Acute lymphoblastic leukemia (ALL) is the most common hematological malignancy in children. 6-Mercaptopurine (6-MP) is a core chemotherapeutic drug for ALL, but its clinical application is limited by poor solubility, low biological availability, and significant side effects. To address these limitations and to investigate the impact of developmental differences between children and adults on drug absorption, this study developed 6-MP-loaded poly (lactic-co-glycolic acid) nanoparticles (6-MPNs) using a double emulsion-solvent evaporation method. We comprehensively applied high-performance liquid chromatography-mass spectrometry (HPLC-MS) and label-free quantitative proteomics, along with GO functional annotation via eggnog-mapper (v2.0) and protein-protein interaction analysis using STRING (v10.5), to systematically evaluate the pharmacokinetics, intestinal distribution, and duodenal protein expression differences of 6-MPNs in animal models of different ages. All animal studies were approved by the Animal Ethics Committee of Capital Medical University (Approval No.: AEEI-2020-072) and conducted in accordance with relevant guidelines. The results showed that 6-MPNs significantly improved the biological availability of 6-MP. Rats at 4 weeks of age group exhibited higher AUC_last (area under curve from time zero to time of last measurable concentration) and C_max (maximum concentration), faster intestinal absorption, more rapid propulsion, and higher transport efficiency from the intestinal epithelium to the portal vein compared to the rats at 16 weeks of age group. Proteomic analysis revealed that the rats at 16 weeks of age group had significantly lower expression of proteins related to fat digestion and transport, pancreatic secretion, membrane lipid metabolism, brush border recruitment, and microvillus adhesion. These findings suggest that the higher biological availability of 6-MPNs in younger animals may result from longer relative intestinal length, higher intestinal wall transport efficiency and more dynamic barrier construction.

Ultra-high performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) was applied for the non-targeted metabolomic characterization of Paeoniae Radix Alba and Paeoniae Radix Rubra decoction pieces. Differential metabolites were identified through multivariate statistical analysis. Interpretable machine learning methods, including random forest (RF), adaptive boosting (AdaBoost), and extreme gradient boosting (XGBoost), were further employed to build classification models, and the Shapley additive explanations (SHAP) algorithm was used to evaluate the contributions of characteristic metabolites. A total of 44 metabolites were identified, including 9 flavonoids, 8 phenolic acids, and other types of metabolites, among which 12 showed significant differences between the two kinds of decoction pieces. In the test set, the RF model exhibited superior performance. SHAP analysis indicated that gluconic acid, quercetin, kaempferol, L-aspartic acid, taxifolin, and oxypaeoniflorin significantly contributed to model discrimination, with L-aspartic acid showing elevated levels in Paeoniae Radix Alba, while the other metabolites were relatively higher in Paeoniae Radix Rubra. The metabonomics-machine learning strategy established in this study provides potential chemical evidence for elucidating the efficacy differences between Paeoniae Radix Alba and Paeoniae Radix Rubra and offers new insights into the identification, origin traceability, and characteristic marker discovery of other easily confused traditional Chinese medicines.

The study aims to establish a method for the content determination of eleven components in Jinchai Qingre Granule by high performance liquid chromatography-diode array detector (HPLC-DAD) through quantitative analysis of multi-components by single marker (QAMS) and verify the feasibility and applicability of the method in its quality control. Agilent ZORBAX SB-C18 column (250 mm × 4.6 mm, 5 μm) was used; acetonitrile-0.1% phosphoric acid aqueous solution was used as mobile phase; gradient elution and the detection wavelength were set at 327, 278, 250 nm; flow rate was 1.0 mL·min^-1; column temperature was 25 ℃. Using puerarin as the internal reference, the relative correction factors of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, daidzin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, baicalin, daidzein and arctiin was established, and the eleven components were determined. Eleventeen components showed good linear relationships within their own ranges (r ≥ 0.999 5), whose average recoveries were 98.05%-100.90% with the RSDs of 0.98%-2.35% (n = 6). The method showed good durability under different conditions such as instruments, columns, column temperature, injection volume, and flow rate. There was no significant difference between the calculated values of eleven components in ten batches of samples by the QAMS and the measured values of the external standard method (P > 0.05). The method established in the study is simple and stable, and can be used for the synchronous quality control of eleven index components in Jinchai Qingre Granule.

