Objective To investigate trinorcycloartane triterpenoids from the stems and leaves of Astragalus membranaceus (AMSL). Methods Comprehensive separation and purification were performed using silica gel, ODS column chromatography, and high-performance liquid chromatography (HPLC). The structures of the compounds were identified by modern spectroscopic techniques. The cytotoxic activities of the compounds against PC12 cells were assessed using the MTT assay. Results Two trinorcycloartane triterpenoids were isolated from the n-butanol fraction of the 70% ethanol extract of AMSL, namely 1α,3β,12β,16β-tetrahydroxy-25,26,27-trinorcycloartan-24-oic acid methyl ester (1) and 1α,3β,12β-trihydroxy-25,26,27- trinorcycloartan-24,16β-olide (2). The MTT assay showed that compounds 1 and 2 exhibited varying degrees of cytotoxicity against PC12 cells within the concentration range of 5—100 μmol/L. Conclusions Compounds1 and 2 are identified as new trinorcycloartane triterpenoids, named cyclastraine K (1) and cyclastraine L (2), respectively. Both compounds 1 and 2 demonstrate cytotoxicity against PC12 cells.

Objective To investigate the chemical constituents and their anti-inflammatory activity of Cynanchum wilfordii. Methods Various chromatographic techniques including silica gel, ODS, and Sephadex LH-20 column chromatography were employed for the isolation and purification of compounds. Their structures were elucidated through comprehensive spectroscopic analysis including HR-ESI-MS, ESI-MS, ¹H-NMR, ¹³C-NMR, HSQC, and HMBC. The anti-inflammatory activity of the compounds was evaluated using the Griess method. Results Two compounds were isolated and identified as (7'R,8'S)-dehydrodiconiferyl alcohol 9'-O-β-D-xylopyranoside (1) and 3β-[(O-β-D-glucopyranosyl)-oxy]-8β,14β-trihydroxycarda-5,20-(22)-dienolide (2). Compounds 1 and 2 inhibited NO production by 55.16% and 15.34%, respectively, with a median inhibition concentration (IC₅₀) of 41.37 μmol/L for compound 1. Conclusion Compounds 1 and 2 are both new compounds, belonging to lignan and cardenolide types, respectively. They have been assigned the names cynawilforin A (1) and wilforcardenoside A (2). In vitro anti-inflammatory assays indicated that compound 1 exhibits certain anti-inflammatory activity.

Objective To investigate the secondary metabolites produced by marine-derived fungi Talaromyces sp. 13-z-02 and their anti-inflammatory activities were studied. Methods The isolation and purification of compounds were performed using silica gel and ODS column chromatography, semi-preparative HPLC. And their structures were elucidated by MS and NMR analyses. The isolated compounds were evaluated for their anti-inflammatory activity in zebrafish. Results One new isochromane and eleven known compounds were isolated and identified as 8-hydroxy-6-methoxy-7-methylisochromane (1) from the fermentation broth of this strain, bacillisporin B (2), macrosporusone D (3), bacillisporin F (4), 1-epi-bacillisporin F (5), macrosporusone E and 1-epi-macrosporusone E (6), bacillisporin A (7), xenoclauxin (8), Sch 725680 (9), aculene C (10), 6-ethyl-2-hydroxy-4-methoxy-3-methylbenzaldehyde (11), 6-ethyl-2,4-dihydroxy-3-methylbenzaldehyde (12). Compounds 1 and 12 significantly reduced the migration of immune cells with the same concentration of 20 μmol/L and exhibited anti-inflammatory activity. Conclusion Compound 1 is a new isochromane, designated as talaroslisochromane. Compounds 1 and 12 showed anti-inflammatory activity.

Objective To investigate the chemical constituents of Mallotus repandus. Methods The compounds were separated and purified using a variety of chromatographic techniques, including silica gel and ODS column chromatography, as well as semi-preparative HPLC. Their structures were established by NMR and mass spectroscopy analysis. The antimicrobial activities of these isolates were evaluated by the broth microdilution method. Results Twenty-nine compounds were obtained from the dichloromethane extract of the twigs and leaves of M. repandus and identified as 3-O-cis-caffeoyl betulinic acid (1) , ergosta-4,6,8(14),22-tetraen-3-one (2), β-sitostenone (3), 5,8-epidioxyergosta-6,9(11),22-trien-3-ol (4), N(N'-benzoyl-S^*-phenylalaninyl)-S^*-phenylalaninolbenzoate (5), aurantiamide acetate (6), isoechinulin A (7), 6-geranylnaringenin (8), questin (9), 1,3,8-trihydroxy-6-methyl-9,10-anthracenedione (10), flavoglaucin (11), isodihydroauroglaucin (12), isoaspergin (13), 5'-cis-isotetrahydroauroglaucin (14), (E)-6-hydroxy-7-(3-methyl-2-butenyl)-2-(3-oxobut-1-enyl)chroman-5-carbaldehyde (15), camellia chlorophyllide Ⅱ (16), 13²(R)-hydroxypheophorbide-a ethyl ester (17), ethyl pheophorbide a (18), ligulariaphytin A (19), (−)-α-tocospirone (20), α-tocospiro B (21), δ-tocopherol (22), α-tocopherolquinone (23), 5-methoxypinosylvin (24), dehydroabietic acid (25), (5E,7E)-9-oxooctadeca-5,7-dienoicacid (26), (10E,12E)-ethyl-9-oxo-octadeca-10,12-dienoate (27), (10E,12Z)-9-hydroxy-10,12-octadecadienoic acid (28) and palmitic acid (29), respectively. Compound 13 has a minimum inhibitory concentration (MIC) of 128 μg/mL against Enterococcus faecalis, 16 μg/mL against methicillin-resistant Staphylococcus aureus (MRSA), and 4 μg/mL against Cryptococcus neoformans. Compound 15 has an MIC of 128 μg/mL against E. faecalis, 64 μg/mL against MRSA, and 8 μg/mL against C. neoformans.Conclusion Compounds 2, 4, 8, 9, 11—19, and 24—28 were reported from Euphorbiaceae plants for the first time, compounds 1, 3, 5, 7, 20, 21, and 23 were reported from the genus Mallotus for the first time, and compounds 6, 10, and 22 were reported from M. repandus for the first time. Compounds 13 and 15 have notable inhibitory effects on E. faecalis, MRSA, and C. neoformans.

Objective To analyze the correlation between the content changes of characteristic components and its antithrombotic effect in different proportions of the Danshen (Salviae Miltiorrhizae Radix et Rhizoma, SMRR)-Honghua (Carthami Flos, CF) herb pair, and optimize the compatibility ratio of SMRR and CF for promoting blood circulation and removing blood stasis. Methods Fingerprint profiles and quantitative determination methods for characteristic components were established for SMRR-CF herb pair at different ratios (1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1, 5:1). The HPLC combined with a comprehensive scoring method was used to determine the optimal extraction process of the SMRR-CF herb pair. The anti-thrombotic activity of the SMRR-CF extracts at different ratios was measured using zebrafish as a model organism. The principal component analysis (PCA) method was used to integrate the content of characteristic components and the intensity of antithrombotic activity, and the optimal ratio of SMRR and CF was comprehensively evaluated from two aspects of composition and efficacy. Results The fingerprint profiles of the SMRR-CF herb pair in nine different ratios collectively identified 17 common peaks, among which nine characteristic components were characterized. These include hydroxysafflor yellow A (peak 2), luteoloside (peak 3), kaempferol-3-O-rutinoside (peak 4), quercitrin (peak 5), salvianolic acid B (peak 8), dihydrotanshinone I (peak 12), cryptotanshinone (peak 14), tanshinone I (peak 15), and tanshinone II_A (peak 17). The optimal extraction process was determined to be maceration with 10 times the amount of 70% ethanol for 1 h by reflux. The content determination showed that the characteristic components changed significantly after the compatibility of SMRR and CF compared with the single herb. When the SMRR-CF ratio was 1:5, decoction yielded the highest concentrations of salvianolic acids and tanshinones derived from SMRR. Conversely, ratios of 3:1, 4:1 and 5:1 produced the greatest amounts of hydroxysafflor yellow A, luteoloside and kaempferol-3-O-rutinoside from CF, indicating that co-decoction facilitates the extraction of bioactive constituents from both herbs. Pharmacodynamic evaluation revealed that the 3:1 ratio exhibited the strongest antithrombotic activity, achieving a thrombosis inhibition rate of 43.7%. Principal-component analysis of the “composition-effect” dataset ranked the comprehensive indices (F-values) as 3:1 > 4:1 > 5:1 > 2:1 > 1:1 > 1:5 > 1:4 > 1:2 > 1:3, confirming that the 3:1 ratio represents the optimal compatibility of the SMRR-CF pair. Conclusion In this study, chemical profiling and in vivo pharmacodynamic evaluation were integrated to elucidate the quantitative relationship between the dynamic changes of characteristic constituents and the antithrombotic efficacy of the SMRR-CF herb pair across different compatibility ratios. An optimal ratio was identified through a “composition-effect”-based comprehensive assessment. These findings not only establish a foundation for subsequent dose-effect investigations of this classic combination, but also offer a rational reference for determining prescription design in latent prescriptions.

