Traditional Chinese medicine (TCM) solid preparations represent the primary dosage form in the TCM industry, and their manufacturing model rapidly transitioning from traditional batch production to continuous manufacturing. The complex composition of TCM ingredients, significant variability in material properties, and highly nonlinear process characteristics pose substantial challenges for online monitoring and dynamic quality evaluation. This paper systematically examines the developmental bottlenecks currently faced by online monitoring and dynamic quality evaluation in the context of continuous manufacturing of TCM solid preparations, focusing on their technical features. It provides a comprehensive review of approaches such as the construction of multidimensional process-quality databases, integration of mechanistic models with data-driven models, application of process analysis techniques at critical stages, and the implementation of digital twin and closed-loop control platforms. Future research should further explore the deep integration of “data-model-control” frameworks to transform passive monitoring into proactive prediction and adaptive regulation, thereby establishing a dynamic quality evaluation system tailored to the unique characteristics of TCM. This will provide theoretical foundations and technical references for standardizing, intelligentizing, and enhancing the quality of TCM solid preparation manufacturing.

Objective To investigate the chemical constituents and their antitumor activities isolated from Cyperus rotundus. Methods The isolates were isolated and purified by silica gel, Sephadex gel, reversed-phase C₁₈ column chromatography, and semi-preparative high-performance liquid chromatography (HPLC). The structures of the obtained compounds were elucidated by spectroscopic methods such as nuclear magnetic resonance (NMR) and mass spectrometry (MS). Finally, all compounds were subjected to MTT assays to evaluate their anti-tumor activities. Results Two new diphenyl ethers were obtained from C. rotundus, specifically as 2,3′-dihydroxy-5,2′,5′-trimethoxy diphenyl ether (1) and 2,3′-dihydroxy-4,2′,5′-trimethoxy diphenyl ether (2). Both compounds exhibited moderate inhibitory effects on human colon cancer cell line HCT116, with median inhibition concentration (IC₅₀) values of (24.20 ± 0.77) and (25.26 ± 0.65) μmol/L, respectively. Conclusion Compounds 1 and 2 are new compounds with anti-tumor potential, named cydipethers A (1) and B (2). Diphenyl ethers were identified from C. rotundus for the first time.

Objective To study the chemical constituents and in vitro antioxidant activity of Euphorbia heterophylla. Methods Comprehensive separation and purification were performed using normal-phase silica gel column chromatography, Sephadex LH-20 gel column chromatography and semi-preparative HPLC. Their structures were elucidated by comprehensive spectroscopic analyses (1D NMR, 2D NMR, HRMS) combined with single-crystal X-ray diffraction. The in vitro antioxidant activities of the isolated compounds were evaluated by DPPH and ABTS methods. Results A total of 18 compounds were isolated from the ethanol extract of E. heterophylla, their structures were elucidated as (3R,5R,8R,9R,13S,14R,17R,18R,21R)-3-hydroxy-21-isopropyl-4,4,9,13,14,17-hexamethyl-cyclopenta[a]chrysen-1(10)-en-2-one (1), β-amyrin ferulate (2), stigmasterol-4-en-3-one (3), isoquercitrin (4), quercetin 3-O-α-L-rhamnoside (5), kaempferol-3-O-α-arabifuranoside (6), kaempferol-3-O-α-L-rhamnoside (7), multiflorin A (8), 4,6-dimethoxyphthalide (9), vanillin (10), p-hydroxybenzaldehyde (11), protocatechuic acid (12), vanillic acid (13), 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl) propan-1-one (14), ferulic acid (15), diethylhexyl adipate (16), 13-propylheptacosane (17) and 1-n-decanoyl hydroxy-benzoic acid (18), respectively. Antioxidant activity evaluation revealed that compounds 5 and 15 exhibited potent DPPH radical scavenging activity, with half maximal inhibitory concentration (IC₅₀) values of (15.88 ± 0.84) μg/mL and (15.95 ± 0.75) μg/mL, respectively. Moreover, compound 15 also exhibited potent ABTS radical scavenging activity, with an IC₅₀ value of (2.54 ± 0.57) μg/mL, outperforming the positive control L-ascorbic acid [(IC₅₀ 12.25 ± 0.82) μg/mL]. Conclusion Compound 1 is a novel fernane-type triterpenoid and named as euheterophyllfernane A, compounds 2—3, 6—11, 14 and 16—18 are isolated from E. heterophylla for the first time.

Objective To investigate the chemical constituents of Fissistigma bracteolatum. Methods The 95% ethanol extract of F. bracteolatum was successively separated by solvent partitioning, macroporous resin, silica gel, MCI, Sephadex LH-20, ODS column chromatography, and semi-preparative HPLC to obtain individual compounds. Their structures were elucidated using modern spectroscopic techniques such as NMR and MS. An in vitro inflammatory model was established using lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages to evaluate the anti-inflammatory activity of the isolated compounds. Results A total of 22 compounds were isolated from F. bracteolatum, comprising of 12 flavonoids, 8 amides, 1 lignan, and 1 lactone. These were determined as: 5,7-dihydroxy-6,8-dimethoxyflavone (1), 5-hydroxy-7,8-dimethoxyflavone (2), quercetin (3), rutin (4), 5,7,8-trimethoxydihydroflavone (5), 5,6,8-trimethoxy-7-hydroxydihydroflavone (6), 6,7-dimethoxy-8-hydroxydihydroflavone (7), (2S)-7-hydroxy-6,8-dimethoxydihydroflavone (8), dihydroquercetin (9), aromadendrin (10), 4,2',6'-trihydroxy-3',4'-dimethoxy- dihydrochalcone (11), 2',6'-dihydroxy-4'-methoxydihydrochalcone (12), N-trans-sinapoylmethoxytyramine (13), N-[2-(4-hydroxy-3-methoxyphenyl)ethyl]-3-phenyl-2-propenamide (14), (E)-3-(4-hydroxy-3-methoxyphenyl)-N-phenethyl-acrylamide (15), N-trans-feruloyltyramine (16), N-trans-p-coumaroyltyramine (17), N-cis-feruloyltyramine (18), cis-N-feruloyl-3-O-methyldopamine (19), cannabisin D (20), 3-caffeoyl-2-methyl-D-erythronic acid-1,4-lactone (21), and (-)-syringaresinol (22). Conclusion Compounds 1, 7, 9—11, 13—15, 17, 19, and 21 were isolated from the genus Fissistigma for the first time.The anti-inflammatory activity results demonstrated that compounds 5 , 9 , 13 , and 16 significantly inhibited the release of nitric oxide in LPS-induced RAW264.7 macrophages.

This study aimed to characterize the chemical profile of Danggui Buxue Decoction (DBD) and investigate the systemic exposure and tissue distribution of its constituents in mice following oral administration, using UHPLC-Q-Exactive Orbitrap high-resolution mass spectrometry (HRMS). Methods A total of 12 healthy male C57BL/6J mice were randomly divided into a blank control group (n = 3), administered purified water, and a DBD-treated group (n = 9), given a DBD extract at 16 g/kg by gavage. Serum and tissue samples were collected at 0.5, 1.5, and 3 h post-dose, pooled, and processed via protein precipitation. Chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column (100 mm × 2.1 mm, 1.7 μm) using a gradient elution with 0.1% formic acid in water and methanol as the mobile phase. Mass spectrometric data were acquired in both positive and negative ion modes via full-scan MS¹/data-dependent MS² scanning. Compound identification was achieved by analyzing relative retention times and MS¹ fragment information using Xcalibur 4.2 software, with confirmation against reference standards, literature data, and databases. Results A total of 158 chemical compounds were identified in DBD, comprising 56 flavonoids, 44 saponins, 24 phthalides, 17 organic acids, 8 coumarins, 2 alkaloids, and 7 other constituents. The number of prototype compounds detected in mice after DBD administration varied over time. The abundance of these migrating components across tissues followed the order: ileum > liver > serum > lung > kidney > spleen > heart > brain. Key constituents identified in the serum and various tissues included 7,3'-dihydroxy-5'-methoxyisoflavone (pterocarpan E), calycosin-7-O-β-D-glucoside, methylnissolin, calycosin, oroxylin A, isomucronulatol, 3-butylidenephthalide, (6αR,11αR)-3,9-dimethoxy-10-hydroxypterocarpan and formononetin. Conclusion This study delineates the comprehensive chemical composition of DBD and in vivo absorption and tissue distribution patterns using HRMS, providing crucial insights for identifying the bioactive constituents of DBD and supporting further tissue pharmacokinetic investigations.

