Search Results (4,595)

Diagnosis of primary biliary cholangitis (PBC) currently depends on invasive liver biopsy or serum markers with inadequate diagnostic performance. This study aimed to identify non-invasive urinary protein biomarkers for PBC detection. Urine specimens from biopsy-verified PBC patients and healthy controls were processed through ultracentrifugation-based protein extraction, enzymatic digestion, and HPLC-ESI-IT/MS proteomic profiling; protein quantification was completed using Spectronaut v14.8. We identified 194 differentially expressed urinary proteins (109 upregulated, 85 downregulated) and screened 10 immune-related candidates through GO and KEGG enrichment. Pearson correlation further filtered three core proteins, osteopontin (SPP1/OPN), RAMP3 and S100A8, that correlated significantly with key PBC biochemical indices (ALP, GGT, AST, ALT, IgM, p < 0.05). Elevated urinary concentrations of OPN, RAMP3 and S100A8 were validated by ELISA in an independent cohort containing 30 PBC patients and 20 healthy volunteers. In summary, urinary OPN, RAMP3 and S100A8 are markedly increased in PBC patients and hold promise as non-invasive diagnostic biomarkers for PBC; however, their diagnostic specificity against other cholestatic and autoimmune liver diseases remains to be evaluated, and further confirmation in larger multicenter cohorts with disease control groups is warranted. Full article

Rubus saxatilis L. (Rosaceae) is used in traditional medicine in Central Kazakhstan, yet the effect of the extraction method on its phenolic profile and biological activity has not been studied systematically. Three extracts were compared: two prepared by ultrasound-assisted extraction differing only in solvent—water (WE) and 50% ethanol (UAE)—and one by microwave-assisted extraction with 50% ethanol (MAE), allowing the effect of solvent (WE vs. UAE) and of extraction technique (UAE vs. MAE) to be assessed separately. Phenolic composition was determined by HPLC-UV-ESI-MS; antioxidant activity in vitro; cytotoxicity for Artemia salina; and antigenotoxic effects using Escherichia coli K12 MG1655 lux-biosensors (pKatG::lux, pSoxS::lux, pColD::lux) and Drosophila melanogaster models. Four phenolics were identified, with gallic acid dominant in the UAE and MAE extracts and hyperoside predominant in WE, while rutin was not detected in WE. MAE showed the strongest radical-scavenging activity (DPPH, IC₅₀ = 7.44 µg/mL), whereas WE had the highest reducing power (FRAP, EC₅₀ = 52.97 µg/mL). All extracts were non-toxic for A. salina and induced neither oxidative stress nor the SOS response; instead, they attenuated dioxidine-induced SOS signalling by up to 57%, suggesting an antigenotoxic (DNA-protective) effect at the bacterial level. In Drosophila, extracts produced moderate extract-dependent genotoxicity: the lowest damage was observed for MAE. The extraction method governs the phenolic profile and biological activity of R. saxatilis, with MAE being most favorable. Full article

Jania rubens (J. rubens), a Mediterranean red seaweed, is rich in bioactive compounds with potential medicinal applications. This study investigates the therapeutic potential of Lebanese J. rubens through in vitro evaluation of its anti-diabetic, anti-coagulant, anti-inflammatory and anti-hemolytic activities. Various extracts were prepared and characterized using GC-MS and HPLC. Biological activities were assessed through α-amylase inhibition assays, effects on prothrombin time and partial thromboplastin time, anti-inflammatory activity and anti-hemolytic activity. Significant α-amylase inhibition was observed with dichloromethane/methanol and lipid extracts, comparable to acarbose. All types of extracts prolonged PT and PTT, with the lipid extract showing the strongest effect at higher concentrations. Additionally, dichloromethane/methanol extracts exhibited potent anti-hemolytic activity. Moreover, all extracts showed strong anti-inflammatory activity, achieving levels of inhibition of heat-induced BSA denaturation comparable to diclofenac. Characterization revealed 11 amino acids in the protein extracts, and significant fatty acids in the lipid profile. The crude extracts contained diverse compounds, including flavonoids, aldehydes, diterpenoids, terpenoids, fatty acids, and sterol esters, which may contribute to the observed biological activities. These results highlight J. rubens as a potential reservoir of bioactive compounds with promising in vitro activities related to diabetes, thrombotic disorders, inflammation, and oxidative stress. Further research is needed to isolate the active compounds, validate their efficacy in vivo, assess safety, and elucidate the underlying mechanisms. Full article

