Search Results (150)

Background/Objectives: Dendrobium officinale (D. officinale), an endangered ornamental and medicinal orchid, displays significant variability in its bioactive compounds depending on geographical and environmental factors. To decipher these influences, we investigated metabolic divergence across three cultivars (GN, LS, DX) cultivated in greenhouse and outdoor conditions using untargeted metabolomics. Methods: Metabolites extracted from stem and leaf tissues were analyzed via UHPLC-Q Exactive Orbitrap MS, and the raw data were processed using XCMS for peak alignment and quantification. Differentially abundant metabolites (DAMs) were identified by multivariate statistical analyses including PCA and OPLS-DA. Metabolic pathways were annotated using KEGG, HMDB, and LIPID Maps databases, with enrichment analysis and visualization performed via TBtools II and Hiplot. Results: Metabolite profiling and multivariate analysis revealed distinct chemotypes. The DX cultivar exhibited anthocyanin enrichment in its stems, correlating with a red pigmentation, while GN accumulated specific amino acid derivatives. Tissue-specific metabolic specialization was evident, with leaves displaying greater flavonoid diversity and stems prioritizing lipid and amino acid metabolism. Outdoor cultivation enhanced flavonoid biosynthesis, whereas greenhouse conditions favored alkaloid accumulation. Functional analysis identified both conserved pathways, like phenylpropanoid biosynthesis, and varietal-specific adaptations in amino acid and secondary metabolism. Notably, alkaloid levels declined sharply during plant defoliation. Conclusions: Our findings demonstrate that environmental factors and geographical origin synergistically shape the metabolic profiles of D. officinale. This provides a scientific basis for optimizing cultivation strategies—through targeted environmental adjustments and varietal selection—to enhance the yield of desired bioactive compounds. Full article

Paeoniae Radix Alba Carbonisata (PRAC), carbonized decoction pieces of the traditional Chinese medicine Paeoniae Radix Alba, has been used in clinical practice for hepatoprotective purposes. Hepatic amyloidosis (HA), a severe complication of systemic AA amyloidosis, is characterized by the deposition of fibrillar amyloid proteins leading to progressive hepatic dysfunction. However, its role in HA remains unclear. Amyloid lysozyme (LYSO-6) was used to induce the NCTC1469 cell injury model and the HA mouse model. The effects of PRAC extract (PRAC-E) on liver injury were then evaluated using biochemical assays, enzyme-linked immunosorbent assay (ELISA), Congo red (CR) staining, Hematoxylin and Eosin (H&E) staining, and immunohistochemical staining. Liver transcriptomics combined with Western blotting was used to analyze the expression levels of key proteins in the cGMP/PKG/ATP2A1 signaling axis. UHPLC-Q-Exactive Orbitrap MS combined with network pharmacology was used to characterize the chemical components of PRAC-E and identify its core active constituents against HA. Quantitative analysis of core components was performed by UHPLC-QTRAP-MS/MS. Molecular docking predicted the binding stability of core components and key targets. The results showed that PRAC-E significantly alleviated HA. Collectively, PRAC-E restored calcium pump activity, corrected calcium homeostasis imbalance, reduced inflammatory factor levels, regulated Phosphodiesterase 5A (PDE5A), and activated the cGMP/PKG/ATP2A1 signaling axis. The main components of PRAC-E were phenolic acids, terpenoids, and flavonoids. Among these, six core components (SCCs) related to HA were Gallate (16.96 mg/g), Paeoniflorin (14.27 mg/g), Albiflorin (7.20 mg/g), Benzoyl paeoniflorin (5.33 mg/g), Methyl gallate (0.78 mg/g), and Catechin (0.09 mg/g). Molecular docking analysis demonstrated that SCCs formed stable complexes (∆G ≤ −6.2 kcal/mol) with ATP2A1. Full article

