Objective Based on the supramolecular “imprinting template” theory, this study investigates the autonomous interaction patterns of the component groups in Wuwei Xiaodu Yin (WXY, 五味消毒饮) during the preparation process from crude herbs to reference samples and elucidates the quantitative transfer law of its quality attributes. Methods Fifteen batches of WXY reference samples were prepared via water decoction. A UPLC fingerprint analysis method was established to identify key chromatographic peaks and determine the contents of chlorogenic acid, esculetin, cichoric acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, glehinnalin, and linarin were determined at 240 nm. Parameters such as transfer rate, extract yield, total statistical moment, and information entropy were calculated for both the herbs and the reference samples. Results The fingerprint similarity of the 15 batches of reference samples was greater than 0.9. Fourteen chromatographic peaks were identified, with 10 characteristic peaks defined. The extract yield ranged from 43.27% to 53.43%. The transfer rates of chlorogenic acid, esculetin, cichoric acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, glehinnalin, and linarin were 34.25%—42.72%, 51.52%—58.80%, 28.21%—32.04%, 56.26%—65.59%, 40.63%—53.89%, 22.35%—31.29%, 38.39%—46.22%, 34.02%—40.82%, 18.64%—20.96%, and 28.55%—32.63%, respectively, with no discrete data observed. The relative standard deviations (RSDs) of the information entropy for the UPLC fingerprints of the five herbs and the reference samples were all less than 10%. The RSDs of mean chromatographic retention time of total quantum (MCRT_T) and information entropy for WXY (crude herbs) were 3.90% and 2.25%, respectively. Conclusion Quality analysis of the reference samples based on the parameters of the supramolecular “imprinting template” provides a scientific basis for the quality control of WXY reference samples and the development of compound formulations.

Thyroid-associated ophthalmopathy (TAO) is an orbital inflammatory disease closely related to thyroid dysfunction and autoimmunity. The pathogenesis of TAO has not been fully elucidated yet. The conventional treatments have problems such as significant adverse reactions and high drug resistance rates. Through a systematic review of the research progress of Tripterygium wilfordii and its active components in the treatment of TAO from five aspects: theory, clinical practice, mechanism, clinical application safety, and nano-formulation, this paper explores the potential of Tripterygium wilfordii and its active components in treating TAO. The theoretical analysis shows that Tripterygium wilfordii’s functions of dispelling wind, removing dampness, relaxing muscles and tendons, and promoting blood circulation and relieving pain are in line with the pathogenesis of TAO, which involves liver depression, spleen deficiency, and blood stasis blocking the meridians. Clinical studies have confirmed that Tripterygium wilfordii alone or in combination with methimazole, prednisone, etc., can effectively improve the clinical symptoms of TAO patients. Mechanism research further reveals that the effects of Tripterygium wilfordii and its active components in treating TAO involve inhibiting inflammatory and immune responses, as well as anti-fibrotic effects, etc. In terms of clinical application safety, adverse reactions can be alleviated by adjusting dosage, formulation, processing methods, and compatibility. In terms of formulation improvement, the nano-delivery system, through targeted strategies, significantly enhances the targeted enrichment and therapeutic selectivity of active components of Tripterygium wilfordii in the lesion site, which is a key approach to achieving clinical enhancement of efficacy and reduction of toxicity. In summary, Tripterygium wilfordii and its active components show good potential in treating TAO in terms of theory, clinical practice, mechanism, and safety. Combined with modern nanotechnology and other formulation improvements, this may provide a promising new strategy for the traditional Chinese medicine prevention and treatment of TAO.

Nanoaggregates of traditional Chinese medicine (TCM) serve as the core vehicle for elucidating the pharmacological basis of TCM formulations and their “multi-component, multi-target” mechanisms of action. This review systematically reviews the latest research advances on the formation mechanisms, influencing factors, characterization techniques and targeted delivery applications of TCM nanoaggregates, and analyzes the current challenges and future prospects of this research field. Studies have shown that TCM nanoaggregates can self-assemble spontaneously through non-covalent interactions such as hydrogen bonding, π-π stacking, electrostatic interactions and hydrophobic interactions. Their particle size, morphology and structural characteristics significantly affect the solubility, stability and bioavailability of drugs. Targeted delivery is the core advantage of TCM nanoaggregates. It enables the precise enrichment of drugs at lesion sites through passive targeting and active targeting strategies, thereby achieving the goal of toxicity reduction and efficacy enhancement. Nevertheless, this field still faces challenges such as complicated preparation processes and insufficient evaluation of stability and safety. Future studies should further elucidate the formation mechanisms and optimize the evaluation system, providing novel perspectives and technical support for the interpretation of the material basis of TCM efficacy and the modernization development of traditional Chinese medicine.

