Traditional Chinese medicine（TCM） and ethnic medicine, as integral components of traditional medicine, exhibit diverse therapeutic effects, complex chemical compositions, intricate physiological processes, and multifaceted mechanisms of action. This makes elucidating the pharmacological basis and mechanisms of action of TCM and ethnic medicine a key research direction in the modernization of TCM. However, challenges such as incomplete pharmacological evaluation models, low correlation with clinical efficacy, and low screening efficiency and throughput remain core obstacles. Artificial intelligence-driven organoid and organ-on-achip technologies offer an innovative research paradigm for the screening of bioactive compounds and the discovery of target mechanisms in TCM and ethnic medicines. By constructing highly biomimetic human organoids/organ-on-a-chip models that precisely simulate the microenvironment and pathological states of human organs, combined with artificial intelligence technologies for in-depth analysis of high-throughput pharmacodynamic data, it is possible to elucidate the synergistic effects of multi-component, multi-target interactions. Systematically screening groups of active components in TCM, quantifying their metabolic kinetic processes, and revealing potential target points and molecular pathways significantly enhances the precision and efficiency of bioactive substance identification. The deep integration strategy of organoids/organ-on-a-chip technology with artificial intelligence provides new technical support for accelerating the modernization of TCM and ethnic medicine research and promoting innovative drug development.

Objective To study the chemical constituents, antioxidant activity and anti-inflammatory activity of the n-butanol fraction from Primulina eburnea. Methods Compounds were separated and purified using chromatographic techniques such as normal-phase and reversed-phase silica gel column chromatography, Sephadex LH-20 gel column chromatography and RP-HPLC. Meanwhile, their structures were identified by spectroscopic techniques including NMR, UV and IR. Subsequently, the antioxidant activities of obtained compounds were evaluated by the DPPH and ABTS assays, while the Griess assay was employed to determine the effect of these compounds on NO production in lipopolysaccharide (LPS)-induced RAW264.7 cells to assess their anti-inflammatory activity. Results A total of 20 compounds were isolated from the n-butanol fraction of P. eburnea and were respectively identified as spathodic acid-28-O-β-D-gentiobioside (1), calceolarioside B (2), 2-(3,4-dihydroxyphenyl)ethanol glucoside (3), salidroside (4), tyrosol (5), 6-hydroxyapigenin-7-O-β-[2-O-β-xyloglucoside] (6), icariol A₂(7), (+)-isolariciresinol (8), 5-methoxy-(+)-isolariciresinol (9), (+)-syringaresinol-4'-O-β-D-glucopyranoside (10), dihydrophaseic acid (11), (6S,9R)-roseoside (12), esculetin (13), p-hydroxybenzoic acid (14), protocatechuic acid (15), vanillic acid (16), syringic acid (17), caffeic acid (18), caffeic acid methyl ester (19) and methyl-3-O-β-D-glucopyranosyl-4-hydroxy caffeic acid (20). Meanwhile, compounds 7, 17 and 18 exhibited significant antioxidant activity. Compounds 3, 5—9 exhibited significant inhibitory effects on the production of NO in LPS-induced RAW264.7 cells. Conclusion Compound 1 is a new triterpenoid glycoside, named eburneoside A. While compounds 4, 6, 9—12, 14, 15 and 20 were isolated for the first time from the genus Primulina. Additionally, compounds 17 and 18 were firstly isolated from P. eburnea. Furthermore, some of these compounds exhibited significant antioxidant and anti-inflammatory activities.

Objective To investigate the chemical constituents and their myocardial cell protective activity from the fruits of Daemonorops draco. Methods Compounds were isolated and purified using a combination of silica gel, RP-C₁₈, Sephadex LH-20 and semi-preparative HPLC. Their structures were elucidated through comprehensive spectroscopic analysis, UV, IR, HRESIMS and NMR. The protective effects of all isolates on the H9c2 myocardial cells injury induced by oxygen-glucose deprivation (OGD) were evaluated. Results Three pairs of new flavans were purified and elucidated as (2S)-2-ethoxy-7-hydroxy-5-methoxy-6-methylflavan (1a), (2R)-2-ethoxy-7-hydroxy-5-methoxy-6-methylflavan (1b), (2S,3S)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (2a), (2R,3R)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (2b), (2R,3S)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (3a) and (2S,3R)-2-ethoxy-3,7-dihydroxy-5-methoxy-6-methylflavan (3b). Compounds 1b and 2b demonstrated protective effects against OGD-induced H9c2 cell injury, which increased cell viability by (6.36 ±1.26)% and (10.78 ±0.76)% at a concentration of 20 μmol/L, respectively. Conclusion Compounds 1a/1b, 2a/2b, and 3a/3b are three pairs of enantiomers, and they are all new compounds, named as dracoropin I₁, dracoropin I₂, dracoropin J₁, dracoropin J₂, dracoropin K₁, dracoropin K₂. Among them, compounds 1b and 2b exhibited protective effects on myocardial cells.

Objective To research the chemical constituents from the fruits of Lycium barbarum. Methods Separation and purification were performed using silica gel, D-101 macroporous resin, MCI, Sephadex LH-20, and semi-preparative HPLC. The structures of the compounds were identified based on spectroscopic data and physicochemical properties. The α-glucosidase inhibitory activity of the selected compounds was evaluated using 4-nitrophenol-α-D-glucopyranoside (PNPG) assay. Results A total of 15 compounds were isolated from the fruits of L. barbarum, which were identified as methyl (2R)-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]-4-methylpentanoate (1), 5-epi-acortatarin A (2), 5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde (3), 4-[2-formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]butanoic acid (4), (2R)-[2-formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]-3-(4-hydroxyphenyl) propanoate (5), 4-[formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]butanoate (6), 3-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl] pentanedioic acid (7), N-trans-feruloyl-3'-O-methyldopamine (8), N-trans-feruloyl-3',4'-dihydroxyphenylethylamine (9), N-cis-p-coumaroyl tyramine (10), N-cis-feruloyl tyramine (11), bungeanoline E (12), N-malonyl-tryptophan (13), 3-hydroxy-4-ethyl ketone pyridine (14), cartorimine (15). Conclusion Compound 1 is a new pyrrole alkaloid, named lycipyrrole A. Compound 2 is a new natural product. Compounds 4, 7 and 12—15 were isolated from L. barbarum for the first time, while compounds 12, 13 and 15 were isolated from this genus for the first time. Notably, compound 9 exhibited significant inhibitory activity against α-glucosidase.

Objective To study the non-saponin chemical constituents of the flower buds of Panax notoginseng and evaluate their antioxidant and anti-inflammatory activities. Methods The extraction was performed by refluxing with 65% aqueous ethanol, followed by successive liquid-liquid extraction with petroleum ether, ethyl acetate (EtOAc) and n-butanol. The compounds were isolated and purified using RP-18, silica gel, MCI gel CHP20P, Sephadex LH-20 column chromatography and semi-preparative liquid chromatography. Structural identification was carried out by comprehensive analysis of nuclear magnetic resonance (NMR) spectroscopy, high-resolution mass spectrometry (HR-MS), ultraviolet (UV) spectroscopy, and computational electronic circular dichroism (ECD) spectroscopy. Some purified compounds were evaluated for preliminary antioxidant activity and anti-inflammatory activity. Results A total of seven non-saponin components were isolated from the EtOAc fraction of P. notoginseng flower buds, including one isocoumarin (1), four simple phenols (2—5), and two loliolides (6, 7), were obtained from the EtOAc fraction. These were identified as isocoumarin F1 (1), ferulic acid (2), evofolin A (3), p-hydroxybenzoic acid (4), 4-hydroxybenzaldehyde (5), (＋)-epiloliolide (6), and (-)-loliolide (7), respectively. Conclusion All compounds (1—7) were isolated from P. notoginseng for the first time. Of which, compound 1 is a new isocoumarin and named isocoumarin F1, while compound 2 exhibited obvious DPPH radical scavenging ability.