Interleukin-15 (IL-15) stimulates the proliferation of CD8⁺ T lymphocytes and activates cytotoxic T lymphocytes, memory CD8⁺ T cells, and natural killer cells, demonstrating broad application prospects in cancer immunotherapy. The activation of effector cells by IL-15 relies on a unique trans-presentation signaling mechanism. This process involves the initial binding of IL-15 to IL-15 receptor alpha (IL-15Rα) to form the IL-15/IL-15Rα complex, followed by binding of this complex to the IL-15 receptor beta/gamma (IL-15Rβ/γ) on target cells, thus exerting its pharmacological effects. The clinical application of recombinant interleukin-15 (rIL-15) faces challenges including its short half-life time and limited availability of endogenous IL-15 receptor alpha (IL-15Rα). Additionally, the immunosuppressive tumor microenvironment impairs its anti-tumor efficacy. In this study, a recombinant IL-15Rα-sushi-HSA fusion protein (rILR-HSA) is designed and expressed, which comprises the IL-15Rα sushi domain fused to human serum albumin (HSA). The co-hitchhiking drug delivery system, rILR-HSA/BPBBT/rIL-15 nanoparticles, prepared by the hydrophobic binding between the HSA component and the photothermal agent molecule BPBBT, followed by the high-affinity binding between IL-15Rα-sushi and rIL-15. Compared with free rIL-15, rILR-HSA/BPBBT/rIL-15 nanoparticles extend the serum half-life time of rIL-15 by 2.6-fold. Simultaneously, the tumor accumulation of rILR-HSA/BPBBT/rIL-15 nanoparticles in the orthotopic CT26-Luc colon cancer-bearing mice facilitates intraoperative near-infrared window-Ⅱ fluorescence image-guided photothermal immunotherapy. The co-hitchhiking drug delivery strategy employs a fusion protein carrier to co-deliver rIL-15 and photothermal agent, providing an approach for safe and effective IL-15-based photothermal immunotherapy of tumor. Animal experiments were conducted with the approval of the Institutional Animal Care and Use Committee of the School of Pharmacy, Fudan University (ethics number: 2018-09-YJ-LW-01).

Volatile oil of traditional Chinese medicine (TCM) is an important intermediate in TCM preparations. However, its strong volatility and poor stability hinder the stable application of solid preparations containing volatile oil. There is an urgent need to develop novel stabilization and sustained/controlled-release technologies. In this study, four porous materials, namely SiO₂, Sylysia 350FCP silica (350FCP-SiO₂), Sylysia 770FCP silica (770FCP-SiO₂), and porous starch, were used to prepare solidified powder of the volatile oil of Chenxiang and Muxiang (CMVO). Comprehensive characterization of the solidified powder was conducted using scanning electron microscopy (SEM), X-ray diffraction (XRD), differential scanning calorimetry (DSC), simultaneous thermal analysis, and electronic nose methods. The pore size, pore characteristics of the materials, and the release behavior of the components before and after solidification of the volatile oil were analyzed to evaluate its sustained-release performance. Finally, multivariate statistical analysis methods were used to evaluate the correlation between the key physicochemical parameters of the carrier materials and the adsorption rate of the essential oil, as well as the sustained/controlled-release performance of the components. The results showed that among the four carriers, 350FCP-SiO₂ exhibited the strongest adsorption capacity for the essential oil, with an adsorption ratio of essential oil to carrier as high as 2.5 mL·g^-1. In terms of sustained-release performance, 350FCP-SiO₂ and 770FCP-SiO₂ exhibited a sustained-release effect on benzylacetone, a strongly volatile component in CMVO. Correlation analysis indicated that the adsorption rate of the material was positively correlated with the BJH cumulative adsorption volume and the total single-point adsorption pore volume. The 24-hour retention rate of benzylacetone was mainly positively correlated with the t-plot micropore volume and t-plot micropore area of the material, while dehydrocostus lactone, a weakly volatile component, was less affected by the carrier material during its release process. This study demonstrates that porous materials with different physicochemical properties and pore characteristics can effectively regulate the sustained-release performance of different volatile components in TCM essential oils. The findings provide theoretical basis and application reference for the solidification and stabilization of TCM essential oils.