Objective To explore the mechanism of Diterpene Ginkgolides Meglumine Injection (银杏二萜内酯葡胺注射液, DGMI) in treatment of acute ischemic stroke (IS) based on platelet generation and activation. Methods Sham group, model group, ibrutinib (10 mg/kg) group, DGMI low-, medium- and high-dose (6.25, 12.50, 25.00 mg/kg) groups were established. A transient middle cerebral artery occlusion (tMCAO) model was prepared in C57BL/6 mice using suture method. Medication was administered continuously for 3 d starting 0.5 h after surgery. The infarct rate was measured using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Motor and neurological function were evaluated using spinning rod experiment and Bederson score. Flow cytometry and ELISA were used to detect the whole blood platelet count and platelet factor 4 (PF4) level in serum. Bone marrow smears and immunohistochemistry were used to observe the proliferation, differentiation, apoptosis and phosphorylation levels of focal adhesion kinase (FAK) in bone marrow megakaryocytes. Immunofluorescence staining was used to detect platelet activation and FAK phosphorylation levels in brain tissue. A combination group of FAK agonist ZINC40099027 (ZIN) and medium-dose DGMI was set up to verify whether DGMI exerts a protective effect by inhibiting FAK activation. Results DGMI significantly reduced cerebral infarction and megakaryocyte generation in tMCAO mice. Among them, the inhibitory effect of medium-dose DGMI on differentiation and proliferation of bone marrow megakaryocytes in tMCAO mice, as well as the improvement of cerebral infarction area, motor coordination ability and neurological function score were the most significant (P < 0.05, 0.01, 0.001). In addition, DGMI could significantly reduce the number of platelets and PF4 level in serum of tMCAO mice (P < 0.05, 0.001), and inhibit the phosphorylation of FAK in bone marrow megakaryocytes and infiltrating platelets in brain (P < 0.01, 0.001). The FAK agonist ZIN significantly reversed the protective effects of DGMI on tMCAO mice (P < 0.05, 0.01, 0.001). Conclusion DGMI exerts neuroprotective effects by inhibiting the excessive generation of megakaryocytes during the acute phase of IS and the phosphorylation of FAK in infiltrating platelets in the brain, thereby suppressing platelet generation and activation.

Objective To identify members of the WRKY transcription factor family in Codonopsis pilosula at the genome-wide level and analyze their characteristics, and explore their potential roles in response to salt stress, thereby laying a foundation for functional studies of C. pilosula WRKY transcription factors. Methods Potential gene family members were screened across the entire genome based on conserved protein domains using the Hidden Markov model (HMM). Protein sequence alignment, conserved domain analysis, phylogenetic tree construction, gene synteny analysis, and cis-acting element analysis in the promoter region were performed using software and online tools including MAFFT, IQ-TREE, MEME, Plant CARE and TBtools. Expression patterns of WRKY genes were analyzed based on transcriptome data from different tissues (roots, stems, leaves, and flowers) and qRT-PCR was used to determine their relative expression levels. In addition, the transcriptome data of C. pilosula treated with 0, 60, and 120 mmol/L NaCl were analyzed to examine the expression changes of WRKY genes under salt stress. Results A total of 50 WRKY transcription factor members were identified in C. pilosula. Motif analysis showed that all members contained the typical WRKY domain with the conserved sequence WRKYGQK. Phylogenetic analysis with WRKY transcription factors from Arabidopsis thaliana and Oryza sativa classified the 50 C. pilosula WRKY transcription factors into three groups, with group II further divided into five subgroups. The amino acid lengths of the identified WRKY proteins ranged from 103 to 986 residues, with isoelectric points (pI) of 4.87−9.99 and molecular weights (MW) of 12 435.14−107 946.28. Gene expression analysis revealed significant differences in the expression levels of CpWRKY26, CpWRKY48, CpWRKY2, and CpWRKY49 among roots, stems, leaves, and flowers of C. pilosula. The results of salt stress treatment indicated that most CpWRKY genes showed differential expression trends under different concentrations of NaCl, and some members cluster phylogenetically with previously reported salt-tolerant WRKY genes, suggesting that they may be involved in the regulation of C. pilosula salt stress responses. Conclusion This study represents the first systematic identification and bioinformatics analysis of the WRKY gene family in C. pilosula at the genome-wide level. By combining tissue expression and salt stress response analyses, the study reveales that the WRKY genes in C. pilosula may be involved in physiological processes such as growth and development, secondary metabolite biosynthesis, and stress responses. This lays a theoretical foundation for further research on the functions of WRKY genes and their roles in regulating medicinal components, and provides gene resources and scientific evidence for molecular-assisted breeding of C. pilosula.

