Objective To study chemical constituents and neuroprotective activities of the branches and leaves of Aphanamixis polystachya. Methods Monomer compounds were obtained through separation and purification using various chromatographic techniques including silica gel column chromatography, reversed-phase ODS column, Sephadex LH-20, semi-preparative HPLC, and recrystallization. Their structures were elucidated by modern spectroscopic methods including nuclear magnetic resonance (NMR), high-resolution electrospray ionization mass spectrometry (HRESIMS), infrared spectroscopy (IR), ultraviolet spectroscopy (UV), and single-crystal X-ray diffraction analysis. Results A total of 21 compounds were isolated from the branches and leaves of A. polystachya, identified as, (2R,5R,8S,9S,10R)-1-[(E)-5-hydroxy-3-methylpent-3-en-1-yl]-4,4,8,10-tetramethyldecahydronaphthalene-2,8-diol (1), (1S,4aS,7R,8R,8aS)-decahydro-8-[(3E)-5-hydroxy-3-methyl-3-penten-1-yl]-4,4,7,8a-tetramethyl-1,7-naphthalenediol (2), 3β,8α,15-trihydroxy-13-labdane (3), 8β,15-dihydroxy-ent-labda-13E-en-3-one (4), labd-13(E)-ene-8α,15-diol (5), labd-13(E)-ene-8α-hydroxy-15-acetate (6), labda-7,13E-dien-15-ol (7), labda-8,13E-dien-15-ol (8), brachytylin D (9), labda-8(17),13E-dien-15-ol (10), selin-11-en-4α-ol (11), (±)-selin-11-en-4α-ol (12), (1S,5R,9R)-10,10-dimethyl-2,6-dimethylenebicyclo[7.2.0]undecane-5-ol (13), caryophyllenol-II (14), ciwujiatone (15), lyoniresinol (16), isolariciresinol (17), (±)-ficusesquilignan A (18), ficusesquilignan B (19), syringaresinol (20), methyl rosmarinate (21). Conclusion Compound 1 is a novel compound designated as labdane diterpenoid Z. Compounds 2-4, 9, 11-15, and 18-21 are isolated from genus Aphanamixis for the first time, which further enriches the chemical component database of the genus Aphanamixis. Furthermore, biological activity assays reveals that compound 21 exhibits neuroprotective activity.

Objective To study the chemical constituents of the dried leaves of Callicarpa nudiflora and discover bioactive compounds with anti-methicillin-resistance Staphylococcus aureus (MRSA) effects. Methods A systematic separation of chemical constituents was conducted by comprehensive chromatographic methods, including silica gel, Sephadex LH-20, ODS column chromatography and other chromatographic techniques. The structures of the isolated compounds were determined on the basis of spectroscopic analysis including IR, HR-ESI-MS, ¹H-NMR, ¹³C-NMR, HSQC, HMBC, COSY, etc. The minimum inhibitory concentrations (MICs) of the compounds against Staphylococcus aureus, MRSA, Escherichia coli, and multidrug-resistant E. coli were determined by the nutrient broth dilution method. Results One monoterpene, named (Z)-2-methyl-6-methyleneocta-2,7-dine-1-yl (E)-3-(4-hydroxyphenyl)acrylate (1), and four sesquiterpenoids, named monasuslunin (2), (1S,5R,9R)-10,10-dimethyl-2,6-dimethylenebicyclo [7.2.0] undecan-5-ol (3), 7-epi-eudesm-4(15)-ene-1β,6β-diol (4), sclareolide (5) were isolated and identified. Compound 1 exhibited strong inhibitory effects against S. aureus 29213 and MRSA T144, with MICs of 8 μg/mL for both. The MICs against E. coli 25922 and E. coli B2 were both greater than 8 μg/mL. Conclusion Compound 1 is a new compound named Callinuene A and compounds 3-5 are first isolated within this species. Compound 1 exhibits significant anti-MRSA activity.

Objective To study the chemical constituents of Salvia miltiorrhiza and their anti-asthma activity. Methods The compounds were isolated and purified by silica gel column chromatography, gel column chromatography and preparative liquid chromatography. Their structures were identified based on spectroscopic data and physicochemical properties. The anti-asthma activity of the obtained compounds was evaluated using β-hexosaminidase release assay and RBL-2H3 cell degranulation model. Results Seven compounds were isolated from the ethyl acetate fraction of S. miltiorrhiza and identified as (S)-3-(3-hydroxy-4-methoxyphenyl)-1-methoxy-1-oxopropan-2-yl (2S,3S)-2-(3,4-dihydroxyphenyl)-4-((E)-3-(((S)-3-(3,4-dihydroxyphenyl)-1-ethoxy-1-oxopropan-2-yl)oxy)-3-oxoprop-1-en-1-yl)-7-hydroxy-2,3-dihydrobenzofuran-3-carboxylate (1), rosmarinic acid (2), methyl linoleate (3), 4-oxononanoic acid (4), (Z)-9-octadecenoic acid (5), (9Z,12Z)-nonadeca-9,12-dienoic acid (6), and n-13,16-nonadecadienoic acid (7). The bioactivity results showed that compound 1 inhibited the release of β-hexosaminidase, reduced the levels of mouse mast cell protease-1 (mMCP-1), β-mast cell tryptase (β-MCT), histamine (HIS) and leukotriene C4 (LTC4), and significantly ameliorated the degranulation of RBL-2H3 cells. Conclusion Compound 1 is a new water-soluble phenolic acid, named 9′ʺ-ethyl salvianolate B, that may exert anti-asthma effect by inhibiting mast cell degranulation.

Objective To develop an integrated strategy for rapid screening of pancreatic lipase (PPL) inhibitors from Rheum palmatum and systematically investigate their potential anti-obesity mechanisms. Methods Fe₃O₄@SiO₂@PPL was synthesized via chemical co-precipitation, Stöber method, and cross-linking, and characterized by Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and X-ray diffraction (XRD). The Fe₃O₄@SiO₂@PPL was applied in ligand fishing experiments on an ethanol extract of R. palmatum, and the captured ligands were identified by UPLC with reference standards. Molecular docking and network pharmacology were integrated to conduct mechanism prediction from multiple dimensions such as molecular interaction, target mapping, and pathway enrichment. Results Material characterization revealed the presence of an enzyme coating layer on the surface, along with characteristic absorption peaks of Fe-O, Si-O-Si, and -NH₂, indicating that pancreatic lipase was successfully immobilized on the carrier surface. Four compounds were specifically captured by fishing experiments, including chrysophanol-8-O-β-D-glucopyranoside, aloe-emodin, rhein, and chrysophanol. Among them, chrysophanol and aloe-emodin exhibited high inhibitory effects against pancreatic lipase, with median inhibition concentration (IC₅₀) values of 67.03 and 85.86 μmol/L, respectively. Molecular docking suggested that the active components can form hydrogen bonds and hydrophobic interactions with key amino acid residues of pancreatic lipase. Network pharmacology identified 150 overlapping targets between the active components and obesity. Screening out the five core targets: epidermal growth factor receptor (EGFR) and protein kinase B1 (AKT1), proto-oncogene tyrosine-protein kinase Src (SRC), heat shock protein 90 alpha family class A member 1 (HSP90AA1), and B-cell lymphoma-2 (BCL2). Pathway enrichment analysis indicated that the HIF-1 signaling pathway and the lipid and atherosclerosis pathway were the main mechanisms of action. Conclusion An integrated approach combining material-based screening, computational validation, and network prediction was established. This strategy provides an efficient tool for high-throughput screening of bioactive compounds from traditional Chinese medicines and offers methodological insights into clarifying the multi-target action mechanism of traditional Chinese medicine from the perspective of systems biology.

