Nanophases of traditional Chinese medicine (TCM) have recently emerged as a novel structural form attracting increasing attention in the modernization of TCM. They are typically formed through the self-assembly of multiple components during decoction or extraction processes, possessing both intrinsic therapeutic activity and imaging potential, thereby providing a new class of carriers for bioimaging applications. This review systematically summarizes the structural basis and signal sources of TCM nanophases in bioimaging, and highlights their advantages, including intrinsic imaging capability, favorable biocompatibility, and multicomponent synergistic responsiveness. Furthermore, their applications are discussed in depth, particularly in the visualization of action mechanisms of TCM formulas, the investigation of tissue distribution related to meridian tropism, the quantitative visualization of TCM syndromes, and imaging-guided theranostics. Meanwhile, key challenges in this field are analyzed from several perspectives, including the correlation between imaging signals and pharmacological effects, the decoding of multicomponent synergistic interactions, the mismatch between imaging resolution and meridian structures, and quality consistency. Looking forward, with the integration of high-resolution imaging technologies and interdisciplinary analytical approaches, TCM nanophases are expected to promote the development of emerging concepts such as panoramic molecular imaging of TCM, facilitating the transition of TCM research from empirical description to visualization and quantification, and providing new technical strategies for the modern scientific interpretation of TCM theories.

Objective To investigate the trace amounts of furanocoumarin dimers from Angelica dahurica var. formosana. Methods Using an LC-MS guided separation method, the furanocoumarin dimers were isolated by silica gel, Sephadex LH-20 column chromatography, and semi-preparative HPLC. The structures of the isolated dimeric coumarins were identified by NMR spectroscopy, and their preliminarily acetylcholinesterase (AChE) inhibitory activities were evaluated. Results Three furanocoumarin dimers were isolated from the ethyl acetate fraction of the ethanol extract of A. dahurica var. formosana, namely 9-{[4-(3-methylbut-2-en-1-yl)-7-oxo-7H-furo[3,2-g]chromen-9-yl]oxy}-4-[(2-methylbut-3-en-2-yl)oxy]-7H-furo[3,2-g]chromen-7-one (1), dahuribiethrin E (2) and dahuribiethrin B (3). Conclusion Compound 1 is a new compound, namely dahuribiscoumarin A. Compounds 2 and 3 are isolated from A. dahurica var. formosana for the first time. Compound 2 exhibits acetylcholinesterase inhibitory activity with a median inhibition concentration (IC₅₀) value of (71.37 ± 1.26) μmol/L.

Objective To investigate the antifungal secondary metabolites from the rice fermentation products of the Valeriana officinalis endophytic fungus Chaetomium elatum FH-3. Methods The one strain many compounds OSMAC (OSMAC) strategy and global natural products social molecular networking (GNPS) molecular networking technology were employed for targeted separation. Chromatographic and spectroscopic methods were used for structural identification. The antifungal activity of the compounds against Colletotrichum gloeosporioides was evaluated using the mycelial growth rate method. Results A total of 12 compounds were targeted and isolated from the ethyl acetate extract of Chaetomium elatum FH-3. These compounds were identified as 4′′-methoxy-asperianas A (1), (3S,4S)-4-hydroxy-6-methoxymellein (2a), (3R,4R)-4-hydroxy-6-methoxymellein (2b), cis-4,6-dihydroxymellein (3), cis-4-hydroxymellein (4), 6-methoxymellein (5), 6,8-dihydroxy-3-methyl-3,4-dihydroisocoumarin (6), 3-methyl-6-hydroxy-8-methoxy-3,4-dihydroisocoumarin (7), xenofuranone B (8), flavipesin B (9), p-hydroxybenzaldehyde (10), and p-hydroxybenzoic acid methyl ester (11). Antifungal testing showed that compounds 1—4 exhibited significantly stronger inhibitory activity against C. gloeosporioides than the positive control carbendazim, with half-maximal effective concentration (EC₅₀) values of 15.02, 25.31, 37.89, and 47.86 μg/mL, respectively. Conclusion Compounds 1 and 2a are new butenolide and dihydroisocoumarin derivatives, named butenolide A and dihydroisocoumarin C respectively, and compounds 8 and 9 are reported for the first time from this strain. Compounds 1—9 demonstrate significant inhibitory activity against C. gloeosporioides.

Objective To investigate the chemical compositions of the ethyl acetate (EtOAc) fraction of Sophora flavescens and evaluate their anti-liver fibrosis activity. Methods The EtOAc extraction of S. flavescens was separated and purified by various column chromatography methods, including silica gel, polyamide, Sephadex LH-20 and C₁₈reversed-phase column chromatography. The structures of the isolated compounds were elucidated based on modern spectroscopic techniques and physical and chemical identification method. Transforming growth factor beta-1 (TGF-β1)-induced LX-2 cells were used as an in vitro model of hepatic fibrosis, and the anti-fibrotic activities of the compounds were evaluated using the CCK-8 assay. Additionally, cell wound scratch assay, real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting, and flow cytometry assays were employed to investigate the anti-liver fibrosis activity and mechanism of compound 14 (kushenol T). Results A total of 28 compounds were isolated from the EtOAc of S. flavescens, and they were identified as follows: ethyl 2,4-dihydroxybenzoate (1), trifohrhizin (2), 2,4- dihydroxybenzoic acid (3), kuraridin (4), kushenol N (5), kurarinol A (6), kushenol C (7), 8-isopentenyl-5-methoxy-7,2′,4′-trihydroxy flavanonol (8), 2′-hydroxy-isoxanthohumol (9), kurarinone (10), methyl 2,4-dihydroxybenzoate (11), sophoflavescenol (12), umbeliferone (13), kushenol T (14), sophophenoside B (15), 6′′-β-D-xylosegenistin (16), 3′,4′-methylenedioxyisoflavone-7-O-β-D-apiofuranosyl-(l→6)-β-D-glucopyranoside (17), ambocin (18), kushenol O (19), 4′-hydroxy-3′-methoxyisoflavone-7-O-β-D-xylopyranosyl-(l→6)-β-D-glucopyranoside (20), 3′-hydroxy-4′-methoxyisoflavone-7-O-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (21), genistein (22), isoxanthohumol (23), 8-isopentenyl-5-methoxy-7,4′-dihydroxyflavanonol (24), 2-(2¢,4¢-dihydroxyphenyl)-5,6-methylenedioxy- benzofuran (25), calycosin (26), daidzein (27), narirutin (28). Bioactivity screening revealed that compounds 9, 11—14, 17, 20, 23—25 and 28 exhibited significant inhibitory effects on LX-2 cells. Among these, 14 demonstrated the most potent activity, markedly downregulating the expression of hepatic fibrosis markers, such as α-smooth muscle actin (α-SMA), fibronectin (FN) and Collagen I, at both the gene and protein levels. Furthermore, 14 promoted cell apoptosis and suppressed TLR4 in the Toll-like receptors 4 (TLR4)/nuclear factor kappa-B (NF-κB) signaling pathway, myeloid differentiation primary response 88 (MyD88), and TGF-β-activated kinase 1 (TGF-β-activated kinase 1), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β). Conclusion Compounds 18, 25 and 28 were isolated from this species for the first time. Compounds 9, 11—14, 17, 20, 23—25 and 28 significantly inhibited LX-2 cell proliferation. Among them, compound 14 had the most significant activity and exerts an anti-liver fibrosis effect through the TLR4/NF-κB signaling pathway.

Objective To systematically analyze the blood-absorbed and brain-absorbed components of the classical formula Kaixin San (KXS) in normal and depression model rats, and preliminarily elucidate the pharmacological material basis of its antidepressant effect. Methods A depression model rat was established using the chronic unpredictable mild stress (CUMS) method, and behavioral verification was performed through open field test, novel object recognition test, sucrose preference test, and forced swimming test. The components in the serum and brain tissue from normal and model rats after intragastric administration of KXS were comprehensively analyzed by UPLC-Q-Exactive Orbitrap MS/MS. Results In the control rats, 143 (including 47 prototypes and 96 metabolites) and 11 components were identified in serum and brain tissue, respectively. In contrast, the overall number in the depression model rats significantly decreased, with only 83 (including 28 prototypes and 55 metabolites) and 4 components detected in serum and brain tissue, respectively. Among them, four prototype components were confirmed to cross the brain in serum and brain tissue samples of the CUMS model rats, including β-asarone, ginsenoside Rg₁, ginsenoside Rb₁, and polygalaxanthone III. Conclusion This study identified four key active components from the blood-absorbed and brain-absorbed components of KXS in normal and depression model rats, providing experimental evidence for screening quality markers of KXS and in-depth analysis its antidepressant mechanism.