Background/Objectives: Natural medicine analysis remains challenging due to chemical diversity. To the best of our knowledge, the comprehensive identification of multiple chemical constituents in Xiaoban Kangfu (XBKF) capsules has not been reported. Therefore, a combined approach utilizing ultra-high-performance liquid chromatography quadrupole-Exactive Orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS) and molecular network analysis needs to be developed to comprehensively characterize the chemical constituents of XBK capsules in heat-clearing and toxin-eliminating granules, thereby enhancing annotation accuracy and enabling visualization. Methods: Firstly, chromatographic and mass spectrometry conditions were optimized to achieve good separation and a rich signal response. Subsequently, the literature searches, database consultations, and reference standards were employed to enhance annotation reliability. Finally, the raw data acquired under optimized conditions were uploaded to Global Natural Products Social (GNPSs), enabling component visualization by linking precursor ions of similar structural features with identical colors. Results: A total of 170 compounds were identified from this medicinal resource for the first time, including 50 flavonoids, 34 phenolic acids, 16 terpenoids, 14 quinones, 14 organic acids, eight coumarins, ive carbohydrates, and 29 other compounds. Conclusions: This study establishes a robust UHPLC-Q-Exactive Orbitrap MS/MS strategy for the comprehensive chemical profiling of XBKF capsules. The use of the presented validated analytical method for the comprehensive quality control of XBKF capsules is highly promising, offering fast, highly sensitive, and reliable analysis. Full article

Background: Low-volume trapping columns are essential for sample enrichment, desalting, and injection profile focusing on nano-LC–MS-based proteomics. They enable higher sample loading, improve chromatographic performance, and protect the analytical column by removing salts and contaminants. Recently, monolithic trap columns with micropillar architecture have emerged as alternatives to conventionally packed traps. This study compares the performance of a packed and a micropillar monolithic trap column for the analysis of tryptic peptides. Methods: A tryptic digest of HeLa cell lysate was analyzed under identical LC–MS conditions using both trap types. Peptides were detected at 214 nm and analyzed by nano-ESI on a Q Exactive Plus Orbitrap. Data were searched against the human UniProt database (February 2023) using FragPipe v20.0, and statistical evaluation of MaxLFQ intensities was performed in Perseus using Welch’s t-test and clustering analysis. Results: Over 2500 proteins were identified with both setups. The packed trap column yielded more total peptides, particularly those with post-translational modifications and higher hydrophilicity, whereas the monolithic column favored peptides of intermediate hydrophobicity. Chromatographic profiles confirmed a slight reduction in the trapping efficiency of hydrophilic peptides by the monolithic trap. Conclusions: Trap column design significantly influences peptide recovery and proteome coverage. Full article

Oxidative stress is a primary driver of diabetic nephropathy (DN), highlighting the urgent need for potent natural antioxidants. This study explored the reno-protective potential and associated mechanisms of Rhinacanthus nasutus aqueous extract (AE). Phytochemical profiling via Q Exactive HF Orbitrap LC–MS/MS and serum pharmacochemistry analysis identified 38 constituents, among which 25 bioavailable constituents (e.g., caffeic acid and naringenin) might be the key bioactive ones. In the L6 myotubes in vitro assays, AE (75 μg/mL) was observed to upregulate the PI3K/AKT and GLUT4 signaling cytokines, coinciding with enhanced glucose uptake, as confirmed by Western blot with insulin as a positive control. Furthermore, in STZ-induced DN rats, AE could reduce MDA levels (0.58 vs. 1.44 nmol/mgprot) and restore T-SOD, CAT, and GSH-Px levels (170.57, 51.93, 63.68 vs. 114.93, 40.84, 50.99 mgprot). The protective effects were accompanied by the modulation of PI3K/AKT/mTOR signaling axis. These findings suggest that AE exerts dual efficacy involving glucose uptake regulation and oxidative stress inhibition. Consequently, Rhinacanthus nasutus represents a promising natural antioxidant resource with potential for the management of DN. Full article