Objective To study the chemical constituents fromdried whole plant of alpine herb Gentiana nanobella, and to conduct preliminary screening for antimalarial activity of the isolated constituents. Methods Ethyl acetate and n-butanol fractions of 75% ethanol extract of the whole plant of G. nanobella were isolated and purified by repeated column chromatography padded with silica gel, RP-18 and Sephadex LH-20, respectively. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. The in vitro inhibitory activity of the testing compound against human Plasmodium falciparum 3D7 strain was achieved with the SYBR Green I method. Results A total of 11 phenol compounds were obtained and identified as 2,3-dihydroxy benzoic acid methyl ester-3-O-β-L-rhamnosyl-(1''→2')-b-D-glucopyranoside (1), 2,3-dihydroxy benzoic acid-3-O-β-L-rhamnosyl-(1''→2')-b-D-glucopyranoside (2), 2,3-dihydroxy benzoic acid methyl ester-3-O-β-D-glucopyranosyl-(1''→6')-β-D-glucopyranoside (3), benzyl β-D-xylopyranosyl-(1''→6')-β-D-glucopyranoside (4), apigenin 5-O-β-D-glucopyranoside (5), salcolin B (6), apigenin-7-O-β-D-glucopyranoside (7), benzoic acid (8), p-hydroxybenzoic acid (9), 3-hydroxy-2-methoxybenzoic acid (10), and isovanillic acid (11). Compound 6 exhibited an in vitro antimalarial activity against Plasmodium falciparum 3D7 strain with IC₅₀ 18.50 μmol/L and inhibition rate of 98.48% at a concentration of 50 μmol/L. Conclusion Compounds 1 and 2 are new compounds, named gentinanoside A and gentinanoside B. Compounds 1—11 were isolated from G. nanobella for the first time. G. nanobella is abundant in phenolic compounds, and some compounds had certain antimalarial activity.

Objective To meet the requirements for high-quality development of food-medicine homology products, taking Xiaoer Xiaoshi Granules (XXG, 小儿消食颗粒) as an example, this study established an intelligent-driven granulation process optimization and quality consistency evaluation method for food-medicine homology granules based on the quality by design (QbD) concept. Methods Firstly, the analytic hierarchy process (AHP) was used to assigned weights to critical quality attributes (CQAs) of XXG, with subsequent risk assessment identifying granulation critical influencing factors. Secondly, following single-factor screening of critical influencing factors, the optimal combination of granulation parameters was selected by comparing the Box-Behnken design-response surface methodology (BBD-RSM) and the genetic algorithm-backpropagation neural network (GA-BPNN). Finally, leveraging the medicine-food homology properties of XXG, a quality consistency assessment was performed targeting both chemical attributes (total flavonoid content) and taste quality attribute based on electronic tongue. Results The excipient properties, ethanol volume fraction of wetting agent, ethanol amount, and excipient-to-drug ratio were determined as critical influencing factors, the optimal granulation parameters for XXG were determined as follows: an excipient system of maltodextrin to mannitol at a 2∶1 ratio, an ethanol concentration of 84%, an ethanol amount of 0.18 mL/g, and an excipient-to-drug ratio of 1.5∶1, yielding a comprehensive score of 103.88. Regarding bioactive constituents, the RSD of total flavonoid content across three batches was as low as 1.95%. For instrumental sensory evaluation, the e-tongue response values demonstrated batch-to-batch consistency with RSDs below 3% across all seven sensors. Conclusion The optimized granulation parameter set demonstrated stability and feasibility, and this process optimization approach can offer valuable insights for developing medicine-food homology products and advancing intelligent manufacturing in traditional Chinese medicine.

Objective To establish HPLC fingerprint profiles of raw Dɑnggui (Angelicae Sinensis Radix, ASR) and ASR processed with wine, and to study the changes in chemical components before and after wine processing by combining multivariate statistical analysis and quantitative determination, providing a reference for the quality evaluation of ASR. Methods HPLC was used to establish the fingerprint profiles of raw ASR and ASR processed with wine, and the contents of 5-hydroxymethylfurfural (5-HMF), chlorogenic acid, vanillin, ferulic acid, and ligustilide were determined. The similarity of the profiles was calculated using the Chinese herbal medicine chromatographic fingerprint similarity evaluation system. Common peaks were calibrated, identified, and assigned.. The peak areas of the common peaks before and after wine processing were used as indicators. Hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were employed to evaluate the intrinsic quality differences between raw and wine-processed ASR and identify the main differential components. Results HPLC fingerprint profiles of ten batches of raw ASR and ASR processed with wine were established. A total of 11 common peaks were identified in raw ASR and 12 common peaks were calibrated in ASR processed with wine. The similarity of the profiles was greater than 0.960. Chemical pattern recognition indicated that peaks 12 (ligustilide), 7, 11, and 6 (ferulic acid) might be the markers of the quality differences before and after wine processing. The content determination results showed that 5-HMF was newly generated after wine processing, while the contents of ferulic acid, chlorogenic acid, and vanillin slightly decreased, and the content of ligustilide increased. Conclusion The established fingerprint profiles and multi-component quantitative determination methods for ASR before and after wine processing are stable and reliable, providing a reference for the quality control, comprehensive utilization, and clinical application of ASR and ASR processed with wine.

Objective To explore the optimal processing method for Dusuanlan (Pleione bulbocodioides) pseudobulbs, this study investigated the effects of scalding method and duration, and drying technique on their morphologies, microstructures, and chemical composition contents. Methods Pseudobulbs of P. bulbocodioides were processed using eight methods: oven direct drying (OD), boiling for 10, 20, 30 min and then drying (B10D, B20D, B30D), steaming for 10, 20, 30 min and then drying (S10D, S20D, S30D), and freeze drying (FD). The contents of 2-O-glucosylbulleyanolide (dactylorhin A), militarine, and batatasin III were quantified by HPLC. By constructing HPLC fingerprints and performing similarity analysis (SA), hierarchical cluster analysis (HCA), and principal component analysis (PCA), the impact of processing methods on the quality of pseudobulbs was evaluated. Structural changes were assessed through examination of macroscopic features, tissue organization, and powdered microscopic characteristics. Using dactylorhin A content, militarine content, batatasin III content, alcohol-soluble extractives, water-soluble extractives, drying rate, moisture content, total ash content, and acid-insoluble ash content as indicators, the effects of eight processing methods on the quality of pseudobulbs were comprehensively evaluated by entropy weight method, analytic hierarchy process (AHP), and technique for order preference by similarity to ideal solution (TOPSIS). Results The values for HPLC fingerprint similarity of the samples obtained from eight methods ranged from 0.681 to 0.994. The freeze-dried (FD) samples exhibited the lowest similarity and were segregated as a distinct cluster in both hierarchical cluster analysis (HCA) and principal component analysis (PCA), while the samples from other seven methods formed a separate cluster. Three common peaks were identified among the 24 common peaks in the HPLC fingerprints: dactylorhin A (peak 7), militarine (peak 9), and batatasin III (peak 15). According to TOPSIS-based comprehensive evaluation, samples steamed for 10 min followed by oven drying (S10D) showed both the highest relative closeness coefficient and the greatest total content of dactylorhin A, militarine, and batatasin III. Conclusion Based on comprehensive multi-index evaluation, steaming for 10 min followed by oven drying (S10D) was identified as an optimal primary processing method for P. bulbocodioides pseudobulbs. This method enhanced the retention of active compounds while being technically straightforward. The study establishes a critical foundation for standardizing processing of P. bulbocodioides pseudobulbs and refining quality standards of Shancigu (Cremastrae Pseudobulbus Pleiones Pseudobulbus) in pharmacopeial monographs.