Objective Novel self-assembled nanoparticles (RMN-SAN) of Dahuang (Rhei Radix et Rhizoma) and Mudanpi (Moutan Cortex) were prepared by the micro-precipitation method, and their characterization, formation mechanism and anti-colorectal cancer (CRC) efficacy were investigated. Methods The process parameters were optimized through single-factor experiments, and characterization methods such as dynamic light scattering, ζ potential and transmission electron microscopy were performed. The core active components were identified by UPLC-Q-TOF-MS, and the self-assembly mechanism of RMN-SAN was revealed by FTIR and UV spectroscopy. The in vitro CT26 cell experiments and in vivo tumor-bearing mouse models were used to verify its anti-CRC efficacy. Results The optimized preparation method for RMN-SAN involves taking 10 g each of Rhei Radix et Rhizoma and Moutan Cortex, adding 10 times their volume of water, soaking for 30 min, and refluxing for 0.5 h, followed by filtration. The residue is then refluxed with 8 times its volume of 70% ethanol for 0.5 h and filtered again. The two extracts are combined, magnetically stirred at 600 r/min and 25 ℃ for 30 min, and the alcohol is removed by rotary evaporation at 60 ℃, concentrating to a final concentration of 0.2 g/mL. The solution is subjected to gradient centrifugation (4 ℃, 6 000, 8 000, and 10 000 r/min, each for 10 min), followed by dialysis (cut-off molecular weight 3 500) for 12 h, yielding the final product. The results showed that the average particle size of the optimized RMN-SAN was (109.30 ± 4.20) nm, the PDI was 0.353 ± 0.009, and the ζ potential was (-24.5 ± 1.3) mV, presenting a uniform spherical structure. Five core components such as rhein, aloe-emodin, sennoside B, paeonol, and paeoniflorin were identified, and their self-assembly was driven by hydrogen bonds and π-π stacking interactions, which was verified by molecular docking. The in vitro experiments confirmed that RMN-SAN could inhibit the proliferation of CT26 cells in a concentration-dependent manner, induce apoptosis and block the cell cycle at the G₁ phase. In the in vivo experiments, the tumor suppression effect of the high-dose RMN-SAN group (6.0 g/kg) was significantly higher than that of the traditional decoction group, and it could regulate the levels of immune and inflammatory factors more efficiently. Conclusion This study enabled the utilization of the therapeutic advantages of traditional Chinese medicine through modern processing, providing new ideas for the optimization of the preparation of self-assembled nanoparticles of traditional Chinese medicine, and also providing an efficient and stable new formulation for the treatment of CRC with traditional Chinese medicine.

Objective Compare the differences in material basis and anti-insomnia and anti-anxiety efficacy between Suanzaoren Decoction (SZRD, 酸枣仁汤) and Suanzao Decoction (SZD, 酸枣汤) in the Jin Gui Yao Lue. Methods This study quantifies 20 chemical ingredients as evaluation indicators to establish the boiling methods of SZRD and SZD, based on ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry (UPLC-Qtrap-MS) technology. Moreover, the ultra-high performance liquid chromatography-orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS/MS) metabolite identification method specific to the Chinese herbal medicine was used to identify and compare the chemical profile of the two decoctions. Lastly, the p-chlorophenylalanine (PCPA) induced insomnia model mice were used to evaluate and compare the efficacy of the two decoctions in anti-insomnia and anti-anxiety. Results The method of boiling Suanzaoren in ceramic-coated iron pot for 25 min and adding other herbs then continuing to boil for 40 min can match well with the original records in Jin Gui Yao Lue. When Suanzaoren and Suanzao fruit were both used with the amount of 2 litters in Han Dynasty, the contents of jujuboside A, jujuboside B, spinosin, 6‴-feruloylspinosin, magnoflorine, pachymic acid and ligustilide in SZRD were over 6-fold higher than those in SZD, whereas the contents of liquiritigenin, isoliquiritin, isoliquiritigenin and timosaponin AIII in SZD were over 2 times than those in SZRD. Among the top 100 compounds with the strongest mass spectrometric signals detected in the two decoctions, 42 compounds exhibited higher contents in SZD and 21 compounds were enriched in SZRD. Among the differential compounds, three isoquinoline alkaloids [(-)-isocorypalmine, cheilanthifoline and asimilobine], one pyrrolidine alkaloid (stachydrine) and one steroidal alkaloid (zhebeinine) were further identified. The relative abundances of cheilanthifoline and asimilobine in SZD were 5.2 times and 4.8 times higher than those in SZRD, respectively; meanwhile, the relative abundances of (-)-isocorypalmine, stachydrine and zhebeinine in SZD only reached 0.3, 0.4 and 0.4 times the corresponding values in SZRD. The anti-insomnia and anti-anxiety efficacies of the two decoctions in the PCPA induced insomnia mice are similar to each other, and no statistically significant intergroup difference in their pharmacological efficacy was observed.. Conclusion Under the original decoction method recorded in Jin Gui Yao Lue, SZRD and SZD show significant differences in material basis, yet their similar anti-insomnia and anti-anxiety effects under specific condition may result from the combined actions of multiple active components.

Objective To explore the self-assembly behavior and molecular association patterns of glycyrrhizic acid (GA) and genkwanin (Gen) after combination, and to analyze the correlation between the incompatibility and the molecular state of Gancao (Glycyrrhizae Radix et Rhizoma)-Yuanhua (Genkwa Flos) from the microscopic perspective. Methods Employ dynamic light scattering (DLS), scanning electron microscopy (SEM), and ultrafiltration to characterize the particle size, morphology and ultrafiltration behavior of GA-Gen mixed solutions. Results When GA and Gen were combined at mass ratios of 55:1 and 45:1, the particle size decreased significantly from 100—150 nm (GA monomer solutions) to approximately 50 nm. After combination, DLS showed a light intensity signal that was not present in the monomer solution at 10—100 nm. SEM images showed that the GA-Gen compatible solutions formed well-defined cubic particles, distinctly different from the globular and long needle-like morphologies of the individual components. When ultrafiltration membranes with molecular weight cut-off (M_w) of 3 000, 10 000, 30 000, 50 000 and 100 000 were used for separation, it was found that with the increase of GA concentration, the transmittance of GA gradually decreased, and the molecular weight at 10% transmittance (T₁₀) and the association coefficient (N) also gradually increased. At the same concentration, the transmittance of GA in the compatibility system was higher than that in the GA monomer system, and T₁₀ and N were lower than those in the monomer system. In addition, the transmittance of Gen in GA-containing system was significantly improved. The decoction system of medicinal materials is consistent with the trends of monomer system. Comparing the SEM of single decoction and co-decoction, the particle size of Glycyrrhizae Radix et Rhizoma single decoction was regular and the size was relatively uniform, and the particle size of Genkwa Flos single decoction was greatly different. The co-decoction failed to reproduce the regular cube of monomer compatibility, but the particle uniformity was between the two single decoctions. Conclusion GA and Gen do not simply coexist in aqueous solution. Instead, Gen is incorporated into GA micelles via molecular substitution, leading to the formation of new self-assembled associates. These findings provide a novel perspective for understanding the mechanism of “reverse” of Glycyrrhizae Radix et Rhizoma-Genkwa Flos herb pair.