Background/Objectives: Breast cancer remains a leading cause of cancer-related mortality in women, with therapeutic resistance frequently arising from tumor heterogeneity and an immunosuppressive tumor immune microenvironment (TIME). While Cynomorium songaricum Rupr. (CS) has been used traditionally in Chinese medicine and exhibits preliminary anti-tumor activity, its bioactive constituents and precise mechanisms against breast cancer remain to be elucidated. Methods: The chemical constituents of CS were systematically profiled using ultra-high-performance liquid chromatography coupled with Q Exactive Orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS/MS). Network pharmacology and functional enrichment analyses were performed to identify immuno-related targets and pathways, followed by molecular docking to prioritize component–target pairs. Molecular dynamics (MD) simulations were conducted to validate the stability of a representative docked complex and to characterize binding stability, interaction persistence, molecular mechanics/(Poisson–Boltzmann) surface area (MM/(P)BSA) energetics, and principal component analysis (PCA)-based conformational landscapes. Results: We identified 1100 compounds, of which 84 satisfied the in silico drug-likeness criteria, including 12 phenylpropanoids, 4 terpenes, 35 flavonoids, 2 quinones, 1 phenol, 3 alkaloids, and other phytochemicals. Network pharmacology analysis revealed 776 overlapping targets associated with both breast cancer and immune regulation. Functional enrichment analysis underscored significant involvement in immune-related pathways, and molecular docking studies supported high-affinity interactions between the components and their targets. MD analyses further supported a stable bound ensemble for the representative SRC–Tomentogenin complex during the equilibrated window, with persistent pocket occupancy, consistent interaction signatures, favorable MM/(P)BSA binding energetics, and a concentrated low-energy basin on the PCA-based free energy landscape. Conclusions: These findings elucidate the chemical basis of CS and uncover its immunomodulatory mechanism against breast cancer, offering a foundation for developing CS-based immunotherapeutic strategies and supporting multi-target drug discovery from traditional medicines. Full article

Broodstock nutrition is a key determinant of reproductive output in marine fishes because lipids support yolk formation, embryonic development, and early larval viability. However, the allocation of lipid classes between fertilized eggs and the egg envelope (chorion) remains poorly characterized for many tropical species. In this study, we performed a comparative lipidomic analysis of dechorionated fertilized egg contents and isolated chorion from three tropical marine fishes (Trachinotus ovatus, Platax teira, and Plectropomus leopardus) using UHPLC–Q Exactive Orbitrap MS/MS. Multivariate analyses revealed clear tissue- and species-specific lipid patterns. Dechorionated eggs were enriched in energy-storage lipids, dominated by triacylglycerols and essential polyunsaturated fatty acids, whereas chorion tissues contained higher levels of structural and signaling lipids, including phosphatidylinositols and sphingolipids. Each species exhibited a distinct lipid signature, with T. ovatus characterized by higher secosteroids and free fatty acids, P. teira by glycerophosphoethanolamines and phosphoinositols, and P. leopardus by abundant triradylglycerols. Pathway enrichment highlighted glycerophospholipid metabolism and sphingolipid signaling as prominent processes during early development. These findings clarify lipid partitioning between dechorionated fertilized egg contents and the chorion and provide a biochemical rationale for optimizing species-specific broodstock diets to enhance egg quality in tropical marine aquaculture. Full article

The complexity of the milk matrix, driven by its lipid-rich composition, complicates pesticide residue analysis. This study developed a simplified and robust analytical procedure for the quantification of 250 pesticides in cow’s milk. Sample preparation involved acidified ethyl acetate extraction followed by centrifugation at 0 °C. A subsequent clean-up step was performed using micro solid-phase extraction (μSPE) in a 96-well format with the enhanced matrix removal-lipid (EMR-lipid) sorbent. Final extracts were analyzed by gas chromatography coupled to high-resolution mass spectrometry (GC-Q-Orbitrap-MS). Method validation demonstrated satisfactory linearity within the 5–100 µg/L range, recoveries between 70.6% and 119.8%, and precision, expressed as relative standard deviation (RSD), was acceptable for both intraday (1.8–19.2%) and interday (1.6–18.5%) conditions. The limit of quantification (LOQ) was set at 10 µg/kg for all compounds. The method was applied to 23 commercial cow’s milk samples, and no pesticide residues were detected above the current European Union (EU) maximum residue limits (MRLs). Full article