Objective Based on the mathematical model of mass transfer, this study aimed to investigate the existence state and transfer principle of chlorogenic acid in the production process of Reduning Injection (RI, 热毒宁注射液) were investigated. Methods Using chlorogenic acid as the detection index, a nanofiltration mass transfer model was established., and choosing the monomer molecular state as the reference, calculated the existence state of chlorogenic acid in the intermediate solution of RI, analyzed the correlation between the change of chlorogenic acid transfer rate and the existence state of chlorogenic acid in the process of production of RI, and explored the intrinsic causes of the difference in the transfer of phenolic acids during the RI concentration process. Results The regression coefficients between the mass transfer coefficients of chlorogenic acid and the power value of solute concentration in the extract solution and the RI intermediate solution were all greater than 0.90, and the nanofiltration mass transfer model was established. chlorogenic acid showed a positive correlation between the ratio of molecular states and the concentration transfer rate in aqueous and ethanol solutions of RI intermediates, and the same trend was observed in solutions of Jinyinhua (Lonicerae Japonicae Flos) and Qinghao (Artemisiae Annuae Herba) intermediates, suggesting that the state of existence of the constituents is the intrinsic cause of the differences in phenolic acid transfer during RI concentration process. In addition, chlorogenic acid was more stable in the molecular state than in the ionic state in the same solution system, and more stable in the ethanol solution than in the aqueous solution in different solvent systems. Conclusion A quantitative calculation method for the molecular state of chlorogenic acid in the intermediate of RI has been constructed, and the positive correlation between its existence state and mass transfer has been initially elucidated, which can provide support for the standardized control of the production process of the preparation.

Objective To prepare sinomenine nanoemulsion (Sin-NE) co-stabilized by Bletilla striata polysaccharide and glycyrrhizic acid, systematically characterize its properties, investigate its stability and drug release behavior, and further evaluate its anti-inflammatory effects through in vitro experiments. Methods The Sin-NE was prepared using the ultrasonic emulsification method. Its morphology, particle size, polydispersity index (PDI), and ζ potential were observed and determined. Optimal formulation parameters were screened via single-factor and orthogonal experiments, followed by formulation validation. pH value, turbidity, viscosity, encapsulation efficiency, and drug loading capacity were measured. Stability was investigated under centrifugation, storage, dilution, and heating conditions was evaluated, and in vitro drug release behavior was investigated. A RAW264.7 cell inflammation model was established to validate the effects of the Sin-NE on nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Results The optimized formulation comprised 3% Coicis Semen oil, 0.5% combined emulsifiers, a 1∶9 mass ratio of Bletilla striata polysaccharide to glycyrrhizic acid, 10 min of ultrasonic treatment, and 325 W ultrasonic power. The resulting Sin-NE exhibited an average particle size of (145.30 ±2.35) nm, ζ potential of (-39.51 ±0.43) mV, PDI of 0.095 ±0.022, and pH value of 3.990 ±0.033. Encapsulation efficiency and drug loading capacity reached 88.78% and 26.14 mg/g, respectively. The nanoemulsion demonstrated excellent stability under centrifugation, storage, dilution, and heating conditions and displayed sustained-release characteristics. In vitro, it significantly inhibited NO release and effectively reduced IL-6 and TNF-α levels in inflamed cells. Conclusion The Sin-NE significantly enhances sinomenine’s bioavailability and anti-inflammatory activity, providing experimental evidence for developing novel traditional Chinese medicine nanoemulsion formulations.

Objective To prepare liposomes co-loaded with shikonin and ligustilide (Lip@Shi/Lig), optimize its preparation process, and to investigate its characterization and transdermal performance in vitro. Methods The CCK-8 assay and Synergy Finder analysis tool were used to determine the optimal combination ratio of shikonin and ligustilide based on cell viability. Lip@Shi/Lig was prepared using the thin-film dispersion method, and the optimal formulation of Lip@Shi/Lig was screened through single-factor investigation using encapsulation efficiency as the evaluation criterion. The quality evaluation of Lip@Shi/Lig was conducted using characterization methods, including appearance and morphology observation, particle size, polydispersity index (PDI), ζ potential determination, X-ray diffraction (XRD) analysis, and Fourier transform infrared spectroscopy (FT-IR) analysis. Finally, the transdermal permeability and dermal retention performance of Lip@Shi/Lig were investigated using the Franz diffusion cell method. Results The optimal combined ratio of shikonin and ligustilide was 1∶1. The optimal conditions were determined as follows: egg yolk lecithin concentration of 10 mg/mL, phospholipid-cholesterol ratio of 4∶1, total drug-phospholipid ratio of 1∶15, and ultrasonication time of 5 min. The resulting Lip@Shi/Lig exhibited regular, spherical vesicle morphology, with encapsulation efficiencies of shikonin and ligustilide at (98.16 ±0.67)% and (97.20 ±0.76)%, respectively, particle size of (88.62 ±0.26) nm, PDI of 0.246 ±0.013, and ζ potential of (-36.57 ±1.65) mV. XRD and FT-IR results indicated that shikonin and ligustilide were successfully encapsulated in the liposomes. The cumulative penetration of shikonin and ligustilide in Lip@Shi/Lig within 30 h were (82.97 ±0.72) μg/cm² and (81.57 ±3.59) μg/cm², respectively, with dermal retention rates of (6.12 ±0.18) μg/cm² and (8.08 ±0.04) μg/cm², respectively, both of which were significantly higher than those of the single drug and the single-drug-loaded liposomes. Conclusion Lip@Shi/Lig was successfully prepared with favorable transdermal properties and stability. It significantly enhanced the transdermal penetration and dermal retention of both shikonin and ligustilide, providing a solid experimental foundation for further in vivo studies and potential clinical applications.

Objective To explore the effect and mechanism of Yinyanghuo (Epimedii Folium) and Nüzhenzi (Ligustri Lucidi Fructus) on Buguzhi (Psoraleae Fructus)-induced liver injury in a rat model of kidney-yin deficiency based on the theory of “heterogeneous medicinals mutual restriction”. Methods The liver injury rats with kidney-yin deficiency model induced by Psoraleae Fructus was taken as a reference, Epimedii Folium and Ligustri Lucidi Fructus were respectively given for intervention, the changes in indicators of kidney-yin deficiency were evaluated by through body weight, anal temperature, fecal water content and cyclic adenosine monophosphate (cAMP)/cyclic guanosine monophosphate (cGMP) ratio in serum. The changes in liver function were evaluated by measuring the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, as well as by staining liver tissue with hematoxylin eosin (HE). Metabolomics, association analysis, ROC analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were combined to elucidate potential mechanisms. Results Epimedii Folium significantly decreased body weight and fecal water content in model rats (P < 0.05, 0.01), significantly increased anal temperature, cAMP/cGMP ratio and activities of ALT, AST (P < 0.05, 0.001), aggravated the infiltration of inflammatory cells in liver and exacerbated liver damage. Ligustri Lucidi Fructus significantly reversed the above changes. Metabolomics identified 17 common metabolites that were differentially regulated between Epimedii Folium and Ligustri Lucidi Fructus. Association analysis and ROC analysis further revealed six differential metabolites with good diagnostic efficacy, including ceramide and taurocholic acid, mainly involving eight pathways such as linoleic acid metabolism and sphingolipid metabolism. In addition, Epimedii Folium could significantly increase the levels of inflammatory factors such as interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) in serum of model rats (P < 0.05), and decrease anti-inflammatory factor IL-10 level. Ligustri Lucidi Fructus reversed the above changes. Conclusion Epimedii Folium and Ligustri Lucidi Fructus may affect the levels of inflammatory cytokines by differentially regulating liver linoleic acid metabolism, leading to differences in toxicity/efficacy in liver of rats with kidney-yin deficiency induced by Psoraleae Fructus. This study provides experimental basis for the theory of “heterogeneous medicinals mutual restriction” in traditional Chinese medicine, and provides reference for the research on reducing toxicity and increasing efficiency of Psoraleae Fructus.

