Hypobaric hypoxia is a major physiological challenge in high-altitude environments, impairing oxygen delivery and utilization, which adversely affects health and work efficiency. Functional foods could provide a practical solution. Here, edestin peptide (EPP) derived from hemp (Cannabis sativa L.) seed is considered a promising target. In this study, the extracted of edestin was verified by SDS-PAGE, and enzymatic digestion to prepare EPP. Freeze-dried EPP showed a homogeneous porous microstructure and was enriched in glutamic acid (19.13%), arginine (14.01%) and aspartic acid (13.28%), and nutritional indices supported high protein quality. Peptidomics defined the peptide composition, and particle size measurements indicated that hydrolysis resulted in smaller particles and a higher polydispersity index. Spectroscopic analyses consistently revealed hydrolysisdriven conformational remodeling toward a looser, random coil enriched state, accompanied by improved interfacial properties and strong in vitro radical scavenging capacity without detectable cytotoxicity. In mice, EPP supplementation significantly prolonged rotarod endurance approximately fourfold (p < 0.01) at 4000 m and increased hematological indices related to oxygen transport under hypobaric hypoxia. In a human trial, EPP markedly improved maximum oxygen uptake by 14.98%, reduced exercise-induced fluctuations in blood oxygen saturation and heart rate, and increased hypoxia tolerance and anti-fatigue capacity by 41.22% and 1.625 times at 3600 m, respectively. This study provides systematic evidence from characterization to animal and human trials, supporting EPP as a functional ingredient to improve hypoxia-related physiological indicators.

Macrophage foaming, characterized by uncontrolled uptake of oxidized LDL (ox-LDL) by macrophages, critically drives atherosclerosis (AS) progression. Although NK cells are found to be atherogenic, their direct impact on macrophage foaming remains unknown. Here we examined the role of NK cells in macrophage foaming and found that NK cells exacerbated macrophage cholesterol accumulation both in the presence/absence of ox-LDL. Under ox-LDL exposure, NK cells promoted cholesterol accumulation via increasing cholesterol influx related gene CD36, and significantly reducing the expressions of cholesterol efflux associated receptors ABCA1 and ABCG1 in the macrophages. When without ox-LDL, NK cells accelerated cholesterol synthesis via the SREBP-2-LDLR/HMGCR pathway, and inhibited cholesterol efflux via the LXR-α-ABCA1/ABCG1 pathway of macrophages. These factors eventually led to the accumulation of cholesterol in macrophages, resulting in the formation of macrophage foam cells. Further, for the first time, we revealed the TIGIT/CD155 signaling pathway as a critical regulatory mechanism for macrophage foam cell formation. Specifically, the downregulation of TIGIT in highly active NK cells altered its interaction with CD155, which influenced macrophage cholesterol metabolism via CD155 and ultimately promoted foam cell formation. Furthermore, direct blockade of CD155 on macrophages exacerbated cholesterol accumulation, thereby establishing CD155 as an important regulator in macrophage-derived foam cell formation. These findings not only confirm NK cells as important drivers of macrophage foam cell formation but also highlight CD155 as a potential therapeutic target for atherosclerosis.

AIMS:To determine whether hypobaric hypoxia-induced alterations of the intestinal microbiota contribute to impaired exercise performance and intestinal dysfunction under simulated high-altitude conditions. METHODS AND RESULTS:A mouse model of exhaustive exercise under simulated 4500 m hypobaric hypoxia was used to assess exercise performance, oxidative stress, intestinal barrier integrity, and gut microbial composition. To evaluate the contribution of the intestinal microbiota, fecal microbiota transplantation (FMT) was performed using microbiota from hypoxia-exposed or normoxic donor mice into antibiotic-treated recipients before exhaustive exercise under hypobaric hypoxia. Compared with normoxic controls, hypoxia-exposed mice exhibited reduced endurance, increased oxidative stress, impaired intestinal barrier function, decreased Ruminococcus abundance, and significant alterations in both α- and β-diversity. Recipient mice receiving microbiota from hypoxia-exposed donors developed similar phenotypes, including greater oxidative stress, reduced barrier protein expression, and microbial community profiles resembling those of hypoxic donors. Comparisons with microbiota-depleted controls further indicated that gut microbial alterations contributed to, but were not sufficient alone to cause, impaired exercise performance in the absence of the hypoxic environment. CONCLUSIONS:Hypobaric hypoxia rapidly remodels the intestinal microbiota, and these microbial alterations contribute to oxidative stress, intestinal barrier dysfunction, and reduced exercise capacity under hypoxic conditions. FMT supports a contributory role of the intestinal microbiota in hypoxia-associated physiological dysfunction while indicating that hypoxic stress remains necessary for the full fatigue phenotype to develop. These findings support further investigation of microbiota-targeted strategies to mitigate exercise impairment during hypobaric hypoxia.