Objective To establish a method for differentiating gelatinous Chinese medicines (GCMs) based on supramolecular sensing technology. Methods An indicator displacement assay (IDA) strategy was employed where test sample solutions of GCMs served as analytes and macrocyclic molecule-dye host-guest pairs with specific fluorescence signals were used as sensing pairs to construct a sensor array. The fluorescence intensity changes resulting from the interaction between different GCMs and the sensing units were detected and combined with pattern recognition methods for differentiation. Results Through screening host-guest sensing units with selective fluorescence responses to GCMs from a supramolecular library consisting of cyclodextrins, cucurbiturils, calixarenes, and biphen[n]arenes hosts combined with dyes, a quaternary supramolecular sensor array was constructed including quaternary ammonium salt calix[5]arene (QAC5A)·eosin Y (EY) (0.4 μmol/L vs 0.5 μmol/L), quaternary ammonium modified azocalix[4]arene (QAAC4A)·EY (1 μmol/L vs 1 μmol/L), dodecyl quaternary ammonium salt azo calix[4]arene (QAAC4A12C)·EY (0.6 μmol/L vs 1 μmol/L), and terphen[3]arene (WTP3)·fluorescein (Fl) (0.3 μmol/L vs 0.6 μmol/L). Principal component analysis (PCA) demonstrated that Ejiao (Colla Corii Asini), Xin’ejiao (Colla Corii Suilli), Biejiajiao (Colla Carapacis Trionychis), Lugujiao (Colla Ossis Cervi), Guijiajiao (Colla Carapacis et Plastri Testudinis) or Lujiajiao (Colla Cornus Cervi) could be significantly distinguished within a 95% confidence interval. Conclusion This study developed a quaternary supramolecular sensor array integrated with pattern recognition analysis which enables rapid discrimination of various GCMs.

Objective To screen for the most effective antimicrobial carbon dots from the traditional Chinese medicines Baiji (Bletilla striata), Cili (Rosa roxburghii), Lingzhi (Ganoderma lucidum) and Tianma (Gastrodia elata), and to systematically investigate the physicochemical properties, photothermal performance, antibacterial activity and mechanism of manganese-doped G. lucidum carbon dots (Mn-GLCDs). Methods Four kinds of herbal carbon dots derived from B. striata, R. roxburghii, G. lucidum and G. elata were prepared by a conventional hydrothermal method. The carbon dots with optimal antibacterial activity were screened out, and the morphology, structure and in vitro photothermal performance of the manganese-doped carbon dots were characterized. The antibacterial performance against Staphylococcus aureus and Escherichia coli under near-infrared (NIR) irradiation was evaluated through zone of inhibition assays, bacterial co-culture experiments, and biofilm inhibition and disruption tests. The antibacterial mechanism was preliminarily investigated by intracellular active oxygen fluorescence detection using a DCFH-DA probe. Biosafety was assessed by hemolysis assay and cytotoxicity test. Intracellular reactive oxygen species (ROS) fluorescence measurement using the 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) probe was adopted to preliminarily elucidate its antibacterial mechanism. Hemolysis assays and cytotoxicity evaluations were conducted to verify their biological safety. Results GLCDs possessed remarkably stronger antibacterial activity than carbon dots prepared from B. striata, R. roxburghii and G. elata carbon dots. Mn-GLCDs had a particle size of approximately 5 nm, and their surfaces were rich in hydrophilic functional groups including -OH, -NH₂, C=O, C-N-C and C-O-C. X-ray photoelectron spectroscopy (XPS) and inductively coupled plasma mass spectrometry (ICP-MS) confirmed that the mass fraction of Mn²⁺ reached 0.83%, and Mn²⁺ was stably present in the carbon dot framework in a coordinated form. The red shift of the UV-Vis absorption peak and the Raman I_D/I_G > 1 indicated that Mn²⁺ doping narrowed the bandgap and increased the density of structural defects. Photothermal performance evaluation showed that Mn-GLCDs could rapidly heat up under 808 nm laser irradiation (1.5 W/cm²), with a photothermal conversion efficiency of 35.03%. The maximum temperature fluctuation in six cycles of heating-cooling stability test was less than 6%, indicating good photothermal stability. Fluorescence performance test showed a fluorescence quantum yield of 20.51% and excitation-dependent fluorescence emission. In vitro antibacterial experiments demonstrated that, compared with undoped G. lucidum carbon dots, Mn-GLCDs exhibited the strongest antibacterial effect against S. aureus and E. coli under near-infrared irradiation, and could significantly inhibit and disrupt bacterial biofilms. Intracellular active oxygen detection showed that upon near-infrared irradiation, Mn-GLCDs elevated the active oxygen levels in S. aureus and E. coli to 6.75-fold and 5.44-fold of the negative control group, respectively. Biosafety evaluation showed that the hemolysis rate of Mn-GLCDs was below 5% within the antibacterial concentration range, and Mn-GLCDs exhibited no obvious toxicity to L929 and RAW264.7 cells. Conclusion This study constructs for the first time Mn-GLCDs, which significantly enhance the photothermal conversion capability and antibacterial activity. The mechanism is related to the induction of intracellular active oxygen. This work provides an experimental basis for the development of novel traditional Chinese medicine-based nanoscale antibacterial agents.

Objective This study focused on the concentration process in the manufacturing of Guizhi Fuling Capsules (GFC, 桂枝茯苓胶囊). By analyzing the correlations among effective components, physical properties, and process parameters, it aimed to identify critical quality attributes (CQAs) and critical process parameters (CPPs), explore quality control indicators and their acceptable ranges, and assess the potential for using physical properties of intermediate to guide its quality control. Methods The variations in process parameters (temperature, relative density at endpoint), physical properties (particle size, polydispersity index, ζ potential, conductivity, pH value, viscosity and refractive index), and effective components and total solid content during the concentration process were systematically investigated. Chemometric methods, including Pearson correlation coefficient, grey relational analysis, and partial least squares regression, were applied. Linear models between effective components and physical properties under different process parameters were established to elucidate the correlations among them. Results Analysis of effective component content and physical properties revealed strong correlations between effective components and ζ potential, conductivity, viscosity, and total solids content. For instance, viscosity was positively correlated with total solids, and the contents of amygdalin and paeoniflorin were both positively correlated with total solid content. Analysis of the active ingredients and physical property parameters of samples under various process parameters revealed that the concentration temperature is a key factor influencing the correlation between active ingredients and physical property parameters. Conclusion Correlation analysis of process parameters, physical properties, and effective components revealed interactive and dynamically linked relationships among these variables. Precise control of the concentration temperature during the manufacturing is essential to improve the quality and batch-to-batch consistency of the extract. To a certain extent, parameters such as viscosity, relative density, refractive index and pH value can reflect the quality of intermediates during concentration, providing a reference for the development of online intelligent monitoring in the concentration process of traditional Chinese medicine extracts.

Objective To explore the mechanism by which Taohong Siwu Decoction (桃红四物汤, THSWD) alleviates cerebral ischemia-reperfusion injury (CIRI) through protecting the integrity and functionality of glycocalyx of brain microvascular endothelial cells. Methods A model of middle cerebral artery occlusion-reperfusion (MCAO/R) was established to simulate CIRI injury. SD rats were randomly divided into sham group, model group, THSWD low-, medium-, high-dose (4.5, 9.0, 18.0 g/kg) groups and nimodipine (20 mg/kg) group, and drugs were given for intervention. After 7 d of modeling, Longa grading system was used for neurological function evaluation, 2,3,5-triphenyltetrazolium chloride (TTC) staining was used to assess cerebral infarction volume. The motion function was evaluated through corner experiments and balance beam experiments. Laser speckle and small animal super-resolution imaging were used to detect cerebral blood flow and cerebral vascular density, Evans blue leak assay combined with Western blotting were to evaluate blood-brain barrier integrity. Glycocalyx morphology was observed using transmission electron microscopy. The expression of glycocalyx-related indicator such as syndecan-1 (SDC-1), hyaluronic acid (HA), heparan sulfate (HS), chondroitin sulfate (CS) and core metabolic enzymes of glycocalyx such as heparanase (HPSE), hyaluronidase 2 (Hyal2), neuraminidase 1 (Neu1), matrix metalloproteinase-9 (MMP-9) were detected by immunofluorescence, immunohistochemistry, Western blotting and ELISA, and the effects of HPSE and MMP-9 were verified by enzyme inhibitors. Results Compared with model group, THSWD significantly reduced the cerebral infarction volume in MCAO/R rats (P < 0.05, 0.001), improved neurological function and limb coordination ability (P < 0.05, 0.01, 0.001), alleviated weight loss, improved cerebral blood flow and microvascular generation in the infarct area, reduced blood-brain barrier leakage (P < 0.01, 0.001), regulated vascular dilation and contraction function (P < 0.01, 0.001). Meanwhile, THSWD could significantly reduce the shedding of core components of glycocalyx (CS, HA, HS, SDC-1) into the serum (P < 0.01, 0.001), and maintain the integrity of glycocalyx by inhibiting the activities of HPSE and MMP-9. Conclusion THSWD reduces the shedding of glycocalyx by inhibiting the activity of HPSE and MMP-9, thereby maintaining the integrity of blood-brain barrier and minimizing stroke damage.