Objective To investigate the effect of dandelion polysaccharide (DP) on lipid metabolism disorder in liver and gut microbiota structure in high-fat diet-induced obese mice. Methods A total of 40 SPF male C57BL/6J mice were randomly divided into control group, model group, orlistat (10 mg/kg) group, DP low-, high-dose (200, 400 mg/kg) groups, with eight mice in each group. Mice were fed with high-fat diet to establish an obesity model, and the model was simultaneously administered with ig for eight weeks. After the end of administration, mice were euthanized by removing their eyeballs and collecting blood. Serum was collected to measure the levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C); Hematoxylin-eosin (HE) staining and Oil red O staining were used to observe the pathological changes of liver and epididymal fat in mice; qRT-PCR was used to detect the expressions of lipid metabolism related genes [carnitine palmitoyltransferase 1 (CPT-1), cytochrome P450 family 7 subfamily A member 1 (CYP7A1), peroxisome proliferator-activated receptor α (PPARα), fatty acid synthase (FAS), peroxisome proliferator-activated receptor γ (PPARγ), stearoyl-CoA desaturase 1 (SCD-1), hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL)] in liver tissue; Feces were collected from each group of mice for microbiome sequencing. Results Compared with control group, the body weight, epididymal fat index, and levels of TC, TG, LDL-C in serum of mice in model group were significantly increased (P < 0.01, 0.001), HDL-C level were significantly decreased (P < 0.01). The structure of liver cells was damaged, the intercellular space was unclear, the arrangement of hepatic cords was significantly disordered, the area of vacuoles and the number of lipid droplets were significantly increased (P < 0.001), and the volume of epididymal fat cells was significantly increased with varying cell shapes and sizes. The expression levels of fat generation related genes FAS, PPARγ and SCD-1 mRNA were significantly increased (P < 0.01), while the expression levels of fat breakdown related genes HSL and ATGL mRNA were significantly decreased (P < 0.01). Compared with model group, DP low-, high-dose groups and orlistat group all reduced the body weight and epididymal fat index of obese mice to varying degrees, and improved the histopathological morphology of liver tissue and epididymal fat. The expression levels of fat synthesis related genes FAS, PPARγ and SCD-1 were significantly reduced in DP high dose group (P < 0.01), while the expression levels of fat breakdown and oxidation related genes HSL, ATGL, CTP-1, CYP7A1, PPARα mRNA were significantly increased (P < 0.01). Meanwhile, DP regulated the species composition of gut microbiota in obese mice to a certain extent, significantly increased the relative abundance of Blautia, s_unclassified_g_Lachnospiraceae_NK4A136_group, Lachnospiraceae_bacterium_28-4 and Helicobacter rodentium, decreased the relative abundance of Erysipelatoclostridium and Ruminococcus_torques_group, and their dominant strains were positively correlated with fat oxidation and decomposition genes HSL, ATGL, CTP-1, CYP7A1 and PPARα, and negatively correlated with fat synthesis genes FAS, PPARγ and SCD-1. Conclusion DP could significantly reduce the body weight of obese mice induced by a high-fat diet. The mechanism may be related to regulating beneficial bacteria to promote fat oxidation and decomposition and inhibit fat synthesis.

Objective To explore the mechanism by which Astragalus polysaccharides (APS) improves the consumption of adipose tissue in cancer cachexia (CC) using lipidomics and transcriptomics. Methods A random number table was used to divide 50 male mice into control group, model group, APS low-, medium-and high-dose (200, 400, 800 mg/kg) groups, with 10 mice in each group. CC model was constructed using abdominal sc CT-26 colorectal cancer cells. The treatment group was given 0.2 mL APS solution by ig daily, while the control group and model group were given an equal volume of physiological saline by ig for four consecutive weeks. The diameter of epididymal adipocytes was evaluated using hematoxylin-eosin (HE) staining; ELISA method was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), cyclic adenosine monophosphate (cAMP) in epididymal adipose tissue and level of free fatty acids (FFA) in plasma. After effectiveness and significance verification, transcriptome and lipidomics sequencing were performed on control group, model group and APS high-dose group to screen differentially expressed genes (DEGs) and differentially expressed lipids (DELs), and enrichment analysis was performed; Western blotting was used to verify the expressions of relevant proteins. Results High-dose APS significantly alleviated inflammatory infiltration in epididymal adipose tissue and reversed adipocyte atrophy (P < 0.01), significantly reduced the levels of TNF-α, IL-6, cAMP in adipose tissue and level of FFA in plasma (P < 0.01, 0.001), indicating that high-dose APS inhibits inflammation and lipolysis reactions in adipocytes. The lipidomics results showed that high-dose APS intervention reduced the expression levels of 66 DELs, including ceramides and their modifications, triacylglycerols, etc. Metabolic pathway enrichment analysis showed that these DELs mainly involved glycerophospholipid metabolism and sphingolipid metabolism pathways. The transcriptomic results showed that the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) pathway was repeatedly enriched in DEGs, indicating that this pathway may be the core pathway for APS to function. In addition, the expressions of 18 DEGs was significantly down-regulated by APS, and the gene interaction network diagram showed that these genes had complex interactions with Akt. Western blotting results confirmed that APS intervention reversed the expression levels of Tribbles homologous protein 3 (TRIB3), phosphorylated Akt (p-Akt), phosphodiesterase 3B (PDE3B), phosphorylated protein kinase A (p-PKA), phosphorylated hormone sensitive lipase (p-HSL) and adipose triglyceride lipase (ATGL) (P < 0.05, 0.01, 0.001). Conclusion APS may alleviate inflammation in adipose tissue, reduce the synthesis of sphingolipids such as ceramides, regulate TRIB3-Akt axis, restore insulin resistance to lipolysis, promote lipid signaling, and improve the consumption of CC adipose tissue.

Objective To investigate the effect of Dendrobium officinale polysaccharide (DOP) on prediabetes (PDM) and explore its mechanism in improving PDMs through the gut short-chain fatty acids (SCFAs)-G protein-coupled receptor 43 (GPR43)-glucagon-like peptide 1 (GLP1) pathway. Methods SD Rats were randomly divided into control group, model group and DOP low-, high-dose (0.1, 0.2 g/kg) groups, with 10 rats in each group. Except for the control group, the remaining rats were fed a high-sugar and high-fat diet for six weeks to establish a PDM model. During the modeling period, the corresponding drugs were administered intravenously, and the control group and model group were given distilled water of equal volume. Changes in general physiological indicators, glucose/lipid levels, glucose regulation function, intestinal/pancreatic histopathology, fecal SCFAs contents, expressions of GPR43 and GLP1 and its receptor in intestinal/pancreatic were analyzed. Results Compared with model group, DOP significantly increased tail microcirculatory blood flow in PDM rats (P < 0.01) and decreased anal temperature (P < 0.01), significantly reduced levels of total cholesterol and low-density lipoprotein cholesterol in serum (P < 0.05, 0.01), significantly decreased the oral glucose tolerance test results, fasting blood glucose, 2 h postprandial glucose, homeostasis model assessment-insulin resistance index, and C-peptide levels in PDM rats (P < 0.05, 0.01), increased the insulin sensitivity index (P < 0.05), and alleviated the pathological damage of pancreas, ileum and colon. Gas chromatography results showed that DOP administration could significantly increase the levels of acetic acid, butyric acid, isovaleric acid and SCFAs in feces (P < 0.05, 0.01). ELISA, Western blotting and qRT-PCR results showed that DOP administration could significantly increase the plasma GLP1 level, as well as the mRNA and protein expressions of GPR43 and GLP1 receptor in ileum and pancreas (P < 0.05, 0.01). Conclusion DOP could improve PDM, and its effect may be related to enhancing gut-derived SCFA production, up-regulating GPR43 expressions in intestine and pancreas, subsequent promoting GLP1 secretion, restoring β-cell function and improving glucose regulation.

