Marine-derived bioactive peptides have attracted increasing attention as value-added functional ingredients. In this study, peptides (<3 kDa) were prepared from yellowfin tuna processing by-products and further fractionated by Sephadex G-25 gel filtration. The major fraction (TBP-MF) exhibited markedly improved compositional homogeneity compared with the unfractionated hydrolysate (TBP), providing a well-defined peptide system for subsequent characterization and biological evaluation. Physicochemical analyses demonstrated that TBP-MF possessed enhanced thermal stability and a more ordered secondary structure, characterized by pronounced β-sheet enrichment, as revealed by TGA/DSC, FTIR, and circular dichroism analyses. Morphological and colloidal characterization further showed that TBP-MF formed relatively uniform lamellar and fibrous assemblies with a narrower particle size distribution and reduced electrostatic stabilization, indicating a higher tendency toward ordered self-association. Peptidomic profiling combined with in silico analysis revealed that TBP-MF was enriched in short peptides with relatively higher PeptideRanker scores and a functional motif distribution containing relatively more neuro-related annotations, although angiotensin-converting enzyme (ACE)- and dipeptidyl peptidase IV (DPP-IV)-related motifs remained predominant in both groups. In differentiated PC12 cells, TBP-MF exhibited excellent cytocompatibility and induced a stable, concentration-dependent increase in the Cell Counting Kit-8 (CCK-8) readout (OD450), indicating enhanced cellular metabolic activity and/or increased cell number. In addition, TBP-MF significantly increased intracellular levels of key neurochemical factors associated with sleep-related regulation, including tetrahydrobiopterin (BH4), serotonin (5-HT), and γ-aminobutyric acid (GABA). Overall, this study highlights yellowfin tuna by-products as a promising marine resource for bioactive peptides and suggests that fractionation-driven structural refinement is associated with neuro-related biological activity in differentiated PC12 cells. These findings support the potential application of marine by-product-derived peptides as functional ingredients in health-related fields.

Pyropia haitanensis polysaccharides have attracted growing attention for their diverse biological activities. In this study, we developed a synergistic extraction approach combining ultrasonic-assisted treatment and enzymatic hydrolysis using cellulase and pectinase. Response surface methodology (RSM) was applied to optimize the extraction conditions, which were determined as follows: 1.48% cellulase, 1.47% pectinase, 180 W ultrasonic power, and 65.9 °C temperature. Under these conditions, the polysaccharide yield reached 10.184 ± 0.27%. The crude extract was then purified through sequential DEAE Sepharose FastFlow and Sephadex G-75 chromatography, resulting in the purified fraction PPHP3. Monosaccharide analysis revealed that galactose, glucose, and glucuronic acid constituted the primary components in a molar ratio of 98.3:0.46:1.24. This polysaccharide exhibited a weight-average molecular weight of 25.208 kDa, a sulfate content of 8.64 ± 0.05%. In hypolipidemic assays using oleic acid-induced HepG2 cells, PPHP3 significantly reduced intracellular triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C), while simultaneously increasing HDL-C levels. These findings highlight the potential of P. haitanensis polysaccharides for hypolipidemic applications and establish a scientific foundation for their development in therapeutic and practical contexts.

The compact muscle architecture and poor aqueous dispersibility of Sthenoteuthis oualaniensis protein limit its use as a food ingredient. This study developed a gel filtration chromatography (GFC)-guided papain–alcalase sequential hydrolysis strategy using target-window peak area (Atarget) as a peptide-distribution response. Box–Behnken optimization identified an enzyme dosage of 1110 U/g, a papain/alcalase mass ratio of 3:5, and 8 h hydrolysis, yielding an Atarget of 0.0648 ± 0.0012 a.u. O-phthaldialdehyde (OPA)-derived degree of hydrolysis (DH) values were 10.72 ± 0.08%, 35.32 ± 0.12%, and 25.03 ± 0.37% for SPH-Pap, SPH-Alc, and SPH-opt, respectively, showing that the highest Atarget did not coincide with the highest DH. The essential-to-total amino acid ratio remained 38.18–39.20%, while lysine and methionine changed modestly. SPH-opt maintained high solubility across a broad pH range and exhibited a peptide profile distinct from those of the single-enzyme hydrolysates. Radical-scavenging capacity was assessed only post-optimization. At 5 mg/mL, SPH-opt showed DPPH and ABTS scavenging rates of 23.75% and 22.84%, respectively; without external standards, these values support only relative within-study comparisons. Overall, Atarget and DH provided complementary information on peptide distribution and bond cleavage, and no causal relationship between Atarget and radical-scavenging capacity was established.