Objective By systematically analyzing the polyphenolic composition content, astringency intensity, astringency mechanism, and their inherent correlations in Hezi (Chebulae Fructus, CF) pieces with different comminution degrees, this study aims to lay a foundation for its further development and research. Methods The CF was processed into six granule sizes (original granule, extra large granule, large granule, median granule, small granule, fine powder) for sampling. HPLC was employed to quantify the contents of chebulagic acid, gallic acid, chebulinic acid, ellagic acid, corilagin, and 1,3,6-tri-O-galloyl-β-D-glucose. The main chemical composition groups and differences of six kinds of particle size CF samples were analyzed by fingerprint. The astringency intensity of the samples was sensorially evaluated using a visual analogue scale (VAS). These data were utilized to analyze the correlations among comminution degree, polyphenolic composition, and astringency intensity. Furthermore, SDS-PAGE technology was applied to elucidate the underlying astringency mechanism through the protein interaction. Results The total content of six kinds of polyphenolic components displayed significant variation across different granule sizes in the following descending order: median granule > extra large granule > fine powder > large granule > small granule > original granule. The results of fingerprint analysis showed that the main chemical composition groups of the six samples were basically the same. Sensory evaluation revealed that different comminution degrees considerably influenced the perceived astringency intensity, which decreased sequentially as: fine powder > small granule > median granule > large granule > extra large granule > original granule. Correlation analysis identified gallic acid and corilagin as key variables linking comminution degree with astringency perception. SDS-PAGE confirmed that the astringency mechanism primarily involves binding interactions between polyphenolic components and salivary proteins, with the fine powder demonstrating the strongest protein-binding capacity. Conclusion Comminution degree serves as a critical processing parameter governing the dissolution of polyphenolic components and astringency intensity in CF. The sample with a particle size of 4.75—6.70 mm demonstrated the highest content of polyphenolic compounds, whereas the sample with a particle size of 0.15—0.18 mm elicited the most intense perception of astringency. Processing CF to a median granule size (4.75—6.70 mm) is therefore identified as the optimal strategy for balancing polyphenolic component dissolution and oral compliance. This study provides key scientific evidence to support the standardization of CF processing techniques and related product development.

Objective To investigate the relationship between the physical properties of Jinzhen Oral Liquid (金振口服液) intermediates and their total solid content (TSC) and the contents of effective components, and to establish a content prediction model, thereby providing a reference for rapid quality evaluation of intermediates. Methods Samples were collected from two types of extracts (mineral-botanical and artificial cow bezoar) and four key manufacturing steps (hot mixing, refrigeration, centrifugation, and filling). Six physical parameters (density, viscosity, surface tension, electrical conductivity, refractive index, and pH value) were measured, alongside the contents of six effective components (baicalin, wogonoside, glycyrrhizic acid, gallic acid, hyodeoxycholic acid, and cholic acid) and TSC. Correlation analysis was performed, and polynomial regression models were developed for the overall process, the mineral-botanical extract, and the artificial cow bezoar extract, respectively. Results Correlation analysis indicated that most physical parameters were significantly correlated with chemical components throughout the entire preparation process. Refractive index showed a strong correlation with TSC (r = 0.845), while density was correlated with TSC and baicalin with coefficients of 0.529 and 0.505, respectively. In mineral-botanical extracts, the correlation coefficient between refractive index and TSC was 0.525. In artificial cow bezoar extracts, refractive index was correlated with TSC, hyodeoxycholic acid, and cholic acid, with coefficients of 0.759, 0.729, and 0.593, respectively. The regression models established based on refractive index performed well in validation: the model for TSC in the full process showed a mean relative error (MRE) of 4.04% and a mean absolute error (MAE) of 1.10%. The MRE values for the baicalin and wogonoside models were both below 10%, and both passed the F-test. The MRE values for the TSC models in both mineral-botanical and artificial cow bezoar extracts were below 6%. Conclusion The refractive index of Jinzhen Oral Liquid intermediates shows significant correlation with both TSC and the contents of effective components. The refractive index method is a feasible approach for predicting TSC and the contents of baicalin and wogonoside during the manufacturing process, as well as TSC in both extracts. This study provides a novel strategy for the rapid quality assessment of Jinzhen Oral Liquid production and offers a technical basis for the practical application of online refractometry.

Objective To optimize the optimal processing technology of different processed products of Huangqin (Scutellariae Radix, SR) [fried Scutellariae Radix (fSR), Scutellariae Radix charcoal (SRc), wine Scutellariae Radix (wSR)], and explore the correlation between sensory traits such as appearance, color, odor, taste, and the main active ingredients. Method The multi-index response surface methodology (RSM) was used, with baicalin, baicalein, wogonoside, wogonin, and appearance trait scores as evaluation indicators. The analytic hierarchy process and entropy weight method were used to calculate the weight coefficients and comprehensive scores of each indicator. Further utilize central combination design (CCD)-RSM to investigate the effects of temperature and processing time (soaking time and the alcohol ratio) on the processing technology. Using electronic sensory technology (electronic eye, electronic nose, electronic tongue) to analyze the response values of samples with different processing levels, and exploring the correlation between sensory traits (color L^*, a^*, b^* value, odor sensor response value, taste sensor response value) and chemical composition through correlation analysis. Results The optimal process parameters were determined as follows: fSR at 205.4 ℃, 6.5 min, SRc at 227.4 ℃, 16 min, and wSR at 180 ℃, 5 min (moistened for 50 min, with a yellow wine dosage of 18.2%). Correlation analysis shows that different degrees of processing of SR have distinguishable sensor response patterns: odor sensors S7 and S10 and taste sensors P6 and P10—P12 in fSR have a strong positive correlation with baicalin and wogonoside; In SRc and wSR, there is a strong negative correlation between multiple sensors and baicalin, while wogonin shows a strong positive correlation with multiple sensors SRc and a strong negative correlation with fSR. The electronic eye color values (L^*, a^*, b^*) are significantly correlated with the degree of processing (ΔE^*_ab > 1.5). Conclusion Optimizing the process is stable and feasible. The quality evaluation system based on electronic sensory technology can objectively distinguish the degree of processing, and the correlation between characteristics and components revealed provides theoretical basis and innovative methods for determining the endpoint of processing and quality control.