Objective To investigate the effect and mechanism of notoginsenoside R₁ (NGR1) on cardiac remodeling after myocardial ischemia-reperfusion injury (MIRI) in mice. Methods A model of cardiac remodeling after myocardial ischemia-reperfusion injury (MIRI) was established by ligating the anterior descending branch of the coronary artery in mice. After three weeks of intervention with NGR1, the cardiac index and heart weight/tibia length of mice were measured. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were measured using a small animal ultrasound instrument. The cross-sectional area of myocardial cells was calculated by wheat germ agglutinin (WGA) fluorescence staining. The collagen volume fraction (CVF) of heart was calculated by Masson staining. The protein expressions of collagen I, collagen III, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), transferrin receptor 1 (TFR1), acyl-CoA synthetase long-chain family member 4 (ACSL4), solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) in myocardial tissue were detected by Western blotting. The possible mechanism of NGR1 inhibiting cardiac remodeling after MIRI was preliminarily explored through network pharmacology. The contents of iron ions (Fe²⁺) and lipid peroxide (LPO) in myocardial tissue were detected using kits. The damage of myocardial mitochondria was observed by transmission electron microscopy. Results Compared with sham group, cardiac index, heart weight/tibia length, LVEDV, LVESV and CVF of mice in model group were significantly increased (P < 0.05, 0.01), protein expressions of collagen I, collagen III, ANP, BNP, TFR1 and ACSL4 in myocardial tissue were significantly increased (P < 0.01), contents of Fe²⁺ and LPO were significantly increased (P < 0.05, 0.01), mitochondrial membrane density was increased and cristae was decreased, SLC7A11 and GPX4 protein expressions were significantly decreased (P < 0.01). Compared with model group, cardiac index, heart weight/tibia length, LVEDV, LVESV and CVF of mice in NGR1 administration groups were significantly decreased (P < 0.05, 0.01), protein expressions of collagen I, collagen III, ANP, BNP, TFR1 and ACSL4 were significantly decreased (P < 0.05, 0.01), contents of Fe²⁺ and LPO were significantly decreased (P < 0.05, 0.01), mitochondrial damage was improved, SLC7A11 and GPX4 protein expressions were significantly increased (P < 0.05, 0.01). Conclusion NGR1 could significantly improve cardiac remodeling in mice after MIRI, and its mechanism may be related to the activation of SLC7A11/GPX4 pathway to inhibit ferroptosis of cardiomyocytes.

Objective To investigate the subcellular distribution patterns of the combination of cantharidin (CTD) and baicalin (BA) based on cell pharmacokinetics and elucidate the synergistic mechanism against hepatocellular carcinoma. Methods Using human hepatoma HepG2 cells as the model, the dynamic concentrations of the drugs in whole cells and organelles (nucleus, mitochondria, endoplasmic reticulum, and lysosomes) over 12 h after administration of single drugs (CTD 6 μg/mL, BA 30 μg/mL) and their combination (CTD 6 μg/mL + BA 30 μg/mL) were quantitatively analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Pharmacokinetic parameters were calculated using a non-compartmental model with Phoenix WinNonlin software. Results At the whole-cell level, the combination increased the intracellular area under the curve (AUC_0—t) of CTD by 48.9% and decreased its clearance (P < 0.05), but did not significantly affect the pharmacokinetics of BA. At the subcellular level, the combination significantly increased the AUC_0—t of CTD and BA in nucleus, lysosome, mitochondria and endoplasmic reticulum by 93.5%, 46.4%, 38.3%, 52.3%, and 68.4%, 40.0%, 41.0%, and 46.7%, respectively (P < 0.05, 0.01). Furthermore, the time to reach peak concentration (t_max) for both drugs in mitochondria was earlier in the combination group, and the mean residence time (MRT_0—t) of CTD in the endoplasmic reticulum was significantly prolonged (P < 0.01), indicating a spatiotemporally synergistic drug delivery optimization. Conclusions The combination of CTD and BA could synergistically optimize the distribution of drugs in key subcellular structures such as nucleus, mitochondria, endoplasmic reticulum and lysosomes, and may enhance the anti-hepatocellular carcinoma effect by inducing DNA damage, accelerating mitochondria-mediated apoptosis and promoting endoplasmic reticulum stress, which provides a theoretical basis for the design of traditional Chinese medicine combinations based on organelle targeting.

Objective To explore “state and target” pathological mechanism of sepsis by comprehensively analyzing the single-cell transcriptomics and transcriptomic datasets related to sepsis, construct the prognostic model for “state and target” of sepsis, and explore potential target Chinese medicines and active ingredients. Methods Differential expression gene analysis and venn diagram analysis were used to identify “state and target” of sepsis related genes. Key module genes were obtained using the random walk algorithm through the connectivity of the protein-protein interaction (PPI) network. Gene ontology (GO) biological process (BP) and Kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the biological functions of key module genes. To explore the role of “state and target” of sepsis related genes by single-cell transcriptome analysis. A prognostic model for “state and target” of sepsis was constructed using gene expression data and clinical survival data from sepsis patients. The expression levels of prognostic genes for “state and target” of sepsis were validated by constructing sepsis mouse model and real-time quantitative polymerase chain reaction (RT-PCR). Through reverse network pharmacology, potential targeted traditional Chinese medicines and their active components were screened, and molecular docking was used to verify the binding performance between active ingredients and prognostic targets. Results A total of 70 genes related to “toxic state” of sepsis, 67 genes related to “stasis state” of sepsis and 54 genes related to “deficiency state” of sepsis were obtained. The key module genes of “toxic state” of sepsis were closely related to inflammation, the key module genes of “stasis state” of sepsis were closely related to coagulation, and the key module genes of “deficiency state” of sepsis were closely related to cell homeostasis. AUCell score showed that monocyte had the highest score for “toxic state” and “deficiency state” of sepsis. Platelet had the highest score for “stasis state” of sepsis. The activities of metabolic and inflammatory related genes and pathways were significantly up-regulated in monocytes with high “toxic state” score. Platelet activation and coagulation/thrombosis related genes and pathways were significantly up-regulated in platelet with high “stasis state” score. Compared with control group, there were a large number of down-regulated genes in monocytes of sepsis patients, which were involved in antigen presentation, cell energy metabolism, cell cycle, protein synthesis, etc. The prognostic model for “state and target” of sepsis constructed based on eight targets [interleukin-1 receptor type 2 (IL1R2), ADP ribosylation factor like GTPase 4C (ARL4C), cytochrome C oxidase subunit 7B (COX7B), eukaryotic translation initiation factor 2 subunit gamma (EIF2S3), immediate early response 3 (IER3), LSM1 homolog (LSM1), paired basic amino acid cleaving enzyme (FURIN) and ankyrin repeat domain-containing protein 9 (ANKRD9)] had good prognostic predictive ability, and the expression levels of these genes were validated using animal experiments and RT-PCR. Based on the bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine (BATMAN-TCM) database and database of constituents absorbed into the blood and metabolites of traditional Chinese medicine (DCABM-TCM), reverse network pharmacology analysis had screened multiple Chinese herbal medicines with effects of clearing heat and detoxifying, activating blood and resolving stasis, and reinforcing healthy qi and their active ingredients entering the blood. Molecular docking showed that representative active ingredients have good binding properties with prognostic targets. Conclusion Activation of inflammatory response driven by monocyte metabolic reprogramming may be the main pathological mechanism of “toxic state” of sepsis. Platelet activation-coagulation/thrombosis may be the main pathological mechanism of “stasis state” of sepsis. Monocyte dysfunction-immunosuppression may be the main pathological mechanism of “deficiency state” of sepsis. A model based on eight genes can serve as a risk prediction model for the prognosis of “state and target” of sepsis.