Objective To explore the effects of different processing methods on the expectorant and antitussive effects of Zhuli (Bambusae Succus, BS). Methods The chemical composition differences of BS prepared by different processing methods were analyzed by GC-MS technology, and a comprehensive evaluation was conducted using principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) to screen for differential and specific components. The microscopic characteristics of bamboo before and after processing were further compared and analyzed. A chronic obstructive pulmonary disease (COPD) mouse model was established to evaluate the expectorant and antitussive effects of BS prepared by different processing methods. Results This study determined the relative contents of guaiacol and syringol and the overall proportion of phenolic components in BS prepared by three processing methods through GC-MS analysis. In the fire treatment method, the proportions were 4.68%, 2.22%, and 14.93% respectively; in the dry distillation method, they were 5.98%, 6.96%, and 24.77% respectively; in the water boiling method, the overall proportion of phenolic components was 3.65%, and the relative contents of the two components were both below the detection limit. PCA and OPLS-DA indicated that the three processing methods had certain differences in chemical composition and screened out 13 characteristic differential components with distinguishing significance, including 2-methylpheno, creosol, eugenol, 2-methoxy-4-propyl-Phenol, (E)-isoeugenol, vanillin, 2-propenal, 2-methoxy-5-methylphenol, 4-ethylguaiacol, maltol, oleamide, 3, 5-dimethoxy-4-hydroxybenzaldehyde, pyrrole-2-carboxaldehyde. Cluster analysis showed that the components of the fire processing and dry distillation methods were relatively similar, while both were significantly different from the water boiling method. Microscopic structure observation revealed that the bamboo structure corresponding to the BS prepared by the water boiling method was similar to that of fresh bamboo, while the fire processing and dry distillation methods had more significant changes in bamboo structure. Pharmacological experiments showed that BS prepared by the three methods all exhibited varying degrees of cough-suppressing and expectorant effects, among which the dry distillation method significantly prolonged the cough latency period (P < 0.05) and increased phenol red excretion (P < 0.01). Conclusion Compared with the fire processing and water boiling methods, the dry distillation method can more effectively retain active and convert components such as guaiacol and syringol. Mechanism studies indicated that the water boiling method, due to insufficient heating temperature, was unable to thermally decompose lignin to generate phenolic acid components, resulting in significant differences in chemical composition compared to the traditional fire processing and dry distillation methods, leading to distinct quality characteristics among the three approaches. The efficacy experiments showed that all three methods exhibited expectorant and antitussive effects to varying degrees; among them, the dry distillation method had a more significant effect in prolonging the cough latency period and increasing the phenol red excretion volume. By studying the effects of different processing techniques on the quality of BS and its efficacy in relieving cough and expectorating phlegm, this research provides a theoretical basis for its clinical application, the development of new dosage forms, and the scientific connotation of dry distillation processing.

Objective Based on a confined mass transfer mathematical model, this study aims to establish a quantitative analysis method for the existing states of matrine in the complex solution environment of traditional Chinese medicine (TCM), and elucidate the influence pattern of the compatibility ratio of Danshen (Salviae Miltiorrhizae Radix et Rhizoma, SMRR) and Kushen (Sophorae Flavescentis Radix, SFR) on its acid-base complexation reaction. Methods Based on the mass transfer differences of component states during nanofiltration (NF) and ultrafiltration (UF), the mass transfer coefficient of matrine was fitted taking the component transmission rate and membrane flux as evaluation indicators. Using the molecular and ionic monomer components as references, a quantitative calculation model for the existing states was constructed. The existing states of matrine in the SFR extract and the SMRR- SFR co-extract were fitted to analyze the occurrence patterns of acid-base complexation reactions under different solution environments. Results Coupling UF and NF enhanced the separation differences between the ionic and molecular states of matrine, thereby improving the accuracy of the quantitative calculation model for the ionic state. The correlation coefficients of the power function equations for quantitatively calculating the existing states of matrine were all greater than 0.97, indicating that the model was successfully established. The proportion of matrine in the molecular state was approximately 1.21%-1.91% in both the SFR extract and the co-extract. In the SFR extract, matrine existed as 36.90% in the ionic state and 61.98% in the associated state. After compatibility and extraction of SFR and SMRR, the existing form of matrine changed significantly. When the compatibility ratio of SMRR was low, the ionic state was dominant, with the increase in the proportion of SMRR, the ionic proportion of matrine gradually decreased from 66.73% to 45.83%, transforming into a complex-structured composite state, and the corresponding composite state proportion rose from 31.87% to 52.90%. Conclusion A quantitative calculation method for the existing states of matrine in different solution environments was successfully constructed, providing technical support for elucidating the states of components in complex solution environments.

Objective To prepare a self-assembled nanoparticles of saikosaponin D (SSD) and cannabidiol (CBD) and preliminarily investigate their anti-hepatoma mechanism in vitro. Methods The SSD-CBD self-assembled nanoparticles were constructed using the nano co-precipitation method at a mass ratio of 3∶1, and their assembly mechanisms were characterized by techniques including X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and ¹H-¹H nuclear overhauser enhancement spectroscopy (¹H-¹H NOESY). Its physicochemical properties were evaluated by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The antitumor activity, apoptosis induction, and preliminary mechanisms were systematically assessed by MTT assay, flow cytometry (Annexin V-FITC/PI staining), and cysteinyl aspartate specific proteinase (Caspase) activity assays. Results The assembly was self-assembled through hydrogen bonding and hydrophobic interactions, with SSD and CBD showing a synergistic effect against HepG2 cells (CI = 0.79). It exhibited pH-responsive drug release, specifically releasing drugs in the tumor microenvironment (pH 6.8) while remaining stable at normal physiological environment (pH 7.4). Mechanistic studies indicated that it primarily induced HepG2 cell death by activating the mitochondrial apoptotic pathway. Conclusion The constructed SSD-CBD self-assembled nanoparticles offer a carrier-free strategy that can simultaneously improve the delivery efficiency of CBD and achieve precise collaborative drug delivery, providing an experimental basis for the development of intelligent nanomedicine based on SSD.

Objective To optimize the optimal processing methods for two different preparations of Huaimi (Sophorae Japonicae, SJ) and establish a method for determining the content of marker components and analyzing their correlation with color values using UPLC. Methods UPLC was employed to determine nine components in SJ (5-hydroxymethylfurfural, protocatechuic acid, rutin, isoquercitrin, quercetin, kaempferol-3-O-rutinoside, narcissin, kaempferol, and isorhamnetin) and alcohol-soluble extract content as evaluation indicators. A single-factor experiment combined with Box-Behnken design-response surface methodology (BBD-RSM) was employed to investigate three factors: roasting power, roasting time, and feedstock quantity. The weighting coefficient of the 10 indicators was calculated using the criteria importance through inter-criteria correlation (CRITIC) method. Both approaches were integrated to analyze and optimize the processing techniques for the two prepared forms of SJ. The IRIS electronic eye was employed to measure color changes in SJ at different processing levels. SPSS 20.0 and Origin 2024 software were used to analyze the correlation between color and indicator components. Results The optimal processing conditions for stir-fried SJ were determined as frying power 900 W, 5 min processing time, and 115 g batch size. For charred SJ, the optimal conditions were frying power 1 600 W, 3 min processing time, and 100 g batch size. Three batches of samples were prepared for validation, yielding average composite scores of 67.33 and 78.98 with RSD of 2.29% and 0.70%, respectively. These values closely matched predicted values, indicating process stability. Correlation analysis between color values and nine components revealed that both L^* and b^* values showed significant positive correlations with rutin, kaempferol 3-O-rutinoside, and narcissin, while exhibiting significant negative correlations with kaempferol, quercetin, and isorhamnetin. The a^* value demonstrated significant positive correlations with protocatechuic acid, isoquercitrin, quercetin, kaempferol, and isorhamnetin, indicating a correlation between color and changes in internal components of SJ. Conclusion The processing methods for stir-fried SJ and charred SJ optimized through BBD-RSM and CRITIC weighting are stable and feasible. Changes in intrinsic components during processing showed significant correlation with color values, providing a simple and intuitive basis for assessing the degree of processing and the relationship between intrinsic component changes in stir-fried SJ and charred SJ, and offering reference for their quality evaluation and clinical application.