Background: Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chow. (ZS) is a valuable plant with diverse economic applications, as all its organs contain bioactive secondary metabolites. The seeds, known as Suanzaoren in traditional Chinese medicine, are utilized as both a medicinal and edible resource, while the fruit pulp and leaves serve as significant raw materials in the food industry. Increasing market demand for Suanzaoren has led to expanded cultivation, though current production practices emphasize seed utilization, resulting in the underutilization of pulp and leaf tissues. In agricultural systems, developing elite varieties is an effective strategy for enhancing crop yield and quality. Breeding initiatives should establish specific objectives aligned with particular end uses, such as seed, pulp, or leaf production. Germplasm serves as the foundational material for breeding programs, so its selection must correspond to intended applications. Evaluating existing germplasm resources based on chemical composition profiles will provide a basis for developing improved ZS varieties. Objective: This study aimed to systematically compare the characteristic chemical composition in the seeds, pulp, and leaves of ZS. By quantifying key chemical components—such as flavonoid glycosides and saponins in seeds, organic acids and phenolic compounds in pulp, and flavonol glycosides and phenolic acids in leaves—we evaluated the quality of ZS germplasm resources. The resulting compositional profiles provide a concrete basis for selecting and breeding elite cultivars tailored to specific end uses, including seed, pulp, or leaf production. Methods: Chemical characterization was performed using ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap MS/MS). Quantitative analysis of chemical composition was conducted using high-performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD). Multivariate statistical analyses—including principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and entropy-weighted technique for order preference by similarity to an ideal solution (entropy-weighted TOPSIS)(EWT)—were employed for comprehensive data evaluation. Results: A comprehensive phytochemical analysis of Ziziphi spinosae (ZS) was conducted, identifying 144 distinct compounds across the seeds, pulp, and leaves. Of these, 114 were found in the seeds, 84 in the leaves, and 79 in the pulp. The seeds were particularly rich in flavonoid glycosides, such as spinosin and 6‴-feruloylspinosin, as well as saponins like jujuboside A and B. The pulp was dominated by organic acids, including citric acid, and phenolic compounds, while the leaves were abundant in flavonol glycosides, including rutin, and phenolic acids such as isochlorogenic acid B. Based on the chemical composition profiles, the ZS germplasms were evaluated for specific applications. ZS24, ZS22, and ZS3 were identified as the most suitable for seed production, ZS3, ZS6, and ZS9 for pulp utilization, and ZS20, ZS3, and ZS18 for leaf-based applications. With respect to the integrated utilization of multiple plant parts (roots, stems, and leaves), ZS6, ZS3, and ZS24 demonstrated the highest potential. Conclusions: The identification of superior germplasm resources provides strategic direction for the breeding of elite ZS cultivars. These findings will enable the comprehensive utilization of ZS plant resources and support the high-quality development of related industries. Full article