Objective To optimize the formulation process of co-loaded nanoparticles (TBF-HQD-SAN NPs) comprising self-assembled nanoparticles of Huangqin Decoction (黄芩汤) (HQD-SAN) and terbinafine (TBF). Methods The HQD-SAN was obtained with high-speed centrifugation combined with dialysis and then TBF was further loaded to prepare TBF-HQD-SAN NPs. Based on the single-factor investigation, taking the concentration of HQD-SAN, the speed of magnetic stirring and the stirring time as the investigation factors, and the drug loading (DL) of TBF, the encapsulation efficiency and the DL of baicalin as the investigation indicators, the formulation and process of TBF-HQD-SAN NPs were optimized by using the 3-factor 3-level Box-Behnken design (BBD)-response surface methodology (RSM). The optimized TBF-HQD-SAN NPs were characterized for morphology, particle size distribution, ζ potential, and drug-loading capacity. Their saturated solubility was determined, and their antifungal activity against Trichophyton rubrum, T. mentagrophytes, and Microsporum canis was investigated. Results The TBF-HQD-SAN NPs process was optimized by BBD. The established quadratic regression model had excellent fit (all R² > 0.99), and the concentration of HQD-SAN, stirring speed, stirring time and some interaction terms had significant effects on the indicators (P < 0.05). The effect surface analysis showed that the drug loading of TBF decreased with the increase of HQD-SAN concentration and first increased and then decreased with the stirring speed/time. The encapsulation rate/drug loading of baicalin increases with the increased of HQD-SAN concentration and decreased with the extension of stirring speed/time. The optimal process for model selection was as follows: HQD-SAN concentration 5.6 mg/mL, TBF 5 mg, distilled water 4 mL, ultrasonic (50 W, 40 kHz) for 30 min, magnetic stirring at 760 r/min for 1.5 h, and filtration through 0.8 μm filter membrane. The measured values of each index in the verification experiment were close to the predicted values (RSD < 5%). The particle size of the NPs was (185.10 ± 1.73) nm, the polydispersity index (PDI) was 0.22 ± 0.01, and the ζ potential was (−15.17 ± 1.40) mV. The encapsulation efficiency and DL of TBF were (99.81 ± 0.33)% and (3.32 ± 0.09)%, respectively, while those of baicalin were (58.59 ± 1.42)% and (6.71 ± 0.15)%, respectively. The equilibrium solubility was (2.27 ± 0.09) μg/mL for TBF alone, (15.70 ± 1.66) μg/mL for the TBF-HQD-SAN physical mixture (PM), and (78.20 ± 2.22) μg/mL for the TBF-HQD-SAN NPs. The antifungal experiment showed that the antifungal activity of TBF-HQD-SAN NPs (with MIC values ranging from 0.15—0.31 µg/mL, equivalent to 4.98—10.13 ng/mL based on TBF content) was significantly superior to that of HQD-SAN (MIC value 1.56—3.13 mg/mL) and TBF alone (MIC value 0.06—0.50 µg/mL) (P < 0.05). Conclusion The BBD successfully optimized the formulation and preparation process of TBF-HQD-SAN NPs. The resulting NPs demonstrated uniform particle size, excellent drug-loading performance, and significantly enhanced antifungal efficacy, laying a foundation for further research.

Objective To investigate the effects of calcination temperature on the phase composition and chemical constituents of Shijueming (Haliotidis Concha, HC), optimize its processing methodology, and conduct comparative analysis and health risk assessment of heavy metal content in HC and calcined HC. Methods Thermal effects during processing at different calcination temperatures were examined via thermogravimetric-differential thermal analysis (TG-DTA). Primary phase transformations were analyzed using X-ray diffraction (XRD), while compositional changes were assessed by Fourier transform infrared spectroscopy (FTIR) to preliminarily screen the key factor “calcination temperature” affecting HC processing. Simultaneously, the optimal processing parameters for HC were determined using the AHP-CRITIC mixed entropy method combined with orthogonal experiments, with CaCO₃ content, crispness, and color as evaluation criteria. Calcined HC were processed according to the optimized method. Inductively coupled plasma mass spectrometry (ICP-MS) was employed to analyze 20 batches of HC and calcined HC for Pb, Cd, As, Hg, and Cu in 20 batches of HC and calcined calcined HC. Using HC powder as a reference, theoretical maximum limits for these five heavy metals and toxic elements were calculated. Daily exposure and target hazard quotient (THQ) were further assessed to evaluate health risks, providing a reference for quality and safety evaluation of HC. Results Preliminary screening of calcination temperatures indicated an optimal range of 400—600 ℃. Using the AHP-CRITIC mixed entropy method, the optimized calcination process for HC was determined as follows: Crush HC to a particle size < 1 cm, place in a muffle furnace, set the temperature to 600 ℃, and calcine for 90 min. After calcination, the organic matter content of HC decreased while calcium carbonate content increased. The calcined HC showed reduced levels of As and Hg, with minimal changes in Cu, Cd and Pb. Health risk assessment indicated that all 20 batches of HC and calcined HC posed no significant health hazards to humans. Notably, the As and Hg levels in the slices were significantly lower than those in the raw material (P < 0.05). Conclusion By integrating phase and compositional changes before and after processing, this study employed the AHP-CRITIC mixed entropy method to scientifically and comprehensively optimize the calcination process for HC. The screening results are scientifically reliable, providing new scientific evidence for quality management and clinical application of HC and calcined HC. Conducting health risk assessments of heavy metals and harmful elements in HC before and after processing provides reference for the safe clinical use of HC and other shell-derived Chinese medicinal materials.

Objective To investigate the therapeutic effects and potential mechanisms of tetramethylpyrazine (TMP) in a high humidity osteoarthritis (HHOA) rat model. Methods The HHOA rat model was established, and rats were given TMP for eight weeks. Safranin O-fast green staining was used to observe morphological changes in cartilage tissue. The levels of matrix metalloproteinase-9 (MMP-9), tissue inhibitors of metalloproteinase-1 (TIMP-1), prostaglandin-endoperoxide synthase 2 (PTGS2), glutathione peroxidase 4 (GPX4), glutathione (GSH), malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were measured. Prussian blue staining was performed to detect iron deposition in cartilage. Immunohistochemistry was used to assess the expressions of GPX4 and acyl-CoA synthetase long-chain family member 4 (ACSL4) in cartilage. Serum untargeted metabolomics was performed to analyze changes in metabolites and their enriched pathways. In vitro, a ferroptosis model of chondrocytes was established and treated with main metabolite N-acetylserotonin. GSH/oxidized glutathione (GSSG) value, MDA and SOD levels were measured, the expressions of GPX4 and ACSL4 were measured by Western blotting. Results Compared with HHOA group, levels of MMP-9, PTGS2 and MDA in serum of rats in TMP group were significantly reduced (P < 0.05, 0.01), while the levels of TIMP-1, GPX4, SOD and GSH in serum were significantly increased (P < 0.05, 0.01). The pathological damage to the articular cartilage was significantly improved, and the deposition of iron ions in articular cartilage was reduced, GPX4 protein expression was increased, and ACSL4 protein expression was reduced. Metabolomics analysis showed that 2 695 differential metabolites were screened between HHOA group and sham group, with significantly more down-regulated metabolites than up-regulated metabolites; A total of 2 846 differential metabolites were screened between TMP high-dose group and HHOA group, with up-regulated metabolites being the main ones. KEGG pathway enrichment analysis results showed that the differential metabolites between HHOA group and sham group were mainly enriched in multiple metabolic pathways such as carbon metabolism and amino acid metabolism, the differential metabolites between TMP group and HHOA group were mainly enriched in multiple pathways closely related to amino acid metabolism and substance transport. Compared with sham group, the levels of γ-glutamylleucine, 3-indoolepropionic acid and N-acetylseletonin in serum of HHOA group rats were significantly reduced (P < 0.05). Compared with HHOA group, the levels of γ-glutamylleucine, 3-indoolepropionic acid and N-acetylseletonin in TMP high-dose group were significantly increased (P < 0.05). The in vitro experimental results showed that the main metabolite N-acetylserotonin significantly increased the GSH/GSSG value and SOD activity in ferroptosis model of chondrocytes (P < 0.05), decreased MDA level (P < 0.05), up-regulated GPX4 protein and down-regulated ACSL4 protein expressions (P < 0.05, 0.01). Conclusion TMP could improve the metabolomic profile of HHOA rats, regulate the tryptophan metabolism pathway represented by N-acetylserotonin, alleviate GPX4/ASCL4 axis mediated lipid peroxidation and iron ion deposition, thereby reducing ferroptosis in chondrocytes and delaying the progression of HHOA.

