Objective Reactive oxygen species (ROS)-responsive co-loaded elemene (Ee) and chlorin e6 (Ce6) liposomes (Ee-Ce6-L) were prepared, characterization of their physicochemical properties, and evaluated for their inhibitory effects on bladder cancer 5637 cells. Methods The synergistic ratio of elemene and Ce6 (Ee + Ce6) was screened using the MTT assay. Ee-Ce6-L was prepared via the thin-film hydration method and characterized for morphology, particle size, encapsulation efficiency, drug loading capacity, stability, ROS generation capacity, and in vitro release rate. MTT assays evaluated the cytotoxicity of Ee-Ce6-L against 5637 cells. Annexin V-FITC/PI double staining was used to study the apoptotic effects of Ee-Ce6-L on cells. Fluorescence microscopy and a fluorometer were employed to examine its impact on ROS generation capacity and mitochondrial membrane potential (MMP). Western blotting analysis was conducted to investigate the expression of apoptosis pathway-related proteins. Results Ee + Ce6 exhibited synergistic effects across the 40:1 to 200:1 concentration range, with optimal synergy observed at 120:1; Ee-Ce6-L exhibited good stability and dispersibility. After irradiation with 660 nm laser, the particle size changed from (194.27 ± 0.20) nm to (390.97 ± 2.76) nm, and the ζ potential shifted from (−19.06 ± 0.07) mV to (−6.30 ± 0.34) mV. In vitro release experiments demonstrated that Ee-Ce6-L exhibits dual functions of sustained release and light-controlled drug release; the encapsulation efficiencies of elemene and Ce6 were (94.02 ± 0.38)% and (96.95 ± 0.70)%, respectively, while the drug loading capacities of Ee and Ce6 were (11.51 ± 0.46)% and (2.32 ± 0.01)%; Ee-Ce6-L exhibited strong singlet oxygen generation capacity; In vitro cell experiments demonstrated that Ee-Ce6-L promoted apoptosis in 5637 cells by increasing ROS production and reducing MMP, as well as by decreasing B-cell lymphoma-2 (Bcl-2) protein expression while increasing Bcl-2-associated X protein (Bax), cleaved poly ADP-ribose polymerase (cleaved PARP), and cleaved cystein-asparate protease-3 (cleaved Caspase-3) protein expression. Conclusion Ee-Ce6-L exhibits ROS-responsive release properties and synergistically inhibits 5637 cell proliferation and induces apoptosis through the combined effects of traditional Chinese medicine and phototherapy.

Objective To elucidate the dynamic changes in moisture, color, and index components during the “sweating” process of Danshen (Salviae Miltiorrhizae Radix et Rhizoma, SMRR) and preliminarily explore the correlation between moisture migration, color evolution, and changes in index components. Methods SMRR samples from Shandong underwent the “sweating” process, low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance imaging (MRI) were employed to analyze moisture migration and distribution characteristics. The high-performance liquid chromatography (HPLC) and colorimeter were used to determine the contents of salvianolic acid B, tanshinone II_A, tanshinone I, cryptotanshinone and chromaticity values (L^*,a^*, b^*, and ΔE) of SMRR at different “sweating” degrees, respectively, followed by Pearson correlation analysis. Results The “sweating” process mainly involved free water loss, with relaxation peak gradually shifting leftward. MRI results indicated that moisture migrated from the core to the exterior of the roots. The cross-sectional color transitioned from the outer to inner layers, changing from yellowish-white to brown and finally to purplish-brown, with the L^* decreasing while a^*, b^*, and ΔE increasing continuously. The contents of salvianolic acid B and tanshinones initially increased and then decreased, reaching the highest values in lightly sweated SMRR (SD3, 47.38 mg/g) and moderately sweated SMRR (SD7, 4.62 mg/g) respectively. The content of salvianolic acid B exhibited a significant correlation with L^*, a^*, b^* and ΔE (P < 0.01). Conclusion By clarifying the characteristics of moisture migration and distribution during the “sweating” process of SMRR, this study scientifically validates the traditional view that “sweating promotes the outward diffusion of internal moisture in medicinal materials”. It reveals the dynamic changes in color and index components during “sweating” and preliminarily explores the role of moisture in driving the color deepening from the outside to the inside and changes in salvianolic acid B content, providing a reference for optimizing the “sweating” process of SMRR.