The chemical composition of the ethanol extract of Acanthopanax senticosus fruit (ASFEE) was systematically characterized using UPLC-MS/MS (Q Exactive Orbitrap), leading to the identification of 45 compounds. Through integrated network pharmacology and molecular docking analyses, the binding affinities between key bioactive constituents—such as eleutheroside E (EE) and quercetin—and core therapeutic targets were predicted and validated. A total of 125 overlapping targets were identified between ASFEE and acute kidney injury (AKI), with significant enrichment observed in critical signaling pathways, including NF-κB, IL-17, and PI3K-Akt. To evaluate the protective effects of ASFEE, both in vitro (HK-2 cells) and in vivo (murine) models of cisplatin (DDP)-induced AKI were employed. Parameters assessed included cell viability, apoptosis, reactive oxygen species (ROS) production, activation of the NF-κB signaling pathway, kidney function, histopathological alterations, and levels of inflammatory cytokines. ASFEE treatment markedly enhanced HK-2 cell viability and reduced cellular apoptosis and ROS generation. In the murine model, DDP administration resulted in significantly elevated serum creatinine (Scr) and blood urea nitrogen (BUN) levels. Both low- and high-dose ASFEE treatments significantly attenuated these increases, improved overall kidney function, and alleviated kidney tubular damage. Furthermore, ASFEE reduced serum levels of pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α. Multi-omics integration analysis enabled the identification of differentially expressed genes and metabolites. ASFEE was found to reverse 4689 DDP-induced gene expression changes and 323 metabolic disturbances, with the uridine diphosphate glucuronosyltransferase (UGT)-mediated ascorbic acid metabolism pathway emerging as the central regulatory axis. Key candidate genes and proteins were further validated via real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. DDP significantly upregulated the expression of inflammatory markers and associated signaling molecules in kidney tissues, while concurrently downregulating UGT family genes and the UGT1A1 protein involved in uronic acid metabolism. Notably, ASFEE treatment effectively counteracted these alterations, confirming its role in enhancing UGT1A1-mediated metabolic processes and suppressing the NF-κB/PI3K-Akt/IL-17 signaling cascade. These mechanisms contribute to improved antioxidant capacity, mitigation of inflammatory responses, and restoration of metabolic homeostasis, thereby conferring protection against DDP-induced AKI. ASFEE exerts a protective effect on AKI caused by DDP by enhancing antioxidant capacity, inhibiting inflammation and restoring metabolic homeostasis, providing an experimental basis for its subsequent development and application. Full article

Kaempferitrin (KAE) is a natural flavonol dirhamnopyranoside with various pharmacological activities, isolated from the antithrombotic fraction of Celastrus orbiculatus Thunb. This study aimed to investigate the antithrombotic activity and “effective forms” of KAE. The results showed that KAE significantly prolonged rabbit plasma recalcification time in vitro. In the FeCl₃-induced rat arterial thrombosis model, KAE demonstrated antithrombotic effects by inhibiting coagulation, platelet aggregation, and fibrinolysis, with a lesser risk of bleeding compared to aspirin. KAE was orally administered to rats, and a total of 192 metabolites were characterized. These included 25 phase I metabolites, 8 hydroxylated and methylated metabolites, 57 sulfated metabolites, 74 glucuronidated metabolites, 26 sulfated and glucuronidated metabolites, and 2 glycosylated metabolites. Twenty-eight compounds were considered the in vivo “effective forms” of KAE for their antithrombotic activity. Network pharmacology, molecular docking, and molecular dynamics simulations collectively predict that these “effective forms” may exert antithrombotic effects by suppressing the SRC/PI3K/AKT pathway. This study provides a foundation for a better understanding of the in vivo “effective forms” and mechanisms underlying KAE’s antithrombotic activity, which is essential for understanding of “hexue” traditional efficacy of C. orbiculatus. Full article

Ziziphi Spinosae Semen (ZSS), a medicinal and edible homologous herbal drug, is commercially available in both raw and fried (FZSS) forms and has been widely used to improve sleep quality. This study aimed to elucidate the differences in chemical composition between the two specifications. A comprehensive metabolomics approach utilizing ¹H NMR, GC–MS, and UHPLC-Q-Orbitrap-HRMS identified a total of 66 potential biomarkers. The results demonstrated that after frying, the content of fatty acids decreased significantly, while the levels of most primary metabolites (e.g., sugars, amino acids) and secondary metabolites (e.g., alkaloids, flavonoids) increased markedly. Targeted quantification of 14 key components validated these trends: the contents of five fatty acids decreased (p < 0.001), whereas the levels of five secondary metabolites (coclaurine, magnoflorine, spinosin, 6‴-feruloylspinosin, and jujuboside A) increased. In contrast, the content of jujuboside B decreased significantly. This study systematically reveals the profound impact of frying on the chemical composition of ZSS, providing a scientific basis for its quality control and processing optimization. Full article