Exercise intolerance is a common issue upon acute exposure to high altitude when a person enters high-altitude environment before adapted. Oxidative stress and intestinal microbiota imbalance are increasingly recognized as important contributors to hypobaric hypoxia-associated fatigue. Selenium is a well-known anti-oxidative nutrient and probiotics are widely accepted as intestinal microbiota regulators. In this study, selenium-enriched Lacticaseibacillus rhamnosus SHA113 (Se-SHA113) was evaluated for its potential to alleviate exercise-related fatigue under simulated high-altitude environment (4500 m). Mice were orally administered Se-SHA113 (0.12 mg Se/kg body weight) for 4 days before hypobaric hypoxia exposure. Se-SHA113 supplementation prolonged exercise performance and was associated with an increased abundance of a taxon annotated as Lactobacillus reuteri in the intestinal microbiota. An intervention with Lactobacillus reuteri ATCC 53608 (LR; 1 & times; 108 CFU/day for 4 days) was also conducted. LR supplementation reproduced part of the beneficial phenotype. Both interventions improved fatigue-related biomarkers, including preservation of hepatic and muscle glycogen, enhancement of intestinal antioxidant enzyme activities, and elevation of fecal short-chain fatty acids, particularly acetate and butyrate. Reduced colonic oxidative stress was also observed after Se-SHA113 treatment. Overall, Se-SHA113 showed potential to alleviate hypobaric hypoxia-associated fatigue under simulated high-altitude environment. These effects were associated with selective intestinal microbiota remodeling, improved glycogen storage, and strengthened intestinal antioxidant defenses, supporting Se-SHA113 as a promising candidate for nutritional intervention under limited-oxygen conditions.

INTRODUCTION:Infections caused by pathogenic fungi such as Candida albicans have led to a continuous increase in disease morbidity and mortality, underscoring the urgent need for safer and more effective antifungal therapies. Current antifungal drugs are limited by toxicity and resistance, highlighting the necessity for innovative discovery strategies. OBJECTIVES:The aim of this study was to identify novel antifungal compounds using a integrative computational approach targeting C. albicans CYP51 (14-α-sterol demethylase). Specifically, we sought to screen the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database for potential antifungal candidates, evaluate their therapeutic potential, and establish a computational-experimental framework to expedite antifungal drug development. METHODS:We employed an integrative computational approach-targeting Candida albicans CYP51 (14-α-sterol demethylase) via molecular docking and pharmacophore modeling-to screen the TCMSP database. The two natural small molecules obtained were subjected to in vivo and in vitro antifungal experiments, and their antifungal mechanisms were analyzed through molecular docking and molecular dynamics simulation. RESULT:This yielded two novel antifungal candidates: α-hederin and elemenin, with minimum inhibitory concentrations (MICs) of 32 μg/mL and 16 μL/mL against C. albicans, respectively. α-Hederin demonstrated superior therapeutic potential, showing low cytotoxicity in 293 T, Raw264.7, and KB cells. It effectively inhibited hyphal formation, biofilm formation, and cell surface hydrophobicity in vitro. In a murine oral candidiasis model, α-hederin outperformed fluconazole by reducing fungal burden, inhibiting hyphal invasion, and preventing tongue adhesion. Molecular dynamics simulations revealed that α-hederin forms a more stable complex with CYP51 than fluconazole, engaging additional hydrogen bonds, alkyl interactions, and carbon-hydrogen bonds. CONCLUSION:This study advances antifungal drug discovery by validating CYP51 as a high-value target for structure-guided screening, identifying α-hederin as a low-toxicity, multi-mechanistic antifungal agent and establishing a computational-experimental framework for rapid antifungal development.