Objective To investigate the effect of catalpol on lipopolysaccharide (LPS)-induced inflammatory injury, extracellular matrix (ECM) metabolic dysregulation and pyroptosis in chondrocytes, and to explore whether these effects involve the AMP-activated protein kinase (AMPK)/nuclear factor-κB (NF-κB)/NOD like receptor family pyrin domain containing 3 (NLRP3) signaling axis. Methods Network pharmacology was used to screening the intersection targets of catalpol, knee osteoarthritis and pyroptosis and enrichment analysis were performed. LPS was used to establish an inflammatory injury and induce pyroptosis model, followed by pretreatment with catalpol (20, 50 μmol/L) for 1 h. Protein expressions of collagen type II alpha 1 chain (COL2A1), aggrecan (ACAN), matrix metalloproteinase 3 (MMP3), MMP13 and key molecules in AMPK/NF-κB/NLRP3 signaling axis were detected by Western blotting. Lactate dehydrogenase (LDH) release assay was performed to assess cell membrane integrity-related injury. The levels of interleukin-1β (IL-1β) and IL-18 in supernatants were detected by ELISA. The mRNA levels of NLRP3, IL-1β, IL-6 and tumor necrosis factor-α (TNF-α) were detected by qRT-PCR. The cystein-asparate protease-1 (Caspase-1) inhibitor VX-765 was used as a positive control, the mechanism was further validated using AMPK inhibitor Compound C. Results Compared with control group, LPS stimulation could induce a decrease in ECM synthesis and an increase in ECM decomposition, significantly activate the NLRP3 related pyroptosis pathway (P < 0.001). Compared with model group, catalpol could significantly upregulate the expressions of COL2A1, ACAN and downregulate the expressions of MMP3, MMP13 (P < 0.05, 0.01, 0.001), while significantly reduce the release of LDH, IL-1β and IL-18 (P < 0.01, 0.001), and inhibit NLRP3, cleaved Caspase-1, N-terminal fragment of gasdermin D (GSDMD-N) and other pyroptosis related indicators (P < 0.05, 0.001). In addition, catalpol could significantly upregulate the expressions of p-AMPK/AMPK and phosphorylated acetyl CoA carboxylase (p-ACC)/ACC (P < 0.05, 0.001), and downregulate the expression of p-p65/p65 (P < 0.001), while downregulate the expressions of NLRP3, IL-1β, IL-6, TNF-α inflammation related genes (P < 0.05, 0.001). AMPK inhibitor Compound C could partially reverse the above-mentioned effects of catalpol (P < 0.05, 0.01, 0.001). Conclusion Catalpol may protect the matrix homeostasis of chondrocytes by activating AMPK and promoting downstream ACC phosphorylation, inhibiting NF-κB/NLRP3 related pathways.

Objective To investigate the mechanism of tannic acid in promoting mitochondrial autophagy in chondrocytes and improving knee osteoarthritis (KOA) in mice based on PINK1/Parkin signaling pathway. Methods A mouse KOA model was established via destabilization of medial meniscus surgery. Sham group, model group and tannic acid low-, high-dose (25, 50 mg/kg) groups were set up, drugs were given for intervention for eight weeks. Joint cartilage pathology was assessed by imaging, safranin O-fast green staining and toluidine blue staining. Immunohistochemistry was used to detect the expressions of collagen II (Col-II), matrix metalloproteinase 13 (MMP13), interleukin-1β (IL-1β), PINK1, Parkin, p62 and microtubule-associated protein light chain 3B (LC3B) in cartilage tissue. ATDC5 mouse chondrocytes were stimulated with IL-1β and then treated with tannic acid. Cell viability was measured by CCK-8 assay. Western blotting and qRT-PCR were used to detect the protein and mRNA expressions of Col-II, MMP13 and IL-1β. Western blotting was used to detect the expressions of mitophagy-related proteins PINK1, Parkin, p62 and LC3-II/I. Changes in reactive oxygen species (ROS) and mitochondrial membrane potential were detected by immunofluorescence. Mitochondrial autophagy level was observed by transmission electron microscopy. And reverse mechanism validation was performed using the mitochondrial autophagy inhibitor Mdivi-1. Results Compared with sham group, the cartilage of mice in model group was severely worn, OARSI score was significantly increased (P < 0.01), the protein expression levels of Col-II, PINK1, Parkin and LC3B in cartilage tissue were significantly reduced (P < 0.01), while the protein expression levels of IL-1β, MMP13 and p62 were significantly increased (P < 0.01). Compared with model group, the cartilage morphology of mice in tannic acid group was improved, OARSI score was significantly reduced (P < 0.05), the protein expression levels of Col-II, PINK1, Parkin and LC3B in cartilage tissue were significantly increased (P < 0.01), while the protein expression levels of IL-1β, MMP13 and p62 were significantly decreased (P < 0.01). The cell experiment results showed that IL-1β-induced chondrocyte viability was significantly reduced (P < 0.01), IL-1β, MMP13 protein and mRNA expression levels were significantly increased (P < 0.01), Col-II protein and mRNA expression levels were significantly reduced (P < 0.01), LC3-II/I, PINK1, Parkin protein expression levels were significantly reduced (P < 0.05, 0.01), p62 protein expression level was significantly increased (P < 0.05), intracellular ROS accumulation was increased (P < 0.01), mitochondrial membrane potential was decreased (P < 0.01), and autophagosome was decreased. Compared with model group, cell viability in tannic acid group was significantly increased (P < 0.05, 0.01), ROS accumulation significantly was decreased (P < 0.01), mitochondrial membrane potential was recovered (P < 0.05), IL-1β, MMP13 protein and mRNA expression levels were significantly decreased (P < 0.05, 0.01), Col-II protein and mRNA expression levels were significantly increased (P < 0.05, 0.01), p62 protein expression level was significantly decreased (P < 0.01), LC3-II/I, PINK1 and Parkin protein expression levels were significantly increased (P < 0.01). After the combined use of mitochondrial autophagy inhibitor Mdivi-1, the therapeutic effect of tannic acid on chondrocytes was reversed (P < 0.05, 0.01). Conclusion Tannic acid alleviates articular cartilage damage and delays the progression of KOA by promoting chondrocyte mitophagy through PINK1/Parkin signaling pathway.

Objective To investigate the mechanism by which tetrandrine regulates the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) signaling pathway to induce ferroptosis in breast cancer cells based on peroxiredoxin 1 (PRDX1). Methods Human breast cancer MCF-7 cells transfected with PRDX1 siRNA were treated with tetrandrine to examine its effects on cell proliferation, apoptosis, migration and invasion. The levels of malondialdehyde (MDA), Fe²⁺, intracellular and mitochondrial reactive oxygen species (ROS), mitochondrial membrane potential, and mitochondrial membrane permeability were measured. Western blotting and qRT-PCR were used to detect the expressions of voltage-dependent anion channel 1 (VDAC1), p53, PI3K/Akt/GSK-3β signaling pathway and nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway related proteins and genes. Results Tetrandrine dose-dependently inhibited MCF-7 cell proliferation, migration and invasion while inducing apoptosis (P < 0.01), increased intracellular and mitochondrial ROS, MDA and Fe²⁺ levels (P < 0.01), aggravated lipid peroxidation, reduced mitochondrial membrane potential (P < 0.01), enhanced mitochondrial membrane permeability (P < 0.01), upregulated HO-1, p53, GSK-3β and VDAC1 expressions (P < 0.01), downregulated Nrf2, PI3K, p-PI3K, Akt and p-Akt expressions (P < 0.01). Knockdown of PRDX1 gene significantly amplified these effects of tetrandrine (P < 0.01). Conclusion Tetrandrine induces mitochondria-associated ferroptosis in MCF-7 cells via PI3K/Akt/GSK-3β signaling pathway, with PRDX1 playing a critical regulatory role.