Objective To investigate the chemical constituents and anti-inflammatory activity of the dried roots of Tibetan medicine Carpesium lipskyi. Methods The ethanol extract of C. lipskyi was isolated and purified using chromatographic techniques including silica gel column chromatography and Sephadex LH-20 gel column chromatography. The structures of the compounds were identified based on their physicochemical properties, spectroscopic methods (MS, NMR), and literature data. The in vitro anti-inflammatory activity of the compounds was evaluated using a lipopolysaccharide (LPS)-induced inflammatory model in mouse macrophage RAW264.7 cells. Results Nine compounds were isolated from the roots of C. lipskyi and identified as (1R,7R,10R,11S)-1-hydroxy-eudesma-3(4),5(6)-dien-11-oic acid methyl ester (1), (1R,4S,5R,7R)-1-acetoxy-11-hydroxy-eremophil-9(10)-ene (2), careudesmane D (3), isoalantolactone (4), 11-hydroxy-eremophil-1(10)-en-2,9-dione (5), carperemophilane A (6), carperemophilane B (7), protocatechuyl aldehyde (8), and umbelliferone (9). Compounds 2 and 7 inhibited LPS-induced NO production in RAW264.7 cells with half maximal inhibitory concentration (IC₅₀) values of (13.62 ± 1.30) μmol/L and (19.47 ± 2.67) μmol/L, respectively. Conclusion Compound 1 is a new eudesmane-type sesquiterpene and designated as carpesiol A, and compound 2 is a new eremophilane-type sesquiterpene and designated as carpesiol B. Compounds 3—7 were isolated from this plant for the first time. Compounds 2 and 7 show potential for development as anti-inflammatory drugs.

High altitude sleep disturbance (HASD) is a common condition occurring in high-altitude regions due to low pressure and low oxygen levels. Its primary manifestations include insomnia, sleep apnea, and significantly reduced sleep quality. Severe cases may also trigger acute high-altitude illnesses such as high-altitude pulmonary edema and high-altitude cerebral edema. Following acute exposure to high altitudes, the incidence of HASD is notably high, posing a serious threat to physical health and quality of life. It has thus become a significant public health concern in plateau regions. Modern medical drug therapies for HASD demonstrate limited efficacy and are associated with side effects such as drowsiness and drug resistance. Therefore, in-depth exploration of its pathogenesis and the pursuit of safer, more effective treatment methods are of significant importance. Tibetan medicine, as a traditional medical system, emphasizes diagnosis and treatment based on the three factors theory (rlung, mkhris-pa, bad-kan). It offers holistic regulatory advantages through multi-dimensional, multi-component, and multi-pathway approaches. Safe and reliable, it regulates bodily functions through multi-target synergistic effects, presenting a potential key pathway for addressing HASD. HASD primarily affects sleep regulation centers across various brain regions. Tibetan medicine's holistic perspective views the human body as deeply interconnected with the natural environment. External changes like hypoxia or atmospheric pressure fluctuations disrupt the body’s “rlung” equilibrium, leading to neurotransmitter imbalances in the brain. This, in turn, triggers emotional disturbances and sleep disorders. This paper explores the application of Tibetan medicine tailored to local conditions for treating HASD based on existing research, summarizing advances in related mechanism studies. It aims to provide more substantial reference for innovative applications of Tibetan medicinal formulas and novel drug development targeting HASD.

Objective To investigate the terpenoid constituents of fruit shells of Euphorbia peplus and evaluate their anti-inflammatory activities. Methods Various chromatographic techniques were applied to isolate terpenoids from the petroleum ether-soluble fraction of the ethanol extract of E. peplus fruit shells. The chemical structures of all isolates were elucidated by comprehensive spectroscopic analyses (1D/2D NMR, HRMS) combined with single-crystal X-ray diffraction. Anti-inflammatory activities were assessed by measuring the inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Results A total of 13 terpenoids were isolated from the petroleum ether fraction of E. peplus fruit shells, including four paraliane-type diterpenoids (1–4), one pepluane-type diterpenoid (5), three jatrophane-type diterpenoids (6–8), two ingenane-type diterpenoids (9 and 10), one abietane-type diterpenoid (11), one ursane-type triterpenoid (12), and one dinor-monocyclic diterpenoid (13). They were respectively identified as (2S,3S,4R,5R,6R,8S,12S,13S,15R)-8b-acetyl-paralianone D (1), (1R,2R,3S,4R,5R,6R,8S,12S,13S,14R, 15R)-1,5,8,14-tetraacetoxy-3-benzoyloxy-15-hydroxy-9-oxo-paraliane (2), (2S,3S,4R,5R,6R,8S,12S,13S,14R,15R)-5,8,14-triacetoxy-3-benzoyloxy-15-hydroxy-9-oxo-parapliane (3), paralianone A (4), (2S,3S,4R,5R,6R,8R,9R,10R,12S,13S,14R,15R)-5,8,9,10,14-pentaacetoxy-3-benzoyloxy-15-hydroxypepluane (5), euphpepluone A (6), euphpepluone D (7), (2S,3S,4R,5R,7S,8R,13S,15R)-5α,7b,8α-triacetoxy-3b-benzoyloxy-15b-hydroxyjatropha-6(17),11E-diene-9,14-dione (8), 20-deoxyingenol (9), 20-O-(2¢E,4¢Z-decadienoyl)ingenol (10), helioscopinolide D (11), ursolic acid (12) and (3R,6R,7E,9E,11E)-3-hydroxy-13-apo-α-caroten-13-one (13). The structure of compound 1 was confirmed by single-crystal X-ray diffraction. In the anti-inflammatory activity screening, only compound 2 exhibited inhibitory activity against NO production, with an IC₅₀ value of (37.6 ±8.2) μmol/L. Conclusion Compounds 1, 2, 10, and 13 were isolated from E. peplus for the first time. Compound 2 demonstrated moderate anti-inflammatory activity. This study enriches the terpenoid chemical diversity of terpenoids from E. peplus and provides an experimental basis for further research on its anti-inflammatory substances.