Objective To screen the optimal purification and drug-loading processes of exosomes (Exo), construct an in vitro blood-brain barrier (BBB) cell model and an Alzheimer’s disease (AD) mouse model, and investigate the efficiency of human embryonic kidney 293T (HEK 293T) Exo-mediated ginsenoside Rg₁ (Rg₁) across the BBB, so as to provide a basis for the treatment of brain diseases. Methods Exosomes were purified using ultracentrifugation, tangential flow filtration, and dual-coupled harmonic oscillation technology. The optimal purification method was selected based on concentration, particle size, and polydispersity index (PDI). On the basis of single-factor screening of voltage, pulse duration, pulse number, and Rg₁ concentration, the electroporation process for constructing Exo-Rg₁ complexes was optimized using orthogonal design with drug encapsulation efficiency as the evaluation index. An in vitro BBB model was established using brain endothelial cell line 3 (bEnd.3). Model integrity was verified by measuring transendothelial electrical resistance (TEER), liquid level permeability, and sodium fluorescein permeability. The Exo-Rg₁ complex group (experimental group), free Rg₁ group (control group), exosome alone group, and empty exosome + free Rg₁ group were set up. The concentration of Rg₁ penetrating the in vitro BBB model at different time points was detected by HPLC, and the permeability efficiency was calculated. In vivo therapeutic effects were evaluated through behavioral tests, histological observation, and detection of protein and inflammatory factor levels in AD model mice. Results The optimal exosome purification process was dual-coupled harmonic oscillation technology, yielding an exosome concentration of 2.9 × 10¹¹ particles/mL, a particle size of (91.8 ± 1.5) nm, and a PDI of 0.21 ± 0.01. The optimized electroporation conditions were voltage of 100 V, pulse duration of 15 ms, pulse number of four times, and Rg₁ concentration of 0.4 mg/mL. Under these conditions, the encapsulation efficiency of the complexes was 61.58%, which was superior to that of the traditional ultrasonic method. The in vitro BBB model using bEnd.3 cells was successfully established, with TEER values stably above 300 Ω·cm². A significant liquid level difference was maintained in the in vitro BBB model group after 4 h. The sodium fluorescein permeability in the model group was significantly lower than that in the blank control group [(41.34 ± 1.83)% vs (98.87 ± 2.09)%, P < 0.01], meeting the requirements for barrier function. In vitro penetration experiments showed that the penetration efficiency of Rg₁ in the experimental group (15.3%) was significantly higher than that in the control group (3.3%) at 8 h (P < 0.05). Animal experiments demonstrated that the complex significantly improved learning and memory abilities in model mice, protected hippocampal neurons, upregulated the expression of choline acetyltransferase (ChAT) and α7 nicotinic acetylcholine receptor (α7 nAChR) proteins, and reduced the levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), with effects superior to those of the free Rg₁ group and the exosome-only group. Conclusion Exosomes can effectively enhance the efficiency of Rg₁ in crossing the BBB, providing an experimental basis for improving the therapeutic effect of brain diseases.

Objective To prepare paeoniflorin-glycyrrhiza protein self-assembled nanoparticles (Pae-GP/SAN) with good bioavailability and investigate its intestinal absorption mechanism. Methods In this study, glycyrrhiza protein (GP) was used as the carrier to prepare Pae-GP/SAN by ultrasonic dispersion method. The formulation and preparation process were optimized using average particle size, polydispersity index (PDI), encapsulation efficiency, and drug loading as evaluation indicators, and the prepared nanoparticles were characterized. A rat in situ single-pass intestinal perfusion model was established to investigate and compare the intestinal absorption behaviors of paeoniflorin solution (Pae/Sol), paeoniflorin-glycyrrhiza protein physical mixture (Pae-GP/PM), and Pae-GP/SAN. Additionally, an aggregation-caused quenching (ACQ) fluorescent probe was used to observe the intestinal absorption of GP/SAN by confocal laser scanning microscopy (CLSM), aiming to initially clarify its penetration-enhancing mechanism. Results The optimized Pae-GP/SAN had an average particle size of (178.2 ± 6.3) nm, a PDI of 0.152 1 ± 0.011 2, a ζ potential of (−14.91 ± 1.13) mV, an encapsulation efficiency of (36.45 ± 2.32)%, and a drug loading of (21.70 ± 1.30)%, with a uniform spherical micromorphology. Intestinal perfusion experiments indicated that the absorption efficiency of Pae-GP/SAN in the ileum was superior to that in the jejunum (P < 0.05, 0.01). The formation of nanoparticles significantly promoted the absorption of paeoniflorin. Moreover, the absorption parameters of high mass concentration Pae-GP/SAN were significantly higher than those of low mass concentration Pae-GP/SAN (P < 0.001), while mass concentration had no significant effect on the absorption of Pae/Sol and Pae-GP/PM. Transporter inhibitor intervention experiments showed that indomethacin and reserpine had no significant effect on the absorption of the three formulations. Verapamil significantly increased the absorption of Pae/Sol and Pae-GP/PM (P < 0.01, 0.001) but had no significant effect on Pae-GP/SAN. CLSM observations confirmed that GP/SAN can be absorbed by the intestine in the form of intact nanoparticles. Conclusion Pae-GP/SAN can be endocytosed and absorbed by the intestine as intact nanoparticles, effectively evading the barrier effect of P-gp efflux protein, thereby significantly improving the oral bioavailability of Pae.

Objective Vitexin liposomes (Vit-Lips), chitosan modified Vit-Lips (CS-Vit-Lips) and deoxycholic acid and chitosan co-modified Vit-Lips (DA/CS-Vit-Lips) were prepared, and their oral pharmacokinetic behavior were compared. Methods Deoxycholic acid-chitosan complex (DA/CS) was synthesized. Film-ultrasonic method was used to prepare Vit-Lips. Encapsulation rate, drug loading and particle size were used as evaluation indexes, single factor investigation combined with Box-Behnken design-response surface methodology (BBD-RSM) were used to investigate the optimal prescriptions of Vit-Lips. CS-Vit-Lips and DA/CS-Vit-Lips were prepared by introducing chitosan and DA/CS, respectively. Transmission electron microscopy (TEM) was used to observe the microstructure of the three kinds of liposomes, and their particle stability in gastrointestinal fluids were compared. Using vitexin as reference, the drug release of Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips in simulate gastrointestinal fluid were compared, and their drug release mechanism were also studied. The absorption of the vitexin and its three kinds of liposomes in different intestinal segments was investigated, and the intestinal absorption parameters were calculated. Blood samples were collected after gastric administration of vitexin and its three kinds of liposomes at a dose of 20 mg/kg (vitexin), respectively. Main pharmacokinetic parameters and relative oral bioavailability were also calculated. Results The optimal formulation of Vit-Lips: phospholipids to cholesterol dose ratio was 7.95:1, lipids to drug dose ratio was 9.63:1 and hydration time was 30.00 min. CS-Vit-Lips and DA/CS-Vit-Lips were prepared using chitosan and DA/CS solution with mass fraction of 0.8%, respectively. Entrapment efficiency of Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips were (82.31 ± 0.96)%, (84.93 ± 1.17)%, (85.15 ± 1.38)%, drug loading were (7.33 ± 0.09)%, (6.24 ± 0.08)%, (6.30 ± 0.11)%, particles size were (197.70 ± 5.07), (233.06 ± 7.19), (237.93±6.96) nm, and ζ potential were (−28.81 ± 0.86), (27.75 ± 1.10), (26.14 ± 1.13) mV, respectively. The appearance of three kinds of liposomes was spherical vesicular, particle stability of CS-Vit-Lips and DA/CS-Vit-Lips was higher than that of Vit-Lips in simulated gastrointestinal fluid. Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips enhanced cumulative release rate to 91.02%, 86.74% and 83.15%, respectively. The drug release process of the three kinds of liposomes conformed to Weibull model. The three kinds of liposomes effectively improved the absorption rate constant (K_a) and apparent absorption coefficient (P_app) of vitexin. Compared to vitexin, Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips increased relative oral bioavailability to 1.10-fold, 3.08-fold and 4.70-fold, respectively. The half-life time (t_1/2) of CS-Vit-Lips and DA/CS-Vit-Lips had significantly prolonged (P < 0.01) and the peak concentration (C_max) was significantly increased (P < 0.01). Conclusion CS-Vit-Lips and DA/CS-Vit-Lips effectively promoted the oral absorption of vitexin, and the advantage of DA/CS-Vit-Lips were more obvious, laying an experimental foundation for further research.