ObjectiveTo investigate the intervention effect and related mechanism of hyperoside on sepsis-associated acute kidney injury (SAKI) in mice. Methods A mouse model of SAKI was established by cecal ligation and puncture (CLP). Sham group, model group, dexamethasone (1 mg/kg) group, hyperoside high-, low-dose (50, 25 mg/kg) groups and hyperoside (50 mg/kg) + nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor ML385 (30 mg/kg) group were set up. After 7 d of drug intervention, the survival curve of mice was plotted. The levels of renal function, inflammatory factors and oxidative stress were detected. The pathological changes of renal tissue were observed by hematoxylin-eosin (HE) staining. TUNEL staining was used to detect apoptosis in renal tissue cells. Immunohistochemical was used to detect Nrf2 expression in renal tissue. Western blotting was used to detect the expressions of Nrf2, PTEN induced kinase 1 (Pink1) and Parkin proteins in renal tissue. Immunofluorescence was used to detect the co-localization of microtubule associated protein 1 light chain 3B (LC3B) and translocase of the outer mitochondrial membrane 20 (TOMM20) in renal tissue. Lipopolysaccharide (LPS) was used in vitro to induce injury in human tubular epithelial cells (HK-2). After intervention with hyperoside, cell viability, apoptosis and reactive oxygen species levels were detected. Mitochondrial ultrastructure was observed by transmission electron microscopy. Flow cytometry was used to detect mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) levels to evaluate mitochondrial function. Western blotting was used to detect the expressions of Nrf2, heme oxygenase-1 (HO-1) and mitochondrial autophagy proteins Pink1, Parkin, p62. Results Compared with model group, hyperoside could significantly improve the survival rate of SAKI mice (P < 0.05), alleviate weight loss and renal pathological damage (P < 0.05, 0.01), improve renal function and oxidative stress response (P < 0.05, 0.01), inhibit inflammatory factor levels (P < 0.05, 0.01), up-regulate the expressions of Nrf2, Pink1, Parkin in renal tissue (P < 0.01), and increase the co-ocalization of LC3B and TOMM20 in renal tissue. In vitro experimental results showed that hyperoside (40 μmol/L) could inhibit LPS-induced apoptosis and oxidative stress in HK-2 cells (P < 0.01), increase MMP and ATP levels (P < 0.01), up-regulate Nrf2, HO-1, Pink1, Parkin protein expressions (P < 0.01), down-regulate p62 expression (P < 0.01), promote mitochondrial autophagy, and alleviate mitochondrial damage. In addition, Nrf2 inhibitor ML385 significantly reversed the protective effect of hyperoside on CLP-induced SAKI mice (P < 0.05, 0.01), while weakening hyperoside induced mitochondrial autophagy in HK-2 cells.Conclusion Hypericin may regulate Pink1/Parkin mediated mitochondrial autophagy by activating Nrf2/HO-1 signaling pathway, alleviate mitochondrial dysfunction, alleviate oxidative stress and inflammatory response in SAKI mice, and thus exert renal protective effects.

ObjectiveTo investigate the protective effect of Rosa roxburghii fermentation broth against acetaminophen (APAP)-induced drug-induced liver injury (DILI) in mice and explore its potential mechanism based on nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway. Methods Taking the R. roxburghii juice as the control, the main active component contents and in vitro antioxidant capacity of R. roxburghii fermentation broth were analyzed. A total of 60 Kunming mice were randomly divided into control group, model group, N-acetylcysteine (NAC, 0.15 g/kg) group, and R. roxburghii fermentation broth low-, medium-, and high-dose (5, 10, 20 mL/kg) groups, with 10 mice in each group. All groups were consecutively administered for 7 d, 1 h after the last administration, except for the control group, all other groups received a single intraperitoneal injection of APAP (400 mg/kg) to establish the acute liver injury model. After 12 h of modeling, the mice were anesthetized and euthanized. The intact liver of mice was immediately removed and observed for any visible damage or lesions. The body weight and liver weight were measured and liver index was calculated. Activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β in serum, as well as activity of superoxide dismutase (SOD) and levels of glutathione (GSH), malondialdehyde (MDA) in liver tissue were detected by ELISA. Hepatic pathological changes were observed by hematoxylin-eosin (HE) staining. The mRNA and protein expressions of Nrf2, heme oxygenase-1 (HO-1), SOD-1 and catalase (CAT) in liver tissue were detected by qRT-PCR and Western blotting. Neutrophil infiltration was observed by Ly6G immunofluorescence staining, and hepatocyte apoptosis was detected by TUNEL staining. Results Compared with R. roxburghii juice, the contents of total acids, total phenols and total flavonoids in R. roxburghii fermentation broth were significantly increased (P < 0.05), while the content of vitamin C was significantly decreased (P < 0.05), and the total amount of major antioxidant active ingredients were increased, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging rate and Fe³⁺ reducing ability were significantly increased (P < 0.01). Compared with control group, model group showed obvious pathological changes such as hemorrhagic injury in liver, liver index and activities of ALT, AST, levels of inflammatory factors in serum and MDA level in liver tissue were significantly increased (P < 0.001), while GSH level and SOD activity in liver tissue were significantly decreased (P < 0.001). Inflammatory cell infiltration in liver tissue, disruption of hepatic cord structure, and increased necrotic area were observed; The mRNA and protein expression levels of Nrf2, HO-1, SOD-1 and CAT in liver tissue were significantly reduced (P < 0.001); Neutrophil infiltration and hepatocyte apoptosis in liver tissue were significantly increased (P < 0.001). Compared with model group, NAC and R. roxburghii fermentation broth all dose groups could improve the above pathological and biochemical indicators to varying degrees, including reducing visible liver damage and lesions, lowering liver index, down-regulating transaminase activity and inflammatory factor levels in serum, reducing MDA level in liver tissue, increasing GSH level and SOD activity, and up-regulating Nrf2/ARE pathway related genes and protein expressions (P < 0.05, 0.01, 0.001), while significantly inhibiting neutrophil infiltration and liver cell apoptosis (P < 0.05, 0.001). Conclusion R. roxburghii fermentation broth has a potential liver-protective effect against APAP-induced DILI in mice, and its mechanism may be related to the activation of Nrf2/ARE signaling pathway, enhancing antioxidant capacity and inhibiting inflammatory responses.

Objective To construct Ginseng polysaccharide-chitosan oligosaccharide nanoparticles (GP-NPs) and investigate whether their combination with radiotherapy could promote the differentiation of natural killer (NK) cells into highly cytotoxic subsets by activating interferon-I (IFN-I)-mediated Janus kinase 3 (JAK3)/signal transducer and activator of transcription 5 (STAT5) signaling pathway, thereby enhancing the antitumor immune response against non-small cell lung cancer (NSCLC). Methods GP-NPs were prepared via the ionic cross-linking method, and their particle size, dispersibility and stability were characterized. A subcutaneous xenograft model of A549 lung cancer cells in nude mice was established, and the mice were divided into PBS group, radiotherapy alone group and GP-NPs combined with radiotherapy group. The antitumor efficacy and mechanism of GP-NPs combined with radiotherapy were comprehensively evaluated by measuring tumor volume, detecting levels of cytokines in serum via ELISA, analyzing NK cells subsets by flow cytometry, detecting the protein expressions of JAK3/STAT5 pathway through Western blotting and performing histopathological analysis. Results GP-NPs combined with radiotherapy significantly inhibited tumor growth, and the effect was significantly better than that of the simple radiotherapy group (P < 0.01). The levels of interferon-γ (IFN-γ), interleukin-15 (IL-15), granzyme and perforin in serum of combined treatment group were significantly increased (P < 0.01). Flow cytometry showed an increase in the proportion of NK cells infiltration within the tumor (P < 0.01), and a significant increase in the proportion of mature cytotoxic subgroups (CD27^-CD11b⁺) (P < 0.01). Western blotting results showed that the combination therapy significantly activated the JAK3/STAT5 signaling pathway (P < 0.01). Organizational analysis showed that the combination therapy group had increased tumor cell apoptosis, aggravated DNA damage and enhanced NK cell infiltration, with no significant toxicity observed in major organs. Conclusion GP-NPs combined with radiotherapy promote the differentiation of NK cells into highly cytotoxic subsets and enhance their antitumor function by activating IFN-I-mediated JAK3/STAT5 signaling pathway. This combination strategy not only significantly improves the efficacy of radiotherapy but also has good biosafety, providing new experimental evidence and strategic directions for the immunocombination therapy of NSCLC.