Objective In response to the lack of objective standards for the nine-steaming and nine-drying process of Huangjing (Polygonati Rhizoma), this study quantified the dynamic changes in traits and components during processing, combined with zebrafish antioxidant activity experiments, to identify the optimal stage for comprehensive quality and provide a scientific basis for establishing modern quality control methods. Methods Using the decoction pieces of Polygonatum cyrtonema as the object of study, a colorimeter, texture analyzer, and electronic tongue were employed to quantify the color, texture, and taste of samples from different steaming cycles (S0—S9). Polysaccharide content, reducing sugars and amino acids was determined by UV-Vis, while 5-hydroxymethylfurfural (5-HMF) were measured using HPLC. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were used to identify patterns in the processing stages. Pearson correlation analysis, the CRITIC objective weighting method, and zebrafish antioxidant experiments were integrated for comprehensive evaluation. Results As steaming progressed, color parameters (E_ab^*, L^* and b^*) continuously decreased, texture parameters (hardness, chewiness, gumminess) significantly declined, and moisture content increased. In terms of taste, bitterness and astringency markedly decreased by the third steaming cycle (S3), while sweetness increased. Chemically, 5-HMF accumulated continuously, reducing sugars peaked at S3 (1.38%), and amino acid and polysaccharide contents stabilized after S3. The correlation network analysis revealed significant relationships among trait indicators, chemical components, and bioactivity (P < 0.05). Multivariate statistical analysis divided the processing into three stages, with the early stage (S1—S3) identified as the critical transition period for quality formation. The comprehensive evaluation using the Criteria importance through intercriteria correlation method indicated that sample S3 achieved the highest overall quality score (65.41). At this stage, the sample was characterized by moderate 5-HMF accumulation, peak reducing sugar conten, and and maintenance of amino acids at a high level. It exhibited blackish-brown color (a^* was 11.13), with a soft and flexible texture (springiness was 0.647), and a disappearance of the numbing sensation. Its taste was sweet with slight bitterness (sweetness value was 4.50, bitterness value was 5.55), accompanied by a significantly enhanced antioxidant capacity. Conclusion This study establishes the third steaming cycle (S3) as the optimized processing endpoint for the nine-steaming and nine-drying process of P. cyrtonema and established amulti-dimensional quantifiable quality control system centered on “appearance properties-intrinsic components-bioactivity”. These findings provide a scientific basis for the standardization and precise quality control of this processing method.

Doxorubicin is an anthracycline antitumor drug widely used in clinical practice, but its therapeutic value is significantly limited due to adverse reactions such as dose-dependent cardiotoxicity and nephrotoxicity, and there is an urgent need to find preventive and therapeutic drugs. Recent studies have found that abnormal activation of NOD-like receptor family pyrin domain containing 3 (NLRP3) is one of the important mechanisms underlying doxorubicin-induced toxicity in different target organs, whereas traditional Chinese medicine (TCM) and its active components can effectively inhibit the NLRP3 inflammasome and alleviate doxorubicin-induced toxicity. By systematically summarizing recent experimental research on how TCM and its active components ameliorate doxorubicin toxicity through regulating the NLRP3 inflammasome, it was found that they can alleviate doxorubicin toxicity in terms of cardiac and renal protection. These studies provide important theoretical evidence and potential therapeutic strategies for using TCM to intervene in the related adverse effects of doxorubicin.