Objective To develop an icariside II nanoemulsion (NE-ICS II) to enhance oral bioavailability and improve efficacy against cerebral ischemia-reperfusion injury (CIRI). Methods NE-ICS II was prepared by ultrasonic emulsification and its physicochemical properties were characterized. Pharmacokinetic parameters and brain distribution were systematically evaluated. Therapeutic efficacy was assessed in rats subjected to middle cerebral artery occlusion (MCAO) by measuring infarct volume, neurological deficit scores, and oxidative stress and inflammatory markers. RNA sequencing was performed to elucidate the underlying mechanisms. Results NE-ICS II showed a mean particle size of (147.59 ± 0.71) nm and an encapsulation efficiency of (84.85 ± 5.47) %. NE-ICS II increased the oral bioavailability of ICS II by 2.4-fold and significantly enhanced its accumulation in the brain. Therapeutic oral administration of NE-ICS II markedly improved neurological function, reduced infarct volume, and attenuated oxidative stress and inflammatory responses in MCAO rats. RNA sequencing revealed that NE-ICS II may downregulate signaling pathways associated with oxidative stress and inflammation. Conclusion NE-ICS II substantially improves the oral bioavailability of ICS II and enables effective treatment of CIRI at a low dose, providing a promising strategy for efficient brain delivery of ICS II and for the treatment of central nervous system diseases.

Objective To establish a multi-index weighted optimization strategy for wine-processed Xiangfu (Cyperi Rhizoma) by integrating CRITIC weighting, Box-Behnken design-response surface methodology (BBD-RSM), and an artificial neural network (ANN), and define a robust operating space based on model prediction, thereby providing a reference for process parameter fixation and quality consistency control. Methods The contents of cyperotundone, α-cyperone, total flavonoids, and total volatile oil were used as comprehensive evaluation indices. Objective weights were assigned using the CRITIC method, and an overall desirability (OD) value was calculated. Based on single-factor experiments, a four-factor, three-level Box-Behnken design (BBD) was conducted with processing temperature, processing time, moistening time, and dosage as independent variables, and a regression model was established. An ANN prediction model was then developed, and the Garson algorithm was applied to interpret the relative contributions of factors. In addition, the Pareto non-dominated solution set was used to screen the process-parameter space to obtain a robust operating interval (model-predicted). Results The CRITIC-derived weights for cyperotundone, α-cyperone, total volatile oil, and total flavonoids were 0.274 9, 0.255 4, 0.253 4, and 0.216 3, respectively. The optimal processing conditions were determined as follows: processing temperature 140 ℃, processing time 19 min, moistening time 6.9 h, and dosage 32 g/L. The OD obtained from validation experiments was 0.655 1, RSD was 2.97%, which was close to the predicted value (0.653 3). The robust parameter interval defined based on ANN prediction and the Pareto non-dominated solution set was as follows: processing temperature 140—150 ℃, processing time 18—22 min, moistening time 6.5—7.5 h, and dosage 30—35 g/L. The model suggested that OD could remain relatively stable under ± 5% parameter fluctuations within this interval. Conclusion The constructed BBD-ANN-Pareto coupling model demonstrated good predictive ability and stability, which may provide a reference for optimizing the processing technology and ensuring the quality consistency of wine-processed Cyperi Rhizoma.

Objective To address the challenges of insufficient standardization in the processing of Orpiment (arsenic disulfide, As₂S₂) and the difficulty in controlling its toxicity, this study elucidated the interactions among “particle size-morphology-porosity-crystal defects-arsenic (As) release”. It analyzed the effects of different processing methods on elemental composition and microstructure, and established a rapid identification model based on multimodal spectral fusion. Methods A multi-dimensional “element-morphology-spectrum” data matrix was constructed. Inductively coupled plasma optical emission spectrometry (ICP-OES) was used to determine elemental content. Scanning electron microscopy (SEM) was employed to observe morphology and analyze surface porosity, combined with dissolution experiments to quantify As release. Furthermore, Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy (RS) data were integrated. Partial least squares-discriminant analysis (PLS-DA) and support vector machine (SVM) discrimination models were built using both low-level and mid-level data fusion strategies. Results The zirconia-ball water-grinding method reduced As content and increased sulfur (S) content in Orpiment. SEM analysis revealed that particles processed by this method exhibited a more concentrated size distribution, their morphology became more rounded and blunt, and surface porosity significantly increased, forming a hierarchical pore structure. In simulated gastric and intestinal fluids, As dissolution decreased by 72.8% and 81.4%, respectively, compared to dry crushing (P < 0.001). The SVM model based on mid-level spectral fusion achieved 100% classification accuracy. Conclusion The water-grinding method achieves detoxification by removing As³⁺ from Orpiment, regulating the particle morphology and pore structure of Orpiment particles, and enhancing the stability of Orpiment crystals. It establishes a multidimensional data matrix integrating elemental composition, microscopic morphology, pore structure, and spectral characteristics, providing efficient and reliable technical support for quality control of the mineral drug Orpiment.

Objective To analyze the structural characteristics of water-eluted fraction of Sanghuangporus vaninii polysaccharides (SHP-W) and explore the mechanism of SHP-W on improving carbon tetrachloride (CCl₄)-induced liver fibrosis in mice by regulating gut microbiota. Methods SHP-W was extracted from S. vaninii using water extraction and ethanol precipitation, followed by purification through weak anion-exchange chromatography. The total sugar, uronic acid and protein contents of SHP-W were determined by phenol sulfuric acid method, m-hydroxybiphenyl method and BCA method. The structural characterization of SHP-W was carried out by high performance size exclusion chromatography (HPSEC), Fourier transform infrared spectrophotometer (FT-IR), ion chromatography (IC) and methylation combined gas chromatography-mass spectrometry (GC-MS). C57BL/6J mice were prepared with a 15% CCl₄ olive oil solution via ip for six weeks to establish a liver fibrosis model. SHP-W (25, 50, 100 mg/kg) or sorafenib (10 mg/kg) were administered for three weeks, and the pathological morphological changes in liver tissues were observed using hematoxylin-eosin (HE), sirius red and Masson staining. A fully automated biochemical analyzer was used to analyze the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acids (TBA), total bilirubin (TBIL) and direct bilirubin (DBIL) in serum. Immunohistochemistry was used to detect the expressions of collagen-I (Col-I) and α-smooth muscle actin (α-SMA) in liver tissue. Western blotting was used to detect the expressions of Col-I, α-SMA and transforming growth factor-β1 (TGF-β1) proteins in liver tissue. 16S rDNA sequencing was used to analyze the gut microbiota, and antibiotic interference was used to verify the role of the microbiota. Results SHP-W was a refined polysaccharide composed of two polysaccharides with different relative molecular weights (1.818 × 10⁴, 2.040 × 10³), with a total sugar content of (88.27 ± 3.76)%, and without uronic acid and protein. The monosaccharide composition results showed that SHP-W was composed of galactose, mannose, fucose and glucose, with a molar ratio of 34.6∶23.0∶22.7∶19.7, and also contained 3-O-methylgalactose. Methylation analysis showed that SHP-W was composed of 14 glycosidic bonds, including 1,3,6-Glcp.The animal experiment results showed that compared with control group, the levels of AST, ALT, TBIL, DBIL and TBA in serum of mice in model group were significantly increased (P < 0.001), the arrangement of liver cell cords was disordered, inflammatory cell infiltration was obvious, and a large amount of fibrous tissue proliferation was appeared, the expressions of Col-I, α-SMA and TGF-β1 protein in liver tissue was significantly up-regulated (P < 0.05, 0.001). Compared with model group, the collagen fiber deposition in pseudo lobules and portal area of liver tissue of mice in each treatment group was improved to varying degrees, and the serum liver function level was significantly reduced (P < 0.05, 0.01, 0.001), the expressions of Col-I, α-SMA and TGF-β1 protein in liver tissue was down-regulated (P < 0.05, 0.01, 0.001). After antibiotic interference, the effect of SHP-W on improving liver fibrosis was significantly inhibited. Conclusion SHP-W were prepared from S. vaninii fruiting bodies, the determination of uronic acid content, infrared spectroscopy and monosaccharide composition showed that SHP-W were neutral polysaccharide. SHP-W could significantly improve serum liver function, inhibit HSCs activation, reduce collagen fiber deposition, and thus improve CCl₄-induced liver fibrosis in mice. Its anti-fibrotic effect depends on the regulation of gut microbiota.