Background:Brassinolide (BR) plays a pivotal role in regulating plant secondary metabolism and can promote the accumulation of bioactive compounds. However, relatively few studies have explored the application of BR in Salvia miltiorrhiza, and the mechanistic basis for its function in this context remains poorly understood.Methods:In this study, S. miltiorrhiza hairy roots were treated with BR at 1 μM and 5 μM. The treated samples were analyzed via high-performance liquid chromatography (HPLC)-based chemical quantification, transcriptomic sequencing, and quantitative real-time polymerase chain reaction (qRT-PCR) validation to systematically elucidate the concentration-dependent and time-dependent molecular mechanisms underlying BR-mediated biosynthesis of tanshinones and salvianolic acids in S. miltiorrhiza.Results:BR treatment at the 1 μM dose level significantly induced the accumulation of rosmarinic acid, salvianolic acid B, and tanshinone IIA, with respective 1.43, 1.82, and 1.78-fold increases in the levels of these compounds compared to controls. Expression of most key biosynthetic genes involved in the salvianolic acid and tanshinone biosynthesis pathways peaked at 1 h after treatment with 1 μM BR. Further transcriptomic analysis identified 15 significantly upregulated transcription factors that may regulate tanshinone and salvianolic acid biosynthesis. These differentially expressed genes were primarily enriched in the phenylpropanoid and terpenoid pathways. qRT-PCR validation confirmed the consistency and reliability of these transcriptomic data.Conclusion:Collectively, these findings reveal that BR treatment enhances secondary metabolism in S. miltiorrhiza by orchestrating changes in the expression of key structural genes and transcription factors, providing a theoretical basis for the breeding of high-quality germplasm resources.

Background/Objectives: Marine microorganism metabolites are structurally unique secondary metabolites possessing therapeutic potential. The current study aims to identify a novel ATPase regulator using a newly established bidirectional activity evaluation system to screen for microbial metabolites that inhibit the activities of Na+-K+-ATPase or Ca2+-Mg2+-ATPase. Methods: A total of 1258 marine microbial strains were isolated from sea mud in Zhoushan, Zhejiang. Results: The extract of strain ZSDH2536 exhibited Na+-K+ and Ca2+-Mg2+-ATPase inhibitory activity and was identified as Sirastachys pandanicola based on morphological and molecular phylogenetic analyses. The secondary metabolite was tentatively identified in the ZSDH2536 strain as a bisindole compound, and named Pandanicoline based on 1H-NMR, 13C-NMR and high-resolution mass spectrometry analysis. The chemical formula of Pandanicoline is C51H68N2O10, with an isotopic mass of 868.4874 Da. The maximum inhibition rate of Pandanicoline on Na+-K+ and Ca2+-Mg2+-ATPase was 36.37% and 37.27%, respectively. Moreover, in silico analysis also showed the binding energy of Pandanicoline with Na+-K+-ATPase was -9.124 kcal/mol and with the Ca2+-Mg2+-ATPase complex was -10.47 kcal/mol. Conclusions: The strain ZSDH2536 represents a promising source of dual inhibitors targeting Na+-K+ and Ca2+-Mg2+-ATPase. Pandanicoline exhibits potential as a lead compound for regulating ion homeostasis, providing new opportunities for further investigation into its mechanism and therapeutic applications.

Marine by-products represent a promising source of bioactive peptides. This study aimed to isolate and characterize a low-molecular-weight peptide fraction with antioxidant activity from Argentine shortfin squid carcass by-products, and to evaluate in vitro its cytocompatibility and protective effects against corticosterone (CORT)-induced oxidative injury in rat adrenal pheochromocytoma (PC12) cells and human astrocyte (hACs) cells. Argentine squid antioxidant peptide (PASN) was obtained by size-exclusion chromatography and fractionation-based screening. PASN exhibited the strongest overall free-radical-scavenging activity and consisted predominantly of components below 1 kDa (211.73–1013.48 Da). Spectroscopic analyses indicated that enzymatic hydrolysis transformed its structure from a rigid triple-helix conformation to a more flexible conformation dominated by β-turns (50.78%) and random coils (17.38%). In addition, thermogravimetric analysis confirmed its excellent thermal stability, with an onset decomposition temperature as high as 244.81 °C, supporting its potential applicability in high-temperature food-processing matrices. In vitro assays demonstrated that PASN exhibited high biocompatibility and promoted proliferation of both PC12 cells and hACs, while significantly improving cell viability under CORT challenge. PASN also reduced lactate dehydrogenase (LDH) leakage (hACs: 38.31%; PC12: 31.17%) in both cell models and restored total superoxide dismutase (T-SOD) activity (hACs: 69.46%, PC12: 66.40%). Immunofluorescence further revealed that PASN rescued the expression of brain-derived neurotrophic factor (BDNF) (hACs: 35.23%, PC12: 12.50%) and glutamate decarboxylase (GAD1/2) (hACs: 102.66%, PC12: 31.31%), key markers associated with synaptic plasticity and GABAergic sleep regulation. Collectively, PASN is a thermally stable squid-derived peptide fraction that exerts antioxidant and cytoprotective effects in neural cell models in vitro and represents a promising sustainable candidate for nutraceutical development.