Inflammatory bowel disease (IBD) is a chronic and relapsing disorder characterised by gut dysbiosis and redox imbalance. This study developed a novel oral micro-nano delivery system (CBs@BAN) based on Clostridium butyricum spores (CBs) and bilirubin-alginate nanoparticles (BAN). Amino-modified bilirubin (BR) was first covalently conjugated with sodium alginate (Alg) to prepare amphiphilic block BR-Alg, which subsequently self-assembled into BAN nanoparticles with a particle size of 163.52 ± 2.71 nm and a zeta potential of -43.2 ± 1.6 mV. These nanoparticles were efficiently loaded onto the surface of CBs via calcium ion cross-linking. In vitro experiments demonstrated that BAN exhibits potent reactive oxygen species (ROS) scavenging capacity, while CBs@BAN maintained spore germination activity. Following review and approval by the Animal Ethics Committee of Zhengzhou University (approval number: yxy11sc20250099), a murine colitis model was induced using 3% dextran sulfate sodium (DSS). Compared with other experimental groups, CBs@BAN demonstrated superior therapeutic efficacy, including accelerated body weight recovery, reduced disease activity index (DAI) scores, ameliorated histopathological damage in colon tissue, effective clearance of colonic ROS, suppression of inflammatory cytokine expression, and improved gut microbiota imbalance. By restoring redox homeostasis and modulating gut microbiota, this delivery system provides a novel and efficient solution for IBD treatment with promising application potential.

Pulmonary fibrosis (PF) is a progressive and irreversible interstitial lung disease of unknown etiology with limited therapeutic options. Bioinformatics enrichment analysis indicates that oxidative stress and metabolic dysregulation of alveolar epithelial type II cells (AECs II) may contribute to PF pathogenesis. With recent advances in molecular biology and genetic engineering, gene therapy has emerged as a promising therapeutic strategy for PF. However, its clinical translation remains challenging due to biological barriers that hinders efficient delivery of therapeutic genes. To address this limitation, we explored main factors affecting transfection efficiency and screened optimal prescription to develop nanocomplexes (NPs@pSIRT3) via delivery of sirtuin 3 plasmid (pSIRT3) to treat PF. The NPs@pSIRT3 exhibited superior transfection efficiency, mediating robust SIRT3 protein expression to achieve therapeutic efficacy. Comparative analysis with excipient-deficient control groups demonstrated that the optimized nanoparticles (NPs) effectively delivered therapeutic genes to fibrotic lesions, enhancing pSIRT3 transfection. The NPs@pSIRT3 significantly attenuated PF progression by modulating oxidative stress normalization of injured AECs II, anti-inflammatory effects, and finally inhibition of pro-fibrotic responses. These findings demonstrate that the engineered NPs@pSIRT3 system represents a promising gene delivery platform for PF therapy, and providing a novel strategy for efficient transfection of gene therapy. All animal procedures were performed in accordance with the guidelines of and approved by the Institutional Animal Care and Use Committee of Jinzhou Medical University (approval number: 240134-5).

Epidemiology shows that alopecia afflicts about half of the adult population worldwide and has become an important health problem affecting the daily lives of the general public. Due to the complexity of the etiology alopecia and the limited number of available treatments, it is important to develop efficient and safe treatment options. In this study, dutasteride (DUT) nanovesicles (NV@DUT) with egg yolk lecithin, oleic acid and vitamin E polyethylene glycol succinate were constructed, aiming to enhance the skin penetration and retention of DUT while having multiple coordinated effects of antioxidation and anti-inflammation, thereby improving the therapeutic effect. NV@DUT has a slightly milky luster, with a particle size of about 141.3 ± 0.45 nm, uniformly dispersed, its polydispersity index (PDI) is 0.17 ± 0.003. After 60 days of placement at room temperature with small changes in particle size and PDI. Compared with free DUT, the transdermal penetration and skin retention of the drug in NV@DUT increased by approximately 1.7 times. In vitro results showed that NV@DUT has antioxidant and anti-inflammatory activity. At 500 μg·mL^-1 NV@DUT, the scavenging rate of radicals was about 50%, and the inhibition rate of nitric oxide production was about 50%, it improves the homeostasis of the hair follicle microenvironment. In addition, at 20 μg·mL^-1 NV@DUT can promote the proliferation and migration of dermal papilla cells (HDPCs) and human umbilical vein endothelial cells (HUVECs), the cell viability were (132.67 ± 4.14)% and (116.64 ± 5.14)%; the migration rates were (84.68 ± 3.31)% and (61.87 ± 4.23)%. A preliminary investigation of its mechanism revealed that NV@DUT up-regulate the mRNA expression of β-catenin, vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1), and down-regulation the mRNA expression of type II 5 alpha-reductase (SRD5A2). NV@DUT can also promote hair regrowth in mice with androgenetic alopecia (AGA), and the weight of the new hair and the thickness of the skin in the mice were higher than those in the model group and the DUT group. The animal experiments were approved by the Laboratory Animal Ethics Committee of Sun Yat-Sen University, and all experiments were conducted in accordance with relevant guidelines and regulations. Approval number: 2022000828. The experimental results show that NV@DUT can treat AGA through multiple mechanisms and pathways, providing a new therapeutic strategy for alopecia.