Objective To investigate the therapeutic effect of icariin on diabetic kidney disease (DKD) and elucidate its molecular mechanism from the perspective of renal inflammatory response and related inflammatory signaling pathways. Methods A DKD model was established using C57BL/KsJ db/db mice, with age-matched db/m mice on the same genetic background serving as control group. The db/db mice were randomly divided into model group, dapagliflozin (1.6 mg/kg) group, icariin high-, medium-, low-dose (120, 60, 30 mg/kg) groups, with 10 mice in each group. Drugs were given for continuously intervention over four weeks, biochemical parameters including body weight, blood glucose, urinary protein, blood lipids and serum creatinine were measured. Renal pathological changes were observed using hematoxylin-eosin staining and transmission electron microscopy. The expressions of extracellular matrix proteins [fibronectin (FN) and collagen type IV (Col-Ⅳ)] in the glomerular mesangium was assessed by immunofluorescence. Levels of inflammatory cytokines [interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α)] and proteins related to high mobility group box 1 (HMGB1)/Toll-like receptors (TLRs)/nuclear factor-κB (NF-κB) signaling pathway were detected by ELISA assay and Western blotting. In vitro experiments were performed to investigate the effect of icariin on extracellular matrix protein expressions in high glucose-induced SV40 MES-13 cells. Results In vivo experiments results showed that high-dose icariin significantly reduced the body weight, urinary protein, blood lipid and serum creatinine levels in DKD mice (P < 0.01, 0.001), alleviated pathological changes such as mesangial expansion, basement membrane thickening and podocyte fusion (P < 0.001), reduced the levels of IL-1β, IL-6 and TNF-α in plasma and renal tissue (P < 0.01, 0.001), decreased the infiltration of macrophages in the kidney, and significantly inhibited the protein expressions of HMGB1, TLR2, TLR4 and p-p65 in renal tissue (P < 0.001). In vitro experiments results showed that icariin inhibited the expressions of FN and Col-Ⅳ in high glucose-induced glomerular mesangial cells. Conclusion Icariin could effectively inhibit HMGB1/TLRs/NF-κB signaling pathway, thereby attenuating renal inflammatory response and abnormal extracellular matrix accumulation, ultimately improving pathological injury in DKD.

Objective To explore the effect of usnic acid on inflammatory injury in mice with intracerebral hemorrhage (ICH) based on high mobility group box 1 (HMGB1)/receptor for advanced glycation end products (RAGE) pathway. Methods C57BL/6 mice were randomly divided into sham group, model group, HMGB1 inhibitor ethyl pyruvate (40 mg/kg) group, usnic acid low-, high-dose (20, 30 mg/kg) groups and usnic acid (30 mg/kg) + rHMGB1 (8 μg/kg) group, with 18 mice in each group. The ICH model was constructed by intracranial injection of 0.5 μL collagenase. After continuous administration for 3 d, the modified neurological severity score (mNSS) and left turn rate of mice in each group were evaluated. The water rate of brain tissue in mice was determined. ELISA was used to measure the levels of interleukin-6 (IL-6), IL-1β and IL-18 in brain tissue. Hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue. Immunofluorescence was used to observe microglia activation in brain tissue. TUNEL staining was used to detect cell apoptosis of brain tissue. Western blotting was used to detect the expressions of HMGB1/RAGE pathway related proteins in brain tissue. Results Compared with sham group, the striatum tissue of mice in model group was severely damaged, mNSS, left cerebral hemisphere water rate, ionized calcium-binding adapter molecule 1 (Iba1) positive cells number, inflammatory factors levels, cell apoptosis rate, HMGB1, RAGE, phosphorylated nuclear factor-κB p65 (p-NF-κB p65)/NF-κB p65 and TNF-α protein expression levels were significantly increased (P < 0.05), and the left turn rate was significantly decreased (P < 0.05). Compared with model group, the damage degree of striatum tissue in usnic acid low-, high-dose group and ethyl pyruvate group was alleviated (P < 0.05), mNSS, left cerebral hemisphere water rate, Iba1 positive cells number, inflammatory factors levels, cell apoptosis rate and HMGB1/RAGE pathway related proteins expression levels were significantly decreased (P < 0.05), and the left turn rate was significantly increased (P < 0.05). rHMGB1 significantly reversed the inhibitory effect of usnic acid on inflammatory injury in ICH mice (P < 0.05). Conclusion Usnic acid may alleviate inflammatory injury in ICH mice by inhibiting HMGB1/RAGE pathway.

Objective To explore the characteristics, medication patterns, and the mechanisms of action of traditional Chinese medicines for treating angle-closure glaucoma. Methods Literature and prescriptions related to the treatment of angle-closure glaucoma with traditional Chinese medicine were retrieved from databases including CNKI, Wanfang, VIP, China Biology Medicine (CBM), and the Ancient and Modern Medical Case Cloud Platform. Through data mining and analysis, core prescriptions were identified. Subsequently, a network pharmacology approach was employed. Active ingredients with oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18 were screened using the TCMSP and BATMAN-TCM 2.0 databases. Targets were uniformly annotated via the UniProt database and intersected with disease targets obtained by integrating and deduplicating data from four major databases (GeneCards, TTD, OMIM, and CTD) to identify pharmacodynamic targets. A “core prescription-active ingredient-pharmacodynamic target” interaction network was constructed using Cytoscape 3.10.2. The pharmacodynamic targets were then imported into the Metascape platform (restricted to human species, with a threshold of P < 0.05) for gene ontology (GO) (biological process, molecular function, cellular component) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis, with the top 20 significant pathways and functions visualized. Additionally, a protein-protein interaction (PPI) network was constructed using the String database, and core targets were screened using the Cytoscape MCODE plugin (cut-off = 0.05) to elucidate the potential molecular mechanisms underlying the core prescriptions in treating angle-closure glaucoma. The binding energy between the active ingredient and the core target was investigated through molecular docking. Sodium dithionite was used to induce hypoxic injury in mouse retinal ganglion cells (RGCs) to establish an in vitro model. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was employed to detect the effects of these active components on mRNA expression of core target genes. ELISA was used to measure the impact of these active components on tumor necrosis factor-α (TNF-α) levels. Immunofluorescence staining was applied to evaluate the effects of these active components on phosphorylated protein kinase B (p-AKT) and nuclear factor-κB (NF-κB) protein expression. Results The top three most frequently used traditional Chinese medicines for angle-closure glaucoma were Fuling (Poria), Danggui (Angelicae Sinensis Radix), and Chaihu (Bupleuri Radix). The core syndromes were mainly liver qi stagnation and liver-kidney deficiency. A core prescription consisted of Mudanpi (Moutan Cortex)-Angelicae Sinensis Radix-Zhizi (Gardeniae Fructus)-Bupleuri Radix-Juhua (Chrysanthemi Flos)-Baizhu (Atractylodis Macrocephalae Rhizoma)-Gancao (Glycyrrhizae Radix et Rhizoma)-Baishao (Paeoniae Radix Alba)-Poria. Cluster analysis yielded three clusters: modified Chaihu Shugan San (柴胡疏肝散), modified Yijing Buyang Huanwu Decoction (益精补阳还五汤), modified Qiju Dihuang Pills (杞菊地黄丸). Network pharmacology analysis showed that the core prescription regulated key targets such as AKT1, TNF-α, NF-κB by modulating signalling pathways such as TNF, phosphatidylinositol-3-hydroxykinase (PI3K)-Akt, and MAPK (mitogen-activated protein kinase), thereby participating in biological processes such as inflammatory response, and oxidative stress, and thus intervening in the pathological process of glaucoma. Quercetin and luteolin were identified as the core components of the core prescription for the intervention of angle-closure glaucoma through molecular docking, and in vitro experiments were conducted for verification. The results showed that all luteolin dose groups (5, 10, 20 μmol/L) upregulated AKT1 mRNA expression (P < 0.01), while the medium and high dose groups downregulated NF-κB and TNF-α mRNA expression (P < 0.05), significantly increased p-AKT protein expression, and decreased NF-κB protein and TNF-α levels (P < 0.05). The medium and high dose groups of quercetin (40, 80 μmol/L) also significantly upregulated AKT1 mRNA expression and downregulated NF-κB and TNF-α mRNA expression (P < 0.05). All low (20 μmol/L), medium, and high dose groups of quercetin increased p-AKT protein expression and decreased NF-κB protein expression (P < 0.05), while the medium and high dose groups significantly reduced TNF-α expression (P < 0.05). Conclusion Data mining techniques were systematically employed to analyze ancient and modern literature and medical records concerning the treatment of angle-closure glaucoma. From multiple dimensions including medication frequency, nature-flavor-meridian tropism, efficacy categories, association rules, and core combinations, the “high-frequency-core-differential” three-dimensional characteristics were progressively analyzed to extract core prescriptions and reveal their compatibility patterns. Network pharmacology analysis further confirmed that the above high-frequency prescriptions synergistically regulate key biological processes such as oxidative stress, inflammatory response, and apoptosis through a “multi-component-multi-target-multi-pathway” mechanism, reflecting the holistic therapeutic advantages of traditional Chinese medicine compound formulas, thereby providing scientific evidence for clinical syndrome differentiation and medication, with certain guiding significance. In vitro experiments simultaneously validated that, in the hypoxic injury model, both luteolin and quercetin significantly upregulated AKT1 mRNA and p-AKT protein expression, inhibited NF-κB and TNF-α mRNA and protein expression, effectively reduced levels of inflammation and apoptosis-related factors, and exerted protective effects on RGCs through a bidirectional mechanism of “anti-inflammatory and pro-survival” effects.