Impaired wound healing is often caused by persistent inflammation, bacterial infection, and insufficient extracellular matrix remodeling. Natural polymer-based hydrogels represent ideal wound dressings but often struggle to balance structural stability and biological activity. Herein, we report a dual-functional network regulation strategy enabled by highly soluble mussel foot protein (HMFP) that acts simultaneously as a structural crosslinking regulator and bioactive effector to fabricate synergistic hydrogels (CS-SH-H) from β-chitosan (CS) and sodium hyaluronate (SH). HMFP homogenizes the porous microstructure, strengthens intermolecular interactions, and significantly improves thermal and structural stability via multivalent non-covalent bonding. In vitro, CS-SH-H shows excellent cytocompatibility, significantly promotes fibroblast proliferation and migration, and exerts potent antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). In a mouse full-thickness skin defect model, the hydrogel dramatically accelerates wound closure, reducing the residual wound area to 25% on day 7, outperforming the control groups. Immunohistochemistry confirms that HMFP suppresses TNF-α-mediated inflammation and enhances Ki-67-positive cell proliferation, leading to accelerated re-epithelialization and collagen deposition. This study establishes HMFP as a promising marine-derived dual-functional network regulator for designing high-performance hydrogel dressings. This strategy is scalable and translatable for treating infected and inflammatory wounds.

The design and optimization of immobilized metal affinity chromatography (IMAC) media are crucial to enhancing the purification efficiency of recombinant proteins. In this study, the agarose-based microspheres are prepared by using a three-factorial Box-Behnken design followed by NTA-Ni2+ agarose-based microspheres (ABM) preparation by the "one-step" crosslinking of epichlorohydrin (ECH)-nitrilotriacetic acid (NTA) to efficiently couple the NTA ligand to the surface of the matrix. After preparation, various sophisticated techniques, including SEM, AFM, DSC, FTIR, and SDS-PAGE, were used to analyze the morphological structure, thermal stability, and chemical composition of NTA-Ni2+ ABM. The optimal conditions are identified as an emulsifier PP concentration of 8.12 wt%, a stirring speed of 1624.46 rpm, and an oil-phase temperature of 53.86 °C, giving a span value (Y) of 0.50684. SEM, AFM, DSC, and FTIR results showed that the fabricated NTA-Ni2+ ABM were structurally stable and had a uniform cross-linking network for up to 8 h of coupling reaction time. The performance results showed that the beads had a high binding capacity for His-tagged proteins (15.2 ± 0.8 mg/mL), and SDS-PAGE results demonstrated the efficient purification ability for target proteins. These findings provide the theoretical basis and a practical solution for the rational design and application of IMAC medium.

Porphyra suborbiculata exhibits strong heat tolerance and has considerable commercial potential under rising sea temperatures; however, its bioactive components remain insufficiently explored. In this study, a heat-tolerant new strain of P. suborbiculata (PS-M4), cultivated by the College of Fisheries, was used as the experimental material. Polysaccharides were extracted using an ultrasound-assisted composite enzymatic method, and extraction conditions were optimized through single-factor experiments and response surface methodology, yielding a maximum extraction yield of 12.45 ± 0.09%. Crude polysaccharides were further purified using a purification apparatus, yielding two fractions, designated PSP-I and PSP-II. Preliminary structural characterization showed that PSP-I possessed a weight-average molecular weight (Mw) of 26.149 kDa, a number-average molecular weight (Mn) of 11.267 kDa, and a polydispersity index of 2.321. Monosaccharide composition analysis indicated that PSP-I was predominantly composed of galactose. Fourier transform infrared spectroscopy (FT-IR) revealed typical polysaccharide functional groups, and scanning electron microscopy (SEM) analysis revealed a porous lamellar morphology. In vitro cell-based assays demonstrated that PSP-I significantly alleviated ultraviolet B (UVB)-induced damage in HaCaT cells by reducing intracellular reactive oxygen species (ROS) levels, enhancing antioxidant enzyme activities, inhibiting apoptosis, and downregulating the expression of matrix metalloproteinases (MMPs). These results suggest that PSP-I has potential as a functional ingredient for mitigating UVB-induced skin damage.