Objective To explore the “medicine-efficacy” associations of traditional Chinese medicine (TCM) in the treatment of sepsis, identify core herbsrelated to reducing mortality and improving key clinical outcomes such as the acute physiology and chronic health evaluation II (APACHE II) score, and provide evidence-based references for TCM-assisted treatment of sepsis. Methods A systematic search of Chinese and English databases was conducted up to September 2024 to collect clinical studies on TCM compound prescriptions for sepsis. A comprehensive sepsis TCM formula database was established. Nine machine learning algorithms were compared using ten-fold cross-validation, and the optimal model for each clinical outcome was interpreted through the Shapley additive explanations (SHAP) method to identify key herbs and their contribution directions. Results The multilayer perceptron showed the best performance in predicting TCM syndrome scores, overall effectiveness, inflammatory and immune indicators, biochemical parameters, organ dysfunction scores, and mortality; logistic regression performed best for blood gas analysis along with gastrointestinal function and intestinal mucosal barrier outcomes; and the support vector machine achieved optimal predictive performance for routine blood tests and APACHE II scores. SHAP analysis revealed that Dihuang (Rehmanniae Radix), Zhishi (Aurantii Fructus Immaturus), Huangqi (Astragali Radix), Fuzi (Aconiti Lateralis Radix Praeparata), Huangqin (Scutellariae Radix), and Houpo (Magnoliae Officinalis Cortex) had positive contributions across outcomes such as mortality, APACHE II score, inflammatory markers, and gastrointestinal function, forming the core nodes of the “medicine-efficacy” network. Conclusion This study established a clinically oriented “medicine-efficacy” association network through multi-model comparison and explainable machine learning analysis. The findings highlight the potential key roles of several core herbs in improving major clinical outcomes of sepsis, providing data support and evidence-based basis for precise syndrome differentiation and medication in TCM, as well as for the research and development of new TCMs.

Objective Based on the Patsnap database, a visual analysis is conducted on the development overview of patent technologies in China’s traditional Chinese medicine (TCM) industry from 2000 to 2024 to reveal its evolution laws and structural characteristics. Methods Employing multidimensional retrieval strategies (title/abstract, IPC classifications, national economic industry classifications, etc.), the study integrates 119 485 TCM-related patents, focusing on four technical domains: Chinese medicinal materials, Chinese herbal decoction pieces, Chinese patent medicine (CPM), and intelligent manufacturing of TCM. Quantitative and visualized analyses were performed on application trends, regional distribution, applicant types, IPC classifications, and legal statuses. Results The number of patent applications in the TCM industry reached a peak of 11 087 in 2017, then declined, and stabilized at 5 000 per year after 2020, reflecting a shift from scale expansion to quality improvement. Shandong, Guangdong, and Jiangsu emerged as innovation hubs, while Anhui leveraged its Bozhou medicinal material market to form a specialized cluster, demonstrating a multi-polar distribution of “eastern dominance with gradual western expansion”. The proportion of patents in the field of CPMs is the highest, while the base in the field of intelligent manufacturing is small but the growth rate is stable. IPC classifications centered on A61K36 (plant-derived preparations) and A61K9 (dosage forms), targeting digestive and anti-infectious diseases. Colleges and universities lead in high-value invention patents, while enterprises have a high number of applications but an authorization rate of less than 20%. Individual applicants were active but showed low conversion efficiency. Invalid patents account for 69.91%, mainly due to insufficient inventiveness; the number of invalid patents in the field of CPM is the largest, but the proportion of valid patents is relatively high. Conclusion TCM patents exhibit “quantitative dominance but qualitative structure imbalance and weak global competitiveness”. Innovations in CPMs focus on process optimization rather than clinical validation. Recommendations include establishing a “quality-transformation-internationalization” synergy mechanism, strengthening frontier technologies like authentic medicinal material DNA barcoding and nano-formulations, and promoting the transformation of TCM patents from “scale-driven” to “value-driven” patent strategies.

Objective To comprehensively analyze the research trends and focuses of Gouteng (Uncariae Ramulus Cum Uncis) from bibliometric and global patent perspectives, providing forward-looking references for its future clinical application and in-depth research. Methods Literature was retrieved from four databases: China National Knowledge Infrastructure (CNKI), Wanfang, VIP, and Web of Science (WOS) with Uncariae Ramulus Cum Uncis as the keyword. Retrieved publications were imported into NoteExpress for deduplication and screening. Software including Excel, Origin, CiteSpace, and VOSviewer were used to visually analyze the current research status and development trends of Uncariae Ramulus Cum Uncis in domestic and international publications, covering publication trends, national distribution, publication institutions, journal of publication, authors, and keywords. The Incopat patent database was searched to analyze global patent applications related to Uncariae Ramulus Cum Uncis, including trends in patent applications, geographical distribution, applicants, and technology fields. Results A total of 1 713 Chinese publications and 594 English publications met the inclusion criteria. China was the primary contributor to publications, with international attention increasing gradually in recent years. Core research teams have emerged in this field, but collaboration among institutions requires strengthening. In recent years, there has been an overlap in the research hotspots of Chinese and English literature in the field of Uncariae Ramulus Cum Uncis, both focusing on the pharmacological components and action mechanisms of Uncariae Ramulus Cum Uncis. Chinese studies specifically emphasize Tianma Gouteng Decoction and its use in hypertension treatment, identifying bioactive compounds and elucidating regulatory mechanisms. English publications focus on phytochemistry, anti-inflammation, antioxidation, and the potential efficacy in neurological diseases. Based on a search of the Incopat patent database, 8 048 patents were initially screened. After merging patent families, 6 251 patents were retained, among which 5 461 were domestic patents, accounting for 87.36% and occupying a dominant position. Conclusion Research in the field of Uncariae Ramulus Cum Uncis shows a trend of continuous expansion and deepening. Overall, it is shifting from traditional Chinese medicine practice to a development path featuring multi-disciplinary collaborative innovation and in-depth analysis of molecular-level mechanisms. Although China dominates international patent applications related to Uncariae Ramulus Cum Uncis, there are key drawbacks: A large number of patents have expired, the degree of transformation and implementation is limited, and market expansion is weak.

Objective To mine CYP gene family members in Coptis chinensis based on genomic data and analyze the evolutionary specificity of CYP719 genes. Methods The CYP gene family members of C. chinensis were identified based on homologous CYP gene sequence from both Arabidopsis thaliana and Oryza sativa. Analyses of the physicochemical properties of proteins, system classification, gene structure, expression profile and cis-acting elements of the identified CYP genes were further conducted by bioinformatics. The CYP719 family members in C. chinensis were identified based on the 23 reported CYP719 genes in other plants, and their collinearity analysis was performed through homologous sequence alignment. Results A total of 308 CYP genes were identified in the C. chinensis genomic data, belonging to 45 families in nine family clans. The range of amino acid numbers for encoded proteins is from 103 to 2 008 aa, with a relative molecular mass ranging from 11 010 to 230 370, and an isoelectric point ranging from 4.85 to 9.79. The number of CYP gene exons in C. chinensis varies between 1 and 38, with a wide range of variation. A total of 308 CYP genes were unevenly distributed on nine chromosomes of C. chinensis and mainly expanded through dispersed duplication（DSD） and proximal duplication（PD）. The expression of CYP genes was tissue-specific, and nine genes were significantly upregulated in rhizomes and fibrous roots. In the promoter regions of these CYP genes, stress response elements were predicted to be the most frequently occurring cis-acting elements. Through genomic collinear analyses between different plants, the C. chinensis CYP719 genes have homologous ones in Aquilegia coerulea, Corydalis tomentella, Papaver somniferum, and Aristolochia debilis, but not in Arabidopsis thaliana, Glycine max, Prunus persica, Chrysanthemum nankingense, Vitis vinifera, Aconitum vilmorinianum, O. sativa, and Amborella trichopoda. Conclusion C. chinensis has a rich CYP supergene family, in which CYP719 gene shows branch-specific evolution, laying the foundation for analyzing the functions of this type of gene.