Objective To investigate the mechanism of total triterpenes from fruits of Chaenomeles speciosa (CST) in alleviating N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and (NIG)-induced human gastric mucosal epithelial cells damage based on microRNA-210 (miR-210)/nuclear factor-κB (NF-κB)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome signaling pathway. Methods MNNG and NIG induced GES-1 cells model were constructed, and then GES-1 cells were treated with CST and/or miR-210 mimic, miR-210 inhibitor, NF-κB siRNA, NLRP3 siRNA. MTT assay was used to detect cell viability, immunofluorescence was utilized to measure reactive oxygen species (ROS) level, flow cytometry was used to test pyroptosis, colorimetric and ELISA methods were utilized to measure the levels of interleukin-4 (IL-4), IL-1β, IL-6, IL-10, IL-18, lactate dehydrogenase (LDH), and tumor necrosis factor-α (TNF-α) in cell supernatant. The colorimetric method was used to determine the levels of catalase (CAT), glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in cells. The dual luciferase reporter assay was employed to verify the targeted intervention between miR-210 and NF-κB. Immunofluorescence was utilized to detect the NF-κB p65 nuclear translocation, the co-localization of NLRP3, apoptosis related spot like protein (ASC) and pro-cystein-asparate protease-1 (pro-Caspase-1). qRT-PCR was used to detect the mRNA expression levels of miR-210, NF-κB, NLRP3. Western blotting was utilized to determine the protein expression levels of cytoplasmic NF-κB, nuclear NF-κB, NIMA-related kinase 7 (NEK7), thioredoxin-interacting protein (TXNIP), NLRP3, ASC, Caspase-1, pro-Caspase-1, gasdermin D (GSDMD), N-terminal domain of GSDMD (GSDMD-N), pro-IL-18 and pro-IL-1β. Results miR-210 was a pro-inflammatory/pyroptosis factor, the dual luciferase reporter assay confirmed that it could bind to 3’-UTR in mRNA of NF-κB. Compared with model group, CST significantly inhibited pyroptosis and LDH release in MNNG and NIG induced GES-1 cells (P < 0.01), reduced levels of IL-1β, IL-6, IL-18, TNF-α in supernatant and ROS, MPO, MDA in cells (P < 0.01), elevated levels of IL-4, IL-10 in supernatant and activities of CAT, GSH, SOD, T-AOC in cells (P < 0.01), suppressed the NF-κB p65 nuclear translocation and the co-localization of NLRP3, ASC and pro-Caspase-1 (P < 0.01), downregulated the mRNA expressions of miR-210, NF-κB, NLRP3 and the protein expressions of nuclear NF-κB p65, TXNIP, NEK7, NLRP3, ASC, pro-Caspase-1, Caspase-1, GSDMD, GSDMD-N, pro-IL-1β, pro-IL-18 (P < 0.01), upregulated the cytoplasmic NF-κB p65 protein expression (P < 0.01). The combination of CST with miR-210 inhibitor, NF-κB siRNA and NLRP3 siRNA transfection could further improve the above indicators (P < 0.01). Conclusion CST has a good preventive and therapeutic effect on MNNG and NIG-induced GES-1 cell injury, and its mechanism is closely related to inhibiting the abnormal elevation of miR-210 and activation of NF-κB/NLRP3 inflammasome signaling pathway, alleviating oxidative stress and subsequent suppressing pyroptosis.

Objective To investigate the effect of andrographolide on head and neck squamous cell carcinoma, and reveal its potential anti-tumor mechanism from the perspective of ferroptophagy through lipidomics. Methods CAL27 cells were cultured in vitro. CCK-8 assay and colony formation assay were used to detect the inhibitory effect of andrographolide on cell proliferation, and flow cytometry was used to detect the cell apoptosis. A patient-derived xenograft (PDX) mouse model of human head and neck squamous cell carcinoma was established. Control group and andrographolide (10 mg/kg) group were set up, and after intervention with administration, the tumor growth was observed dynamically, and the expression of proliferation-related index Ki67 in tumor tissues was detected. Lipidomics technology was used to analyze the metabolic profile changes of CAL27 cells after andrographolide treatment, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. Western blotting was used to detect the expressions of ferroptophagy-related proteins in cells. Results The in vitro experimental results showed that andrographolide could significantly inhibit the proliferation of CAL27 cells and induce cell apoptosis (P < 0.01, 0.001), and showed a dose-dependent effect. The results of in vivo PDX model experiment showed that andrographolide could significantly inhibit the growth of tumors in vivo (P < 0.001), and the positive expression of Ki67 in tumor tissues was significantly reduced compared to the control group (P < 0.001). The lipidomics analysis and KEGG pathway enrichment results showed that the differential metabolic pathways of cells treated with andrographolide were enriched in ferroptosis and autophagy-related pathways. Western blotting results confirmed that andrographolide could significantly regulate the expressions of ferroptophagy-related proteins (P < 0.05, 0.01, 0.001), indicating that andrographolide could effectively induce ferroptophagy in CAL27 cells. Further validation with the nuclear receptor coactivator 4 (NCOA4) targeted degrader and ferroptosis inhibitor demonstrated that both agents markedly reversed the effects of andrographolide (P < 0.05, 0.01, 0.001), verifying that its antitumor effect depends on the ferroptophagy signaling pathway. Conclusion Andrographolide could significantly inhibit the proliferation and induce apoptosis of head and neck squamous cell carcinoma in vitro and in vivo. Its anti-tumor mechanism is related to inducing ferroptophagy in tumor cells.