Introduction: PKMYT1 is a WEE-family G2/M cell cycle checkpoint kinase commonly overexpressed in a broad spectrum of human malignancies. WEE1 exclusively phosphorylates CDK1 at Tyr15, whereas PKMYT1 targets both Thr14 and Tyr15. Unlike WEE1 inhibition, PKMYT1 suppression triggers synthetic lethality with CCNE1. Nearly all disclosed PKMYT1 inhibitors so far fall into structural analogs originating from RP-6306, making the discovery of PKMYT1 inhibitors with chemotypes distinct from RP-6306 crucial. Methods: The compounds were structurally optimized using CADD, synthesized, and characterized by ¹H NMR, ¹³C NMR, HRMS, and HPLC. They were then assessed for kinase binding affinity via the LanthaScreen^TM Eu kinase binding assay, for cellular activity using the CCK-8 assay, and for cell-cycle distribution by flow cytometry, along with investigations into related mechanisms. Results: This study yielded 24 compounds of 2-aminopyrimidine through substituent derivatization of the pyrimidine scaffold. Among these derivatives, MS13 exhibited potent kinase binding affinity against PKMYT1 (IC₅₀ = 0.86 nM) and demonstrated strong anti-proliferative activity against CCNE1 high-amplification OVCAR3 cells and HCC1569 cells (IC_50-OVCAR3 = 1.52 μM, IC_50-HCC1569 = 0.66 μM). Additionally, it showed some selectivity towards A549 and HEK293T cells, with SI values of 4.31 (A549/OVCAR3), 9.92 (A549/HCC1569), 2.04 (HEK293T/OVCAR3), and 4.70 (HEK293T/HCC1569). Compound MS13 dose-dependently suppressed clonogenicity and triggered S-phase cell cycle blockade. Pharmacokinetic studies showed moderate hepatic microsomal stability (t_1/2 = 32.2 min). Molecular dynamics simulations indicated a favorable binding mode between compound MS13 and PKMYT1 (docking score: −9.322 kcal/mol). Conclusions: MS13 is a promising highly potent tool compound that provides a clear direction for the future optimization of PKMYT1 inhibitors. Full article

Background: Rosmarinic acid (RA) is a major phenylpropanoid constituent of many medicinal plants belonging to the Nepetoideae subfamily of the Lamiaceae family and contributes significantly to their biological activities. Traditionally, RA-containing extracts have been prepared using organic solvents or hot-water extraction. Methods: In addition to conventional extraction techniques, the phenylpropanoid composition of medicinally important tissues from 13 Nepetoideae species was investigated following incubation in an aqueous medium at room temperature. Phenylpropanoids in the differently prepared extracts were identified by extensive HPLC-UV-HR-MS/MS and NMR analyses. The antioxidant and cytostatic activities of isolated phenylpropanoids were evaluated using DPPH and Alamar Blue assays, respectively. Results: In tissues of all investigated species, RA underwent rapid enzyme-catalyzed conversion during aqueous incubation through oxidative decarboxylation, accompanied by the transient accumulation of nepetoidins A and B. The RA-related phenylpropanoid salvianolic acid K was identified in the roots of three Salvia species, while a previously undescribed isomer, designated salvianolic acid O, was discovered in the roots of Salvia verticillata. Both compounds underwent enzymatic transformations analogous to those of RA in aqueous medium, yielding two previously undescribed phenylpropanoids, salvianolic acids P and Q. Optimized aqueous incubation conditions enabled the high-yield accumulation of these conversion products. Together with their parent compounds, seven phenylpropanoids were isolated from selected plant tissues and were found to exhibit structure-dependent antioxidant and metabolic inhibitory activities in vitro. Conclusions: Aqueous incubation of Nepetoideae plant tissues markedly alters their phytochemical composition and provides access to previously unavailable bioactive phenylpropanoids. Full article