Objective Taking the glycosyltransferase in Platycodon grandiflorus as the research object, the bioinformatics analysis, gene cloning and recombinant plasmid construction, protein expression and purification, catalytic function validation, enzymatic properties, and substrate promiscuity of the glycosyltransferase gene PgUGT72B21 were investigated. Methods The key glycosyltransferase PgUGT72B21 was screened from the transcriptome data of P. grandiflorus and subsequently cloned. A prokaryotic expression vector, pET-28a-PgUGT72B21, was constructed using gene recombination technology and transformed into Escherichia coli BL21（DE3） competent cells. Protein purification was performed according to the instructions of the His-tagged protein purification kit, and protein expression was detected by SDS-PAGE electrophoresis. The enzymatic reaction products were analyzed using HPLC and LCMS systems. Results P. grandiflorus glycosyltransferase gene Pg UGT72B21 obtained through cloning had an open reading frame（ORF） of 1 407 bp, encoding 468 amino acid residues with a relative molecular mass of approximately 51 000. A highly conserved plant secondary product glycosyltransferase（PSPG） motif was identified at the C-terminus. Phylogenetic analysis indicated that this glycosyltransferase belongs to the UGT72 family. In vitro enzymatic assays demonstrated that PgUGT72B21 could catalyze the glycosylation of the C3 hydroxyl group of quercetin to produce isoquercitrin. After heterologous expression and purification, the recombinant protein was obtained, and its enzymatic properties were analyzed. The optimal pH and temperature for the catalytic reaction were determined to be 6.0 and 60 ℃, respectively, with the highest substrate conversion rate achieved after 2 h of reaction. The kinetic parameters for quercetin catalysis were K_m = 390.10 μmol/L and kcat = 11.10/min. Further substrate promiscuity studies revealed that PgUGT72B21 could not only catalyze flavonols such as quercetin but also flavonoids such as apigenin. Conclusion The newly discovered glycosyltransferase PgUGT72B21 holds significant importance for enriching the glycosylation tool enzyme library and provides a foundation for further elucidating the glycosylation process of flavonoid glycosides in P. grandiflorus.

Objective To explore the key enzyme genes of the pentacyclic triterpene esters biosynthesis pathway in Balanophora subcupularis, which lays a certain research idea and theoretical foundation for its biosynthesis mechanism and the development and utilization of medicinal plant resources in the genus Balanophora. Methods Transcriptome sequencing and analysis was performed on the three tissues of the flowerstalk, rhizome and umbel of B. subcupularis, then searching the candidate enzyme genes involved in triterpene ester biosynthesis by PfamScan, constructing the yeast expression vector of BsubOSCs gene which was introduced into the Saccharomyces cerevisiae lanosterol synthase gene（erg7）-deficient mutant strain GIL77 for functional characterization. Results A total of 87 candidate genes involved in pentacyclic triterpene esters biosynthesis were identified from the transcriptome of B. subcupularis, most of them were highly expressed in umbel, suggesting that umbel may be the main site of synthesis of pentacyclic triterpene esters. The enzyme function of two BsubOSC genes were characterized, which was found that BsubOSC2 could catalyze 2, 3-oxidosqualene to form β-amyrin, but no catalytic product of BsubOSC1 was detected. Conclusion Based on transcriptome analysis, the key enzyme genes involved in pentacyclic triterpene esters biosynthesis were found in B. subcupularis, and BsubOSC2 is a β-amyrin synthetase gene, and is one of the key enzyme genes for the biosynthesis of balanophorin A. This study provides gene resources for further analysis of the biosynthesis pathway of triterpene ester compounds such as balanophorins, and facilitate the research on molecular breeding of B. subcupularis.

Objective To reveal the changes of morphological characteristics and amino acid metabolism in fruit of Sinopodophyllum hexandrum at different ripening stages. Methods The morphological characteristics, amino acid content, and related genes expression were determined and analyzed in fruit of S. hexandrum at different ripening stages（S1 to S9）. Results There were significant differences in morphological characteristics（e.g., fruit color, length, and diameter）, amino acid content, and related genes expression during fruit ripening. The fruit length, diameter, fresh weight, and dry weight showed a 2.2, 2.18, 31.42, and 49.05 fold increase at the S6 compared with S1; the content of amino acid were greater at the S5 and S9 than other stages; the 10 candidate genes related to amino acid metabolism [e.g., arginine decarboxylase 2（ADC2）, arogenate dehydratase 6（ADT6）, and asparagine synthetase（AS）] showed a down-regulation to some extent at S4 to S8 compared with S2. Conclusion There were dynamic differences in morphological characteristics, amino acid metabolism and related gene expression levels with fruit ripening. These finding will provide useful references for the development and utilization of S. hexandrum fruits.

Objective To develop the determination methods for the content of vanillic acid, isovanillic acid, rhamnetin, luteolin, quercetin, diosmetin, oleanolic acid, ursolic acid, anemarrhenasaponin A2, gracillin, prosapogenin A, dioscin, protogracillin, β-sitosterol, stigmasterol, total ash and acid insoluble ash in Smilax riparia, and evaluate the quality of S. riparia by combining multivariate statistical analysis and entropy weight TOPSIS model. Methods HPLC was launched on an Agilent Symmetry C₁₈ chromatographic column（250 mm×4.6 mm,5 μm） by gradient elution with a mobile phase of acetonitrile-0.3% phosphoric acid at a flow rate of 1.0 mL/min, detection wavelength were 280 nm（for detecting vanillic acid and isovanillic acid）, 345 nm（for detecting rhamnetin, luteolin, quercetin, diosmetin） and 210 nm（for detecting oleanolic acid, ursolic acid, anemarrhenasaponin A2, gracillin, prosapogenin A, dioscin, protogracillin, β-sitosterol, stigmasterol）, column temperature was 30 ℃, and an injection volume was 10 μL. The contents of total ash and acid insoluble ash were detected according to the method of Chinese Pharmacopoeia 2025 edition. Based on the content measurement value, the quality of 18 batches of S. riparia samples was comprehensively evaluated by using the multivariate statistical analysis and entropy weight TOPS model. Results A total of 15 chemical components had good linear relationships in their respective mass concentration ranges, and the average recovery rates were 96.99%—100.14%. The contents of vanillic acid, isovanillic acid, rhamnetin, luteolin, quercetin, diosmetin, oleanolic acid, ursolic acid, anemarrhenasaponin A2, gracillin, prosapogenin A, dioscin, protogracillin, β-sitosterol, stigmasterol, total ash and acid insoluble ashin in 18 batches of S. riparia were 0.338—1.213, 0.074—0.200, 0.197—0.360, 0.423—0.676, 0.620—1.814, 0.496—1.007, 0.192—0.410, 0.095—0.187, 1.360—2.281, 0.379—0.651, 0.711—1.221, 2.120 —3.657, 0.965—1.652, 0.113—0.275, 0.063—0.123 mg/g, 3.6%—9.5%, 1.4%—3.5%, respectively. Multivariate statistical analysis classified 18 batches of S. riparia into three categories, principal component analysis yielded two principal components, with a cumulative variance contribution rate of 88.149%. A total of eight components such as dioscin, quercetin, luteolin, anemarrhenasaponin A2, oleanolic acid, vanillic acid, protogracillin and β-sitosterol were screened by VIP analysis, which could be used as differential markers of S. riparia in different producing areas. The results of entropy weight TOPSIS method showed that the quality of 18 batches of S. riparia was significantly different, and the comprehensive score was 0.230 6—0.706 2. The quality of samples numbered S16 and S15 was ranked in the front. Conclusion The quality evaluation model based on chemometrics and entropy weight TOPSIS is convenient and effective, and can be used for the quality evaluation of S. riparia.