Objective To investigate the molecular mechanism by which Linggui Zhugan Decoction (苓桂术甘汤, LGZGD) ameliorates heart failure (HF) after myocardial infarction (MI) via regulating mitophagy mediated by mitochondrial calcium uniporter (MCU)-mitochondrial calcium (mtCa²⁺) signaling pathway. Methods Rat models of post-MI HF and hydrogen peroxide (H₂O₂)-induced H9c2 cardiomyocyte injury models were established. For in vivo experiments, rats were assigned to sham group, model group, LGZGD group and captopril group. For in vitro experiments, H9c2 cells were divided into corresponding groups with additional interventions of MCU knockdown or overexpression. Echocardiography, Masson staining and transmission electron microscopy were performed to evaluate rat cardiac function, myocardial fibrosis and mitochondrial ultrastructure, respectively. The mRNA and protein expression levels of genes related to mitophagy and mitochondrial function were detected by qRT-PCR and Western blotting. The levels of reactive oxygen species (ROS) and mtCa²⁺ were measured, myocardial injury markers were quantitatively analyzed, and changes in cellular autophagic flux were observed using confocal laser scanning microscopy. Results Compared with sham group, rats with post-MI HF exhibited significantly impaired cardiac function, aggravated myocardial fibrosis and mitochondrial damage, dysregulated transcription of mitophagy-related genes, and markedly upregulated MCU transcription level (P < 0.001). In H₂O₂-stimulated H9c2 cells, ROS and mtCa²⁺ levels were significantly increased, autophagic flux was blocked, and myocardial injury marker levels were notably elevated (P < 0.01).Compared with model group, LGZGD treatment significantly improved cardiac function, alleviated myocardial fibrosis and mitochondrial damage, downregulated MCU expression (P < 0.01), regulated the mRNA expressions of autophagy and mitophagy-related genes in post-MI HF rats (P < 0.05, 0.01). At the cellular level, LGZGD reduced ROS and mtCa²⁺ levels in H₂O₂-injured H9c2 cells, restored mitochondrial morphology and autophagic flux, and decreased myocardial injury marker levels (P < 0.01). MCU knockdown enhanced the cardioprotective effect of LGZGD, whereas MCU overexpression exacerbated cellular injury. LGZGD effectively reversed MCU overexpression-induced cellular damage and ameliorated the dysregulation of oxidative stress-related enzymes (P < 0.05, 0.01). Conclusions LGZGD alleviates oxidative stress injury and improves post-MI heart failure by restoring MCU-mtCa²⁺ homeostasis and promoting mitochondrial autophagic flux.

Objective To investigate the effect and underlying mechanism of Prunella vulgaris-derived exosomes loaded with orlistat (PVENs-Orl) on lipid metabolism and malignant biological behaviors of hepatoma cells. Methods PVENs were extracted by ultracentrifugation. After characterization by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA), a PVENs-Orl drug delivery system was constructed. The inhibitory effects of PVENs-Orl on the proliferation, migration and invasion of HepG2 cells were evaluated through CCK-8, cloning, scratch and Transwell assays. The free fatty acids (FFA) uptake, fatty acid synthase (FASN) activity, triglyceride (TG) content, lipid peroxidation indicators and mitochondrial membrane potential and morphology were detected. The expressions of genes related to lipid metabolism and ferroptosis [acyl-CoA synthetase long-chain family member 4 (ACSL4), glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), B-cell lymphoma-2 interacting protein 3 (BNIP3L), lysophosphatidylcholine acyltransferase 3 (LPCAT3), transferrin receptor (TFRC)] was detected by qRT-PCR. Results The PVENs were successfully extracted and characterized, and the constructed PVENs-Orl could be effectively internalized by HepG2 cells. PVENs and PVENs-Orl exhibited dose-dependent inhibition of the proliferation, migration and invasion of HepG2 cells (P < 0.001). In terms of lipid metabolism phenotype, PVENs-Orl could inhibit FASN activity (P < 0.01), and simultaneously cause intracellular accumulation of FFA and TG (P < 0.01). PVENs-Orl significantly increased the levels of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) (P < 0.01, 0.001), and increased the release of lactate dehydrogenase (LDH) (P < 0.001). PVENs-Orl significantly induced a decrease in mitochondrial membrane potential (P < 0.01) and triggered mitochondrial dysfunction (P < 0.001). At the molecular mechanism level, PVENs-Orl upregulated the expressions of genes related to lipid peroxidation and ferroptosis promotion (P < 0.001), and downregulated the expressions of genes related to antioxidant and ferroptosis inhibition (P < 0.001). Conclusion PVENs could effectively deliver orlistat, disrupt lipid homeostasis by inhibiting FASN activity, induce mitochondrial dysfunction and lipid peroxidation, and synergistically regulate the expressions of key genes involved in ferroptosis, thereby inhibiting the growth of liver cancer cells. This provides experimental evidence for targeted lipid metabolism therapy based on natural exosomes.

Objective To explore the therapeutic effect and related mechanism of citronellol on methicillin-resistant Staphylococcus aureus (MRSA)-induced skin infections in mice via NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome pathway. Methods The minimum inhibitory concentration (MIC) of citronellol against standard MRSA strains was determined using the microbroth dilution method. A full-thickness skin MRSA infection model was established in female C57BL/6 mice, control group, model group, positive control (2% mupirocin) group, citronellol low-, medium- and high-dose (0.75%, 1.50%, 3.00% citronellol gel) groups were set up. Local administration was performed for 7 d. Wound healing rates were dynamically monitored, hematoxylin-eosin (HE) staining was used to observe histopathological changes in skin tissues, and ELISA was used to detect levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD) and malondialdehyde (MDA) in skin tissues. Masson staining was used to observe collagen fiber deposition in skin tissues. RNA-seq transcriptome sequencing, qRT-PCR and Western blotting were employed to analyze the expressions of NLRP3 inflammasome pathway-related genes and proteins in skin tissues, while molecular docking was used to predict the binding affinity of citronellol to relevant inflammatory targets. The lactate dehydrogenase (LDH) release assay was used to evaluate the effect of citronellol on pyroptosis in MRSA-infected immortalized keratinocytes. Results Citronellol exhibited significant antibacterial activity against MRSA, with an MIC ranging from 0.312 5—0.625 0 mg/mL. In vivo experiments revealed that compared with model group, citronellol significantly accelerated wound healing, reduced inflammatory cell infiltration and tissue edema, decreased IL-1β and TNF-α levels in skin tissues (P < 0.05, 0.01, 0.001), markedly increased collagen fiber deposition at wound sites (P < 0.05, 0.01), and inhibited the activation of NLRP3 inflammasome-cell pyroptosis pathway (P < 0.05, 0.01, 0.001). Molecular docking results indicated that citronellol could bind to target proteins such as IL-1β. In vitro experiments demonstrated that citronellol significantly suppressed LDH release in MRSA-infected HaCaT cells (P < 0.05, 0.01). Conclusion Citronellol exerts significant antibacterial, anti-inflammatory and wound repair-promoting effects on MRSA-induced skin infections in mice. Its mechanism may be related to inhibiting NLRP3 inflammasome-cell pyroptosis pathway, reducing the release of pro-inflammatory cytokines, improving the local immune microenvironment, and promoting collagen deposition.

Objective To evaluate the clinical efficacy and safety ofKushen Gel combined with oral fluconazole in the treatment of vulvovaginal candidiasis (VVC). Methods A multicenter, randomized, double-blind, placebo-controlled trial was conducted. Patients with VVC who visited four hospitals between June 2024 and October 2025 were recruited and randomly assigned in a 1∶1 ratio to the placebo group or the Kushen Gel group. Both groups received a single oral dose of fluconazole 150 mg. Additionally, the Kushen Gel group received intravaginal Kushen Gel, while the placebo group received a placebo gel with identical appearance and properties, both for 14 consecutive days. The primary efficacy endpoint was the clinical response rate at 7—10 days after drug discontinuation. Secondary efficacy endpoints included the proportion of patients with normalization of at least four vaginal microecological indicators, time to complete resolution of vulvar pruritus, pain, and abnormal discharge, vaginal cleanliness, and mycological recurrence rate at 25—31 days after drug discontinuation. Adverse events were monitored to evaluate safety. Results A total of 93 patients were randomized. At the 7—10 days post-treatment follow-up visit, the clinical response rate in the Kushen Gel group was significantly higher than that in the placebo group (87.50% vs 65.00%, P = 0.033 9). The proportion of patients achieving normalization of at least four vaginal microecological indicators was significantly higher in the Kushen Gel group compared with the placebo group [97.50% vs 77.50%, P = 0.0143]. The times to resolution of vulvar pruritus, pain, and abnormal discharge in the Kushen Gel group were all shorter than those in the placebo group, but the differences were not statistically significant (P > 0.05). At the 25—31 days post-treatment follow-up, the mycological recurrence rate was 10.53% (2/19) in the Kushen Gel group, which was lower than that in the placebo group (23.81%, 5/21). The incidence of adverse events was low in both groups, with no statistically significant difference between groups (P > 0.05), and no serious adverse events occurred. Conclusion The combination of Kushen Gel with oral fluconazole can alleviate clinical symptoms, enhance short-term clinical efficacy, and may help reduce the recurrence rate, as well as, the recovery of vaginal microecology in patients with VVC. The treatment is safe and possesses significant clinical value.