Manganese dioxide (MnO₂), as an intelligent drug carrier responsive to the tumor microenvironment (tumor microenvironment, TME), has recently garnered significant attention due to its unique physicochemical properties. However, the low specific surface area of conventional MnO₂ materials severely limits their drug-loading capacity. In this study, three-dimensional urchin-like MnO₂ nanomaterials were successfully synthesized through an optimized hydrothermal process, and a composite MnO₂/Au nanoparticles (GNPs) system was constructed by immobilizing Au NPs onto the MnO₂ surface via an impregnation method. Structural characterization revealed that the as-prepared urchin-like MnO₂ exhibits a distinctive hollow-radial architecture with a specific surface area of 35.75 m²·g^-1, significantly higher than that of conventional MnO₂ materials (14-30 m²·g^-1). Under near-infrared laser irradiation (808 nm, 2.5 W·cm^-2), the composite demonstrated exceptional photothermal performance, achieving a temperature rise to 55 ℃ within 5 minutes and a calculated photothermal conversion efficiency of 61.85%. Drug-loading evaluations indicated a high loading efficiency of 81.3% for 5-fluorouracil, along with pH-responsive release characteristics, achieving 86.4% cumulative drug release within 2 h under simulated TME conditions (pH 5.0). By integrating morphology engineering and noble metal hybridization strategies, this work establishes a multifunctional nano-platform that combines high drug-loading capacity, controllable release, and efficient photothermal conversion, providing critical theoretical and technical foundations for developing synergistic chemo-photothermal therapeutic systems.

The treatment of gliomas faces significant challenges due to limitations imposed by the blood-brain barrier and the immunosuppressive tumor microenvironment. This study developed a perillyl alcohol-based nanoemulsion (POH-NE), which utilizes the reversible BBB-opening property of POH to efficiently deliver the STING agonist MSA-2 into the brain, aiming to achieve synergistic immunomodulation and antitumor therapy. ^MSA-2POH-NE exhibits uniform particle size (87.6 ± 2.3 nm) and neutral surface charge (-0.17 ± 0.1 mV), demonstrating system stability. In vivo distribution experiments demonstrated that POH-NE increased drug accumulation in the brain to 4.6 times that of conventional nanoemulsions. Mechanistic studies revealed that POH-NE reversibly reduced transendothelial electrical resistance and downregulated the expression of tight junction proteins Claudin-5 and ZO-1, while exerting no significant effect on the non-tight junction protein CD31. This suggests that POH may facilitate controllable BBB opening through coordinated paracellular and transcellular pathways. In an orthotopic glioma model, ^MSA-2POH-NE significantly activated the STING signaling pathway, promoted type I interferon production, enhanced the infiltration and activation of dendritic cells and cytotoxic T lymphocytes within tumors, and cooperatively reversed the immunosuppressive microenvironment. All animal experiments had been approved by the Experimental Animal Ethics Committee of the Institute of Laboratory Animals Science, Chinese Academy of Medical Sciences (No: SXD23001). In conclusion, this study proposed a novel combinatorial strategy for glioma immunotherapy based on reversible BBB opening and STING pathway activation, providing an experimental foundation for subsequent translational research.