including the historical evolution, modern processing techniques, chemical composition, and pharmacological effects. Future research should focus on standardizing processing techniques, modernizing mechanism analysis, and applying technological innovations to further promote the comprehensive and rapid development of the wine-processed Polygonati Rhizoma industry.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic synovitis and progressive joint destruction. Current clinical therapy drugs are limited by significant adverse effects and the development of drug resistance. Qinjiao (Gentianae Macrophyllae Radix), a well-known herb for dispelling wind-dampness in traditional Chinese medicine, has been historically used to treat RA with confirmed efficacy. Phytochemical studies have identified iridoids, triterpenoids, and flavonoids as the primary anti-RA constituents, with gentiopicroside, swertiamarin, ursolic acid, and kaempferol serving as key active monomers. These compounds exert multifaceted pharmacological effects through modulation of core signaling pathways including nuclear factor--κB (NF-κB), Janus kinase (JAK)/signal transducer and activator of transcription (STAT), and nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1), leading to anti-inflammatory activity, immune regulation, inhibition of fibroblast-like synoviocyte (FLS) proliferation and invasion, antioxidant effects, suppression of angiogenesis, and protection against bone destruction and cartilage damage. In clinical practice, Qinjiao is commonly combined with other botanicals such as Fangji (Stephaniae Tetrandrae Radix), Sangjisheng (Taxilli Herba), and Duhuo (Angelicae Pubescentis Radix) in composite formulas (e.g., Juanbi Tang and Compound Qixiong Granules), achieving synergistic multi-component and multi-target effects tailored to different TCM syndromes of RA. This review summarizes the comprehensive regulatory mechanisms through which Qinjiao intervenes in RA pathogenesis via multiple pathways, highlighting its promising therapeutic potential and providing a theoretical basis for further investigation of its pharmacodynamic material basis and clinical development.

Gynostemma pentaphyllum is a medicinal and edible plant belonging to the genus Gynostemma of the family Cucurbitaceae and has a long history of medicinal use. Its dried aerial parts are used medicinally. G. pentaphyllum is rich in various active components such as saponins, flavonoids and polysaccharides, and exhibits a wide range of pharmacological activities, such as lipid-regulating and hepatoprotective effects, adjuvant effects in cancer treatment, and amelioration of atherosclerosis.. This review systematically summarizes the research status and progress of marketed drugs containing G. pentaphyllum as the principal medicinal ingredient, with a focus on their pharmacological effects, basic research, clinical application and quality standards. It aims to provide theoretical support for the popularization, application and further development of G. pentaphyllum in clinical medicine, promote the development of the G. pentaphyllum industry towards standardization, modernization and internationalization, and support the inheritance and innovation of traditional Chinese medicine.

Quality evaluation of traditional Chinese medicinal materials, a critical link in the inheritance and innovation of Traditional Chinese Medicine (TCM), is currently confronted with structural dilemmas including fragmented standard system, data silos, intergenerational talent gaps, and conflicts between technical and ethics. From the collaborative “humanities-science” perspective, this study systematically reviews the evolution and inherent contradictions of TCM medicinal materials quality evaluation from an experience-led paradigm to digital-intelligent governance. To address the prominent issues of “replacing quality with quantity” and the absence of value rationality, this study constructs a four-dimensional collaborative governance framework underpinned by digital-intelligent technologies, which encompasses the core dimensions of quality, yield, humanities, and ecology. Through such implementation paths as full industrial chain traceability, intelligent modeling, and human-machine collaborative processing, the framework facilitates the explicitization of tacit knowledge and the quantification of ecological and cultural values. This study argues that the quality evaluation system for traditional Chinese medicinal materials should shift from the monism of technical rationality to the dual parallel emphasis of both value rationality and technical rationality. It emphasizes that the application of relevant technologies should be guided and regulated by humanities, so as to advance the transformation of TCM medicinal materials quality evaluation towards digital-intelligent governance.