Objective To investigate the medication regularity of traditional Chinese medicine (TCM) for preventing and treating alcohol-related liver and brain injuries, and to explore the intervention mechanisms of Jiejiu Formula (JJF) with homology of medicine and food on alcoholic liver disease (ALD) and alcohol-related brain injury (ARBI) through network pharmacology and experimental validation. Methods Herbal formulas targeting ALD and ARBI were collected from TCM classics of the Tang, Song, Yuan, and Ming dynasties. Frequency analysis was performed on the constituent herbs to identify high-frequency ingredients. Their properties, flavors, meridian affinities, and dosages were analyzed. Association rule mining and cluster analysis were then applied to identify a core herbal combination for the JJF. Network pharmacology was used to screen the active components and potential targets of the JJF against ALD and ARBI. Core targets were subjected to gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses. Molecular docking and molecular dynamics simulations were employed to validate the predicted mechanisms and binding stability. Subsequently, an alcohol-induced liver and brain injury mouse model was established to verify the findings in vivo. Results A total of 301 formulas containing 263 distinct herbs were identified. Herbs with a warm nature, pungent flavor, stomach meridian affinity, and heat-clearing functions were the most frequently used. The combination of “Gancao (Glycyrrhizae Radix et Rhizoma)-Chenpi (Citri Reticulatae Pericarpium)-Ganjiang (Zingiberis Rhizoma)” demonstrated the highest frequency and strongest association, and all three herbs are classified as medicinal food ingredients. Therefore, this combination was designated as the JJF. Network pharmacology identified 289 active components and 254 potential targets for the JJF against ALD/ARBI. Protein-protein interaction (PPI) network analysis revealed five core targets. KEGG enrichment analysis highlighted pathways related to endocrine resistance. Molecular docking suggested that 2-[(3R)-8,8-dimethyl-3,4-dihydro-2H-pyrano[6,5-f]chromen-3-yl]-5-methoxyphenol and phaseolinisoflavan have good binding affinity and stability with protein kinase B1 (AKT1), which was further confirmed by stable binding in MD simulations. In vivo experiments showed that the JJF alleviated alcohol-induced damage in liver and brain tissues to varying degrees. Conclusion JJF exerts preventive and therapeutic effects against ALD and ARBI. It potentially acts on key targets such as Src proto-oncogene tyrosine-protein kinase (SRC), phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA), and AKT1, and modulates pathways like endocrine resistance. Furthermore, the JJF may accelerate alcohol metabolism, enhance anti-oxidative stress capacity, upregulate mRNA expression within the phosphatidylinositol-3-hydroxykinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) signaling pathway, and inhibit mitogen-activated protein kinase 3 (MAPK3) expression, thereby regulating cell proliferation, apoptosis, and inflammatory responses to achieve its protective effects.

Objective To comprehensively sort out and visually analyze the research status, hotspots and trends of knowledge graph technology in the field of traditional Chinese medicine (TCM) from the perspectives of bibliometrics and global patents, provide references for in-depth research and development in this field. Methods The relevant Chinese and English literature published between January 1, 2012, and December 31, 2025, was systematically retrieved from the CNKI, Wanfang, VIP, and Web of Science databases. Using tools such as CiteSpace, VOSviewer, and Excel, we performed bibliometric and visual analyses of publication trends, institutional and author collaboration networks, journal sources, and keyword co-occurrence and evolution. Additionally, we analyzed global patent applications from the Incopat database, focusing on application trends, geographical distribution, applicants, and technical themes. Results A total of 745 valid publications were included (618 in Chinese, 127 in English) and 432 patent families. Research on knowledge graph technology in TCM is in a phase of rapid development. Current hotspots focus on refining knowledge extraction techniques, developing KG-assisted diagnosis and treatment systems, intelligent question-answering, and synergistic applications with large language models. The field is moving toward deeper interdisciplinary integration and clinical application . China occupies a dominant position in both research output and patent applications (96% share), with established collaboration networks. However, cross-regional and interdisciplinary cooperation requires further strengthening. Conclusion Research on knowledge graph technology in the field of traditional Chinese medicine is deeply evolving from fundamental construction to intelligent applications. Despite China’s quantitative advantage in patents, challenges persist, including a relative lack of high-value patents, technical homogenization, and insufficient international competitiveness.

Objective To analyze the knowledge evolution context, patent genealogy characteristics and prospective innovation directions of Euphorbia plant research, and clarify the efficiency of achievement transformation and the global competitive pattern, this study adopts bibliometric and knowledge graph analysis methods, combined with patent information lineage analysis technology to carry out systematic research. Methods The relevant literature in the CNKI, Wanfang, VIP and Web of Science Core Collection were systematically searched with Euphorbia pekinensis and Euphorbia as the subject words, imported into NoteExpress literature management software for deduplication, screening and format conversion, and CiteSpace and VOSviewer software were used to visualize the annual publication trends, the publishing country, institution, author, journal distribution, discipline distribution and other dimensions of Chinese and English literature. At the same time, based on the Incopat patent database, the application and distribution status of patents related to Euphorbia plants were systematically analyzed from aspects such as the number of patent applications and geographical distribution patterns. Results After the screening procedure, a total of 1 682 Chinese literature and 1 339 English literature were incorporated into the study, indicating that the research of Euphorbia plants was at the international stage. A total of 98 countries had issued documents, among which China had published the most achievements. The author with the largest number of publications in both Chinese and English was Zhang Li. The core publishing institutions of the Chinese and English literature were Nanjing University of Chinese Medicine and the United States Department of Agriculture. The cooperation connections between these institutions were still relatively scattered, but some authors had formed stable research teams. The Chinese and English journals with the highest number of articles published were China Journal of Chinese Materia Medica and Journal of Ethnopharmacology. The English literature mainly involved plant science and extended to pharmacology, medicinal chemistry and other fields. The Incopat patent database retrieved 8 350 patents related to Euphorbia plants, and 5 603 domestic patents, accounting for 67.1% of the total, which was in a dominant position. Conclusion Research on Euphorbia plants shows a pattern dominated by China, coexisting with global participation. The fundamental research system in this field has become increasingly well-established, with remarkable progress in the modernization of traditional medicinal applications and broad prospects for industrial transformation, but there are still problems such as regional development imbalance. In the future, it should deepen the basic theory, promote interdisciplinary research, optimize the patent layout, and realize the efficient transformation and sustainable development of resource value.

Objective To clone the cycloartenol synthase (CAS) gene PcCAS from Polygonatum cyrtonema, perform bioinformatic and recombinant protein expression analyses, and determine its subcellular localization etc, thereby providing essential resources for further elucidating the pivotal role of PcCAS in saponin biosynthesis. Methods Based on the transcriptome data of P. cyrtonema, the PcCAS gene was screened and cloned, and bioinformatics approaches were employed to analyze the physicochemical properties, secondary structure, and tertiary structure of its encoded protein. qRT-PCR was used to analyze the tissue-specific expression of PcCAS. A prokaryotic expression vector was constructed and transferred into Escherichia coli for prokaryotic expression analysis. Yeast expression vectors were constructed and transformed into Pichia pastoris for subsequent yeast expression analysis. A subcellular-localization vector was built and infiltrated into Nicotiana benthamiana leaves for the analysis of heterologous expression in tobacco. Results The open reading frame (ORF) of PcCAS is 2 286 bp in length and encodes 759 amino acids. The protein is hydrophilic and lacks a signal peptide. The total saponin content and PcCAS expression level in the rhizome are significantly higher than those in the stem, leaf, and flower, indicating the expression level of PcCAS is positively related with the total saponin content. Yeast-expressed recombinant protein matched the expected molecular weight of 86 400. Subcellular-localization assays in tobacco leaves demonstrated that the PcCAS protein is predominantly localized to the endoplasmic reticulum. Conclusion PcCAS protein belongs to the OSC superfamily of enzymes and is localized in the cytoplasm, positively regulating the total saponin biosynthesis.