Objective To explore the mechanism of Astragalus bhotanensis (AB) against chronic pharyngitis based on network pharmacology, molecular docking, in vitro and in vivo experiments. Methods The antibacterial activity of AB extract against Staphylococcus aureus (SA), β-hemolytic streptococcus (HS), Escherichia coli (EC) and Diplococcus pneumoniae (DP) was assessed in vitro. A chronic pharyngitis rat model induced by HS was established and the pharmacological effects of AB extract was investigated. The interaction network between AB extract and chronic pharyngitis was constructed using databases such as PubChem, GeneCards, and STRING, and core targets were screened using Cytoscape software, followed by gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. Molecular docking was performed using Autodock vina software, and the predicted results were verified by Western blotting. Results AB extract showed certain antibacterial activity against SA, HS, EC and DP. Compared with model group, AB extract significantly reduced the levels of inflammatory factors in serum of rats with chronic pharyngitis (P < 0.05, 0.01, 0.001), and effectively reversed the pathological damage of pharyngeal tissue. Network pharmacology analysis suggested that AB extract may act on targets such as heat shock protein 90 alpha family class B member 1 (HSP90AB1), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), MAPK1, MAPK3, epidermal growth factor receptor (EGFR) and estrogen receptor 1 (ESR1) by regulating signaling pathways such as mitogen activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), thereby exerting therapeutic effects on chronic pharyngitis. The molecular docking results showed that oleic acid, 7β-hydroxysitosterol and astribhotin A had good binding ability with MAPK protein. Western blotting results showed that AB extract significantly reduced TLR4 protein expression and phosphorylation levels of nuclear factor-κB p65 (NF-κB p65) and p38 MAPK in pharyngeal tissue of chronic pharyngitis rats (P < 0.05, 0.01, 0.001). Conclusion AB extract alleviates HS-induced chronic pharyngitis in rats by regulating TLR4/p38 MAPK/NF-κB p65 signaling pathway.

Objective To explore the mechanism of galangin-induced apoptosis in hepatocellular carcinoma (HCC) based on network pharmacology, molecular docking and cell experiment. Methods Potential targets of galangin were predicted using SwissTargetPrediction and PharmMapper databases, HCC-related targets were collected from GeneCards, TTD and OMIM databases. The intersection of these targets was used to construct a protein-protein interaction (PPI) network via STRING database, followed by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of intersected targets using DAVID platform. Molecular docking of galangin with core targets was validated using AutoDock Vina. Human hepatocellular carcinoma PLC/PRF/5 cells were cultured in vitro, and the effects of galangin on cell viability and apoptosis were assessed using CCK-8 assay and flow cytometry. Western blotting was used to detect the expressions of epidermal growth factor receptor (EGFR)-phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway and apoptosis-related proteins. Results Network pharmacology analysis identified 142 common targets between galangin and HCC. PPI network analysis revealed that proteins such as EGFR, Akt and cystein-asparate protease-3 (Caspase-3) may serve as core targets. GO and KEGG analyses suggested that galangin primarily influenced cellular processes by regulating PI3K/Akt signaling pathway. Molecular docking results indicated galangin had strong binding affinity with targets such as EGFR. In vitro experiments confirmed that galangin significantly inhibited PLC/PRF/5 cells viability and induced apoptosis (P < 0.01, 0.001), significantly down-regulated p-EGFR and p-Akt protein expressions (P < 0.05), and significantly up-regulated cleaved Caspase-3 and Caspase-9 protein expressions (P < 0.05, 0.01). Conclusion Galangin may exert its anti-HCC effects by inhibiting EGFR-PI3K/Akt signaling pathway and inducing apoptosis in HCC cells.

Objective To investigate clinical efficacy of Jiaotai Wan (JTW, 交泰丸), and selective serotonin reuptake inhibitors (SSRIs) in patients with depression, and to explore their underlying mechanisms. Methods Based on biological sample data from a previous multicenter, randomized, active-controlled parallel clinical trial, a total of 83 depression patients were divided into three groups: JTW group (n = 25), JTW combined with SSRIs group (n = 29), and SSRIs alone group (n = 29). A mixed linear model was used to analyze changes in short-chain fatty acids (SCFAs), neuroinflammatory markers [cyclooxygenase-2 (COX-2), prostaglandin E₂ (PGE₂)] and brain-derived neurotrophic factor (BDNF) before and after treatment in each group. Spearman correlation and structural equation modeling were used to explore the relationships between post-treatment biomarkers and clinical symptoms [Hamilton depression rating scale (HAMD) scores], and potential mediating pathways. Results HAMD scores decreased significantly from baseline in all three groups (P < 0.001), but there was no significant differences between the groups. The JTW group showed a significant increase in butyrate levels (P = 0.002) and maintained higher levels of acetate and propionate. The SSRIs group was associated with increased levels of isovalerate, isobutyrate, and hexanoate. After treatment, the JTW + SSRIs group showed significant increases in isobutyrate, isovalerate, and isohexanoate levels, and a significant decrease in acetate levels. The combined group had significantly higher BDNF levels than the other two groups (P < 0.05). The PGE₂ levels in the combined group were also higher than in the JTW group both at baseline and after treatment (P < 0.05). Correlation analysis showed a strong positive correlation between PGE₂ and BDNF in the combined group after treatment (r = 0.609, P < 0.001). In the SSRIs group, COX-2 was positively correlated with HAMD score (r = 0.451, P = 0.014). In the JTW group, isohexanoate was positively correlated with BDNF (r = 0.445, P = 0.026). Mediation analysis suggested a potential indirect effect in the combined group, where PGE₂ influenced HAMD scores through BDNF (indirect effect = −0.065, P = 0.093). Conclusion JTW alone, SSRIs alone and JTW combined with SSRIs all can effectively alleviate depressive symptoms. However, they showed different patterns in regulating gut SCFAs, neuroinflammation, and neurotrophic factors. JTW combined with SSRIs may modulate branched-chain fatty acid metabolism and promote a positive association between PGE₂ and BDNF, thereby creating a distinct pathway that differs from single therapies. These findings provide preliminary evidence for understanding the differential mechanisms of various antidepressant strategies, suggesting that personalized treatment should consider patients’ specific biological characteristics.

Objective To explore the medication patterns of medicinal and food homologous traditional Chinese medicines (TCMs) in treatment of vascular dementia (VD), to elucidate their potential mechanisms using network pharmacology, and to validate these findings experimentally, providing a scientific basis for the prevention and treatment of VD. Methods Clinical literature on TCM prescriptions for VD was retrieved from China National Knowledge Infrastructure (CNKI), Wanfang, China Science and Technology Journal Database (VIP), and the China Biology Medicine databases. After comparing with the list of TCMs that are both food and medicine, we obtained a set of VD-related TCM prescriptions. We analyzed the medication patterns and screened for core herbs. Using network pharmacology, we identified the active components of the core herbs and their target points in VD treatment, followed by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses. Molecular docking was performed on core targets and major active components. Animal experiments were conducted to verify the therapeutic effects of the core herbs. Results Data mining yielded 531 TCM prescriptions that are both food and medicine, revealing three high-frequency and strongly associated herbal pairs. Huangqi (Astragali Radix), Danggui (Angelicae Sinensis Radix), Taoren (Persicae Semen), and Shanzhuyu (Corni Fructus) were selected as the core herbs for treating VD. The anti-VD mechanisms involve regulating pathways such as interleukin‑17 (IL-17) and tumor necrosis factor (TNF). The main active components, quercetin and kaempferol, showed good binding affinity with key VD targets including tumor protein p53 (TP53), protein kinase B1 (AKT1), TNF, and IL-6. Animal experiments demonstrated that the TCM prescriptions improved learning and memory in VD rats, reduced mitochondrial and synaptic damage, and downregulated the mRNA expression levels of TP53, TNF, and IL-6 in the hippocampus, as well as the positive expression of TP53. Conclusion Astragali Radix, Angelicae Sinensis Radix, Persicae Semen, and Corni Fructus are core herbs in the category of TCMs that are both food and medicine for treating VD. Their anti-VD mechanisms are related to the regulation of targets such as TP53, TNF, and IL-6, which help alleviate synaptic injury and neuroinflammation. This study provides valuable insights into the use of these herbs for VD prevention and treatment.