Objective To prepare an amino acid-based deep eutectic gel dressing and study in vitro performance and the in vivo therapeutic effect on oral ulcer. Methods Deep eutectic solvents (DESs) were screened using lysine, proline, arginine, and glycine as hydrogen bond donors, and lactic acid, citric acid, glycerol, mannitol, and malic acid as acceptors, with curcumin solubility identifying the optimal system. Gel dressings were prepared by optimizing the formulation process based on the composite matrix of gelatin methacryloyl/chitosan methacryloyl (GelMA/CSMA) in combination with 3D printing technology. Its mechanical properties, adhesiveness, swelling, and biocompatibility were evaluated, and in vivo efficacy was verified in an SD rat oral ulcer model. Results The optimal DES system (proline-lactic acid, 1:12) increased curcumin solubility over 700-fold (0.789 mg/mL) compared to its solubility in water. The optimal printing formulation and process parameters were as follows: 1% CSMA, 5% GelMA, 0.1% lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), 0.02% tartrazine, 20% DES content, 100 μm layer height, and a layer-by-layer exposure time of 25 s. The obtained amino acid-based gel dressing had excellent mechanical properties, with a 2.5 N adhesion force at 25 min and an approximately 500% swelling ratio at 6 h. In vitro biocompatibility showed that the survival rate of L929 cells was all above 90%, and the DES system had no significant cytotoxicity. It also showed high in vitro biocompatibility (L929 viability > 90%, non-cytotoxic). In vivo, the gel achieved localized, sustained curcumin release in SD oral ulcer model rats, reducing ulcer area by 40% with decreased neutrophil density and inflammatory infiltration. Conclusion Successfully prepared an amino acid-based DES gel dressing loaded with curcumin, which exhibits excellent wound adaptability and precise drug release behavior, offering an efficient and safe therapeutic strategy for oral ulcers.

Objective To evaluate the quality of Fagopyri Dibotryis Rhizoma Tablets (FDRT) produced by different companies. Methods A total of 18 batches of FDRT produced by company A and 20 batches of FDRT produced by company B were collected, respectively. The fingerprints of 38 batches FDRT were determined using HPLC, the common peaks were assigned and the constituents of these peaks were identified by quadrupole time-of-flight mass spectrometry (Q-TOF-MS/MS). Using quantitative common peak areas as variables, pattern recognition analysis of the fingerprint was conducted using hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). The content of the nine constituents confirmed by comparison with reference substances and have good separation in samples were determined. Results There were 33 common peaks in the fingerprint of 38 batches of FDRT, and a total of 53 constituents were identified by Q-TOF-MS/MS, including 21 tannins, 12 phenolics, seven flavonoids, seven phenylpropanoid glycosides, two amino acids, two organic acids, one alkaloid, and one terpenoid, among them, three phenolics (gallic acid, protocatechuic acid, and protocatechualdehyde), five tannins (procyanidin B1, B2, B3, C1, C2), four flavonoids (catechin, epicatechin, epicatechin gallate, and rutin) were confirmed by reference substance. The separation degree of nine constituents except for procyanidin C2, B2, and rutin was relatively good. The average content of these nine constituents in FDRT produced by company B was significantly higher than that in FDRT produced by company A (P < 0.01). The results of HCA, PCA, and OPLS-DA were consistent, clearly distinguished the FDRT produced by company A and company B into two categories. The constituents corresponding to common peaks 30, 29, 16, 17, 2, 21, 10, 11, 18, 20, and 27, such as procyanidin C2, B2, C1, B1, B3, gallic acid, epicatechin, etc., are the marker constituents responsible for the quality differences in FDRT produced by two enterprises. Conclusion There is a significant quality difference in FDRT produced by two companies.

Objective To explore the mechanism of morin, an active ingredient of Sangzhi (Mori Ramulus), against gastric cancer based on network pharmacology, molecular docking and in vitro experiments. Methods Potential targets of Mori Ramulus and gastric cancer-related targets were screened by network pharmacology, and the intersection of targets was obtained. Protein-protein interaction (PPI) network topological analysis, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, and GEO dataset validation were used to screen core targets. Molecular docking and molecular dynamics simulation were applied to verify the binding ability of morin (the active ingredient of Mori Ramulus) to phosphatidylinositol 3-kinase regulatory subunit 1 (PIK3R1) and the stability of the formed complex. In in vitro experiments, human gastric adenocarcinoma AGS cells were treated with 100, 200, 300, 400 μmol/L morin alone or combined with phosphatidylinositol 3-kinase (PI3K) agonist 740Y-P. Cell proliferation ability was detected by CCK-8 assay and plate clone formation assay. Cell apoptosis rate and cell cycle distribution were detected by flow cytometry. Expressions of PI3K/protein kinase B (Akt) pathway, apoptosis and cycle related proteins were detected by Western blotting. Results A total of 178 potential targets of Mori Ramulus, 13 100 gastric cancer-related targets, and 159 intersecting targets were identified. Enrichment analysis showed that the key pathway was the PI3K/Akt pathway. Morin had strong binding affinity with PIK3R1, was significantly highly expressed in gastric cancer tissues, and the constructed morin-PIK3R1 complex maintained excellent stability. The in vitro experiment results showed that morin could inhibit AGS cells proliferation in a dose-dependent manner, induce G₀/G₁ phase arrest, and promote cell apoptosis (P < 0.05, 0.01). Meanwhile, morin downregulated the expressions of PI3K/Akt pathway, B-cell lymphoma-2 (Bcl-2), cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6) protein (P < 0.05, 0.01), and upregulated the expressions of Bcl-2 associated X protein (Bax) and p21 (P < 0.01). Combined treatment with 740Y-P significantly reversed the effects of morin on proliferation, apoptosis and cell cycle (P < 0.05, 0.01). Conclusion Morin can exert anti-gastric cancer activity by targeting and stably binding to PIK3R1, inhibiting the activation of PI3K/Akt pathway, inducing cell cycle arrest, promoting apoptosis, and inhibiting proliferation in gastric cancer cells.