Minicitrus (Fortunella hindsii) is a wild kumquat with potential nutritional and medicinal value, but its tissue-specific bioactive metabolites, antioxidant activity, and endophytes remain poorly characterized. Here, we integrated LC-MS profiling, HPLC quantification, chemical and cellular antioxidant assays, 16S rRNA gene sequencing, bacterial isolation, and treatment experiments across five tissues. Among all the metabolites determined by HPLC, 13 were quantified using authentic standards and 24 were semi-quantified as linarin equivalents. Leaves, stems, and fruits accumulated abundant flavone glycosides, with phloretin-3′,5′-di-C-glucoside reaching 1823.19 ± 278.71 μg/g FW in leaves and 847.11 ± 27.28 μg/g FW in fruits, while roots and seeds showed distinct coumarin- and furanocoumarin-rich profiles, respectively. Leaf extracts showed the strongest chemical and cellular antioxidant activities, followed by fruit extracts. Isoorientin 2″-O-rhamnoside and diosmin exhibited strong chemical antioxidant capacity, and several flavone glycosides showed protective effects in cellular assays. Endophytic bacterial communities differed markedly among tissues, with fruit harboring a distinct community dominated by Weissella and Pantoea, which accounted for 65.64% and 19.13% of the relative abundance, respectively. Among two culturable fruit-associated isolates, Weissella sp. SJG-1 treatment was associated with 32.92–104.45% increases in six HPLC-quantified metabolites and higher antioxidant activity of fruit extracts. These findings highlight minicitrus as a source of bioactive metabolites and suggest a potential association between fruit-associated bacteria and fruit functional properties. Full article

Background/Objectives: Vancomycin remains a cornerstone antibiotic for Gram-positive infections, but its narrow therapeutic window and marked pharmacokinetic variability in critical patients complicate dosing. This study aimed to analyze survival and identify factors associated with mortality in intensive care unit (ICU) patients receiving vancomycin under therapeutic drug monitoring (TDM). Methods: A single-center, retrospective cohort study was conducted at City Clinical Hospital named after I.V. Davydovsky (Moscow, Russia) between December 2021 and January 2026 (IRB protocol VANCO-2021). A total of 190 adult patients from surgical, therapeutic, and cardiac ICUs receiving vancomycin for at least 3–5 days were analyzed; trough concentrations were determined by HPLC-MS/MS. Cox proportional hazards, spline Cox, and segmented (piecewise) Cox models were applied to identify mortality predictors, with bootstrap subsampling assessing threshold robustness. Results: Three independent predictors of mortality were identified: ln(minimum vancomycin concentration) (HR = 1.644, p = 0.028), age (HR = 1.019, p = 0.037), and serum creatinine (HR = 1.004, p < 0.001). A data-driven threshold trough concentration (Ctrough) of 25.5 µg/mL was identified, above which mortality increased substantially (63.0% vs. 26.4%; p < 0.05). The adjusted hazard ratio for Ctrough ≥ 25.5 µg/mL was approximately 1.88 (p = 0.054); bootstrap subsampling yielded an uncertainty interval of 23.2–33.6 µg/mL. Conclusions: Elevated minimum vancomycin concentrations exceeding 25.5 µg/mL are independently and statistically associated with increased mortality in ICU patients, likely reflecting altered drug exposure and illness severity rather than a direct causal effect. Systematic TDM is essential for maintaining vancomycin within the therapeutic range. Full article

The persistence of antibiotics like ciprofloxacin (CIP) in aquatic environments necessitates the development of efficient, low-cost wastewater treatment technologies. This study investigates a low-temperature fabrication strategy for mixed-phase copper oxide (CuO/Cu₂O) coatings synthesized via co-precipitation and immobilized using electrophoretic deposition (EPD). A critical finding was that a 25 min precursor aging time (T25) preserved a metastable mixture of CuO and Cu₂O phases, which is highly advantageous for creating heterojunction interfaces that enhance charge separation. In contrast, extended aging (T35) promoted phase consolidation toward bulk CuO, reducing catalytic surface area. During the EPD process, applied voltage acted as an influence to the relative phase composition and deposition behavior of the deposited coatings; 1.0 V was identified as the optimal condition, balancing high phase fidelity with enhanced crystallinity (average crystallite size of 30.6 nm) and mechanical stability. Photocatalytic experiments demonstrated significant CIP degradation, with the 0.9 V and 1.0 V films outperforming the 1.2 V film, possibly due to more favorable surface chemistry and phase diversity. While the 0.9 V film achieved the highest mineralization efficiency (18% TOC removal), the 1.0 V film offered the best balance between photocatalytic activity, structural stability, and phase selectivity for practical applications. High-Performance Liquid Chromatography-Mass Spectrometry HPLC-MS analysis suggested that degradation proceeds through oxidative pathways involving piperazine ring cleavage and defluorination. Full article