Objective To systematically review the overall landscape of research on hepatotoxicity associated with traditional Chinese medicine (TCM) using bibliometric methods, identify key research areas and emerging trends, and conduct a multidimensional analysis encompassing material basis, mechanisms of action, research methodologies, toxicity reduction strategies, and clinical evaluation, thereby providing reference for TCM safety research and rational application. Methods Chinese and English literature on hepatotoxicity of TCM was retrieved from the China National Knowledge Infrastructure (CNKI) and Web of Science (WOS) databases. The search period spans from January 1982 to July 2025. The CiteSpace bibliometric tool was employed to analyze the temporal distribution of published studies, core authors, key research institutions, high-frequency keywords, and emerging terms, thereby revealing research frontiers and hotspots in this field. Results A total of 552 Chinese-language articles were identified from the CNKI database, while 181 English-language articles were retrieved from the WOS database. The authors with the highest number of publications in both Chinese and English literature are Xiao Xiaohe and Wang Jiabo. The research institutions with the highest number of publications in both Chinese and English literature are Beijing University of Chinese Medicine and Shanghai University of Traditional Chinese Medicine. Bibliometric analysis indicates a steady annual increase in publications on TCM-induced hepatotoxicity. Research hotspots primarily focus on high-risk herbs (e.g., Polygonum multiflorum, Rheum officinale, Tripterygium wilfordii), toxic constituents (e.g., anthraquinones and pyrrolizidine alkaloids), and their molecular mechanisms. Further analysis revealed that research on hepatotoxicity caused by TCM has progressively established a systematic research chain spanning from fundamental mechanism analysis to clinical safety evaluation. Regarding material basis and mechanisms of action, research have focused on high-risk herbs and their active components, revealing multi-pathway synergistic mechanisms of liver injury mediated by metabolic activation, oxidative stress, immune inflammation, and apoptosis. Regarding research methodologies, research have evolved from traditional animal experiments and pathological observations to multidisciplinary approaches including cellular and molecular biology, systems toxicology, network pharmacology, and emerging organoids and liver-on-a-chip technologies. Regarding toxicity reduction strategies, a systematic toxicity reduction system has been established through the approaches of herbal processing, rational combination, formulation optimization, and personalized interventions guided by pharmacogenomics. Regarding clinical evaluation, the continuous refinement of case reporting, causality determination systems, and real-world studies has propelled TCM safety research toward evidence-based and international standards. Conclusion Current research on hepatotoxicity of TCM demonstrates a scientific progression from qualitative descriptions to quantitative analysis, and from empirical judgments to evidence-based conclusions. Future studies should deepen the elucidation of material bases and action mechanisms while strengthening the application of emerging technologies and big data mining. This will enhance the TCM safety evaluation system, providing support for rational TCM use and its modernization.

Objective To explore food and medicine homologous prescriptions for heat-clearing and detoxifying, with Lugen (Phragmitis Rhizoma) as the core, evaluate their antipyretic effect, and develop compound Phragmitis Rhizoma herbal tea. Methods A total of 181 Chinese patent medicine and formulations containing Phragmitis Rhizoma were retrieved from the Yaozh™ database. After screening, 97 formulations were included for data mining. The core composition of the herbal tea was identified through frequency statistics, cluster analysis, association rule analysis, and core complex network. The antipyretic effect and underlying mechanism were investigated using a dry yeast-induced febrile rat model. Results Data mining revealed that herbs used in Phragmitis Rhizoma-based formulations were predominantly sweet in taste and cold in property, primarily acting on the lung and stomach meridians. Eight traditional Chinese medicine combinations with high confidence and strong associations with Phragmitis Rhizoma were identified. Guided by traditional Chinese medicine theory, the core formulation was finalized as: Phragmitis Rhizoma, Gancao (Glycyrrhizae Radix et Rhizoma), Bohe (Menthae Haplocalycis Herba), Dazao (Jujubae Fructus), Maidong (Ophiopogonis Radix), Jiegeng (Platycodonis Radix), Jinyinhua (Lonicerae Japonicae Flos), and Korla fragrant pear. Animal studies showed that administration of low, medium, and high doses of the compound Phragmitis Rhizoma herbal tea significantly reduced the body temperature of febrile rats (P < 0.01). All dose groups significantly decreased serum tumor necrosis factor-α (TNF-α) levels, as well as hypothalamic prostaglandin E₂(PGE₂) and cyclic adenosine monophosphate (cAMP) levels (P < 0.01). Serum levels of interleukin‑6 (IL-6) were significantly reduced in the all groups (P < 0.05, 0.01). Conclusion The compound Phragmitis Rhizoma herbal tea can significantly reduce the body temperature of febrile rats. Its antipyretic mechanism is likely associated with decreasing serum levels of pyrogenic factors IL-6 and TNF-α, as well as reducing hypothalamus levels of pyrogenic, pro‑inflammatory, and algesic factors PGE₂ and cAMP content, there by causing a downward shift of the thermoregulatory set point.

ObjectiveTo systematically analyze the TIFY gene family members in Gastrodia elata f. glauca and explore their potential molecular mechanisms in the interaction between G. elata. f. glauca and Armillaria gallica. Methods Based on the whole-genome data of G. elata f. glauca, bioinformatics methods were used to analyze the system evolution, gene structures, and chromosome distribution. The gene expression characteristicswere detected by qRT-PCR, and the jasmonic acid (JA) content was detected by mass spectrometry. The correlation between gene expression and JA content was further analyzed. Results A total of 30 TIFY family members were identified in G. elata f. glauca, belonging to four subfamilies: TIFY, JAZ, PPD, and ZML. They have highly conserved protein motifs and similar exon/intron structures, and are unevenly distributed on seven chromosomes. The number of conserved homologous genes in G. elata f. glauca is more than that in Arabidopsis thaliana, indicating a specific expansion of the TIFY family. The cis-acting element analysis revealed that the promoter regions of the TIFY family members in G. elata f. glauca are enriched with a large number of hormone response, stress response, and growth and development-related elements. Transcriptome analysis showed that A. gallica infection induced differential expression of 19 TIFY family members in G. elata f. glauca. In the mother tubers, GeJAZ4-3, GeJAZ8, and GeJAZ1-1 were significantly upregulated, and the expression levels of multiple members were significantly positively correlated with JA content, especially the expression level of GeJAZ4-3. Conclusion The TIFY gene family in G. elata f. glauca shows specific expansion. GeJAZ4-3, as a key regulatory factor, mediates the JA signaling pathway to regulate the balance of the interaction between G. elata f. glauca and A. gallica, laying an important foundation for the analysis of the symbiotic mechanism of G. elata and facilitating disease-resistant breeding.

ObjectiveTo identify the members of the ERF gene family (TkERFs) of Trichosanthes kirilowii based on transcriptome sequencing data, and to analyze their biological functions in the growth and development of T. kirilowii, so as to provide a theoretical basis for further studying the mechanism of sex differentiation in T. kirilowii. Methods Bioinformatics methods were used to systematically identify the ERF gene family members of T. kirilowii, and the key candidate gene for sex differentiation, TkERF2, was screened for functional elucidation. The tissue expression patterns of the tissues were analyzed by real-time fluorescence quantitative PCR (qRT-PCR). The TkERF2 overexpression vector was constructed and its biological function was explored through subcellular localization and genetic transformation experiments of Arabidopsis thaliana, combined with phenotypic observation, related gene expression detection and ethylene content determination. Transcriptome and small RNA omics data were integrated to screen miRNAs that target the regulation of TkERFs and improve their regulatory networks. Results Based on transcriptome data, a total of 54 TkERF genes (TkERF1—TkERF54) were identified by open reading frame (ORF) and conserved domain analysis. It encodes a protein sequence length of 101—389 amino acids, a molecular weight of 11 580—43 310, and a theoretical isoelectric point (pI) of 4.66—10.60. Subcellular localization prediction showed that 74% of TkERFs were localized in the nucleus and 26% were distributed in chloroplasts and cytoplasm. Protein conservation motif analysis identified 10 motifs. The phylogenetic tree divides the family into eight subfamilies. TkERF2, TkERF20 and TkERF45 were identified by specific miRNA targeting. Transgenic A. thaliana experiments showed that TkERF2 significantly regulated ethylene synthesis pathway, and the expressions of auxin synthesis genes (AtYUC2, AtYUC6), auxin response factor (AtARF8) and auxin transport gene (AtPIN2) were significantly higher than those of wild type (P < 0.05), confirming that TkERF2 was involved in developmental process by positively regulating auxin synthesis and transport. Conclusion This study has completed the systematic identification and preliminary exploration of the ERF gene family, which provides a theoretical basis for analyzing the mechanism of sex determination and genetic improvement of T. kirilowii.