Objective FAD enzymes introduce double bonds at specific positions in fatty acids, converting saturated fatty acids into unsaturated fatty acids, playing crucial roles in plant growth, development, and stress responses. This study aimed to systematically identify the FAD gene family in Panax ginseng and verify the function of PgFAD2-56, so as to elucidate the functions and stress response mechanisms of ginseng FAD genes. Methods Using bioinformatics approaches, 98 PgFAD genes were identified from the P. ginseng T2T genome, and their physicochemical properties, phylogenetic relationships, chromosomal localization, gene structures, conserved motifs, cis-acting regulatory elements, collinearity, and expression patterns under stress conditions were analyzed. Molecular docking was performed to predict the binding affinity of highly expressed FAD2 proteins with oleic acid, and the function of PgFAD2-56 was verified using a yeast heterologous expression system. Results The results showed that the 98 PgFAD genes could be classified into six subfamilies (ADS, SLD, FAD3/FAD7/FAD8, FAD6, FAD2, and FAB) based on amino acid sequence similarity, which was consistently supported by their gene structures and conserved motifs. Numerous hormone- and stress-responsive cis-acting elements and transcription factor binding sites (TFBSs) were identified in the promoters of PgFAD genes. Collinearity analysis revealed significant expansion of the FAD2 subfamily in P. ginseng. Gene expression analysis demonstrated that some PgFADs displayed significant differential expression under stress conditions. Molecular docking indicated that FAD2-56 exhibited the lowest binding energy with oleic acid (−6.5 kcal/mol) among the highly expressed FAD2 proteins. Heterologous expression in yeast confirmed that PgFAD2-56 altered fatty acid composition by producing linoleic acid and enhanced cold stress resistance. Conclusion This study demonstrates that the ginseng FAD gene family exhibits strong subfamily specificity and plays a vital role in stress resistance, with the expansion of the FAD2 subfamily and functional characterization of PgFAD2-56 providing potential targets for stress-resistant molecular breeding in P. ginseng.

Objective In order to study the relationship between the morphology of Angelica dahurica root system and the accumulation of coumarin-like components at different developmental stages under different light intensity treatments, to provide reference and basis for improving the quality of A. dahurica herbs and for efficient and reasonable cultivation and production. Methods The morphological indexes of A. dahurica root system were determined in 50% shade, natural light, and supplemental light treatment groups; High-performance liquid chromatography (HPLC) and correlation analysis were used to determine and screen the coumarin-like components that were strongly associated with the morphology of A. dahurica root system; The morphology of the root system of A. dahurica was observed by the application of exogenous coumarin, and the expression level of coumarin biosynthesis-related genes in the root system was determined based on the RT-qPCR technique. Results Root morphology indexes of A. dahurica (dry weight of primary root, number of lateral roots, diameter of primary root, root-crown ratio) were positively correlated with light intensity; the overall change of the total coumarin content in the different light intensity treatment groups showed an “S” - shaped curve. Umbelliferolide was significantly negatively correlated with primary root length, number of lateral roots, and root-crown ratio, and significantly positively correlated with primary root diameter; exogenous application of umbelliferolide significantly promoted the increase of the morphological parameters of A. dahurica root system (total surface area of the root, the length of the total root, and the number of root branches), and the content of the four complex coumarins (oxypeucedanin, imperatorin, phellopterin, and isoimperatorin), and significantly up-regulated the expression of key genes of coumarin biosynthesis pathway, including phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL) and other key genes expression. Conclusion A. dahurica root indexes and coumarin content varied in different light intensities and developmental periods. The exogenous application of umbelliferolide affected the establishment of dahurica root system and the accumulation of coumarin-like components in dahurica roots by regulating the expression of genes related to the coumarin synthesis pathway in dahurica roots.

Objective To establish a fingerprinting method and multi-component quantification approach for Rehmannia glutinosa leaves from different geographical origins, and to evaluate the quality of these samples using chemometric analysis. Methods The HPLC fingerprint of R. glutinosa leaves was developed and analyzed for similarity using the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012.130723). Chemical constituents were identified and quantified by comparison with reference standards. Hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed using Origin 2025, IBM SPSS Statistics 27.0, and SIMCA 14.1 software to evaluate the quality of R. glutinosa leaves samples and screen for differential markers. Results A characteristic HPLC fingerprint of R. glutinosa leaves from different origins was developed. In this fingerprint, 19 common peaks were calibrated, and 13 components were unequivocally identified using reference standards. The similarity evaluation of the 23 batches yielded results ranging from 0.879 to 0.997. The cluster heatmap analysis demonstrated a clear classification, dividing the samples into two groups. Principal component analysis (PCA) yielded five principal components with a cumulative variance contribution rate of 83.545%, and the score plot revealed that samples originating from Henan province clustered separately, suggesting their superior quality. Subsequently, OPLS-DA was employed, which successfully screened four compounds-catalpol, acteoside, luteolin, and apigenin-as critical markers for quality discrimination. The quantitative analysis showed that the contents of these markers were 25.474—61.784 mg/g, 11.633—47.462 mg/g, 0.096—0.848 mg/g, and 0.100—1.268 mg/g, respectively. Conclusion The developed method, integrating HPLC fingerprinting and multi-component assay, proved to be simple, accurate, and reproducible. It can systematically reflect the differences among samples of R. glutinosa leaves from different producing areas, thereby providing a reference for their quality control.

Objective To evaluate the quality of Lycium barbarum leaves from various production areas in Ningxia, an HPLC fingerprinting technique combined with chemometrics and quantitative determination of multiple alkaloids constituents with LC-MS/MS was used to construct the alkaloidal fingerprints of L. barbarum leaves (S1—S50) from five bases in three production areas in Ningxia. Methods The HPLC method and Similarity Evaluation System for Chromatographic Fingerprinting in Chinese Medicine (version 2012) were used to generate fingerprints, and similarity evaluation and common peak confirmation were carried out. Meanwhile, LC-MS/MS was used to establish a method for the determination of the contents of seven alkaloids constituents (choline, betaine, trigonelline, lyciumin A, lyciumin B, N-trans-feruloyltyramine, N-feruloyl-3-methoxytyramine) to evaluate the leaves of L. barbarum from different origins in Ningxia. Results The obtained fingerprints had a total of 18 common peaks. The similarity between the alkaloid fingerprints of 50 batches of L. barbarum leaves. The control fingerprints ranged from 0.692 to 0.981. Zhongning and Guyuan origin L. barbarum leaves fingerprints have higher similarity to the control fingerprints, while Yinchuan origin L. barbarum leaves fingerprints have lower similarity to the control fingerprints. Hierarchical clustering analysis (HCA) clustered the 50 batches of samples into four categories; principal component analysis (PCA) screened out four principal components, with a cumulative variance of 89.014%; orthogonal partial least squares-discriminant analysis (OPLS-DA) screened out the components with larger contributions to inter-group variance, which were, in the order of the size of the VIP, peak 5 > peak 3 > peak 10 > peak 11 > peak 13 > peak 2 > peak 4 > peak 9; content determination showed that seven alkaloids from Guyuan origin had the highest content. The results of content determination showed that the seven alkaloids of Guyuan origin had the highest content. Conclusion A stable HPLC fingerprint of L. barbarum leaves of different origins and a quantitative LC-MS/MS method for the determination of seven alkaloids were constructed, which can be used for the analysis of the difference in the origins of L. barbarum leaves and quality evaluation by combining with chemometrics.

The treatment of wastewater from traditional Chinese medicine (TCM) pharmaceutical production is crucial for the industry’s green transformation and ecological conservation. It reduces removes high-concentration pollutants, prevents soil and water contamination, safeguards ecological and human health, and supports sustainable industry development. However, due to the complex composition of TCM raw materials and diverse extraction processes, the resulting wastewater contains substantial amounts of organic matter, suspended solids, and refractory substances. It exhibits characteristics such as high organic pollutant concentration, significant water quality fluctuations, and strong toxicity and unpleasant odors. Traditional treatment processes consume high energy, exhibit low degradation efficiency, generate substantial carbon emissions, and impose greater environmental burdens, making treatment significantly more challenging than for conventional industrial wastewater. Guided by the “dual carbon” policy and focusing on the two core objectives of “reducing energy consumption and enhancing energy reuse,” this study reviews pharmaceutical wastewater treatment technologies. It summarizes low-carbon treatment strategies and techniques for TCM pharmaceutical wastewater, analyzes their technical pathways and application essentials, and explores future development trends. The aim is to provide practical insights for the green transformation and low-carbon development of the TCM pharmaceutical industry.