Dysregulation of the microbiota–gut–brain axis has been implicated in Parkinson’s disease (PD), and modulation of the intestinal microbiota may offer a complementary nutritional approach. This study evaluated the preventive attenuation of MPTP-induced PD-like abnormalities by Lactiplantibacillus plantarum SHOU LAC-1 and examined associated microbiota–gut–brain axis-related changes. Forty male C57BL/6J mice were allocated to Control, MPTP, L-DOPA, and SHOU LAC-1 groups (n = 10 per group). SHOU LAC-1 was orally administered at 1 × 109 CFU per mouse per day for 42 days, with MPTP administered during the final 14 days; L-DOPA was administered during the MPTP-treatment period as a positive control. Motor performance and defecatory parameters, nigrostriatal TH and α-SYN immunoreactivity, neurotrophic-support-related gene expression, inflammatory markers, glial-cell-related gene expression, Nrf2-related antioxidant gene expression, colonic histopathology and tight-junction-related gene expression, gut microbiota composition, and serum metabolic profiles were assessed. Compared with MPTP-treated mice, SHOU LAC-1-treated mice showed better motor and defecatory performance, partial preservation of nigrostriatal TH immunoreactivity, reduced α-SYN immunoreactivity, increased neurotrophic-support-related gene expression, lower central and colonic inflammatory markers and glial-cell-related gene expression, increased Nrf2/HO-1/NQO1-related antioxidant gene expression, and comparatively better-preserved colonic morphology with higher tight-junction-related gene expression. 16S rRNA sequencing and untargeted serum metabolomics further showed that SHOU LAC-1 administration was accompanied by changes in gut microbial diversity and composition and serum metabolic profiles. Overall, under this preventive experimental design, SHOU LAC-1 administration was associated with attenuation of multiple MPTP-induced PD-like abnormalities, accompanied by changes in gut microbial and serum metabolic profiles. These preclinical findings are predominantly associative; causal relationships among microbial, metabolic, intestinal, and neurological changes remain to be established, and the translational relevance of SHOU LAC-1 requires further validation.

Although blood–brain barrier (BBB) models are of great value in investigating neurological diseases, the structural complexity and intricate function based on cell–cell interactions of the BBB bring various limitations to the applications of existing models. In this study, a novel BBB micro-organoid model was established by culturing neurovascular unit (NVU) cells on a decellularized squid mantle scaffold (DSMS) film to reconstitute a more authentic and reliable NVU microenvironment for in vitro research. The DSMS applied was obtained from squid mantle scaffolds via decellularization, followed by defatting, and showed good biocompatibility with no cytotoxicity. The DSMS film was finally prepared by lyophilization. The lyophilized film exhibited a void ratio and pore size suitable for the adhesion and growth of endothelial cells (hCMEC/D3) and astrocytes (hACs), which led to the formation of a BBB-like spatial structure. The BBB micro-organoid model exhibited functional barrier properties, including an effective transendothelial electrical resistance (TEER) of approximately 230 Ω/cm2, restricted permeability to macromolecules—with apparent permeability coefficients (Papp) of 6.3 × 10−7 cm/s for 10 kDa and 2.7 × 10−7 cm/s for 70 kDa FITC–dextran—and expression of tight junctional complex (TJC) proteins such as vascular endothelial cadherin (VE-cad) and Zonula Occludens-1 (ZO-1). Furthermore, low-density lipoprotein receptor-related protein 1 (LRP1), a key receptor stably expressed in these two NVU cell types, was utilized as a critical indicator to assess the integrity of the BBB micro-organ model and its responsiveness to pathophysiological stimuli, particularly under thrombotic conditions. This study not only validates the feasibility of constructing a functionally competent BBB micro-organ model using DSMS films integrated with NVU cells but also provides a promising in vitro platform for subsequent studies on the BBB-related pathological mechanisms and the evaluation of drug permeability across the BBB.