The use of natural killer (NK) cell-based immunotherapy has been extensively explored in clinical trials for multiple types of tumors and has surfaced as a promising approach in tumor immunotherapy. Interleukins (ILs), a vital class of cytokines, play a crucial role in regulating several functions of NK cells, thereby becoming a focal point in the advancement of NK cell-based therapies. Nonetheless, the use of ILs as single agents is significantly constrained by their short half-life, limited efficacy, and adverse reactions. Currently, nanomaterials are being progressively employed in the delivery of ILs to enhance NK cell-based immunotherapy. However, there is currently a lack of comprehensive reviews summarizing the design of NK-cell-targeted nanomaterials and related systems for delivery of ILs. Furthermore, certain nanomaterials, either alone or in conjunction with other therapeutics, can also promote the secretion of ILs, representing a promising avenue for further exploration. Accordingly, this review begins by outlining various types of ILs and subsequently discusses the advancements in applying nanomaterials for IL delivery. It also examines the potential of nanomaterials to enhance IL secretion from other immune cells, thereby influencing the NK cell functionality. Lastly, this review addresses the challenges associated with using nanomaterials in these contexts and offers perspectives for future research. This study aims to provide valuable insights into the development of NK cell immunotherapy and innovative nanomaterial-based drug delivery systems.

Cross-conjugated flavonoids exhibited higher bioactivities in certain aspects compared to others. Applying them in food and medical industries could promote human health. Conjugated porous polymer (CPP) served as an excellent adsorbent for N,N,N ',N '-tetramethylethylenediamine (TEMED) due to its porous properties, large specific surface areas, and highly delocalized it-it conjugated skeleton. The constructed TEMED-CPP distinguished between flavonoids with or without cross conjugation. This method showed good linearity (R2 >= 0.979), low detection limits (0.19-1.84 mu g/mL) and acceptable intraday and interday precision (<= 9.97 %). Density functional theory and molecular planarity calculations indicated that degree of conjugation, planarity, and energy band gap of flavonoids collectively influenced their response sensitivity to TEMED-CPP. Additionally, CPP could be recycled and subsequently reloaded with TEMED for the next detection after its initial use. The rapid identification of cross-conjugated flavonoids could guide the extraction and enrichment processes of highly bioactive flavonoids from natural sources, which improved extraction efficiency and economic benefits.

Cholesterol is an essential component of the cell membrane which plays a critical role in the survival of immune and tumor cells. Reprogramming of cholesterol metabolism in both tumor cells and immune cells can impact tumor progression and anti-tumor immune responses. Strategies aimed at modulating cholesterol metabolism have been demonstrated to be effective in hindering tumor growth and boosting anti-tumor immune functions. This review provides a thorough analysis of intracellular cholesterol homeostasis regulation in cells, focusing on key genes and signaling pathways. It particularly emphasizes the regulatory mechanisms and importance of the cholesterol presence state (esterified/free), levels of cholesterol, and its metabolites in immune and tumor cells. Additionally, the review thoroughly explores how cholesterol metabolism and sources (endogenous/exogenous) in the tumor microenvironment (TME) contribute to the interplay among tumor cells, immune suppressor cells, and immune effector cells, promoting cancer progression and immune evasion. It also delves into current insights on the influence of cholesterol metabolites and related drugs in regulating tumor development or immunotherapy. Finally, it presents an overview of recent advancements in clinical and preclinical trials investigating the efficacy of targeted cholesterol metabolism treatments and combination therapies in cancer management, while proposing potential future research directions in tumor immunity. This review is poised to offer fresh perspectives and avenues for examining the potential of cancer immunotherapy centered on cholesterol metabolism regulation.