Objective To investigate the existing states of Panax notoginseng total saponins in complex solution systems and elucidate the micellar association behavior of total saponins based on their ultrafiltration separation characteristics. Methods A total saponins solution of P. notoginseng was used as the research subject, with notoginsenoside R₁ and ginsenoside Rg₁, Rb₁, Rd selected as representative analytes. Ultrafiltration rejection rate, permeation rate, and micellar compositional ratio were employed as evaluation indices to analyze how key parameters—including inorganic salt concentration, temperature, and saponin concentration—affect saponin molecular states and ultrafiltration behavior. Results Owing to their amphiphilic structures, the saponins readily form micelles in aqueous solution, and their ultrafiltration performance is regulated by multiple environmental factors. Inorganic salts modulate micellar stability through ionic effects and salting-out actions: the divalent anion SO₄²⁻ disrupts hydration layers and strengthens hydrophobic interactions, thereby promoting micelle formation and precipitation, with salting-out intensity ranked as Na₂SO₄ > MgSO₄ > KCl. Monovalent salts such as KCl facilitate micellar association at low concentrations via charge shielding, whereas high concentrations intensify salting-out. Dammarane-type saponins, particularly Rb₁ and Rd, showed higher participation in salt-induced micelles, indicating a structure-driven association preference. Increasing temperature enhances molecular motion, weakens hydrophobic interactions and intermolecular hydrogen bonding within micelles, thereby promoting micelle dissociation, increasing permeation rates, and reducing rejection. Elevating saponin concentration strengthens hydrophobic and aggregative interactions among solute molecules, facilitating micelle assembly. By fitting the relationship between ultrafiltration rejection and membrane pore size at different solute concentrations, the molecular weight of micelles at 90% rejection and the internal compositional ratio of characteristic saponins were estimated. The results showed that at 10—20 mg/mL, micelles mainly consisted of diol-type saponins, but their association degree remained limited at low concentrations. At 30—60 mg/mL, the micellar molecular mass increased markedly, indicating that higher solute concentrations promote the formation of hybrid micelles. Conclusion This study elucidates the mechanisms by which pharmaceutical processing parameters regulate the micellar structure and ultrafiltration efficiency of P. notoginseng total saponins, providing theoretical and practical guidance for quality control and purification process optimization in saponin‑based traditional Chinese medicine formulations.

Objective To optimize the honey-frying process of Pipaye (Eriobotryae Folium, EF) and establish a quality differentiation system between raw and processed products, providing scientific evidence for standardized production and quality control of honey-fried EF. Methods The effects of five factors-honey-to-water ratio, solid-to-liquid ratio, moistening time, stir-frying temperature, and stir-frying duration were investigated through single-factor experiments. The comprehensive weights of maslinic acid, corosolic acid, oleanolic acid, ursolic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, and hyperoside were determined using the analytic hierarchy process (AHP) combined with the criteria importance through intercriteria correlation (CRITIC) objective weighting method. Key process parameters (honey-to-water ratio, stir-frying temperature, and stir-frying duration) were optimized via Box-Behnken design coupled with response surface methodology (BBD-RSM) to establish the optimal honey-processing conditions for EF. Ultra-performance liquid chromatography/high-performance liquid chromatography (UPLC/HPLC) was employed to develop fingerprint profiles of raw and honey-processed EF. Component differences were compared using similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA). Additionally, correlation analysis between colorimetric values and indicator components was conducted. Results The optimized honey-processing parameters for EF were determined as follows: honey-to-water ratio of 1:1.06, solid-to-liquid ratio of 2.0:1, moistening time of 2.5 h, stir-frying temperature of 157.9 ℃, and stir-frying duration of 11.1 min. In the established fingerprint profile of triterpenic acids in EF, 12 common peaks were identified in the raw products, while nine common peaks were identified in the processed products. Among these, four peaks were assigned as maslinic acid (peak 6), corosolic acid (peak 7), oleanolic acid (peak 11), and ursolic acid (peak 12), all of which showed increased content after honey-processing. In the fingerprint profile of organic acids, 11 common peaks were identified in the raw products, and 12 common peaks were found in the processed products. Four of these peaks were identified as neochlorogenic acid (peak 1), chlorogenic acid (peak 3), cryptochlorogenic acid (peak 4), and hyperoside (peak 11)-all of which decreased in content after honey-processing. The colorimetric values (L^*, a^*, b^*) of the honey-processed loquat slices showed a significant positive correlation with the content of triterpenic acids, and a significant negative correlation with the content of organic acids and flavonol glycosides. Conclusion The AHP-CRITIC weighting method combined with BBD-RSM can effectively balance multiple indicators. The honey-frying process for EF is stable and feasible. Integrating fingerprints with chemometric methods precisely characterizes quality changes before and after processing, offering evidence for enhanced quality control standards.