The bacterial Quorum Sensing system refers to the recognition of signaling molecules called autoinducers produced by bacteria when a certain cell density is reached in the environment. Those cell-density-dependent autoinducers regulate and coordinate diverse functional processes, such as bioluminescence, biofilm production, sporulation, and even the expression of some virulence factors, among others. There is a wide variety of autoinducers, and for Gram-negative bacteria, the canonical autoinducers are the N-acyl-homoserine lactones (AI-1). Presently, the production of autoinducers in Leptospira has not been described; therefore, the objective of this study was to detect and identify autoinducers in this bacterial genus. We report here the expression of AI-1 in cultures ≥2.4 × 10⁸ of Leptospira meyeri. Ethyl acetate extracts of Leptospira culture supernatants were capable of activating the β-galactosidase system in the biosensor Agrobacterium tumefaciens strain NTL4. Partial identification of the leptospiral supernatant extracts was done by thin-layer chromatography (TLC), showing a similar retention factor to the synthetic standard N-Octanoyl-DL-homoserine lactone (C8-AHL) in the Leptospira supernatant extracts. In addition, infrared spectroscopy (IR) analysis showed peaks corresponding to the lactone and amide groups in both the C8-AHL standard and the Leptospira meyeri culture extracts. Moreover, High-Performance Liquid Chromatography–Mass Spectrometry (HPLC-MS/MS) confirmed the same retention time (10.7 ± 0.1 min) in both the Leptospira meyeri supernatant extracts and the C8-AHL standard. These results show that Leptospira meyeri synthesizes N-acyl homoserine lactone family autoinducers, particularly the N-Octanoyl-DL-homoserine lactone, and lay the groundwork for future research on Quorum Sensing systems in Leptospira. Full article

Clusia grandiflora Splitg. (Clusiaceae), popularly known as “cebolão” or “orelha-de-onça”, is a Neotropical species recognized as a rich source of bioactive secondary metabolites, particularly prenylated benzophenones. This study investigated the chemical composition and antileishmanial potential of the ethanolic extract (EBCG), dichloromethane fraction (FDCG), and hexane fraction (FHex) obtained from C. grandiflora fruits. Chemical profiling by HPLC-ESI-IT-MS/MS revealed a metabolome dominated by prenylated benzophenones, terpenes, and flavonols. Biological evaluation included activity against promastigote and axenic amastigote forms of Leishmania (L.) amazonensis, cytotoxicity toward RAW 264.7 macrophages, assessment of cell death phenotypes, nitric oxide (NO) production, and molecular docking against Leishmania infantum trypanothione reductase (TRLi). FDCG and FHex exhibited moderate activity against promastigotes (IC₅₀ = 20.34 and 85.86 µg mL⁻¹, respectively), whereas EBCG was inactive. FHex showed the highest activity against axenic amastigotes (IC₅₀ = 7.0 µg mL⁻¹), while EBCG and FDCG displayed moderate activity (IC₅₀ = 15.92 and 26.43 µg mL⁻¹, respectively). Cell death occurred predominantly through a necrosis-like phenotype in promastigotes and an apoptosis-like phenotype in axenic amastigotes, and all samples significantly reduced NO production. Molecular docking indicated that several putatively annotated metabolites from C. grandiflora can be favorably accommodated within the TS2 substrate-binding cavity of Leishmania infantum trypanothione reductase (TRLi). Garcinielliptone I and Guttiferone E exhibited the most favorable predicted binding affinities, whereas xanthocymol displayed an interaction pattern involving the catalytic residues Cys52 and Cys57. These findings highlight selected putatively annotated metabolites as promising candidates for future biochemical investigation. Overall, these findings highlight C. grandiflora fruits as a promising, previously underexplored source of prenylated benzophenones with antileishmanial activity and the ability to reduce nitric oxide production in LPS-stimulated RAW 264.7 macrophages. Full article