Objective To provide theoretical guidance for the phylogenetics, genetic diversity, and species identification of Dioscorea by integrating chloroplast genomics, codon usage bias, DNA barcoding, and machine learning technologies. Methods The chloroplast gene structures of 11 plant species were compared via IRscope and a phylogenetic tree was constructed. Codon usage bias was analyzed using tools such as CodonW and CUSP. Dioscorea materials were collected, and DNA barcode sequences were amplified and sequenced. Molecular identification was performed based on the maturase K gene (matK), photosystem b a protein gene-transfer RNA-Histidine intergenic spacer (psbA-trnH), and ribulose-bisphosphate carboxylase/oxygenase large subunit gene (rbcL) barcodes using machine learning algorithms. Results The chloroplast genomes of Dioscorea were found to be conserved and stable. Codon usage showed a significant A/T bias, with the third codon position favoring A/U endings. Natural selection was the primary factor influencing codon bias, and the common optimal codons identified were GGA and UCA. All three barcodes successfully discriminated species. Single-barcode identification using the BLOG algorithm achieved 100% success rate, while the SMO and NaïveBayes classifiers in WEKA demonstrated high identification accuracy. Conclusion The chloroplast genome of Dioscorea is conserved, and natural selection dominates its codon usage pattern. This study provides a basis and guidance for research on gene expression regulation, species identification, and resource conservation of Dioscorea.

Objective To evaluate the quality and differences among different varieties of Mume Flos by analyzing their fingerprints using chemical pattern recognition techniques, combined with quantitative analysis of characteristic components and antioxidant activity assays. Methods Ultra-high performance liquid chromatography (UPLC) was used to establish the fingerprints of different varieties of Mume Flos, combined with the chemical pattern recognition technology to explore the differences between different varieties of Mume Flos, screened for the characteristic chemical components that lead to the differences between different varieties of Mume Flos, and quantitatively analyzed them. The antioxidant activity of different varieties of Mume Flos was detected by DPPH free radical scavenging method. Results A total of 22 common peaks were identified in the fingerprints of 33 batches of Mume Flos.The similarity within the varietal group was above 0.869, while the similarity between the varietal groups was relatively low. Hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to classify the 33 batches of Mume Flos into two major groups and three subgroups, among which the green calyx group could be distinguished from the rest of the samples. The combination of OPLS-DA indicated that 12 components, including chlorogenic acid, rutin, hyperoside, isoquercitrin, isochlorogenic acid B, quercitrin, and naringenin, might be the characteristic markers leading to the differences in different varieties of Mume Flos. The results of the content determination showed that the samples from the green calyx variety group were dominant in the content of the components except naringenin. The results of antioxidant activity showed that the antioxidant capacity of green calyx variety group, the vermilion variety group, and DHH were relatively higher, and the antioxidant activities of various varieties of Mume Flos had strong correlations with chlorogenic acid, hyperoside, isoquercitrin, isochlorogenic acid B. Conclusion The content of components and antioxidant activity of different varieties of Mume Flos have some variability, among which the green calyx variety group is relatively stable. These results provide data support for exploring the variability of different Mume Flos varieties and the preference of “green calyx” for Mume Flos.

ObjectiveTo establish the method of multi-index quantification and quality difference evaluation of Zhenzhutougucao (S. tuberculata) from different producing areas. Methods HPLC method was used to establish the content determination method of chlorogenic acid, p-coumaric acid, ferulic acid, vanillic acid, amentoflavone, diosmetin, luteolin, kaempferol, quercetin, lonicerin, rutoside, β-sitosterol and stigmasterol in S. tuberculata. The mobile phase was acetonitrile-0.4 % phosphoric acid, the flow rate was 1.0 mL/min , the detection wavelength was 320 nm and 210 nm, the injection volume was 10 μL, and the column temperature was 30 ℃, water-soluble extract, total ash and acid-insoluble ash were also examined. Principal component analysis and orthogonal partial least squares discriminant analysis were performed on 18 batches of S. tuberculata by SPSS 26.0 and SIMCA14.1 software, and the quality difference markers were determined by the values of variable importance for projection. The CRITIC-TOPSIS method was used to construct the comprehensive quality evaluation model of S. tuberculata. Results The methodological investigation showed that each component showed a good linear relationship and good accuracy.The results of quantitative analysis showed that the contents of 13 chemical components in S. herba from different habitats were (0.867 ± 0.198), (0.586 ± 0.187), (1.454 ± 0.324), (0.347 ± 0.070), (8.262 ± 1.486), (3.247 ± 0.547), (2.397 ± 0.545), (4.464 ± 0.744), (3.224 ± 0.979), (0.266 ± 0.101), (1.153 ± 0.216), (0.706 ± 0.164), (0.079 ± 0.011) mg /g, respectively. The contents of water-soluble extract, total ash and acid-insoluble ash were (25.2 ± 6.4) %, (12.0 ± 2.5) % and (2.2 ± 1.1) %, respectively. The results of multivariate statistical analysis showed that 18 batches of S. herba were obviously divided into three categories. Eight quality difference markers with great contribution were screened and mentioned. According to the contribution degree, they were amentoflavone, quercetin, kaempferol, ferulic acid, luteolin, diosmetin, chlorogenic acid and β-sitosterol. The results of CRITIC-TOPSIS analysis showed that the comprehensive quality of samples S6 (from Jiangsu), S5 (from Henan), S2 (from Shandong), S4 (from Henan), S3 (from Shandong), S1 (from Shandong) and S7 (from Jiangsu) was relatively good. Conclusion There are differences in the quality of S. herba from different producing areas. The established multi-index quantification, chemometrics and CRITIC-TOPSIS method provide experimental basis for improving the quality evaluation system of S. tuberculata.

In recent years, protoplast technology has widely applied in the fields of plant genetic transformation, breeding, and plant regeneration. Related research on model plants (such as tobacco and Arabidopsis thaliana) has been in-depth, and its application in crops like forage grass, rice, and tomatoes has also become more and more mature. With the development of the economy and the increase in health demands, the market demand for medicinal plants has been continuously growing. However, the research on their breeding and genetic transformation is still insufficient, and protoplast technology can just provide a new path for this. This paper focuses on the preparation and isolation techniques of protoplasts of medicinal plants and their application scenarios, reviews their research status and challenges, and provides a reference for in-depth research on protoplasts of medicinal plants in fields such as genetic transformation, variety improvement, and plant regeneration.