Objective To investigate the different effects of carboxymethyl starch sodium (CMS) and polyvinylpyrrolidone (PVPP) on the structure, free surface energy, and key direct compression properties of composite particles produced by co-spray drying based on traditional Chinese medicines (TCM) extracts. Methods This study selected extracts of Fuling (Poria), Danggui (Angelicae Sinensis Radix), Huangqi (Astragali Radix), Danshen (Salviae Miltiorrhizae Radix et Rhizoma), and Jinyinhua (Lonicerae Japonicae Flos) (PC、AS、AM、SM、LJ) as model drugs, and CMS and PVPP as modifiers. The composite particles of TCM extracts (CP-PC/AS/AM/SM/LJ-CMS and CP-PC/AS/AM/SM/LJ-PVPP) were prepared via co-spray drying. The properties of the co-spray dring liquid, and structure, free surface energy and direct compression properties of composite particles were comprehensively characterized. The different effects of CMS and PVPP on these properties were analyzed by statistical analysis. Results The application of different modifiers led to differential variations in the properties of the co-spray dring liquid, as well as in the structural characteristics, free surface energy, and direct compression properties of the composite particles. Compared to PVPP, co-spray dried liquid containing CMS exhibited increased solid content and viscosity, and lower surface tension and hydrodynamic size (except for CP-SM/LJ-CMS); All composite particles exhibited spherical structures, and those containing CMS showed smoother surfaces and larger particle size than those containing PVPP; The composite particles containing CMS brightness values (except for CP-AM-CMS), hardness (except for CP-AS/AM-CMS), elasticity, and rebound property (except for CP-AS/AM-CMS) were lower; The cohesion (except for CP-PC/LJ-CMS), cohesion work (except for CP-SM/LJ-CMS), free surface energy (except for CP-SM/LJ-CMS), and polarity index (except for CP-SM -CMS) of composite particles containing CMS were relatively high; The Carr’s index, Hausner ratio, angle of repose (except for CP-SM-CMS), and disintegration time (except for CP-LJ-CMS) of composite particles containing CMS were all smaller, while composite particles containing PVPP demonstrated higher tensile strength. Conclusion The properties of TCM extract co-spray drying solutions, structures, free surface energy, and direct compression performance of composite particles containing CMS and PVPP were exhibited different effects. These findings provide valuable insights for advancing co-spray drying technology in TCM extracts and for preparing composite particles suitable for direct powder compression.