Natural killer (NK) cells play an important role in antitumor and viral resistance. However, the mechanism of impaired NK cell function in microgravity remains unclear. Cholesterol metabolism, a new research hotspot, plays a critical role in NK cells function. This study found that simulated microgravity downregulate NK cell membrane cholesterol levels by disrupting cholesterol biosynthesis and transport to the membrane, leading to the obstruction of activated immune synapse formation, which inhibit the release of NK cell cytotoxic particles and ultimately decreasing NK cell immune function. Most importantly, this study identified a new target for regulating NK cell function, LEPR (leptin receptor). LEPR affected NK cell membrane cholesterol levels by influencing the CAMKK-SREBP1-HMGCR cholesterol endogenous synthesis pathway and regulating the expression of NPC1 and NPC2 genes, which ultimately influencing NK cell cytotoxic function. The study is significant for understanding the mechanism of NK cell activity in microgravity and offers new targets for clinical immunotherapy of NK cells.

Hepatocellular carcinoma (HCC) ranks among the most prevalent types of tumors globally. There is no effective cure for HCC in clinical practice, which emphasizes the need to conduct in-depth research into mechanisms that can facilitate the development of effective treatments for HCC. Numerous studies have demonstrated that the mechanical factors within an HCC microenvironment, such as matrix stiffness and fluid shear stress, have significant effects on the different stages of development. And these mechanical factors are also involved in the immune-evasion process of HCC, which affects the progression of HCC. However, current reviews related to HCC mainly focus on discussing the influence of mechanical factors on various cells in the tumor microenvironment, with less consideration of the impact of mechanical factors on each stage of tumor development. Therefore, this review discussed the influence of these different mechanical factors on the various stages of HCC development, including the initiation, proliferation, angiogenesis, migration, invasion, metastasis, drug resistance and immune escape. This review provides a theoretical basis for future in-depth investigations into the relationship between mechanical factors and HCC.

Development of efficient cell factories that can compete with traditional chemical production processes is complex and generally driven by case-specific strategies, based on the product and microbial host of interest. Despite major advancements in the field of metabolic modeling in recent years, prediction of genetic modifications for increased production remains challenging. Here, we present a computational pipeline that leverages the concept of protein limitations in metabolism for prediction of optimal combinations of gene engineering targets for enhanced chemical bioproduction. We used our pipeline for prediction of engineering targets for 103 different chemicals using Saccharomyces cerevisiae as a host. Furthermore, we identified sets of gene targets predicted for groups of multiple chemicals, suggesting the possibility of rational model-driven design of platform strains for diversified chemical production.

Inflammatory bowel disease (IBD) is a chronic inflammation with a high incidence rate. Many probiotics, including Lacticaseibacillus rhamnosus (L. rhamnosus), have shown promise in IBD treatment. The therapeutic effects of most probiotics are greatly decided by the available live cells in the disease lesion, which is compromised as they pass through the gastric juice and intestinal tract, resulting in a loss of activity. To improve probiotic delivery efficiency in the intestinal tract, broken Ganoderma lucidum spore shells (bGLS) were explored as a carrier to enhance the intestinal tract delivery of L. rhamnosus SHA113, a probiotic that has been verified to have capability to treat IBD. It was found the bGLS treated with iturin A and hydrochloric acid (IH-bGLS) had much higher affinity to probiotic cells than the untreated ones. This is possibly due to the enhancement of hydrophobic and positive charge of bGLS. Furthermore, IH-bGLS demonstrated an 81% loading efficiency for L. rhamnosus SHA113 and 2.2% for Escherichia coli. More importantly, loading in IH-bGLS greatly enhanced the delivery of L. rhamnosus SHA113 cells to the colon and prolonged their retention time from 48 to over 120 h (P < 0.01). The mechanisms might be related to the enhancement of probiotic cell adhesion to the gastrointestinal mucosa, increase of mucus secretion and the upregulated expression of tight junction proteins, occludin and ZO-1, in the colon. The results of the animal experiment showed that the therapeutic effects of L. rhamnosus SHA113 on IBD were greatly enhanced when they were loaded with IH-bGLS. The novelty of this research is in the development of probiotic carriers from bGLS, which has significance in the improvement of intestinal delivery efficiency and the therapeutic effects of probiotics on IBD. This system may have attractive application in the enhancement of probiotic delivery efficiency in the intestinal tract, which is important to ensure and enhance the beneficial effects of probiotics.