Objective To conduct a bibliometric analysis of Chinese and English literature on the prevention and treatment of knee osteoarthritis (KOA) by traditional Chinese medicine (TCM) from 1995 to 2025, and visualize its research status, development context, cooperation network, hot topics and frontier trends. Methods Relevant Chinese literature from January 1995 to December 2025 were retrieved from CNKI, Wanfang and VIP, and relevant English literatures from January 1998 to December 2025 were retrieved from Web of Science (WOS). CiteSpace and VOSviewer software were used to visually analyze the trend of publication, countries, institutions, authors, keywords, etc., and draw a knowledge map. Results A total of 6 093 literature were included, including 5 242 Chinese literature and 851 English literature. The overall number of publications showed a fluctuating upward trend, and Chinese research output and cooperation in this field were dominant. The core research institutions are represented by Guangzhou University of Chinese Medicine, Fujian University of Traditional Chinese Medicine and Beijing University of Chinese Medicine. Keyword analysis showed that the current research hotspots focused on traditional therapies such as clinical efficacy, mechanism of action, traditional Chinese medicine compound, acupuncture and massage. Emergent words and cluster analysis showed that the frontier trend was turning to the mechanism exploration and application research of interdisciplinary and new drug delivery systems such as network pharmacology, metabolomics, programmed cell death, intestinal flora-articular axis, self-assembled nanocarriers and metal-organic frameworks. Conclusion Research on TCM for KOA prevention and treatment has entered a phase of rapid development and deep integration. While continuing to validate clinical efficacy, it is increasingly converging with modern technologies, shifting the research paradigm toward systems biology, precision medicine, and intelligent drug delivery. Through the integration of multi-omics, molecular subtyping, and the development of innovative formulations, TCM holds promise for playing a more pivotal role in the prevention, early intervention, and disease-modifying treatment of KOA.

Objective To conduct a visual analysis of hotspots and development trends in Curcumae Radix research domestically and internationally using bibliometric software CiteSpace and VOSviewer, and provide a reference for in-depth research and application of Curcumae Radix. Methods The China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Platform, Web of Science (WOS) Core Collection, and PubMed databases were searched as data sources. The search period was set from January 1, 2016 to December 12, 2025. Analyses of publication volume, authors, and keywords were performed on the selected literature using CiteSpace6.4.R1 and VOSviewer1.6.20 softwares. Results A total of 1 354 Chinese literature and 77 English literature were included. Shi Zhenggang was identified as the core author of Chinese publications with 21 papers, while Ji De was the core author of English publications with six papers. Chinese research has formed a bidirectional knowledge network of “clinical experience-computational mechanism” represented by “data mining” and “network pharmacology”. English research focuses on “Curcumae Radix” and “molecular docking”, demonstrating an in-depth path of “computational prediction-disease focus”. The research hotspots are rapidly evolving from early-stage clinical efficacy observation to data-driven and computational pharmacological mechanistic elucidation. Conclusion Research on Curcumae Radix has evolved from simple efficacy observation into a systematic exploration that equally emphasizes “clinical problems” and “mechanistic elucidation”, while deeply integrating computational simulation and systems biology methods. Future efforts should focus on establishing a closed-loop research system of “computational prediction-experimental verification”, promoting high-quality evidence-based transformation, thereby driving the transformation and upgrading of this field toward precision medicine and modernized pharmacology.

Objective To integrate the correlation between the administration routes of traditional Chinese medicine essential oils and diseases, as well as their core biomarkers in disease treatment, analyze their pathways and mechanisms of action on the body, and promote the rational application and innovative development of traditional Chinese medicine essential oils. Methods China National Knowledge Infrastructure(CNKI), VIP Information(VIP), Wanfang Data, Web of Science were used to analyze the distribution of administration routes for traditional Chinese herbal essential oils and the types of diseases treated. The correlation between different administration routes and disease types was examined, along with frequency statistics of biomarkers involved in the therapeutic process. After identifying the most closely associated biomarkers, their mechanisms of action in disease treatment by traditional Chinese herbal essential oils were analyzed. Results Among all administration routes, oral administration accounted for 66.23%, inhalation for 21.37%, topical application for 8.18%, intraperitoneal injection for 3.17%, and intravenous injection for 1.06%. The disease types most strongly associated with oral and inhalation routes were neurological and psychiatric disorders, while those most associated with topical application were skin diseases and immune-related conditions. Intraperitoneal injection showed the strongest association with tumor diseases. Among biomarkers appearing more than 10 times, those related to oxidative stress regulation included malondialdehyde (MDA), nitric oxide (NO), and superoxide dismutase (SOD); those involved in inflammatory response regulation included tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, and IL-8; those associated with apoptosis regulation included B-cell lymphoma-2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), and cysteinasparate protease-3 (Caspase-3); and those involved in neural regulation included 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), norepinephrine (NE), and acetylcholinesterase (AchE). Conclusion Traditional Chinese medicine essential oils enter the human body through three primary routes: inhalation, transdermal absorption and oral administratio to exert their biological effects. They can exert effects via by regulating multiple mechanisms such as oxidative stress, neurotransmitter, inflammatory responses, and apoptosis, demonstrating significant advantages in disease prevention and treatment.

Objective By conducting a whole-genome identification and functional analysis of auxin response factors (ARFs) family members in Gentiana rigescens, this study investigate their potential roles in the thermosensitive corolla opening-closing movement of its corolla. Methods Based on whole-genome data and transcriptome data, bioinformatics methods were used to analyze the physicochemical properties, conserved motifs, gene structure, cis-acting elements, and evolutionary relationships of the G. rigescens ARFs (GrARFs) gene family. Real-time quantitative reverse transcription PCR (RT-qPCR) technology was employed to detect the expression patterns of GrARFs genes under different opening and closing states. Additionally, subcellular localization experiments were conducted to verify the protein localization of GrARF9, so as to preliminarily analyze the biological functions of the GrARFs gene family. Results A total of 21 GrARFs were identified in the G. rigescens genome, encoding 612—1 065 amino acids. Their relative molecular weights ranged from 68 082.29 to 118 117.97, and their isoelectric points spanned 5.52—8.43. The secondary structure was dominated by random coils. Phylogenetic tree analysis classified the ARFs in G. rigescens into four subgroups, namely Group A, Group B, Group C, and Group D. Most GrARFs contained Auxin-resp and B3 DNA-binding domains. The promoter cis-acting elements of GrARFs were rich in hormone-responsive and light-responsive elements. The 21 GrARFs were unevenly distributed across eight chromosomes. Collinearity analysis revealed that the GrARFs in G. rigescens were relatively conserved among dicotyledonous plants and shared a considerable number of collinear pairs with the functionally characterized ARFs from Rosa hybrida. All 21 GrARFs genes exhibited differential expression during the thermosensitive process of G. rigescens corollas. Results of subcellular localization experiments showed that the GrARF9 protein was localized in the nucleus, displaying the nuclear localization characteristic of transcription factors. Conclusion The results of this study preliminarily clarify the expression pattern of GrARFs in the thermosensitive movement of G. rigescens, and lay a foundation for further research on the function of GrARFs in the corolla movement of G. rigescens.