Adonis aestivalis L. is a member of the genus Adonis, traditionally associated with cardiotonic compounds; however, its polyphenolic composition and anti-inflammatory potential warrant further phytochemical and pharmacological evaluation. The aim of this study was to characterise the metabolic profile of A. aestivalis extracts, with a focus on flavonoid-containing components, and to evaluate their anti-inflammatory activity in LPS-stimulated RAW 264.7 macrophages. Aqueous, 50% ethanolic, and absolute ethanolic extracts were analysed by HPLC–ESI–QTOF-MS/MS in positive and negative ionisation modes. The ethanolic extracts exhibited the most diverse chromatographic profiles, with a predominance of flavonoids, including derivatives of kaempferol, quercetin, and apigenin, as well as C-glycosylated flavones such as orientin, isoorientin, vitexin, and isovitexin. Phenolic acids, fatty acid derivatives, organic acids, and adonitol were also tentatively identified. The occurrence of glycosylated, acylated, and glucuronidated flavonoid derivatives indicates marked structural diversity within the flavonoid fraction. In biological assays, the 50% ethanolic extract showed no cytotoxicity towards RAW 264.7 cells at concentrations up to 50 μg/mL and produced a statistically significant but moderate reduction in LPS-induced NO production. Furthermore, it attenuated the release of pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α, to varying degrees. The data obtained indicate that A. aestivalis is a promising source of flavonoid-containing extracts with anti-inflammatory potential and may provide a foundation for further studies aimed at developing standardised phytochemical preparations with a defined polyphenolic profile. Further quantitative phytochemical analysis, bioactivity-guided fractionation, mechanistic studies, safety assessment, and in vivo evaluation are required to confirm the active constituents and therapeutic relevance of the extracts. Full article

Nursery-plant grading relies on morphological traits, but leaf metabolic variation associated with nursery-plant grade remains unclear. In this study, untargeted HPLC-QTOF-MS metabolomics was applied to mature and young leaves of Grade I, II, and III apple nursery plants to explore grade-associated metabolic variation and its relationship with growth performance. After quality filtering, 198 positive-ion features were retained from mature leaves, while 259 positive-ion and 8 negative-ion features were retained from young leaves. Multivariate analysis showed clear grade-associated separation in both leaf types, with stronger discrimination in young leaves. A combination of orthogonal partial least squares (OPLS) modeling and trend analysis identified 20 and 28 differential features in mature and young leaves, respectively. Cross-model prioritization further highlighted key metabolites putatively annotated as methyl nicotinate and 1-palmitoyl-sn-glycero-3-phosphocholine in mature leaves, and methyl nicotinate, nicotinamide riboside, cis-jasmone, and methyl (9Z,14Z)-12,13,16-trihydroxyoctadeca-9,14-dienoate in young leaves. These key metabolites were potentially associated with NAD precursor metabolism, membrane lipid remodeling, and oxylipin-related signaling. Correlation analysis showed that they were associated with both initial grading traits and post-transplant growth performance. These metabolites represent candidate molecular correlates of grade-associated physiological variation and growth performance in apple nursery plants. Full article

Background/Objectives: Niaprazine is widely used for the management of sleep disorders in pediatric and geriatric patients; however, no commercially available oral liquid formulation is currently available in Italy, making extemporaneous compounding necessary. In routine practice, the beyond-use date (BUD) is often limited to 30 days, potentially affecting therapeutic continuity. This study aimed to evaluate the chemical, physical, and microbiological stability of a compounded niaprazine syrup to support evidence-based BUD and to assess the suitability of selected ready-to-use compounding vehicles for preparing alternative niaprazine oral liquid formulations. Methods: A niaprazine syrup (3 mg·mL⁻¹) was prepared in a sucrose-based vehicle acidified with tartaric acid and preserved with potassium sorbate. Chemical stability of niaprazine was evaluated by high-performance liquid chromatography coupled with diode array detector (HPLC-DAD), whose specificity was confirmed by forced degradation studies. Stability was monitored for up to 9 months at 4–8, 25, and 40 °C and confirmed after 12 months by HPLC-DAD and mass spectrometry (MS). Physical stability of the formulation was monitored by pH and visual inspection up to 12 months. Microbiological quality was assessed for 2 months at 4–8 and 25 °C according to the European Pharmacopoeia. In parallel, four selected ready-to-use compounding vehicles were evaluated for their suitability to prepare stable niaprazine oral liquid formulations. Results: Niaprazine concentrations in the syrup remained within pharmacopeial acceptance limits (±10%) at all temperatures, although a decrease was observed under accelerated conditions (40 °C). pH remained stable (≤0.5-unit variation) and the formulation stayed clear and homogeneous throughout the study, with only minor visual changes after prolonged storage at 40 °C. Statistically significant differences (p < 0.05) were observed in both the HPLC-DAD and HPLC-MS datasets. Microbiological testing confirmed compliance up to 2 months at 4–8 and 25 °C (TAMC ≤ 10³ CFU/mL; TYMC ≤ 10² CFU/mL; Escherichia coli absent). Comparable chemical and physical stability was observed for three of the four formulations prepared with the ready-to-use vehicles over at least two months. Conclusions: The compounded niaprazine syrup demonstrated chemical, physical, and microbiological stability under refrigerated and room-temperature storage, supporting evidence-based beyond-use dating of up to two months under the tested conditions. Ready-to-use vehicles may represent a practical complementary approach, offering standardized alternatives for the preparation of niaprazine oral liquid formulations. Full article