Psychrotrophic bacteria are major contributors to spoilage in refrigerated foods, posing significant challenges for food safety and quality management. Consequently, the development of rapid, on-site detection methods in specific food matrices is essential for controlling the proliferation of these microorganisms and reducing economic losses. This study aimed to develop a portable dual-color test strip platform based on recombinase polymerase amplification (RPA) and CRISPR/Cas12a technology for the simultaneous detection of Pseudomonas fluorescens (ATCC 17397) and Bacillus cereus(ATCC 14579). The platform integrates RPA, CRISPR/Cas12a-specific recognition and cleavage, and dual-color latex microsphere-labeled lateral flow immunoassay. The system was evaluated using purified bacterial DNA to assess sensitivity and specificity. The results demonstrated high sensitivity, with a detection limit of 10 copies/mu L for both target bacteria, no cross-reactivity against five common foodborne pathogens, and a complete assay time of within 45 min. This portable platform offers a sensitive, specific, and user-friendly solution for monitoring psychrotrophic bacteria in food matrices, facilitating timely intervention and contributing to improved food safety management.

Bacilus cereus and Pseudomonas fluorescens are major foodborne psychrotrophic bacteria posing global health and economic risks. B. cereus has a 23.8% food prevalence worldwide. P. fluorescens is a leading cause of spoilage in refrigerated products. Their rapid detection is crucial for food safety. However, existing detection methods often rely on open-tube operations, risking aerosol contamination. In this study, we developed two independent one-tube RPA-CRISPR/Cas12a visual detection assays for B. cereus and P. fluorescens. Using a physical separation design, the recombinase polymerase amplification (RPA) and CRISPR/Cas12a detection were pre-assembled in a single reaction tube. After incubation, a brief centrifugation combined the components for enclosed detection. This step is compatible with portable mini-centrifuges. The assays can be completed within 40 min at 37 °C, with results visualized directly under blue light. Both assays demonstrated good specificity against six common non-target pathogens. The visual detection limits were 5.1 × 101 copies/μL for B. cereus and 2.1 × 101 copies/μL for P. fluorescens. Each assay was applied to 14 types of real-world food samples (naturally contaminated and uncontaminated, confirmed by PCR), achieving 100% concordance with conventional PCR. The one-tube assays are tailored for psychrotrophic bacteria in refrigerated foods. They minimize aerosol contamination risk and provide a reliable solution for on-site cold-chain food safety monitoring.

Controlled drug delivery systems not only augment the therapeutic efficacy of drugs but also mitigate their adverse effects. Methotrexate (MTX) is a prominent first-line therapeutic agent in the management of rheumatoid arthritis (RA), yet it is characterized by its limited aqueous solubility, pronounced hemolytic activity, and propensity for off-target binding, which collectively contribute to its significant toxicity profile. To address these issues, a temperature-responsive controlled release system for MTX based on the analogous base pairing rule was designed for the RA treatment. Magnetic ferroferric oxide nanoparticles (Fe3O4 NPs) coated with polydopamine (PDA) were modified with thymidine-1-acetic acid (TAA) to prepare nanocarrier (FPT NPs). Furthermore, the thymine molecules within the FPT NPs possessed the strong ability to specifically bind with the pteridine structure present in MTX, thereby enabling the fabrication of a nanomedicine (designated as FPT-MTX NPs) through the base pairing rule. Under the influence of an applied magnetic field, the FPT-MTX NPs demonstrated a remarkable capacity for precisely targeting the joint tissue in a mouse model of RA. Analogous to DNA, where double-strand breaks occurred due to heating-induced disruption of base pairing, MTX was released from FPT-MTX NPs following near-infrared irradiation-induced light-to-heat conversion. This mechanism facilitated the achievement of satisfactory therapeutic outcomes in the RA treatment.

Viral pneumonia poses a major global public health challenge, where excessive inflammatory responses contribute to tissue damage and respiratory failure. Inflammation-responsive nanoparticles can target inflamed areas, improving drug delivery while minimizing side effects. Chitosan, a biocompatible polysaccharide with anti-inflammatory and immunomodulatory properties, gains enhanced antioxidant and anti-inflammatory capabilities when combined with selenium. This study developed selenium-chitosan nanoparticles loaded with Moringa A (MA), a natural antiviral compound from Moringa oleifera seeds. These nanoparticles target lung inflammation, releasing MA to suppress viral replication and infection while reducing inflammatory responses. Additionally, selenium-chitosan nanoparticles mitigate oxidative stress, regulate immunity, and inhibit PANoptosis-a cell death pathway that exacerbates inflammation. By blocking core proteins in this pathway, they further curb inflammatory factor release. This approach offers a promising therapeutic strategy for viral pneumonia, combining targeted drug delivery, antiviral action, and inflammation control with reduced side effects.