Objective To identify members of the cycloartenol synthase (CAS) gene family (PgCAS) using the Panax ginseng transcriptome database, and analyze the expression patterns of PgCAS04-5 in overexpressed and RNAi-positive hair root materials), as well as explore the function of this gene in ginsenoside biosynthesis. Methods Bioinformatics approaches were adopted to characterize the chromosomal localization, conserved motifs, phylogenetic relationships and expression profiles, and the correlations with the gene expression of key saponin synthesis enzymes and the content of monomer saponin of PgCASs. The protein structure and subcellular localization of PgCAS04-5 were predicted. Meanwhile, the expression levels of key enzyme genes and variations in ginsenoside contents were detected in overexpression and RNAi-positive hairy root lines. Results A total of 20 PgCAS transcripts were identified from the P. ginseng transcriptome database. Chromosomal mapping showed that PgCAS genes were unevenly distributed on five chromosomes of P. ginseng. Phylogenetic tree analysis divided the PgCAS family into three subfamilies, and most PgCAS members belonged to subfamily Ⅱ and subfamily Ⅲ. The PgCAS genes displayed obvious spatiotemporal specificity in expression across different cultivars, growth years and tissues. Additionally, the PgCAS family formed an interaction network with key genes involved in ginsenoside biosynthesis, and their content was significantly correlated with the accumulation of monomeric ginsenosides. Subcellular localization prediction demonstrated that PgCAS04-5 was localized to chloroplasts. Moreover, the overexpression of this gene was inversely regulated with the expression of four genes including PgCAS04-5, PgDS-1, PgSE2-4 and PgCYP137 in RNAi positive materials, with significant differences. Conclusion A total of 20 members of the PgCAS gene family were identified in this study, and functional differentiation exists among PgCAS family members. PgCAS04-5 positively regulates the biosynthesis of ginsenosides Rg₂and Rf, and negatively modulates the synthesis of ginsenosides Rb₂and Rd. The results provide a valuable reference for further research on the PgCAS gene fami

Objective To clone the full-length cDNA sequence of the CoMYB62 gene from Cornus officinalis, and analyze the structural characteristics, phylogenetic relationships, subcellular localization, and tissue expression patterns of its encoded protein, thereby providing a molecular basis for elucidating the potential roles of MYB transcription factors in the growth, development, and environmental adaptation of C. officinalis. Methods Based on transcriptome data of C. officinalis, the CoMYB62 gene sequence was screened and its full-length cDNA sequence was cloned using RT-PCR. Bioinformatics analyses were conducted to predict the physicochemical properties, conserved domains, secondary and tertiary structures, and phylogenetic relationships of the CoMYB62-encoded protein. A pCAMBIA1300-CoMYB62-GFP fusion expression vector was constructed and transiently expressed in tobacco leaves, and the subcellular localization of the CoMYB62 protein was observed using a laser confocal inverted microscope. In addition, quantitative real-time PCR (qRT-PCR) was performed to analyze the expression patterns of CoMYB62 in four tissues, including root, stem, leaf, and fruit. Results The full-length cDNA of the CoMYB62 gene was successfully cloned. Sequence analysis indicated that the encoded protein belonged to a typical R2R3-MYB transcription factor and contained a highly conserved DNA-binding domain at the N-terminus, which was predicted to be mainly localized in the nucleus. Phylogenetic analysis revealed that CoMYB62 shared high evolutionary conservation with MYB homologous proteins from various plant species. Laser confocal microscopy further confirmed that the CoMYB62 protein was localized in the nucleus. The qRT-PCR results showed that CoMYB62 was expressed in all examined tissues of C. officinalis, although the expression levels varied significantly among different tissues. Conclusion CoMYB62 is a conserved MYB transcription factor in C. officinalis with tissue-specific expression characteristics, which may participate in the regulation of plant growth, development, and stress responses to provide a foundation for further biological functional analysis of CoMYB62 in C. officinalis.

Objective To explore the chemical profile variations of Glycyrrhiza uralensis from different origins and screen origin-specific characteristic markers by using UPLC fingerprinting integrated with chemometrics. This approach provides a scientific basis and reference for the quality evaluation of G. uralensis with unified attributes. Methods This study established UPLC fingerprints for 30 batches of G. uralensis collected from five origins and validated the analytical method. The quality consistency of the samples was preliminarily analyzed through similarity evaluation. Subsequently, chemometric methods such as principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to identify the patterns and differences of G. uralensis samples from different origins by SIMCA 14.0 software. Further, the key discriminant markers were identified based on the variable importance projection (VIP) values. Finally, the contents of the five characteristic markers were determined. Results UPLC fingerprints of 30 batches of G. uralensis samples were established. The similarities between each sample batch and the reference fingerprint varied but all exceeded 0.849. This indicates that while the overall chemical profiles of the samples were generally consistent, quantitative differences existed. Chemometric analysis showed clear clustering trends for samples from different origins, and the consistent results of PCA and HCA confirmed the presence of chemical differences among the origins. The OPLS-DA model effectively distinguished different origins, and the permutation test confirmed the reliability of the model. Five key characteristic markers were selected based on VIP > 1. The content determination results indicated that the contents of the five characteristic markers in G. uralensis from different origins varied significantly, further confirming the differences in chemical components among the origins. Conclusion The UPLC fingerprint combined with chemometrics (PCA, HCA, OPLS-DA) analysis method established in this study can effectively distinguish G. uralensis from different origins, and glycyrrhizin, glycyrrhizic acid, liquiritin apioside, glycyrrhizin B, and violanthin can be used as key characteristic markers for origin discrimination. This analytical system provides a reliable strategy and data support for objectively evaluating the quality differences of G. uralensis from different origins, identifying their origin sources, and improving future quality standards.

Objective To investigate the composition and differential characteristics of volatile organic compounds (VOCs) in Fritillariae Cirrhosae Bulbus (FCB) from six legally authorized origins, and provide a reference for origin identification and quality evaluation of FCB. Methods Headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) was employed to analyze the volatile components in dried bulbs of Fritillaria cirrhosa, F. unibracteata, F. przewalskii, F. delavayi, F. taipaiensis and F. unibracteata var. Wabuensis. Qualitative matching was performed using the built-in NIST and IMS database of GC-IMS Library Search. Principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and hierarchical cluster analysis (HCA) were applied to evaluate the differences in volatile components among samples from different origins. Results A total of 90 volatile organic compounds were preliminarily identified, mainly including aldehydes, alcohols, ketones, esters, carboxylic acids, pyrazines, and sulfur-containing compounds. Among them, aldehydes, alcohols, and ketones were the major categories. PCA and OPLS-DA results indicated that FCB samples from the six origins could be well distinguished. Based on variable importance in projection (VIP) values, 17 potential differential markers were screened out, including ethyl acetate, 2-propanone, α-phellandrene, 3-hydroxy-2-butanone, methyl acetate, 2-propanethiol, 2-isobutyl-3-methoxypyrazine, propyl acetate, 3-methylbutanal, 1-butanol, 2-methyl-2-propenal, hexanal, 3-methyl-1-pentanol, 2-methylpropanoic acid, 3-butenenitrile, benzaldehyde, and 2-octanone. Cluster heatmap analysis further demonstrated that the above differential compounds could reflect the volatile metabolic characteristics of FCB from different origins. Conclusion HS-GC-IMS combined with chemometrics can effectively characterize the differences in volatile organic compounds among FCB from different origins, providing technical reference for origin identification, quality evaluation, and characteristic component screening of FCB.

Postmenopausal osteoporosis (PMOP) results from a combination of multiple pathological mechanisms, including osteogenic impairment and osteoclast activation triggered by estrogen deficiency-induced chronic low-grade inflammation and mitochondrial dysfunction, as well as systemic inflammation and metabolic dysregulation caused by gut microbiota imbalance. Faced with this intricate and multidimensional pathological network, conventional single-target therapeutic strategies exhibit obvious limitations. Traditional Chinese medicine (TCM), with its theoretical advantages of “holistic view” and “syndrome differentiation-based treatment,” demonstrates potential for systematic intervention through multi-target and multi-pathway coordinated regulation. Current studies indicate that various TCM formulas and active components can effectively restore the homeostasis of the bone-immune microenvironment and balance between gut microbiota and bone metabolism by comprehensively modulating these multidimensional pathological networks. Based on this, this review systematically summarizes the multidimensional mechanisms of PMOP, highlighting the multi-target regulatory effects of TCM within this pathological network, thereby providing theoretical support and research ideas for in-depth elucidation of PMOP pathogenesis and for advancing integrated prevention and treatment strategies combining TCM and western medicine.