Objective To investigate the mechanisms by which Compound Danshen Dropping Pills (复方丹参滴丸) ameliorate myocardial ischemia-reperfusion injury (MIRI) in mice using single-cell and spatial transcriptomics. Methods C57BL/6 mice were randomly divided into sham group, model group, Compound Danshen Dropping Pills low-, medium-, and high-dose (61.43, 122.85, 245.70 mg/kg) groups, with 15 mice in each group. Mice were continuously given drug intervention for 7 d, and a MIRI model was constructed using left anterior descending artery ligation reperfusion method 30 min after the last administration. Cardiac function was assessed by echocardiography 24 h after surgery, and myocardial histopathology was evaluated by hematoxylin-eosin (HE) and Masson staining. Hearts from sham group, model group, and Compound Danshen Dropping Pills medium-dose group were subjected to single-cell and spatial transcriptomic analyses. Single cell transcriptomics was used for cell type identification, cardiomyocyte subpopulation typing, differential gene expression analysis, pseudo temporal analysis, differential gene expression analysis and enrichment analysis. Spatial transcriptomics analysis was used to identify the spatial transcriptomic features of MIRI and the characteristic genes of injury boundary region. Western blotting was performed to detect the expression levels of nuclear receptor subfamily 3 group C member 1 (NR3C1), pyruvate dehydrogenase kinase 4 (PDK4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) in myocardial tissue of mice. Results Compared with sham group, the left ventricular ejection fraction (LVEF) of mice in model group was decreased (P < 0.05), pathology showed significant necrosis of myocardial cells and increased collagen fibers. Compared with model group, Compound Danshen Dropping Pills significantly improved the cardiac function of MIRI mice (P < 0.05), reduced myocardial injury and decreased collagen fibers. The results of single-cell transcriptomics indicated that Compound Danshen Dropping Pills could increase the proportion of myocardial cells and reduce the proportion of fibroblasts and inflammatory cells. Spatial transcriptomics revealed key genes involved in the spatial transcriptomic characteristics and damage boundary regions of MIRI. Western blotting results showed that compared with sham group, the expression levels of NR3C1, PDK4 and ACSL4 proteins in myocardial tissue of mice in model group mice were significantly increased (P < 0.05), while the expression level of GPX4 protein was significantly decreased (P < 0.05). Compared with model group, the expression levels of NR3C1, PDK4 and ACSL4 proteins in myocardial tissue of mice in Compound Danshen Dropping Pills group were significantly reduced (P < 0.05), while the expression level of GPX4 protein was significantly increased (P < 0.05). Conclusion Compound Danshen Dropping Pills may alleviate MIRI in mice by modulating myocardial metabolism and attenuating ferroptosis.

Objective To explore the effect differences and biological mechanisms of Huanglian (Coptidis Rhizoma) combined with sour herb Wumei (Mume Fructus) and pungent herb Wuzhuyu (Evodiae Fructus) in treating cholestasis based on “liver desires dispersion” theory from “five zang-organs preferences and aversions in tonification and purgation” in traditional Chinese medicine, thereby enriching the scientific connotation of “sour purgation” and “pungent tonification”. Methods A total of 36 male SD rats were randomly divided into control group, model group, Coptidis Rhizoma group, Coptidis Rhizoma-Mume Fructus group, Coptidis Rhizoma-Evodiae Fructus group and ursodeoxycholic acid group. A cholestasis model was induced using α-naphthylisothiocyanate (ANIT). After drug intervention, the anti-cholestatic effects were evaluated by measuring serum biochemical markers sun as alkaline phosphatase (ALP), γ-glutamyl transferase (GGT), total bilirubin (TBIL), total bile acid (TBA) and histopathological examination. The anti-inflammatory effects were assessed by measuring levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 in serum. The anti-oxidative stress was evaluated by measuring activity of superoxide dismutase (SOD) and levels of malondialdehyde (MDA), reactive oxygen species (ROS) in serum. The lipid-regulating effects were evaluated by measuring levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in serum. LC-MS-based untargeted metabolomics was performed on fecal samples to analyze metabolites, identify differential metabolites, and conducting Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis and gene set enrichment analysis (GSEA). Results Compared with model group, the serum biochemical index levels of each treatment group were significantly reduced (P < 0.05, 0.01, 0.001), the phenomena of liver tissues inflammatory cell infiltration, liver cell swelling and focal necrosis were significantly alleviated. Compared with Coptidis Rhizoma group, Coptidis Rhizoma-Mume Fructus group demonstrated significantly enhanced anti-inflammatory and anti-oxidative stress effects (P < 0.05, 0.01), while Coptidis Rhizoma-Evodiae Fructus group showed significantly enhanced lipid-regulating effects (P < 0.05). Metabolomics analysis indicated that the enhanced anti-inflammatory and anti-oxidative stress effects following the combination of Coptidis Rhizoma with Mume Fructus were associated with the modulation of primary bile acid biosynthesis pathway, the enhanced lipid-regulating effect following the combination of Coptidis Rhizoma with Evodiae Fructus was linked to the regulation of linoleic acid metabolism pathway. Conclusion Coptidis Rhizoma, whether combined with Mume Fructus or Evodiae Fructus, exerts definite interventional effects on cholestatic rats, demonstrating the differential characteristics of “purgation with sour herbs” and “tonification with pungent herbs”. The modulation of primary bile acid biosynthesis pathway and linoleic acid metabolism pathway represents the potential mechanism underlying these differential effects.

