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Objective To investigate whether menaquinone-4 （MK-4） can exert a protective effect against carbon tetrachloride （CCl4）-induced acute liver injury （ALI） in mice by alleviating ferroptosis. Methods After adaptive feeding， adult male ICR mice， aged 8 weeks， were divided into Control group， MK-4 group， CCl4 model group （6-hour， 12-hour， and 24-hour）， and MK-4+CCl4 group （6-hour， 12-hour， and 24-hour）， with 6 mice in each group. The mice in the Control group were given intraperitoneal injection of an equal dose of corn oil； the mice in the MK-4 group were given intraperitoneal injection of 40 mg/kg MK-4 solution， followed by an equal dose of corn oil after 1 hour； the mice in the MK-4+CCl4 group （6-hour， 12-hour， and 24-hour） were given intraperitoneal injection of 40 mg/kg MK-4 solution， and after 1 hour， the mice in this group and the CCl4 model group （6-hour， 12-hour， and 24-hour） were given intraperitoneal injection of 0.3 mL/kg CCl4 solution， with samples collected at 6， 12， and 24 hours. HE staining was used to observe the pathological changes of mouse liver； Prussian blue staining was used to observe iron accumulation in liver tissue； a biochemical analyzer was used to measure the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）； related kits were used to measure the levels of tissue iron content and the oxidative stress indices malondialdehyde （MDA） and glutathione （GSH） in liver homogenate； RT-PCR was used to measure the expression levels of ferroptosis marker genes （acyl-CoA synthetase long-chain family member 4 ［ACSL4］， prostaglandin-endoperoxide synthase 2 ［PTGS2］， and glutathione peroxidase 4 ［GPX4］） and iron metabolism-related genes （hemojuvelin ［HJV］， transferrin receptor 1 ［TFR1］， and ferroportin ［FPN］）， and Western blot was used to measure the protein expression level of GPX4. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. Results In the aging study， compared with the Control group， the CCl4 model group （6-hour， 12-hour， and 24-hour） had significant increases in liver weight coefficient and the serum levels of ALT and AST （all P<0.05）， and HE staining also showed that liver injury gradually aggravated over time. Meanwhile， compared with the CCl4 model group （6-hour， 12-hour， and 24-hour）， the MK-4+CCl4 （12-hour） group had significant reductions in liver weight coefficient and the serum levels of ALT and AST （all P<0.05）， with a reduction in the necrotic area of liver tissue， and therefore， 12-hour mouse tissue samples were used for detection in the following study. Compared with the Control group， the CCl4 group had a significant increase in MDA and a significant reduction in GSH （both P<0.05）， and compared with the CCl4 group， the MK-4+CCl4 group had a significant reduction in MDA and a significant increase in GSH （both P<0.05）. Compared with the Control group， the CCl4 group had significant increases in the key ferroptosis indices ASCL4 and PTGS2 and a significant reduction in GPX4 （all P<0.05）； compared with the CCl4 group， the MK-4+CCl4 group had significant reductions in the mRNA expression levels of ASCL4 and PTGS2 and a significant increase in the mRNA expression level of GPX4 （all P<0.05）. Western blotting showed that compared with the Control group， the CCl4 group had a significant reduction in the protein expression level of GPX4 （P<0.05）， and compared with the CCl4 group， the MK-4+CCl4 group had a significant increase in the protein expression level of GPX4 （P<0.05）. Prussian blue staining showed that compared with the Control group， the CCl4 group had a significant increase in iron accumulation； after MK-4 intervention， compared with the CCl4 group， the MK-4+CCl4 group had a significant reduction in iron accumulation. As for the measurement of iron metabolism genes in mouse liver， compared with the Control group， the CCl4 group had a significant increase in iron content， significant reductions in the mRNA expression levels of FPN and HJV， and a significant increase in the mRNA expression level of TFR1 （all P<0.05）； after protection with MK-4， there was a significant reduction in iron content， significant increases in the mRNA expression levels of FPN and HJV， and a significant reduction in the mRNA expression level of TFR1 （all P<0.05）. Conclusion MK-4 intervention in advance can alleviate CCl4-induced ALI in mice， possibly by inhibiting ferroptosis and improving the expression of iron metabolism-related genes in mouse liver. [ABSTRACT FROM AUTHOR]