Objective To investigate the pharmacodynamic effect and mechanism of casticin on improving colitis-associated colorectal cancer (CAC). Methods A CAC mouse model was established using azoxymethane (AOM)/dextran sulfate sodium (DSS) induction, followed by intervention with casticin. Therapeutic efficacy and safety of casticin were evaluated by comparing body weight changes, disease activity index, colonic tumor burden, tumor-bearing survival rates, organ indices, pathological alterations and serum liver function indicators. Intestinal microbiota structural alterations were analyzed using 16S rRNA sequencing. Differentially expressed proteins and key signaling pathways were screened through proteomic techniques. Western blotting was performed to validate key protein expressions. Kaplan-Meier survival analysis based on TCGA database was conducted to evaluate the prognostic value of target genes, and molecular docking was employed to predict the binding modes between casticin and target proteins. The interaction between gut microbiota and host targets was revealed through correlation analysis. Results Compared with model group, casticin significantly alleviated body weight loss in CAC mice, improved intestinal inflammatory responses, reduced colonic tumor volume and tumor burden (P < 0.05), and improved survival rates of tumor-bearing mice. Safety evaluation showed no significant abnormalities in serum liver indicators and histomorphology of major organs in all treatment groups. 16S rRNA sequencing demonstrated that casticin effectively reversed CAC-induced intestinal dysbiosis, significantly downregulated the relative abundance of pro-carcinogenic phyla Fusobacteriota and Patesbacteria, while enriched the anti-inflammatory genera Lachnospiraceae_NK4A136_group and Prevotellaceae_UCG-001 that produce short-chain fatty acids. Proteomic analysis identified the complement and coagulation cascade pathway as the core responsive pathway, with dose-dependent restoration of microenvironment remodeling-related molecule serine protease inhibitor 1 (Serpine1) and immune recognition receptor integrin alpha M (Itgam) expression. Western blotting validation demonstrated that compared with model group, the protein expression levels of Itgam and Serpine1 in colonic tissues were significantly downregulated in casticin group (P < 0.01, 0.001), consistent with the proteomic trend analysis. Integrated analysis further revealed that the abundance of beneficial bacteria such as Lachnospiraceae_NK4A136_group was significantly negatively correlated with the expression of microenvironment remodeling molecule Serpine1, while the abundance of pro-carcinogenic bacteria such as Fusobacteriota was significantly positively correlated with the expression of immune recognition receptor Itgam. Conclusion Casticin can reshape the intestinal microbiota structure, regulate key molecular responses in the complement and coagulation cascade pathway, thereby synergistically blocking the inflammatory-carcinogenic transformation process of CAC.

Objective To explore the mechanism of tubeimoside II (TUB-II) in inhibiting the growth and metastasis of colorectal cancer. Methods Human colorectal cancer cell lines RKO and SW620 were used as research subjects, the effect of TUB-II on viability of RKO and SW620 cells was detected. The cell proliferation ability was detected by colony formation assay. The cell migration ability was detected by scratch assay. Western blotting was used to detect the effect of TUB-II on expressions of lethal (2) giant larvae homolog 2 (LLGL2) and epithelial-mesenchymal transition (EMT)-related proteins in RKO and SW620 cells. Co-immunoprecipitation was used to detect the effect of TUB-II on ubiquitination of LLGL2. Subcutaneous tumor model and colorectal cancer lung metastasis model in vivo were constructed to investigate the effect of TUB-II on tumor growth and lung metastasis. Results TUB-II significantly inhibited the proliferation and metastasis of colorectal cancer both in vitro and in vivo (P < 0.05, 0.01, 0.001). TUB-II significantly upregulated the expression of LLGL2 (P < 0.05, 0.01, 0.001) and inhibited the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway (P < 0.05, 0.01, 0.001), thereby suppressing tumor cell proliferation and metastasis. At the same time, TUB-II inhibited the interaction between murine double minute 2 (MDM2) and LLGL2 to suppress the ubiquitination of LLGL2, thereby stabilizing its expression and exerting an anti-colorectal cancer effect. Conclusion TUB-II inhibits the occurrence and development of colorectal cancer, and its mechanism may be related to the inhibition of LLGL2 ubiquitination and PI3K/Akt signaling pathway.

Objective To study the protective effect and potential mechanism of soybean isoflavones (SI) on ischemic stroke. Methods SI were isolated from soybeans and components were analyzed. An oxygen glucose deprivation/reperfusion (OGD/R)-induced PC12 cells model was constructed to investigate the effect of SI on cell survival, migration ability, apoptosis, intracellular Ca²⁺, reactive oxygen species (ROS), lactate dehydrogenase (LDH), cysteine aspartate protease-3 (Caspase-3) levels, and expressions of Kelch-like ECH associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway related proteins. A rat model of global cerebral ischemia-reperfusion injury was established using bilateral common carotid artery ligation. The effect of SI on neurological function score, brain tissue water content, proportion of cerebral infarction volume, pathological changes, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), LDH activities, malondialdehyde (MDA) level, and expressions of Keap1/Nrf2 pathway related proteins in brain tissue were investigated. Results In vitro experiments showed that compared with model group, SI significantly increased cell survival rate and migration rate (P < 0.05, 0.01), decreased cell apoptosis rate and intracellular Ca²⁺, ROS, LDH, Caspase-3 levels (P < 0.05, 0.01), upregulated Nrf2, heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQO1) protein expressions (P < 0.01), and downregulated Keap1 protein expression (P < 0.05, 0.01). In vivo experiments, compared with model group, SI significantly improved neurological deficits, reduced brain edema and infarct volume in rats (P < 0.01), repaired brain tissue pathological damage, reduced MDA level and LDH activity in brain tissue (P < 0.01), increased SOD, CAT and GSH-Px activities in brain tissue (P < 0.01), upregulated Nrf2, HO-1 and NQO1 protein expressions in brain tissue (P < 0.01), and downregulated Keap1 protein expression in brain tissue (P < 0.01). Conclusion SI have a significant neuroprotective effect on ischemic stroke, which may be related to the activation of Keap1/Nrf2 pathway, enhancing the antioxidant capacity of body, inhibiting oxidative stress damage, reducing neuronal apoptosis and alleviating brain tissue damage.

Objective To explore the potential active components, targets and molecular mechanism of Tianma Gouteng Yin (天麻钩藤饮) in improving vascular endothelial function in hypertension based on network pharmacology, molecular docking and experimental verification, and to clarify whether it exerts effects via phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) signaling pathway. Methods Active components and targets of Tianma Gouteng Yin were screened through HERB and SwissTargetPrediction databases. Hypertension-related targets were obtained from OMIM and GeneCards databases. A protein-protein interaction (PPI) network was constructed, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. Molecular docking was used to verify the binding affinity between core components and key targets in PI3K/Akt/eNOS pathway. Human umbilical vein endothelial cells (HUVECs) injured by angiotensin Ⅱ (Ang-Ⅱ) were used as the cell model. CCK-8, qRT-PCR, Western blotting, reactive oxygen species (ROS) and nitric oxide (NO) assays were applied to evaluate the effects of Tianma Gouteng Yin on cell viability, oxidative stress, inflammatory response and PI3K/Akt/eNOS pathway. Results Network pharmacology identified 228 active components and 969 overlapping hypertension-related targets of Tianma Gouteng Yin. Core targets included serine/threonine kinase 1 (AKT1) and PI3K family members [phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta (PIK3CB), etc.]. KEGG enrichment indicated that PI3K/Akt pathway was the key pathway. Molecular docking confirmed that core components such as kaempferol and apigenin showed strong binding affinity to PI3K/Akt pathway targets. The results of cell experiment showed that 25-100 mg/mL Tianma Gouteng Yin had no cytotoxicity in normal HUVECs, and dose-dependently attenuated Ang-Ⅱ-induced reduction in cell viability (P < 0.001), significantly inhibited the expressions of angiotensin-Ⅱ type 1 receptor (AT1R) and NADPH oxidase 4 (NOX4), as well as ROS production (P < 0.001), upregulated the levels of antioxidant enzymes including superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase 1 (GPX1) (P < 0.05, 0.01, 0.001), reduced the expressions of inflammatory factors including interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α), intercellular adhesion molecule 1 (ICAM1) and vascular cell adhesion molecule 1 (VCAM1) (P < 0.01, 0.001), increased NO production and suppressed endothelin-1 (ET-1) expression (P < 0.01, 0.001), upregulated the protein expressions of p-PI3K, p-Akt and p-eNOS (P < 0.001). After administering the PI3K specific inhibitor Wortmannin, the activation effect of Tianma Gouteng Yin on PI3K/Akt pathway was significantly weakened, indicating that its endothelial protective effect depends on PI3K/Akt/eNOS signaling pathway. Conclusion Tianma Gouteng Yin may activate PI3K/Akt pathway through multi-target synergistic effects of core components such as kaempferol and apigenin, improve oxidative stress and inflammatory response, regulate the balance of NO and ET-1, and protect endothelial function in hypertension.