In recent years, significant changes have taken place in the variety type of Citrus cultivars in China, which has profoundly impacted the Chenpi (Citri Reticulatae Pericarpium) medicinal materials. Except for Citrus reticulata ‘Chachi’, the cultivated varieties of the primary botanical source of traditional Citri Reticulatae Pericarpium have seen a severe decline in cultivation, leading to an insufficient supply of Citri Reticulatae Pericarpium medicinal materials. Consequently, peels derived from Citrus aurantium, Citrus sinensis, and other related species, collectively referred to as “Za Chenpi”, have entered the market and become the predominant commodity. This shift has a serious impact on the safety of clinical medication and the healthy development of the Citri Reticulatae Pericarpium industry. Citri Reticulatae Pericarpium are widely distributed and serve both medicinal and culinary purposes. The development of Citri Reticulatae Pericarpium industry plays an important role in driving and leading the development of China’s traditional Chinese medicine industry and rural revitalization. However, except for C. reticulata ‘Chachi’ which has successfully transformed from a single agricultural product into a billion-yuan industry, the broader Citri Reticulatae Pericarpium industry currently faces multiple challenges, including a broken chain in the cultivation sector, the use of “Za Chenpi” as a substitute in manufacturing, insufficient exploration of its health value as a food-medicine dual-purpose substance, a lack of innovation in product development, and inadequate expansion of the service sector. This article provides a systematic review of the historical origin and research progress of Citri Reticulatae Pericarpium. In response to the challenges facing the industry, and using the development model of C. reticulata ‘Chachi’ as a benchmark, this study propose strategic recommendations for Citri Reticulatae Pericarpium industry from multiple perspectives, including base construction, quality assurance, technology empowerment, product innovation, brand building, and integrated whole-industry-chain development. The aim is to provide decision-making support for the high-quality development of Citri Reticulatae Pericarpium industry.

Herbal textual research, through systematic collation and analysis of historical texts, clarifies the nomenclature and historical applications of medicinal materials, and serves as a key factor in ensuring their quality and clinical efficacy. This study primarily conducted a comprehensive textual investigation into the nomenclature, botanical origin, efficacy, harvesting and collection, processing and geo-authentic producing areas of Chaihu (Bupleuri Radix) as recorded in historical herbal literature. Simultaneously, it systematically reviewed modern ecological research on the quality formation of geo-authentic Bupleuri Radix. The study identified three major shifts in the documented efficacy of Bupleuri Radix, ultimately establishing its current recognized functions of “dispelling pathogenic factors to relieve fever, soothing the liver to alleviate depression, and elevating yang qi” through three major revolutions. Furthermore, the geo-authentic producing areas of Bupleuri Radix have remained relatively stable historically, with their distribution centered on the adjoining region of Shaanxi and Henan provinces and gradually expanding outward along the watersheds of the Yellow River and the Wei River. Concurrently, modern research, from the perspective of interactions between ecological factors and microorganisms, elucidates the “stress effect” and the multidimensional regulatory network underlying the quality formation of Bupleuri Radix, thereby providing a theoretical basis for understanding the causes of its geo-authenticity. This study interprets the scientific connotation of dao-di herbs through the cross-verification of “traditional textual research” and “scientific inquiry”. It identifies ecological factors—ranging from water, heat, and light to microorganisms—as the core elements driving the formation ofthe geo-authentic quality of Bupleuri Radix. This provides a theoretical foundation for enhancing the quality of Bupleuri Radix, promoting in-depth resource development, and guiding rational clinical application.

Diabetic nephropathy (DN) is one of the most common complications of diabetes, which may eventually lead to end-stage renal disease. Astragalus polysaccharides (APS) are the main active components of Huangqi (Astragali Radix) and have been proven to possess multiple functions such as antioxidative stress, anti-inflammation and immunomodulation in recent years. Based on the multi-dimensional pharmacological effects of scavenging reactive oxygen species and regulating adenosine monophosphate activated protein kinase signaling pathway, inhibiting the release of inflammatory factors, and regulating immune responses, APS can reduce the renal injury caused by oxidative stress in patients with DN, reduce the inflammatory response of the kidneys, improve the immune microcirculation, exhibit renal protective effects, and ultimately delay the progression of DN. However, APS are macromolecular mixtures by acidic heteropolysaccharides and dextran, with poor membrane permeability and low oral bioavailability, which limit their clinical application. This article reviews the mechanism of action of APS in treatment of DN based on their multi-dimensional pharmacological effects and summarizes the research on the formulations of APS, which provide reference and guidance for the application of APS in DN.

With the rapid development of the traditional Chinese medicine industry, a large amount of waste is produced during the processing of traditional Chinese medicine, especially Chinese herb residues, which lead to severe resource waste and environmental pressure. Meanwhile, due to its advantages of high customizability, design flexibility, and material efficiency, fused deposition modeling 3D printing technology is gaining widespread adoption across various fields. However, commonly used printing materials are often limited by high brittleness, susceptibility to cracking and denting, and lack of functionality. Containing approximately 40% cellulose, Chinese herb residues can serve as a natural reinforcing phase to enhance the mechanical propert ies of printing materials, thus enabling high-value utilization of resources. This article provides a comprehensive review of the classification and material composition of Chinese herbal residues, as well as the transformation of their disposal methods from traditional to green and environmentally friendly approaches, highlights the advantages of new Chinese herbal medicine residue composite materials, the key points for performance optimization, the preparation process, and their application potential in the development of 3D printed products. This article summarize the current challenges and future development directions in this interdisciplinary field, aiming to provide theoretical insights for advancing the high-value utilization of waste and the development of novel green printing materials.

Zhizichi Decoction (栀子豉汤) firstly recorded in the Treatise on Febrile Diseases written by Zhang Zhongjing, was included in the Catalogue of Ancient Classical Prescriptions (second batch) published by the National Administration of Traditional Chinese Medicine, which is a classic prescription for treating “restlessness due to deficiency” and “inner distress”. The formula of ZhiziChi Decoction is simple. Modern pharmacological studies have found that the active components of Zhizichi Decoction have the effects such as sedation and sleep induction, hypoglycemic action, regulation of lung tissue damage, protection of liver and myocardial cells. This classic formula is used to treat agrypnia and annoyance with heat, clinically, it has been used for treatment of neurological disorders and gastrointestinal diseases. This paper systematically summarizes and sorts out the pharmacological effects, modern clinical applications and modern research of Zhizichi Decoction, which can provide a theoretical basis for the future research and development of ZhiziChi Decoction.

Zebrafish are a critical model organism for investigating liver disease mechanisms and drug discovery because of their embryonic transparency and genetic manipulability, as well as their possession of a structurally and functionally conserved liver highly homologous to mammals. This review systematically summarizes methodologies for establishing zebrafish models of liver injury, hepatic fibrosis, and hepatocellular carcinoma, and focuses on exploring the therapeutic effects of traditional Chinese medicine (TCM) components in ameliorating liver injury, counteracting fibrogenesis, and inhibiting tumorigenesis,with the aims of providing novel perspectives and methodologies for advancing the application of zebrafish models in hepatopathology research and promoting TCM-based interventions for liver diseases.

The scale-up of traditional Chinese medicine (TCM) extraction processes is a crucial step in transitioning from laboratory research to industrial production. With the rapid development of the TCM industry, the scale-up of TCM extraction processes faces numerous key challenges, such as parameter fluctuations caused by multi-factor coupling, instability in cross-scale amplification due to unclear kinetic mechanisms, and quality variations resulting from the failure of traditional scaling-up theories. In recent years, with the rapid advancement of process analytical technology (PAT), its application in the scale-up of TCM extraction processes has garnered increasing attention. PAT enhances the efficiency and stability of TCM extraction process scale-up by real-time monitoring and analyzing key parameters during the extraction process, optimizing extraction techniques, establishing precise models, and implementing dynamic regulation and feedback control. This provides robust data support for process scale-up. This review summarizes the application of PAT in extraction process scale-up, systematically elaborates on the current research status of data acquisition, process modeling, and dynamic feedback techniques, and analyzes the current problems and challenges faced by extraction process scale-up. It aims to provide insights for promoting the application of PAT in the TCM manufacturing industry, therebyfacilitating the intelligent transformation, upgrading, and high-quality development of the TCM industry.

Ferns exhibit a wide variety of species in our country, with some possessing significant medicinal value. However, the overall development of medicinal fern research is insufficient, and the status of resources remains unclear. This article summarizes the current state of medicinal ferns in our country, as well as the research progress on artificial cultivation and propagation techniques, thoroughly analyzes the impacts of soil substrates, light, temperature, diseases and insects pests on the growth of medicinal ferns and the accumulation of their medicinal components, and systematically reviews the current status of processing methods and product research and development of ferns. The standardized cultivation and in-depth utilization of medicinal ferns warrant further attention, as they offer new prospects for the development and utilization of traditional Chinese medicine resources.