Oxidative damage is harmful to human health, which becomes serious in high-altitude areas due to the low oxygen partial pressure there. However, which organs are mostly influenced and the mechanisms of these are not clear. This study found that the liver was more susceptible to oxidative damage than other organs after mice entered a hypobaric hypoxia condition (4000 m). Only after 6 h of hypobaric hypoxia exposure, obvious oxidative damage was observed in hepatic tissues. The key role of Piezo1 mediated iron overload in the cause of such damage by using Piezo1 inhibitors and activators. Piezo1-mediated iron influx and the inhibition of iron efflux by hepcidin activation contributed to this process. Selenium accumulation in the liver coincided with this iron overload, suggesting a stress response. Oral administration of Lactobacillus rhamnosus SHA113-synthesized biological selenium significantly enhanced the selenium content, inhibited Piezo1 mediated iron overload and protected against oxidative damage to the liver under hypobaric hypoxia. These findings underscore that Piezo1mediated iron overload was a key contributor to the oxidative damage to the liver in high-altitude environment; selenium accumulation was a stress response to this damage; oral supplementation of biological selenium in advance might be an efficient way to protect the liver and human health in hypobaric hypoxia.

Peptides are recognized in functional foods for their biological activities. This study investigated the amino acid composition, physicochemical properties, in vitro antioxidant activity, and physiological effects in vivo of bovine collagen peptide (BCP) under hypoxic conditions. Amino acid analysis showed glycine, proline, and glutamic acid as the predominant components. Dynamic light scattering characterized its particle size distribution, while fourier transform infrared spectroscopy (FT-IR) confirmed its peptide structure. In vitro assays demonstrated significant antioxidant activity of BCP. A two-stage approach was used to examine BCP's effects on hypoxia tolerance and fatigue reduction under hypobaric hypoxic (HH) conditions. In the first stage, mice were given BCP for 5 days and tested using the rotarod assay at a simulated altitude of 4000 m. The BCP group exhibited an 8-fold increase in rotarod time compared to the control group (p < 0.05), indicating improved hypoxia tolerance and anti-fatigue capacity. In a human trial at a simulated altitude of 3600 m, 5 days of BCP supplementation led to a 4.7% increase in resting SpO2 (p < 0.05) and a 15.45% decrease in heart rate (HR) (p < 0.05). Submaximal exercise test revealed a 61.54% improvement in hypoxia tolerance (p < 0.05) and a twofold increase in anti-fatigue capacity (p < 0.001). This study provides both animal and human evidence supporting the benefits of BCP in HH conditions, suggesting its potential as a functional ingredient for athletes, fitness enthusiasts, and high-altitude travelers.

The antimicrobial efficacy of silver nanoparticles (AgNPs) in food safety applications is increasingly compromised by rapid bacterial resistance evolution through virulence upregulation. A biofunctionalized nanohybrid (Iturin A-AgNPs) was engineered to synergistically combine the amphiphilic lipopeptide iturin A with AgNPs to counteract resistance mechanisms in Escherichia coli (E. coli). Transcriptomic and phenotypic analyses revealed that PVP-AgNPs triggered bacterial adaptation via overexpression of outer membrane vesicle (OMV) biogenesis genes (e.g., MlaA/C/E), flagellar assembly proteins (FliC/D/F/G/I), and suppression of energy metabolism (atpC/G/H). In contrast, Iturin A-AgNPs suppressed these resistance-driving pathways by (1) downregulating flagellar assembly proteins to impair bacterial motility, (2) breaking the “dormant mode” of reduced energy metabolism, and (3) overriding the PVP-AgNPs resistance phenotype mediated by Mla system upregulation. This multi-target mechanism effectively prevented the emergence of resistant phenotypes, as evidenced by a reduction in the minimum inhibitory concentration (MIC) against AgNPs-resistant E. coli. These findings highlight the potential of biofunctionalized nanohybrids to combat antimicrobial resistance through coordinated genetic and metabolic interference, offering a template for engineering next-generation antibacterial agents.