Objective To explore the differences in the in vivo drug metabolism of raw and processed Dahuang (Rhei Radix et Rhizoma) in a rat model of middle cerebral artery occlusion and reperfusion (MCAO/R). Methods The MCAO/R rat model was established by the suture method. The successfully modeled rats were randomly divided into model group, raw Rhei Radix et Rhizoma (2.50 g/kg) group and processed Rhei Radix et Rhizoma group, with six rats in each group. Rats were administered continuously for 7 d. 12 h before the last administration, the rats were fasted, and 1 h after administration, blood and brain tissues were collected. Ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap linear ion trap mass spectrometry (UPLC-Orbitrap Fusion Lumos Tribrid-MS) was used to characterize the prototypes and metabolites of raw and processed Rhei Radix et Rhizoma in plasma and brain tissues of MCAO/R rats. Multivariate statistical analysis was used to screen the differential components, and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) technique was combined to visualize the in situ spatial distribution of the differential components of raw and processed Rhei Radix et Rhizoma in brain tissue. Results A total of 109 prototype components and metabolites were identified in MCAO/R rats. According to variable important in projection (VIP) > 1 and P < 0.05, 23 and eight differential components of raw and processed Rhei Radix et Rhizoma were screened from the plasma and brain tissues, respectively. In the plasma of processed Rhei Radix et Rhizoma group, anthraquinone glycosides were decreased, while in the brain tissue, aglycones and metabolites (such as gallic acid, M31) were increased, and were mainly distributed in hippocampus, midbrain and cortex of brain tissue. The common differential components gallic acid and metabolite M31 were significantly increased in the processed Rhei Radix et Rhizoma group and were specifically enriched in the striatum. Conclusion This study characterized and spatially located the differential components of raw and processed Rhei Radix et Rhizoma in MCAO/R rats and found that processed Rhei Radix et Rhizoma increased the exposure levels of active aglycones and metabolites in the brain, enhancing its protective effect on brain tissue, providing a scientific basis for the “different treatment for raw and processed” processing theory.

Objective This study aims to systematically analyze the intellectual property landscape of traditional Chinese medicine (TCM) globally, with a focus on comparing the patent layout characteristics and competitive dynamics in five key countries: China, the United States, Japan, South Korea, and Germany. The goal is to provide strategic references for promoting the high-quality development and internationalization of China's TCM industry. Methods Leveraging the Patyee database, this research retrieved and obtained TCM-related patent data globally and from the aforementioned five countries. Employing a combination of quantitative and qualitative analysis methods, an in-depth examination was conducted from multiple dimensions, including patent application trends, geographical distribution, applicant types, core technological topics, and international layout strategies. Results The research findings indicate that despite a vast number of domestic TCM patent applications (approximately 360 000), the proportion of international applications filed via the Patent Cooperation Treaty (PCT) is relatively low (only 1.37%), revealing a significant deficiency in overseas layout. Regarding the distribution of technological topics, patents related to TCM quality control and standardization technologies are relatively scarce globally (including in China), leaving a gap in meeting the high standards of quality controllability required for modern industrialization and internationalization. In terms of applicant types, 44% of TCM patents in China are held by individuals, while enterprises account for only 34%, reflecting a pattern of individual-led inheritance with enterprise-driven R&D yet to be enhanced. In contrast, TCM-related patent activities in the United States, Japan, and Germany are predominantly driven by multinational pharmaceutical companies or consumer goods giants with complete industrial chains. Conclusion To achieve high-quality internationalization of TCM intellectual property, China urgently needs to establish a systematic overseas patent layout strategy, forming a patent protection system covering the entire industrial chain of R&D, quality control, production, and marketing. The core task is to transform domestic technological advantages into internationally recognized technical standards and to promote the transformation of TCM from a traditional empirical model to a modern, evidence-based innovation paradigm by strengthening collaborative innovation among industry, academia, research, and users.

Objective To investigate the targets and mechanisms of Huachansu in alleviating oxaliplatin-induced peripheral neurotoxicity using bioinformatics and network pharmacology. Methods The targets of Huachansu were predicted via SwissTarget Prediction, PharmMapper, and SuperPred databases. Targets associated with oxaliplatin-induced peripheral neurotoxicity were retrieved from GeneCards, OMIM, and Harmonizome 3.0 databases. Combined with differential genes after oxaliplatin treatment from the GEO database, common targets were obtained through intersection analysis. The DAVID database was used for gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment. Core targets were screened via the PPI network and machine learning. Single-cell analysis was performed to explore the specific expression landscape of the core targets. AutoDock and Gromacs were employed for molecular docking and molecular dynamics simulation, with visualization assisted by PyMOL, QtGrace 2.6 and Python 3.7. Results A total of 37 active components of Huachansu were identified (e.g., telocinobufagin, hellebrigenol). Forty-five common targets were obtained. CCND1, PDGFRB, and other core targets were jointly determined by PPI network and machine learning. Molecular docking showed that telocinobufagin and other components exhibited binding energies of < -5 kcal/mol with Cyclin D1 (CCND1) and platelet-derived growth factor receptor beta (PDGFRB). Molecular dynamics simulation confirmed the stable binding between PDGFRB and telocinobufagin. Single-cell analysis suggested that Huachansu targets PDGFRB in stromal cells to reshape the neuro-supportive microenvironment. Conclusion Huachansu exerts neuroprotective effects at the acute OIPN stage by targeting PDGFRB⁺ neural stromal cells and activating the phosphatidylinositol-3-hydroxykinase (PI3K)-protein kinase B (Akt) signaling pathway, providing a theoretical basis for the early prevention of chemotherapy-induced neurotoxicity.

Objective A comprehensive analysis of the research hotspots and prospective trends of Croton tiglium from the perspectives of knowledge graphs and global patents is conducted to provide theoretical basis and reference direction for clinical practice and subsequent in-depth exploration of C. tiglium. Methods C. tiglium was used as the keyword to search four databases: CNKI, Wanfang, VIP, and Web of Science (WOS), and the literature was imported into the NoteExpress literature management software for duplication checking and screening. Excel, CiteSpace, VOSviewer and other softwares were used to conduct a visual analysis of the current status and development trends of C. tiglium research at home and abroad from the dimensions of publication trends, publishing countries, institutions, authors, and keywords. The Incopat patent database was used to search and analyze C. tiglium-related global patent applications from aspects such as patent application trends, global geographical distribution, patent applicants and patent technology fields. Results A total of 1 320 Chinese literature and 1 528 English literature that met the criteria were retrieved. After a preliminary search, a total of 11 004 patents were obtained in the Incopat patent database. After the merger process, 5 196 patents were finally retained. Among them, there are 2 346 domestic patents, accounting for 45.15% of the total, occupying a leading position. In general, the research heat in the field of C. tiglium continues to rise, the number of Chinese and English publications is comparable, and the academic influence at home and abroad is balanced. Among them, the United States has the highest number of publication volume in the world, and China is in the forefront, which are important research force in this field. In terms of scientific research cooperation, Chinese medicine colleges and affiliated medical institutions are the main body at home, with a single research structure and loose institutional cooperation, and there are obvious research barriers. International research relies on transnational scientific research subjects, with close cooperation networks and mature collaboration systems. In terms of research content, the domestic focus is on the traditional pharmacology and clinical application of C. tiglium, with an emphasis on the inheritance of traditional medicinal properties. The international focus is on fundamental and cutting-edge research such as active ingredients and mechanisms of action On the whole, C. tiglium research continues to break through the traditional category, relying on multidisciplinary cross to achieve innovative development, forming a development trend of inheritance and innovation. Conclusion Research in the field of C. tiglium is constantly expanding and deepening. Its research paradigm is undergoing a significant transformation, gradually shifting from the application of traditional Chinese medicine to multi-disciplinary interdisciplinary research and in-depth analysis of molecular mechanisms. China dominates the patent layout among various countries in the C. tiglium field. However, its patent system has significant weaknesses, mainly manifested in the premature expiration of a large number of patents, low technology industrialization, and insufficient commercial expansion capabilities. The research hotspots in Chinese literature focus on the anti-inflammatory pharmacological effects and processing techniques for reducing toxicity of C. tiglium, gradually extending to agricultural applications. Meanwhile, English research hotspots are concentrated on antioxidant activity, chemical taxonomy, and natural product chemistry, with antioxidant research showing significant growth in recent years.