Severe asthma is a heterogeneous condition often resistant to conventional corticosteroid therapy, necessitating the identification of novel biomarkers and therapeutic targets. This study investigates immunological, transcriptional, and proteomic biomarkers in severe asthma patients from the Brazilian ProAR cohort. Cytokines were measured using a multiplex technology and the differential sputum cell count was performed by cytospin preparations. Sputum transcriptomics was performed by RNA-seq using Ion S5 next-generation sequencing platform. The proteomic study of sputum was performed by liquid chromatography–tandem mass spectrometry (LC-MS/MS) using Q Exactive Orbitrap technology. Compared to mild-to-moderate asthma (MMA) and treatment-controlled severe asthma (SAC), the treatment-resistant severe asthma (SAR) group exhibited increased sputum neutrophilia, eosinophilia, and elevated IL-6 and TNF levels, correlating with impaired lung function. Transcriptomic and proteomic analyses revealed a Th2-independent molecular signature characterized by downregulation of the Hippo signaling pathway and upregulation of JAK–STAT inflammatory cascades. Distinctive microRNA profiles suggest regulatory involvement in inflammatory and proliferative processes. These findings align with prior studies, reinforcing the presence of an IL-6- and TNF-high severe asthma phenotype across diverse populations. Our results highlight key inflammatory pathways that may underlie corticosteroid resistance, offering potential targets for personalized therapeutic interventions in severe asthma. Full article

An integrative approach combining network pharmacology, molecular docking, and cellular assays was used to elucidate the potential mechanisms by which the n-butanol extract of Biebersteinia heterostemon ameliorates type 2 diabetes mellitus (T2DM). Chemical constituents of the n-butanol extract were identified via ultra-high-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry. Active compounds and T2DM-related targets were retrieved from public databases, and intersecting targets were identified. Protein–protein interaction (PPI) networks were constructed using the STRING database, while Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed via the DAVID database. A comprehensive “drug–compound–target–disease–pathway” network was established, and molecular docking was conducted to evaluate binding affinities of key compounds to core targets. Functional validation was performed in insulin-resistant cell models. Network pharmacology analysis identified 37 active constituents within the extract and 222 overlapping targets associated with T2DM. GO enrichment indicated involvement in protein phosphorylation, MAPK cascade activation, and negative regulation of apoptosis. Key signaling pathways included PI3K/AKT and lipid and atherosclerosis pathways. Molecular docking revealed strong binding affinities (binding energies ≤ −9.3 kcal·mol⁻¹) between core compounds—such as cheilanthifoline, glabridin, acetylcorynoline, skullcapflavone II, liquiritigenin, and dinatin—and pivotal targets including GAPDH, AKT1, TNF, SRC, EGFR, and PPARγ. In vitro experiments demonstrated that the extract significantly enhanced glucose uptake and glycogen synthesis in insulin-resistant cells, while suppressing oxidative stress and the expression of pro-inflammatory mediators such as TNF-α, MMP9, and IL-6. Collectively, B. heterostemon shows potential as an effective intervention for T2DM by targeting key molecular pathways, improving insulin sensitivity, and mitigating oxidative stress and inflammation in insulin-resistant cells. Full article

Camellia oleifera Abel shell is an abundant lignocellulosic byproduct of the Chinese woody oil industry, which is currently underutilized. To achieve its high-value utilization, this study developed an innovative cascade process integrating ultrasound-assisted extraction and macroporous resin purification for the efficient preparation of purified polyphenols from the shell (P-CPCS). The major constituents were identified by quadrupole/Orbitrap high-resolution mass spectrometry (HPLC-Q-Exactive-MS: Biotech Pack Co., Ltd., Beijing, China) analysis. The optimized process significantly enhanced the polyphenol yield (40.05 ± 0.58 mg GAE/g dw) and purity (57.72%), surpassing conventional methods. P-CPCS exhibited exceptional multifunctional bioactivities, including potent antioxidant capacity (with low IC₅₀ values against DPPH, ABTS⁺·, and ·OH radicals), effective tyrosinase inhibition (whitening effect), and significant bacteriostatic effects against various pathogens. Furthermore, P-CPCS notably suppressed the LPS-induced inflammatory response in RAW264.7 macrophages by reducing NO overproduction. This work highlights a novel and efficient strategy for upcycling agricultural waste into a high-performance natural antioxidant, positioning P-CPCS as a promising ingredient for applications in functional foods, cosmetics, and biomaterial stabilization. Full article