Objective To investigate the mechanism of Xuezhikang Capsules (血脂康胶囊) in intervening coronary microvascular dysfunction (CMD) in ApoE^−/− mice based on non-targeted metabolomics. Methods After seven weeks of high-fat diet feeding, SPF-grade ApoE^−/− mice were injected with sodium laurate via tail vein to establish CMD model. The mice were then randomly divided into model group, atorvastatin (6 mg/kg) group, Xuezhikang low-, medium-, and high-dose (90, 180, 360 mg/kg) groups, another 12 C57BL/6J mice were selected as the control group. After eight weeks of drug intervention, the morphology and pathological changes of myocardial tissue and myocardial microvessels in mice were detected using hematoxylin-eosin (HE), Carstairs and Heidenhain staining. Cardiac functional injury was evaluated by echocardiography. Oil red O staining was used to evaluate the degree of lipid accumulation in the aorta. Lipid levels, myocardial injury and endothelial function-related indicators were detected by biochemical assays, ELISA, immunohistochemistry and immunofluorescence. The changes in fecal metabolites were analyzed using non-targeted metabolomics, and differential metabolites and significantly enriched metabolic pathways were identified. Western blotting was used to detect silent information regulator 1 (SIRT1), peroxisome proliferator activated receptor γ coactivator-1α (PGC-1α), p-p65 and p65 protein expressions in myocardial tissue. Results Compared with model group, Xuezhikang Capsules significantly reduced the levels of triglycerides, total cholesterol, low-density lipoprotein cholesterol, as well as the degree of myocardial and endothelial injury in serum of mice (P < 0.05, 0.01), and significantly improved the degree of microthrombus, myocardial ischemia and heart function decline in mice (P < 0.01). Non-targeted metabolomics identified 14 differentially expressed metabolites and 15 enriched pathways (P < 0.05). Western blotting results showed that compared with model group, Xuezhikang Capsules significantly up-regulated the protein expressions of SIRT1 and PGC-1α in myocardial tissue (P < 0.01), and down-regulated the protein expression of p-p65/p65 (P < 0.01). Conclusion Xuezhikang Capsules could regulate blood lipids, improve endothelial dysfunction and inflammatory response, thereby effectively improving cardiac function and reducing myocardial structural damage in CMD model mice. Its mechanism may be related to the regulation of arginine-proline-α-ketoglutarate metabolic axis and eNOS/SIRT1/PGC-1α signaling pathway.