Protamine is a promising marine-derived bioactive compound that is highly arginine-rich and has demonstrated unique advantages in medical and biological research. This study, for the first time, investigates the molecular mechanisms underlying the immunomodulatory effects of Salmon Protamine Sulfate (SPS), Symplectoteuthis oualaniensis Protamine (SOP), and its polyethylene glycol (PEG) derivative (SOP-PEG) on RAW264.7 macrophages. The results demonstrate that both SOP and SOP-PEG significantly enhance the proliferation of RAW264.7 cells by promoting the secretion of pro-inflammatory cytokines and nitric oxide (NO), increasing ROS production, and improving antioxidant capacity, in comparison to SPS. Elevated ROS levels play a crucial role in enhancing macrophage immune activity, while the enhanced antioxidant defense mechanisms help maintain redox homeostasis and protect against oxidative stress-induced cellular damage. A Western blot analysis reveals that SOP and SOP-PEG notably regulate the expression of key proteins associated with the PI3K/Akt signaling pathway and anti-apoptotic mechanisms. Furthermore, a flow cytometry analysis indicates a significant increase in the G2/M-phase cell population in the treatment groups, which is corroborated by Western blot data showing alterations in critical regulatory proteins. Notably, SOP-PEG exhibits the strongest effects in regulating macrophage immune activity, which can be attributed to the enhanced stability and prolonged bioactivity resulting from the PEGylation of SOP. This comprehensive study reveals how SOP and SOP-PEG enhance macrophage immune function through multiple mechanisms, including PI3K/Akt activation, redox regulation, and cell cycle modulation. It provides valuable insights and a theoretical foundation for their potential applications in immunotherapy and immune regulation.

Microbial exopolysaccharides from extreme environments are increasingly becoming valuable candidates for drug development. In this study, four fractions named XL-1, XMRS-1, XL-1-D, and XMRS-1-D were isolated and purified from the hadal bacterium Psychrobacter pulmonis by column chromatography. The structural features of these fractions were characterized by molecular weight, monosaccharide composition, Fourier transform infrared (FTIR) spectrum, amino acid analysis and NMR. The results showed that XL-1 and XMRS-1 were mainly composed of mannose, glucose, and glucosamine, while XL-1-D and XMRS-1-D were mainly composed of mannose. In vitro bioactivity assays demonstrated that all four fractions significantly enhanced RAW264.7 macrophage proliferation and phagocytosis, stimulated nitric oxide (NO) and reactive oxygen species (ROS) production, and induced the secretion of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and the expression of inducible nitric oxide synthase (iNOS) mRNA. Moreover, plate cloning tests, cell scratch tests, and apoptosis assays, along with RT-qPCR analysis, demonstrated that the four fractions significantly inhibited A549 cells’ proliferation. Specifically, XMRS-1 and XMRS-1-D upregulated Bax, Caspase-3, Caspase-8, and Caspase-9, while downregulating Bcl-2, suggesting transcriptional activation of apoptosis-related pathways. These results offered a reference for the further development and utilization of this hadal bacterium in the future.

A supramolecular drug delivery system is fabricated here based on the assembly of β-cyclodextrin-modified hyaluronic acid (HACD) and ferrocene (Fc) with pH-sensitive polyhistidine (PHIS). The system demonstrates efficient encapsulation of hydrophobic anticancer drugs with high and stable drug loading capacity. The drug-loaded nanoparticles utilize HACD as the hydrophilic corona, enabling active tumor targeting through CD44 receptor-mediated endocytosis and exhibiting dual stimuli-responsive release properties (ROS and pH). In the tumor microenvironment characterized by elevated ROS levels and acidic pH, the nanoparticles undergo structural disassembly, switching from controlled release to rapid drug liberation. Blank nanoparticles exhibit excellent biocompatibility, while drug-loaded formulations demonstrate selective cytotoxicity with significantly reduced toxicity toward normal cells HFF-1 and enhanced therapeutic efficacy against HeLa cancer cells. This nanoplatform significantly improves the aqueous solubility and biocompatibility of hydrophobic drugs, achieving intelligent delivery and double-modal stimulus-responsive release.