Ochratoxin A (OTA), a carcinogenic mycotoxin produced by Aspergillus and Penicillium species that contaminates food crops and threatens public health. Although ergosterol and its synthetic enzymes are important antifungal targets, their regulatory roles and mechanisms in OTA production remain unclear. Therefore, elucidating the roles of ergosterol synthase genes erg3 (C-5 sterol desaturase) and erg24 (C-14 sterol reductase) in oxidative stress response and OTA biosynthesis in Aspergillus carbonarius is of critical importance. Herein, we employed homologous recombination to knockout and overexpress ergosterol synthase gene erg3 and erg24 in A. carbonarius. We identified two homologous erg3 (erg3-1 and erg3-5) and one erg24 in A. carbonarius. Δerg24 significantly reduced ergosterol levels whereas Δerg3-1 markedly increased it. Notably, only mutant with erg24 knockout, including single (Δerg24), double (Δerg3-5Δerg24) and triple knockout (Δerg3-1Δerg3-5Δerg24) strains showed significantly reduced OTA production, colony diameter, conidial formation and germination rates. Furthermore, transcriptomic analysis revealed that Δerg24 significantly downregulated expression of OTA biosynthetic genes pks and hal, while genes associated with antioxidant defense mechanisms regulating ROS levels were upregulated. Enzyme assay confirmed that catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH) were enhanced, accompanied by reduced ROS levels. This study provides new insights into the regulatory role of ergosterol synthase in OTA synthesis and potential targets for developing innovative antifungal strategies.

The presence of live cells in the intestinal tract is crucial for the beneficial effects of probiotic products containing lactic acid bacteria. However, many bacterial cells lose viability during production, storage, and gastric passage. Protein-based hydrogels and polysaccharide-based capsules effectively enhance the viability of live probiotic cells in products, but the underlying mechanisms and in vivo effectiveness are unclear. In this study, a complex was developed by self-aggregating walnut protein, tea polyphenol, and Lactobacillus rhamnosus cells under acidic conditions, followed by alginate coating. Complex formation was facilitated by hydrogen bonding and hydrophobic interactions. The complex had a low thermal diffusion coefficient of 4.9 x 10- 7 m2/s, making the bacterial cells resistant to temperature fluctuation. Cell survival rate was 75.5% after 12 h at 55 degrees C and 92.8% after freeze-drying. Shelf-life predictions were 14 years at 4 degrees C. and 2.3 months at 25 degrees C, with a live cell count exceeding 106 CFU/g. The product demonstrated a 60% increase in live bacterial cells compared to free cells in simulated gastric fluid, and achieved complete release in simulated intestinal fluid within 120 min. This method was successfully applied to L. rhamnosus, L. casei and L. plantarum. Oral administration of the L. rhamnosus product significantly increased the abundance of Lactobacillus and L. rhamnosus in the colon and cecum of mice. These findings present a new method for producing highly active probiotic products using plant materials and provide insights into the underlying mechanisms from a thermodynamic perspective, contributing to the industrialization and advancement of probiotics.

ABSTRACT Candida albicans is responsible for conditions ranging from superficial infections such as oral or vaginal candidiasis to potentially fatal systemic infections. It produces pathogenic factors contributing to its virulence. Iturin A, a lipopeptide derived from Bacillus sp., exhibits a significant inhibitory effect against C. albicans . However, its exact mechanism in mitigating the pathogenic factors of C. albicans remains to be elucidated. This study aimed to explore the influence of iturin A on several pathogenic attributes of C. albicans , including hypha formation, cell membrane permeability, cell adhesion, biofilm formation, and therapeutic efficacy in an oral C. albicans infection model in mice. The minimal inhibitory concentration of iturin A against C. albicans was determined to be 25 µg/mL in both YEPD and RPMI-1640 media. Iturin A effectively inhibited C. albicans hyphal formation, decreased cell viability within biofilms, enhanced cell membrane permeability, and disrupted cell adhesion in vitro . Nonetheless, iturin A did not significantly affect the phospholipase activity or hydrophobicity of C. albicans . A comparative study with nystatin demonstrated the superior therapeutic efficacy of iturin A in a mouse model of oral C. albicans infection, significantly decreasing C. albicans count and inhibiting both fungal hypha formation and tongue surface adhesion. High-dose iturin A treatment (25 µg/mL) in mice had no significant effects on blood indices, tongue condition, or body weight, indicating the potential for iturin A in managing oral infections. This study confirmed the therapeutic potential of iturin A and provided valuable insights for developing effective antifungal therapies targeting C. albicans pathogenic factors.