Current quality evaluation of traditional Chinese medicine (TCM) primarily focuses on chemical composition analysis, neglecting the interactions among components. The phase of TCM is an ordered combination formed by different or identical components through intermolecular forces, serving as a crucial form for TCM efficacy and a vital supplement to its quality assessment. It reflects the characteristic of multiple components synergistically exerting pharmacological effects. This article systematically reviewed the importance of TCM phase in quality evaluation. We summarized the formation mechanisms and research methods of TCM phase, as well as their roles in reducing toxicity and enhancing efficacy. Furthermore, we discussed the research strategies for screening active phases, characterization approaches, and potential challenges, providing a theoretical basis for transitioning TCM quality evaluation from a composition-based perspective to a “chemistry-structure-function integrated” model.

High-altitude sleep disturbance (HASD), a common physiological disorder caused by hypobaric hypoxia, clinically manifests as insomnia, sleep apnea, and sleep fragmentation. Its pathogenesis involves hypoxia-inducible factor activation, oxidative stress, and neuroendocrine dysregulation, with severe cases potentially leading to irreversible neurological damage. The gut-brain axis, a bidirectional communication network between the central nervous system and the gastrointestinal tract, plays a key role in HASD progression. Gut microbiota, as a critical interface between intestinal and cerebral functions, may modulate sleep disorders through neurotransmitter regulation, short-chain fatty acid production, and hormonal pathway modulation. Traditional Chinese medicine (TCM) and traditional Tibetan medicine (TTM) offer unique therapeutic advantages for HASD management. TCM theory suggests that “gastric imbalance disrupts sleep”, while TTM attributes insomnia to “tri-humoral dysregulation”. Both systems align with the modern gut-brain axis theory, emphasizing sleep quality improvement through spleen-stomach system regulation. This study examines the theoretical foundations of TCM and TTM for preventing and treating HASD, summarizing relevant classical prescriptions, monomeric drugs, active ingredients, and novel formulations. Additionally, it explores the pharmacological mechanisms by which TCM and TTM improve HASD by regulating the gut-brain axis, providing safe and effective strategies for individuals entering high-altitude regions.

The comorbidity of depression and cognitive impairment is increasingly prevalent in clinical practice, particularly among the elderly population, which faces a higher risk of onset. Given the severity of the current comorbidity situation and the guiding significance of the traditional Chinese medicine (TCM) theory of “treating different diseases with the same method,” it is essential to systematically explore the underlying mechanisms of this comorbidity and conduct an in-depth analysis of TCM’s multi-target intervention strategies. According to TCM theory, the core pathogenesis of this comorbidity involves visceral deficiency, insufficiency of qi and blood, and phlegm turbidity clouding the orifices and disturbing the spirit, collectively leading to impaired mental functioning. Modern medical research, on the other hand, indicates that the pathological basis of this comorbidity arises from the interplay of multiple systemic mechanisms, including vascular pathologies, reduced brain-derived neurotrophic factor, hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis, neuroinflammation, and gut microbiota dysbiosis. TCM demonstrates unique advantages in holistic intervention by multi-target regulation of neurotransmitters, suppression of neuroinflammation, enhancement of neuroplasticity, and modulation of the gut-brain axis. Through a systematic review of the mechanisms underlying the comorbidity of depression and cognitive impairment and the progress in multi-target intervention research in TCM, this study aims to provide new insights and references for scientific research and clinical practice in this field.

Astragalus polysaccharides (APS) are the principal bioactive constituents of Huangqi (Astragali Radix), exhibiting diverse pharmacological activities. However, their clinical application is limited by challenges including high molecular weight, low solubility, and consequently poor bioavailability. Selenization modification has emerged as an effective strategy to improve the physicochemical properties and biological functions of APS, leading to the formation of selenized APS (Se-APS) and APS-selenium nanoparticles (APS-SeNPs). This review systematically summarizes the preparation methods, structural characteristics, biological activities, and structure-activity relationships of selenium-containing APS derivatives, and discusses their application prospects. Primary preparation techniques include biological transformation and chemical synthesis approaches. Following selenization, the resulting products demonstrate significantly improved solubility, stability, and bioavailability, and can form uniformly sized nanoparticles. Research indicates that selenium-containing APS derivatives exhibit superior pharmacological efficacy compared to native APS, including enhanced antioxidant, immunomodulatory, antitumor, and hepatoprotective activities. Furthermore, APS-SeNPs can serve as effective drug delivery carriers and show synergistic effects when combined with other bioactive compounds. Structure-activity relationship studies suggest that their bioactivity is closely associated with selenium content, molecular weight, and nanostructural attributes. Nevertheless, several challenges remain in this field, such as insufficient characterization of selenization sites and a lack of systematic toxicological evaluation. Future research should focus on optimizing preparation processes, strengthening safety assessments, and promoting clinical translation to advance their applications in the pharmaceutical and functional food industries.

Rehmannioside D is an indicator component of Dihuang (Rehmanniae Radix), a traditional Chinese medicine that shares the same origin in both medicine and food. Rehmannioside D, a natural iridoid glycoside, exhibits a broad spectrum of pharmacological activities, including nourishing yin and blood, cooling blood and stopping bleeding, providing analgesia, acting as an antidepressant, and preventing osteoporosis, thereby meriting intensified investigation. This paper systematically reviews the recent advances in rehmannioside D research, detailing the optimization of determination methods and the application of new technologies for rehmannioside D, as well as the content determination in different dosage forms. The principles of content variation in different producing areas, processing methods, and tissues of Rehmanniae Radix are summarized. Furthermore, it focuses on elucidating the pharmacological effects of rehmannioside D in the endocrine and cardiovascular systems, nervous system, and other aspects. In addition, this paper critically discusses the existing challenges in its research. The aim is to provide a scientific basis for the further development and clinical application of rehmannioside D, as well as the quality identification and comprehensive utilization of Rehmanniae Radix.

Transmembrane transport, as a core process in drug absorption, distribution, and metabolism, directly determines the bioavailability and target tissue distribution of active ingredients in traditional Chinese medicine (TCM). It is a key limiting factor for their systemic exposure and exertion of drug efficacy. The structural diversity and complex mechanisms of TCM components generally lead to low oral bioavailability, which severely restricts their full clinical therapeutic effects and has become a bottleneck in the development of modern TCM formulations. This article systematically reviews the transmembrane transport mechanisms of major active ingredients in TCM, such as flavonoids, alkaloids, saponins, terpenoids, polysaccharides, and organic acids, with a focus on elucidating their structure-transport relationships and the complex regulatory networks involving various transport proteins (e.g., P-glycoprotein, multidrug resistance-associated proteins, organic anion transporting polypeptides, etc.). Given that traditional research has long focused on qualitative and quantitative analysis of components while relatively neglecting the exploration of transmembrane transport mechanisms, hindering mechanism-guided formulation design and optimization. This review further delves into novel enhancement strategies based on transport mechanisms. These include advanced delivery systems, structural modification, efflux protein inhibition, gut microbiota-mediated metabolic enhancement, and synergistic compatibility of TCM ingredients. The aim is to overcome the in vivo delivery barriers of active TCM components, improve the bioavailability of TCM formulations, and ultimately provide insights for achieving precise clinical medication.

The health status of soil environment directly determines the quality of medicinal plants. Soil physical, chemical, and biological properties are key indicators for evaluating soil health. Their individual or synergistic effects exert profound influences on the growth and development, nutrient absorption and metabolism, as well as bioactive component accumulation of medicinal plants. By clarifying the connotation of soil health, this paper focuses on the impacts of soil health indicators (soil physical, chemical, and biological properties) on the quality development of medicinal plants. It systematically reviews the response mechanisms of medicinal plants in terms of morphology, physiology, biochemistry, and bioactive component accumulation under different soil conditions, and expounds the comprehensive effects of the interactions among soil health indicators on the quality formation of medicinal plants. The purpose is to summarize the interaction mechanism between soil environment and medicinal plants for the growth and quality improvement of Chinese medicinal materials, so as to provide a solid theoretical basis for optimizing the cultivation environment of medicinal plants and promoting the technical application of increasing their yield and quality.