Objective To clarify the comprehensive clinical value, advantages, characteristics and therapeutic positioning of Ningmitai Capsules (宁泌泰胶囊) in the treatment of chronic prostatitis (damp-heat accumulation syndrome), and to provide references for decision-making of national medical and health administrative departments and rational clinical drug use. Methods An evaluation system was constructed based on the “6 + 1” dimensions of safety, effectiveness, economy, innovation, suitability, accessibility, and traditional Chinese medicine (TCM) characteristics. Multi-source evidence including questionnaire surveys, official website data, secondary evaluation of literature, and real-world data was integrated. The multi-criteria decision analysis (MCDA) model was used to measure each dimension, and the total value score was calculated based on TCM clinical evidence using software CSCv2.0. Results Ningmitai Capsules were rated Grade A for both safety and effectiveness (no serious adverse reactions, significant effects in anti-infection, relieving pain and alleviating urinary abnormalities); Grade B for economy, innovation, suitability, accessibility, and TCM characteristics (supported by economic evidence, obtained 1 invention patent, high acceptance among target populations, good resource accessibility, and a unique Miao medicine formula); the comprehensive evaluation of clinical value was Grade A. Conclusion Ningmitai Capsules have significant clinical value in the treatment of chronic prostatitis. It is recommended to further conduct pharmacological mechanism research and high-quality evidence-based research, optimize the production process, improve the drug insert, and give full play to its clinical advantages.

Objective This study investigated the regulatory mechanism by which methyl jasmonate (MeJA) modulates triterpenoid saponin biosynthesis in Jixuecao (Centella asiatica). Methods Plants were treated with 200 μmol/L MeJA, and temporal changes in leaf asiaticoside and madecassoside contents were quantified by high-performance liquid chromatography (HPLC). Transcriptome data were analyzed using weighted gene co-expression network analysis (WGCNA) to identify modules significantly correlated with asiaticoside accumulation, followed by Mfuzz clustering to refine gene subclusters. Expression patterns of candidate key genes were validated by quantitative real-time PCR (qRT-PCR). Results MeJA treatment markedly promoted asiaticoside and madecassoside accumulation, increasing their contents by 18.28% and 14.58%, respectively. WGCNA revealed 33 co-expression modules, with the darkred module displaying a highly significant positive correlation with asiaticoside content (P < 0.01). Mfuzz clustering subdivided the 286 genes in this module into four subclusters; Notably, 239 genes in Clusters 1-3 were significantly enriched in the terpenoid backbone biosynthesis, sesquiterpenoid biosynthesis, and triterpenoid biosynthesis pathways. Twenty-eight structural genes directly involved in asiaticoside synthesis were identified, exhibiting expression profiles closely aligned with saponin accumulation. qRT-PCR confirmed significant MeJA-induced upregulation of 11 key structural genes associated with triterpenoid backbone synthesis and modification, consistent with the transcriptome findings. Conclusion MeJA enhances triterpenoid saponin biosynthesis in C. asiatica through coordinated upregulation of these 11 key structural genes within the darkred module. By integrating multi-omics profiling with experimental validation, this work systematically elucidates the molecular mechanisms underlying MeJA-induced triterpenoid saponin accumulation, offering valuable candidate genes for future functional characterization and genetic enhancement of C. asiatica.

Objective To systematically identify the R2R3-MYB transcription factor family in Xuanmugua(Chaenomeles speciose) based on transcriptomic and metabolomic data, and to screen key candidate genes potentially involved in the regulation of flavonoid biosynthesis. Methods Transcriptome and metabolome databases were constructed using different tissues of C. speciosa. The R2R3-MYB transcription factor family was systematically identified based on full-length transcriptome data. Subsequently, the sequence characteristics, phylogenetic relationships, and tissue-specific expression patterns of these genes were analyzed. Results A total of 31 R2R3-type CsMYB genes were identified from the transcriptome data of C. speciosa. Based on phylogenetic analysis with Ninanjie(Arabidopsis thaliana) and Yueji (Rosa chinensis), we classified these genes into 11 subfamilies. Members within the same subfamily showed similar conserved motifs and gene structures. Flavonoids, including catechin and epicatechin, were significantly enriched in the pulp and peel. The 31 CsMYB genes exhibited differential expression patterns across various tissues of C. speciosa, among which 14 CsMYB genes, such as CsMYB1, CsMYB21, and CsMYB25, were highly expressed in the peel or pulp. Further correlation analysis revealed that six candidate genes, including CsMYB1, CsMYB2, and CsMYB14, were significantly and positively correlated with the accumulation of multiple flavonoids, such as catechin and epicatechin. These results were further validated by RT-qPCR. Conclusion This study provides the first systematic identification of the R2R3-MYB transcription factor family in C. speciosa and screens key CsMYB genes potentially involved in the regulation of flavonoid biosynthesis. These findings provide a reference for subsequent functional verification of candidate genes and lay a foundation for elucidating the molecular mechanisms underlying the quality formation of C. speciosa.

Objective This study aims to clone the LbbHLH94-like gene from Gouqi (Lycium barbarum), analyze its spatiotemporal expression, stress and hormone response patterns, and clarify its regulatory function in carotenoid biosynthesis, so as to provide a theoretical basis for revealing the molecular mechanism of goji berry fruit quality formation. Methods Using Lycium barbarum cv. ‘Ningqi 1’ as material, we employed techniques such as PCR and qRT-PCR to clone and analyze the expression patterns of LbbHLH94-like in different tissues, fruit developmental stages, abiotic stresses and exogenous hormone treatments. The overexpression vector was constructed and transformed into goji plants, and the expression of key genes, enzyme activities and total carotenoid content in carotenoid metabolism were determined. Results The LbbHLH94-like gene was successfully cloned, with an 888 bp open reading frame encoding 296 amino acids, and the sequence was identical to the genome reference sequence. This gene showed obvious spatiotemporal specificity, with the highest expression level at fruit ripening stage. It responded to high and low temperature, salt, drought stresses as well as 6-BA, IBA and GA₃ induction, showing a general pattern of promotion at low concentration and inhibition at high concentration. Overexpression of LbbHLH94-like significantly up-regulated the expression of PDS, enhanced the enzyme activities of PSY and PDS, inhibited the expression and enzyme activity of CCD4, and remarkably increased total carotenoid content in fruits. Conclusion LbbHLH94-like is a key positive regulator of carotenoid biosynthesis in goji berry, and also participates in abiotic stress response and hormone signaling. The results provide important gene resources and theoretical support for quality improvement and precise molecular breeding of Lycium barbarum.