Ancient classical prescriptions, with their long clinical application experience and clear therapeutic effects, have become an important direction for the research and development of new traditional Chinese medicine drugs. With the continuous advancement of related research, it has not only provided a new path for explaining the scientific connotations of traditional prescriptions, but it has also significantly promoted the technological innovation of traditional Chinese medicine compound preparations. This has profound significance for the inheritance and innovative development of traditional Chinese medicine. In recent years, under the guidance of national policies, the development of classic prescription traditional Chinese medicine compound preparations has received much attention. Among them, solid preparations, due to their good stability, controllable dosage, and convenience in administration, have become the main dosage form in the development of classic prescription preparations and have gained wide application and market recognition. This article takes traditional Chinese medicine compound preparations as the entry point, systematically reviews the application status, problems and challenges, and solutions of solid preparation technology in the development of classic prescriptions, and discusses the prospects of solid preparation technology in the development of classic prescriptions, providing certain references for the future development of classic prescription traditional Chinese medicine compound preparations.

As a classic method of traditional Chinese medicine (TCM) processing, wine-processing has been employed for toxin reduction and preservation of medicinal properties, efficacy enhancement and medicinal property modification, or purposely guiding the therapeutic effects of herbal medicines. This review retrieved literature from PubMed, Web of Science, China national knowledge infrastructure (CNKI), and Wanfang databases using keywords including “wine-processing” “medicinal properties” and “efficacy-enhancing mechanisms”. Findings indicate that traditional wine-processing predominantly utilizes HuangJiu, with distinctive techniques such as wine rinsing, wine stir-frying and wine steaming remaining in current practice. Modern processing technology emphasizes optimization of critical parameters. Wine-processing facilitates component transformation through physical dissolution, enhanced cellular permeability, and synergistic enzymatic/non-enzymatic reactions. Mechanisms involving multi-target networks and microbiota-targeted metabolic regulation, enable directional modulation of herbal cold/hot properties alongside efficacy enhancement and toxicity reduction. Future research should prioritize establishing digitalized process quality control standards, conduct interdisciplinary investigations into wine-processing effects on efficacy-related component groups, and elucidate directional regulation mechanisms of medicinal properties. These advancements are anticipated to promote standardization and modernization of wine-processing techniques while scientifically interpreting the theoretical foundations of TCM processing.

Myocardial fibrosis (MF) is a key pathological feature following myocardial infarction (MI). This condition leads to myocardial stiffening and impaired cardiac pumping, ultimately resulting in complications including arrhythmias, heart failure, and cardiac rupture. Currently, interventions such as chemical drugs and surgery play an important role in the treatment of post-MI MF. However, they are still limited by suboptimal prognoses and high medical costs. Meanwhile, traditional Chinese medicine (TCM) has demonstrated potential value in this area, owing to its holistic regulation and multi-target therapeutic effects. This review provides a systematic overview of the pathological mechanisms underlying MF after MI and the preventive and therapeutic strategies of TCM for this disease. It aims to provide references for future clinical research and guide drug development efforts.

Danggui (Angelicae Sinensis Radix), as a bulk genuine medicinal material from Gansu, its quality and safety directly affect clinical efficacy and public health. In recent years, pesticide use during cultivation has led to increasingly prominent residue issues, which have become key risk factors constraining the sustainable development of the industry. Although traditional chromatography-mass spectrometry techniques offer high accuracy, they are limited by heavy reliance on specialized instruments dependency and long analysis cycles, making it difficult to meet the demands for on-site rapid screening and high-throughput detection of large-scale samples. Therefore, the development of fast, sensitive, and convenient novel detection technologies has become a current research priority. This paper systematically reviews the main types of pesticide residues in Angelicae Sinensis Radix, as well as advancements in detection technologies. It focuses on evaluating the principles, characteristics, and application status of emerging technologies based on nanomaterials, biosensing, and molecular imprinting, and discusses the challenges of interference and adaptability in the complex matrix of Angelicae Sinensis Radix. Furthermore, this paper emphasizes that quality control of traditional Chinese medicine must balance “safety” and “effectiveness”, analyzes the synergistic relationship between pesticide residue detection and active component analysis, and puts forward technical pathways for their integrated detection. Finally, it outlines the development trends of Angelicae Sinensis Radix pesticide residue detection technologies toward integration, intelligentization, and whole-process traceability, aiming to provide a reference for constructing a fast, accurate, and high-throughput quality monitoring system suitable for the complex matrix of traditional Chinese medicine.

Natural product hydrogels, self-assembled from active components of traditional Chinese medicine (TCM) such as steroids, triterpenoids, flavonoids, alkaloids, and quinones, represent a significant class of functional systems. Leveraging their intrinsic amphiphilic structures, these compounds spontaneously form stable three-dimensional networks through intermolecular interactions including hydrophobic effects, π-π stacking, hydrogen bonding, and coordination bonds. These hydrogels possess the core “drug-carrier integration” feature, offering unique advantages for drug delivery and disease therapy. Key properties such as bioadhesion, targeted localization, stimuli-responsive release, and in situ gelation enable precise drug delivery and enhanced therapeutic efficacy. However, significant challenges persist in this field: the self-assembly pathways and dynamic evolution mechanisms of many natural products remain incompletely elucidated, hindering the precise construction of functional materials. Furthermore, existing evaluation systems often fail to comprehensively reflect their in vivo pharmacological effects, resulting in a notable lag in clinical translation. Future research should, therefore, prioritize the systematic analysis of multi-level self-assembly mechanisms. This involves exploring the intrinsic relationships between molecular structure, assembly kinetics, and macroscopic properties, alongside their applications in intelligent drug delivery and therapy. Such efforts are expected to provide a theoretical basis for the rational design of natural product-based functional materials and to inform new directions for developing innovative TCM formulations and precision medicine.

Aristolochic acid analogues (AAAs) are a class of nitrophenanthrene derivatives predominantly occurring in Aristolochiaceae plants. Over the past decades, academic research has mostly focused on its strong adverse reactions such as nephrotoxicity and carcinogenicity. In contrast, their potential pharmacological activities—including anti-inflammatory, analgesic, and antitumor effects—have remained relatively underexplored, resulting in an imbalanced understanding of this compound class. In the face of the dual characteristics of toxicity and efficacy of this type of component, how to balance its toxicity and medicinal value is the key. Methods such as processing, compatibility, and structural modification may reduce adverse reactions, which could provide guarantees for the safe use of such components and traditional Chinese medicine. This review integrates current global advances in AAAs research, with a particular focus on their pharmacological activities and detoxification strategies, to provide references for the clinical application of drugs containing AAAs.

Pugongying (Taraxaci Herba) is a well-established medicinal and edible herb with a long history of use in traditional Chinese medicine, known for its heat-clearing, detoxifying, anti-inflammatory, and swelling-reducing properties. The Chinese Pharmacopoeia defines Taraxaci Herba as the dried whole plant of Taraxacum mongolicum, Taraxacum borealisinense, or several other closely related species within the genus Taraxacum (Asteraceae), making it a typical multi-origin herbal medicine. Taraxaci Herba comprises numerous species with highly similar interspecific morphological characteristics. Coupled with diverse ecological conditions across producing areas, this has led to mixed botanical origins in the marketplace and inconsistent quality of medicinal materials, thereby compromising the efficacy and safety of clinical use. This review summarizes recent advances in molecular identification technologies and quality evaluation methods for Taraxaci Herba, outlines the current status of quality control and comprehensive quality assessment research, and proposes future research perspectives. The aim is to provide a scientific reference for the standardization of medicinal materials, the selection of high-quality germplasm resources, and the high-quality development of the Taraxaci Herba industry.