The safety of traditional Chinese medicines (TCMs) is of paramount importance. Sulfur fumigation, a traditional processing and preservation technique for TCMs, serves to prevent mold during storage and improve product appearance. However, excessive sulfur fumigation leads to sulfur dioxide (SO₂) residues in the herbs, posing potential health risks. Currently, the SO₂ residue detection methods suffer from limitations including long analysis time, high cost, and operational complexity, making them inadequate for meeting the market demand for rapid testing. Fluorescent probe technology, with its advantages of high sensitivity, portability, simple preparation, and rapid response, has been widely applied in pharmaceutical development, biotechnology, and food safety. In recent years, the use of highly selective and sensitive fluorescent probes for the quantitative or semi-quantitative detection of SO₂ residues in TCMs has emerged as a significant research focus. This review systematically summarizes fluorescent probes developed in recent years for detecting SO₂ residues in TCMs, categorizing them based on their recognition mechanisms or structural features. It focuses on application progress of these probes in practical detection scenarios, analyzes the current technical challenges, and discusses future development directions. This review aims to provide theoretical foundations and technical references for enhancing the medicinal safety and quality control of TCMs.

Psycho-cardiological disease refers to a clinical syndrome involving the coexistence of cardiovascular diseases and psychological disorders. It maintains a consistently high prevalence and seriously affects patients’ quality of life as well as long-term prognosis. Treatments with Western medicine present limitations including risks of drug interactions and a shortage of psychological intervention resources. Huanglian Wendan Decoction (黄连温胆汤, HLWDT), derived from Liu Yin Tiao Bian of the Qing dynasty, has the effects of clearing heat and resolving phlegm, regulating the stomach and benefiting the gallbladder, and tranquilizing the mind. It conforms to the core pathogenesis of psycho-cardiological disease: phlegm-heat disturbing the heart. By systematically reviewing mechanistic studies on HLWDT against psycho-cardiological disease in recent years, this paper summarizes the active components of HLWDT and its mechanisms in preventing and treating psycho-cardiological disease. It is found that this prescription exerts effects through multiple pathways, such as ameliorating immune inflammatory responses, regulating neurotransmitter metabolism, upregulating the expression of brain-derived neurotrophic factor, alleviating oxidative stress injury, and inhibiting hyperactivity of the hypothalamic-pituitary-adrenal axis, thus illustrating its advantages in multi-component and multi-target improvement of related psycho-cardiological diseases including coronary heart disease, arrhythmia, heart failure and hypertension. This provides a reference for the clinical promotion and in-depth research of HLWDT in treatment of psycho-cardiological disease.

The forkhead box protein 3 (FoxP3)/regulatory T cell (Treg) axis is a key regulatory network that maintains peripheral immune tolerance and immune homeostasis, and its functional imbalance is closely associated with immune-related diseases, including autoimmune diseases, cancer, and allergic diseases. Traditional Chinese medicine (TCM), with its characteristics of multi-components, multi-targets, and multi-pathways, shows unique advantages in regulating the FoxP3/Treg axis. This article systematically reviews the multidimensional mechanisms by which TCM and its active components regulate FoxP3 expression and Treg cell differentiation, stability, and immunosuppressive functions. These mechanisms include direct regulation of key signaling pathways and cytokine networks, indirect influence on the gut microbiota-immune axis, and mediation of FoxP3 epigenetic modifications. It also summarizes the characteristics of bidirectional regulation, treating different diseases with the same method and treating the same disease with different methods observed in immune regulation. This provides important reference for the precise and modern application of TCM in treatment of immune-related diseases.

Yupingfeng Powder (玉屏风散) was first recorded in Jiu Yuan Fang of the Southern Song Dynasty. It is composed of three herbs—Huangqi (Astragali Radix), Baizhu (Atractylodis Macrocephalae Rhizoma), and Fangfeng (Saposhnikoviae Radix)—and possesses the therapeutic effects of tonifying qi, consolidating the exterior, and arresting sweating. Existing reviews have primarily focused on its chemical constituents, pharmacological activities, or therapeutic progress in a single disease, whereas a systematic integration of the tripartite relationship among “clinical evidence-mechanism of action-material basis” remains insufficient. From a “multi-component, multi-target, multi-pathway” perspective, this article synthesizes the common regulatory networks of Yupingfeng Powder across different diseases, elucidates the scientific basis of its principle of “treating different diseases with the same method,” and proposes concrete research pathways across four dimensions: evidence-based validation, clinical therapeutic patterns, bioactive substances, and mechanisms of action. By constructing a four-in-one research framework, this review aims to advance the precise clinical application and in-depth development of Yupingfeng Powder.

The incidence and mortality of pulmonary diseases remain persistently high. Conventional therapeutic modalities encounter restrictions, including drug resistance and notable side effects, underscoring the urgent need for novel treatment strategies. Nucleic acid drugs, with their strong target specificity and long-lasting effects, provide a promising new direction for treating pulmonary diseases. Meanwhile, traditional Chinese medicine (TCM), with its extensive theoretical and practical foundation the prevention and treatment of pulmonary diseases, offers innovative intervention strategies through its derived nucleic acid components, which can function via trans-kingdom regulatory mechanisms. This paper explores the potential of TCM-derived nucleic acid drugs as an emerging trans-kingdom therapeutic strategy. Guided by TCM theory, it systematically elaborates drug, type, mechanisms of action, and advances in areas such as drug stability and delivery efficiency, as well as the unique advantages of these agents. Furthermore, the article reviews the application status of nucleic acid drugs in various pulmonary diseases, including chronic obstructive pulmonary disease, asthma, pneumonia, lung cancer, acute lung injury, pulmonary fibrosis, and acute respiratory distress syndrome. Finally, the discussion then addresses the current challenges and future prospects to inform the theoretical foundation and research ideas for the systematic development of this drug class.

Triple-negative breast cancer (TNBC) is a subtype of breast cancer characterized by the absence of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 expression. It exhibits high invasiveness, elevated rates of recurrence and metastasis, poor prognosis, and currently lacks effective targeted therapeutic approaches. The Coptidis Rhizoma-Euodiae Fructus herb pair is a classic Traditional Chinese Medicine (TCM), its major active components include alkaloid compounds such as berberine, coptisine, evodiamine, and rutaecarpine. In recent years, Coptidis Rhizoma-Euodiae Fructus herb pair has demonstrated tremendous potential in the field of anti-tumor therapy, with increasing research focusing on the pharmacological effects of their alkaloid components in ameliorating TNBC. Accordingly, this review briefly summarizes the alkaloid components of Coptidis Rhizoma and Euodiae Fructus, emphasizing their mechanisms of action in enhancing TNBC treatment, which include inhibiting cell invasion and metastasis, regulating the cell cycle, and inducing apoptosis. This aims to provide references for the further development and utilization of the Coptidis Rhizoma-Euodiae Fructus herb pair and to offer new insights and scientific evidence for the application of TCM in the prevention and treatment of TNBC.

The direct incorporation of crude drug powders into traditional Chinese medicine preparations represents a distinctive approach. While this method has unique advantages in preserving the full spectrum of herbal constituents and embodying the principle of “drug-excipient combination”, it also presents challenges such as microbial contamination, poor formulation formability, low dissolution rates of active ingredients, and difficulties in quality control. This paper statistically analyzes the distribution of crude drug powder formulations in the 2025 edition of the Chinese Pharmacopoeia, examines the application characteristics of crude drug powders in pharmaceuticals, and systematically reviews existing key issues. Furthermore, it explores solution strategies from dimensions including sterilization technology, powder properties modification, ultrafine grinding (particle design), and online detection. It also outlines future research directions, aiming to provide references for process optimization and quality enhancement of crude drug powder formulations.