Objective To establish an accurate and efficient molecular identification method for specific DNA Signature Sequence Tags (DSS) of Tianshanxuelian (Saussurea involucrate) and its adulterants Shuimuxuetuzi (S. medusa) and Miantouxuetuzi (S. laniceps). Methods The Chloroplast Genome Information Resource (CGIR) were used to obtain the complete chloroplast genome sequences of Saussurea DC, the IdenDSS software was used to screen for specific DNA signature sequence tags (DSS) of the S. involucrata. Five sets of DSS markers were selected, and the corresponding specific primer pairs were designed using IdenDSS. Through DNA extraction from the sample, PCR amplification, Sanger sequencing and sequence alignment verification, the presence of DSS or its reverse complementary sequence of the target species S. involucrata in the test sample was detected for original identification. Results By comparing the selected five pairs of DSS sequences with the authentic source and its adulterants sequences of S. involucrata, all five sets of DNA Signature Sequence Tags could be used for identification of S. involucrata were finally determined. The above sequences were uploaded to the standard database of the Traditional Chinese Medicine Molecular Identification Platform (www.herbsdna.com), and combined with the DSS identification function of the database, accurate and efficient identification of S. involucrata origin can be achieved. Conclusion The five selected DNA Signature Sequence Tags had strong specificity and high accuracy, which can effectively solve the problem of original identification of S. involucrata.

Objective To establish a fingerprint and multi-indicator component content determination method for Juemingzi (Cassiae Semen), and combine it with chemometrics to evaluate the quality of Cassiae Semen from different origins. Methods Waters HSS T3 C₁₈ (150 mm × 3.0 mm, 1.8 μm) was used as the chromatographic column, acetonitrile-methanol (3∶1) was used as mobile phase A, 0.1% phosphoric acid solution was used as mobile phase B, and gradient elution was performed. The flow rate was 0.40 mL/min, the column temperature was 40 ℃, the detection wavelength was 285 nm, and the injection volume was 2 μL. Traditional Chinese medicine fingerprint similarity evaluation software was used for similarity evaluation, chemometrics was applied to analyze Cassiae Semen from different origins, and the content of 11 indicator components in Cassiae Semen was determined. Results The established fingerprint identified a total of 26 common peaks, and 13 components were identified through reference standards. Through cluster analysis and principal component analysis (PCA), different Cassiae Semen samples can be clustered into 4 categories, and through the application of orthogonal partial least squares-discriminant analysis (OPLS-DA) method with VIP > 1 as the criterion, peaks 3 (emodin-1-O-β-D-glucoside), 6 (cassiaside), 7 (rubrofusarin-6-O-β-gentiobioside), 8 (aurantio-obtusin-6-O-D-glucoside), 9 (cassiaside C), 10, 17 (aurantio-obtusin), 21, and 23 (rubrofusarin) were selected as differential markers for distinguishing Cassiae Semen from different origins. The content ranges of isorhamnetin-3-O-β-D-glucopyranoside, cassiaflavone B2, cassiaflavone, isorhamnetin-6-O-β-D-glucopyranoside, chrysin-6-O-glucoside, cassiaflavone C, cassiaflavone B, chrysin, chrysin-3-O-β-D-glucopyranoside, isorhamnetin, and chrysophanol in 18 batches of Cassiae Semen were 0.425-1.315, 4.129-8.790, 0.453-1.849, 2.617-4.739, 0.792-2.372, 1.424-3.533, 0.496-1.439, 0.630-1.359, 0.483-2.298, 2.130-9.516, and 2.006-3.841 mg/g, respectively. According to the methodology, each component exhibited a good linear relationship. Conclusion The established fingerprint of Cassiae Semen, combined with the chemical pattern recognition, can effectively distinguish Cassiae Semen from different origins. The multi-component content determination method is stable and reliable, providing a reference for the quality evaluation and control of Cassiae Semen.

Spectrum-effect relationship research in traditional Chinese medicine (TCM) serves as a crucial technical bridge between traditional therapeutic efficacy and modern scientific research. By elucidating the intrinsic correlations between the chemical spectra of TCM and their pharmacological activities, this approach enables accurate identification of pharmacodynamically active substances and elucidation of their mechanisms of action. It thereby provides an innovative foundation for scientific quality control of TCM. However, inherent characteristics of TCM and current research limitations present significant challenges in this field. Centered on framework of “spectrum-efficacy, spectrum-effect relationships and achievement transformation”, this review systematically summarizes the key challenges facing spectrum-effect research in TCM, analyzes their underlying causes, and offers recommendations for standardizing future studies and industrializing research outcomes. This work aims to advance spectrum-effect research toward greater scientific rigor, systematicity, and practical value.

Plant-associated microorganisms, as important components of the plant holobiont, affect the health and survival of plants and thus have attracted much attention. The growth and development of medicinal plant Salvia miltiorrhiza, the accumulation of active ingredients, and the process of disease control are closely related to the interaction with root microorganisms. Previous studies have shown that rhizosphere and endophytic microorganisms can promote the synthesis of phenolic acids and tanshinones, secondary metabolites, by regulating plant hormone levels, signal pathways, and the activity of key metabolic enzymes, and enhance the plant stress resistance and medicinal material quality. Arbuscular mycorrhizal fungi, plant growth-promoting bacteria, and antagonistic fungi play significant roles in alleviating continuous cropping obstacles, inhibiting soil-borne pathogens, and maintaining rhizosphere ecological balance. Based on a systematic review of relevant research literature, this article focuses on summarizing the composition types, functional roles, and interaction mechanisms between S. miltiorrhiza and root microorganisms, and discusses their application potential of these microbes in S. miltiorrhiza cultivation. Finally, this paper discusses the existing problems in the current research and the future development directions, aiming to provide references for the microecological regulation of S. miltiorrhiza cultivation and the development and utilization of functional microorganisms of S. miltiorrhiza.

This paper reviews the traditional application basis and modern research progress of Kushen (Sophorae Flavescentis Radix), and performs a comprehensive analysis focusing on its chemical constituents, shared common pharmacological mechanisms, and safety profiles, thereby providing a scientific reference for its research and standardized application. Using keywords such as Sophorae Flavescentis Radix, alkaloids, total flavonoids, immune-inflammatory regulation, and safety evaluation, this review collected relevant domestic and foreign relevant literature published in the past decade was retrieved from databases including Wanfang, China National Knowledge Infrastructure, PubMed, and Web of Science. The retrieved studies were systematically analyzed to summarize the traditional medical understanding and modern pharmacological research, with special emphasis on the pharmacodynamic material basis and clinical application mechanisms of Sophorae Flavescentis Radix. Sophorae Flavescentis Radix contains multiple classes of chemical components, predominantly alkaloids, while flavonoids, polysaccharides, and other constituents also contribute to its holistic efficacy. Modern pharmacological studies have revealed that Sophorae Flavescentis Radix exerts its overall effects through immune-inflammatory regulation as the core mechanism, specifically by targeting desmoglein-2 for barrier repair and bidirectionally modulating immune polarization (M1/M2). Its material basis exhibits clear structure-synergy patterns, non-alkaloid components can regulate the toxicity threshold of alkaloids by inhibiting the reactive oxygen species/NOD-like receptor family pyrin domain containing 3 pathway, demonstrating a significant “efficacy-toxicity” balance feature. Meanwhile, adverse reactions related to Sophorae Flavescentis Radix are closely associated with its alkaloid components, dosage, and administration routes, suggesting that the safety risks are manageable to a certain extent. Modern research on Sophorae Flavescentis Radix has, to some extent, validated its traditional efficacy basis. However, critical issues remain, such as unclear relationships among components, insufficient integration of mechanisms, and limited systematic safety evaluations. Future research should strengthen studies on multi-component synergistic mechanisms and safety risk control to promote the rational development and standardized clinical application of Sophorae Flavescentis Radix.