Objective To investigate the therapeutic effect of ethanol extract of bran-fried Cangzhu (Atractylodis Rhizoma) (EBAR) on bile duct ligation-induced liver fibrosis in mice and explore the mechanism of its treatment for liver fibrosis from the perspective of regulating hepatic metabolites. Methods C57BL/6 mice were subjected to bile duct ligation to establish a liver fibrosis model and treated with EBAR for 14 d. Levels of liver function markers and inflammatory cytokines in serum were measured. The expressions of α-smooth muscle actin (α-SMA), collagen type I (COL1A1) and COL4A2 in liver tissues were detected by immunohistochemistry, qRT-PCR and Western blotting. JS1 cells were induced with transforming growth factor-β (TGF-β) to establish a hepatic stellate cell activation model. After EBAR intervention, the expressions of α-SMA, COL1A1, COL4A2 and tissue inhibitor of metalloproteinases 1 (TIMP1) were examined by immunofluorescence, qRT-PCR and Western blotting. Network pharmacology, molecular docking, molecular dynamics simulation and metabolomics were employed to investigate the potential mechanisms of EBAR against liver fibrosis, and key mechanisms were validated by Western blotting. Results In the liver fibrosis mouse model, EBAR significantly reduced liver index, serum liver function indicators and inflammatory cytokine levels (P < 0.05, 0.01), significantly inhibited the expressions of α-SMA, COL1A1 and COL4A2 in liver tissues (P < 0.05, 0.01), significantly alleviated hepatic fibrous tissue hyperplasia and collagen deposition and improved pathological damage in liver tissue. Network pharmacology analysis revealed that Atractylodis Rhizoma exerts anti-hepatic fibrotic effects primarily by regulating signaling pathways such as adenosine 5’-monophosphate-activated protein kinase (AMPK). Metabolomic analysis showed that EBAR affected the generation of various metabolites in liver tissues of liver fibrosis mouse model, particularly metabolites related to the citric acid cycle and fatty acid metabolism. Further mechanistic studies demonstrated that EBAR exerted anti-hepatic fibrotic effects by promoting AMPK phosphorylation (P < 0.05, 0.01) and activating AMPK/silent information regulator 1 (SIRT1)/peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) signaling pathway. Conclusion EBAR exerts anti-fibrotic effects by activating AMPK/SIRT1/PGC-1α signaling pathway and regulating hepatic energy metabolism.

Objective To investigate the effect and mechanism of Yaobitong Capsules (腰痹通胶囊, YBT) on deep vein thrombosis (DVT). Methods Multi-scale evaluation of the effect of YBT was conducted using a rat lower limb deep vein ligation model, a zebrafish blood stasis model induced by phenylhydrazine, and an oxygen glucose deprivation (OGD)-induced human umbilical vein endothelial cell (HUVEC) injury model. In the rat lower limb deep vein ligation model, the ratio of thrombus weight to length and hematoxylin-eosin (HE) staining were used to evaluate the pathological changes of thrombus. Six coagulation parameters, plasma tissue factor (TF), endothelin-1 (ET-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), malondialdehyde (MDA), hypoxia inducible factor-1α (HIF-1α) and other indicators were detected. In the zebrafish blood stasis model, the stasis of red blood cells and platelet aggregation were assessed via o-dianisidine staining and confocal microscopy, respectively. qRT-PCR was employed to measure the expressions of coagulation-related genes (fga, fgb, fgg, f2). In the OGD induced HUVEC cell injury model, qRT-PCR, Western blotting and immunofluorescence were used to detect the expressions of TF, VCAM-1, ET-1, HIF-1α, as well as the phosphorylation and nuclear translocation of nuclear factor-κB (NF-κB) p65. Results In the rat model of deep vein ligation of the lower limbs, YBT could reduce the ratio of thrombus weight to length, improve histopathological changes (P < 0.05), prolong activated partial thromboplastin time (APTT) (P < 0.001), reduce fibrinogen (FIB), D-dimer, fibrinogen and fibrin degradation products (FDP), TF, ET-1, sVCAM-1, IL-6, IL-1β, TNF -α, MDA and HIF-1α levels (P < 0.01, 0.001), and increase SOD activity (P < 0.05, 0.01). In the zebrafish blood stasis model, YBT could significantly increase cardiac erythrocyte staining intensity and circulating platelet number (P < 0.001), and down-regulate the expressions of coagulation related genes (P < 0.01, 0.001). In the OGD induced HUVEC cell injury model, YBT could down-regulate the mRNA expressions of TF, VCAM-1, ET-1 and HIF-1α (P < 0.05, 0.01, 0.001), down-regulate the protein expressions of TF and HIF-1α (P < 0.05, 0.01, 0.001), inhibit the phosphorylation and nuclear translocation of NF-κB p65 (P < 0.05, 0.01). Conclusion YBT may effectively inhibit DVT by inhibiting HIF-1α/NF-κB p65/TF axis (down-regulating HIF-1α expression, inhibiting NF-κB p65 phosphorylation and nuclear translocation, thereby reducing TF overexpression), alleviating the hypoxia inflammation cascade, protecting endothelial function, and improving hypercoagulability.