Bioactive food ingredients that regulate the gut microbiota are promising dietary strategies for supporting systemic health, but their roles in skin photoaging remain insufficiently defined. Qiongyu Gao (QYG), a classical food–medicine homologous formula composed of Rehmanniae Radix, Panax ginseng, and Poria cocos, was evaluated as an oral functional food candidate for UV-induced skin photoaging. QYG was chemically characterized by HPLC and UPLC–QTOF–MS/MS. Young and aged mice were subjected to D-galactose plus UVA/UVB exposure and orally administered QYG, followed by skin transcriptomics, gut microbiota sequencing, serum metabolomics, and validation in UVB-injured primary dermal fibroblasts. QYG alleviated wrinkle formation, epidermal thickening, oxidative stress, inflammation, extracellular matrix degradation, collagen disorganization, and hyaluronic acid loss. Multi-omics analysis showed that QYG selectively remodeled gut microbiota, enriching Lactobacillus-, Bifidobacterium-, and Akkermansia-associated taxa, regulated serum metabolites related to energy and lipid metabolism, and enriched mitochondrial energy metabolism-related pathways in photoaged skin. In fibroblasts, QYG-containing serum restored mitochondrial membrane potential, reduced ROS accumulation and cellular senescence, and regulated AMPK/PGC-1α-associated markers. These findings support QYG as a promising food–medicine homologous functional food candidate for skin health maintenance through gut microbiota-associated systemic metabolic regulation. Full article

Background/Objectives: This study engineered and evaluated a targeted, folate-functionalized chitosan nanoparticle (CS-FA NP) delivery system to enhance the therapeutic efficacy of standard quercetin and Coriandrum sativum seed extract against breast cancer. Methods: Phytochemical profiling confirmed a 14% crude yield for the methanolic extract, with gas chromatography–mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) identifying quercetin as the principal bioactive agent. The synthesized CS-FA NPs exhibited a core size of 7–20 nm, an average hydrodynamic diameter of 150–160 nm, a stable zeta potential of −55 mV, and high encapsulation efficiencies (87.2% for quercetin and 80.5% for coriander). Kinetic assessments confirmed a biphasic, diffusion-controlled release matching Higuchi matrix kinetics. Anticancer activity was evaluated in vitro using MTT cytotoxicity, Annexin V-FITC/PI apoptosis analysis, RT-qPCR, and ex vivo rat aortic ring assays, followed by validation in a syngeneic 4T1 mammary tumor mouse model. Results: In vitro, folate-receptor-targeted quercetin nanoparticles (T4) demonstrated superior, selective cytotoxicity, particularly against triple-negative MDA-MB-231 cells, while sparing normal fibroblasts. Annexin V-FITC/PI apoptosis profiling and ex vivo aortic ring assays revealed profound, cell-line-dependent programmed cell death and up to 90% inhibition of microvessel sprout outgrowth. Mechanistically, RT-qPCR verified that nano-formulations induced complete transcriptional silencing of NANOG, MMP-1, VEGFA, TSPAN8, TWIST, EMMPRIN, and CDK1, alongside marked upregulation of P27^KIP1 and P21^CIP1. In vivo, these nano-formulations successfully improved tumor-associated pathological features, reduced aggressive tumor spindle-cell proliferation, and suppressed elevated serum CA15-3 and arginase biomarkers. Conclusions: Folate-functionalized chitosan nano-formulations significantly enhanced the anticancer efficacy of quercetin and Coriandrum sativum seed extract through improved targeted delivery, potent antiproliferative, anti-angiogenic, and pro-apoptotic activities, together with favorable modulation of multiple molecular pathways associated with breast cancer progression. These findings support their potential as promising targeted nanotherapeutic strategies for breast cancer treatment. Full article