Honey-roasted Huangqi (Astragali Radix) is a widely used processed product of Astragali Radix in clinical practice. After honey-roasting, the effect of tonifying qi and invigorating spleen is significantly enhanced. Its processing technology and quality evaluation system are very important to ensure clinical efficacy and promote modernization process of traditional Chinese medicine processing. Therefore, this paper systematically summarized the historical evolution of processing of honey-roasting Astragali Radix, as well as the differences between national and local processing standards, by consulting the ancient herbal monographs, processing standards at all levels and modern research results at home and abroad. The effects of pretreatment, honey selection and refining, honey content, honey-water ratio and other key process links on quality of finished products and the status of process optimization were described. The changes of polysaccharides, saponins and flavonoids during honey roasting and the potential mechanism of processing synergistic efficiency were discussed. The development of multidimensional quality evaluation indices and analysis technologies is summarized. On this basis, the key problems such as inconsistent process standards, unclear processing mechanisms and imperfect quality evaluation system were analyzed, and the corresponding countermeasures were put forward, aiming to provide a scientific evidence for process standardization and quality evaluation of honey-roasting Astragali Radix.

With the booming development of artificial intelligence (AI), machine learning, deep learning and transfer learning have gained growing applications in healthcare, bringing new perspectives and strategies to antidepressant research. Existing antidepressants present slow onset, low efficacy and marked inter-individual variability, with their underlying mechanisms yet to be fully clarified. Traditional research is hampered by small sample sizes, substantial data heterogeneity and poor cross-modal integration capacity. As a pivotal AI technology, transfer learning realizes cross-domain, cross-task and cross-modal knowledge transfer, and mitigates issues such as small-sample modeling, data distribution discrepancy and the curse of dimensionality. This review systematically elaborates the current status and future trends of transfer learning in antidepressant efficacy evaluation and the exploration of their action mechanisms. We analyze the characteristics and performance of transfer learning models construction strategies and application paradigms, and discuss the challenges, technical limitations and clinical translation barriers in practical application. This study provides novel ideas for exploring depression pathogenesis and optimizing clinical antidepressant use, and further promotes the innovative and standardized application of AI in clinical medicine.

Genuine medicinal materials (Dao-di medicines) are renowned for their superior quality and proven efficacy, and serve as a core marker for assessing and ensuring the quality of traditional Chinese medicine. However, due to the combined effects of the widespread adoption of modern cultivation techniques and market demand, the boundaries of traditional Dao-di producing regions are becoming increasingly blurred. Consequently, the accurate identification of Dao-di medicines is becoming ever more important. Significant progress has been made in the identification research of Dao-di medicines. Building upon traditional empirical identification methods, research in this field is now exhibiting a trend towards diversification. Through strategies such as assessment of characteristic morphological features, analysis of key constituents, molecular marker identification, multi-indicator combined analysis, and digital and intelligent recognition, Dao-di medicines have been comprehensively identified from various dimensions. In this article, we systematically reviewed various identification strategies and research progress regarding Dao-di medicines, including macroscopic morphological identification, microscopic identification, physicochemical identification, molecular identification, and digital and intelligent identification, thereby providing foundational reference materials for research into the identification of Dao-di medicines.

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline. Its pathological mechanisms are complex, involving multiple key links such as beta-amyloid protein (Aβ) deposition, excessive phosphorylation of Tau protein, neuroinflammation and synaptic dysfunction. Traditional Chinese medicine (TCM) categorizes AD under conditions like “dementia” and “forgetfulness”, with the core pathogenesis being “deficiency at the root and excess at the surface”, where kidney essence deficiency is the root cause and phlegm turbidity and blood stasis obscuring the orifices are the surface manifestations. As a distinctive TCM therapy, aromatic orifice-opening drugs, with their pungent, fragrant and penetrating nature that can reach and regulate meridians, follow the principle of “awakening the orifices and refreshing the mind, resolving phlegm and dispelling foulness”, demonstrating unique advantages in the prevention and treatment of AD. Representative drugs include Shichɑngpu (Acori Tatarinowii Rhizoma), Yuanzhi (Polygalae Radix), Shexiang (Moschus), Bingpiɑn (Borneolum Syntheticum), etc. Modern pharmacological research has revealed their multi-target intervention potential, including inhibiting Aβ generation and aggregation, reducing Tau protein phosphorylation, anti-neuroinflammation, anti-oxidative stress, and regulating neurotransmitters and promoting neurogenesis. Commonly used TCM formulas with the effect of aromatic orifice-opening such as Angong Niuhuang Pill (安宫牛黄丸), Suhexiang Pill (苏合香丸) and Changpu Yujin Decoction (菖蒲郁金汤) have shown good efficacy in improving patients’ cognitive function, memory, and daily living abilities through compatibility and synergy. This review systematically summarizes the research and application status of aromatic orifice-opening drugs in the prevention and treatment of AD, providing theoretical basis and practical references for the intervention of AD by aromatic orifice-opening TCMs.

Berberine (BBR) is an isoquinoline alkaloid extracted from traditional medicinal plants such as Coptis chinensis and Phellodendron amurense, with a long history of clinical application. Recent studies have demonstrated that BBR exhibits multifaceted pharmacological activities, including blood glucose lowering, lipid regulation, anti-tumor effects, cardiovascular protection, anti-inflammation, and neuroprotection. However, existing research findings are relatively scattered and lack systematic integration. This article aims to systematically review the main pharmacological effects of BBR and its related molecular mechanisms, intending to elucidate the common regulatory network underlying its multifaceted effects and provide important theoretical support for its clinical translation and the development of innovative drugs. The results indicated that the multifaceted pharmacological effects of BBR are mainly achieved through the coordination of three key pathways: inhibiting the nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways to block inflammatory cascade reactions; activating the nuclear factor erythroid 2-related factor 2/antioxidant response element (Nrf2/ARE) pathway to enhance antioxidant defenses; and suppressing the assembly of the NOD-like receptor family, pyrin domain containing protein 3 (NLRP3) inflammasome to reduce cell pyroptosis. This study not only provides a clear molecular map for an in-depth understanding of the systemic pharmacological mechanisms of BBR, but also lays a scientific foundation for the development of drug targets and multi-pathway synergistic treatment strategies for diseases related to inflammation, oxidative stress, and programmed cell death, possessing significant theoretical value and translational prospects.

The “Special Provisions for the Supervision and Administration of Traditional Chinese Medicine Production” is China’s first regulatory document specifically designed to systematically regulate the oversight of traditional Chinese medicine (TCM) production. It signifies a deepening shift from general requirements to a specialized system that reflects the unique characteristics of TCM manufacturing. This development helps to enhance the standardization of TCM production and the stability of its quality, modernize regulatory approaches, and provide institutional support for building a drug regulation system tailored to TCM’s features, thereby promoting the high-quality development of the TCM industry. Grounded in the inherent characteristics of TCM-such as the natural origin of its raw materials, long quality formation chains, and complex influencing factors-the provisions establish a regulatory framework covering the entire TCM production lifecycle. This framework addresses key areas including source control of Chinese medicinal materials, production process management, testing and release, traceability systems, pharmacovigilance, and regulatory oversight. The institutional design emphasizes adherence to TCM principles, strengthens the subject responsibility of the marketing authorization holder, highlights source control, rigorous process management, and risk-based classification supervision. Furthermore, it provides detailed requirements for critical issues such as raw material homogenization, extract yield management, contract manufacturing, production resumption, and the oversight of TCM injections. By analyzing the background of its issuance, institutional logic, main content, key innovations, and implementation pathways, this paper aims to facilitate a deeper understanding of the significance of implementing these Special Provisions among the industry.