Intracerebral hemorrhage is a kind of hemorrhagic stroke resulting from bleeding in the brain parenchyma. SUMOylation is one of post-translational protein modifications based on covalent binding of small ubiquitin-like modifier (SUMO) peptide. Although SUMOylation has been reported to play neuroprotective roles in ischemic stroke, the role of SUMOylation in the pathology of intracerebral hemorrhage remains unclear. Here we addressed the relationship between SUMOylation and pathogenesis of intracerebral hemorrhage. For in vivo study, hemorrhage was induced in the striatum of male ICR mice by local injection of type VII collagenase. After induction of hemorrhage, there was a significant increase in the levels of SUMO2/3 protein conjugation in the brain, without accompanying significant changes in the levels of free SUMO2/3 protein and Sumo2/3 mRNA. When intracerebroventricular injection of TAK-981, an inhibitor of SUMOylation, was performed 3 d before hemorrhage induction, it delayed recovery of body weight, recovery of neurological functions and hematoma clearance. TAK-981 also suppressed CD36 expression in hematoma-associated microglia/macrophages. For in vitro study, phagocytic activity of microglial BV-2 cells was examined using erythrocytes obtained from mouse blood. We found that both TAK-981 and small interfering RNA-mediated knockdown of SUMO2/3 inhibited erythrophagocytosis by BV-2 cells. These results suggest that SUMOylation by SUMO2/3 promotes hematoma clearance via regulation of CD36 expression in microglia and contributes to the recovery from pathology of intracerebral hemorrhage., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 International Brain Research Organization (IBRO). Published by Elsevier Inc. All rights reserved.)

Oxygen therapy after acute lung injury can regulate the inflammatory response and reduce lung tissue injury. However, the optimal exposure pressure, duration, and frequency of oxygen therapy for acute lung injury remain unclear. In the present study, after intraperitoneal injection of lipopolysaccharide in ICR mice, 1.0 atmosphere absolute (ATA) pure oxygen and 2.0 ATA hyperbaric oxygen treatment for 1 hour decreased the levels of proinflammatory factors (interleukin-1beta and interleukin-6) in peripheral blood and lung tissues. However, only 2.0 ATA hyperbaric oxygen increased the mRNA levels of anti-inflammatory factors (interleukin-10 and arginase-1) in lung tissue; 3.0 ATA hyperbaric oxygen treatment had no significant effect. We also observed that at 2.0 ATA, the anti-inflammatory effect of a single exposure to hyperbaric oxygen for 3 hours was greater than that of a single exposure to hyperbaric oxygen for 1 hour. The protective effect of two exposures for 1.5 hours was similar to that of a single exposure for 3 hours. These results suggest that hyperbaric oxygen alleviates lipopolysaccharide-induced acute lung injury by regulating the expression of inflammatory factors in an acute lung injury model and that appropriately increasing the duration and frequency of hyperbaric oxygen exposure has a better tissue-protective effect on lipopolysaccharide-induced acute lung injury. These results could guide the development of more effective oxygen therapy regimens for acute lung injury patients., (Copyright © 2024 Copyright: © 2024 Medical Gas Research.)

Rationale: The newly emerging synthetic cannabinoids (SCs) 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201 have been observed to produce effects by activating cannabinoid type 1 (CB1) receptors. Nevertheless, the pharmacological effects and potential for abuse of these three substances remain to be studied. These substances have yet to be regulated in many countries., Objectives: We investigated the safety, pharmacological effects, rewarding effects, and cannabinoid withdrawal of 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201., Methods: This study evaluated the drug safety and the cannabinoid-specific pharmacological effects of the three substances through acute toxicity experiments (in which the LD 50 of each substance was obtained) and tetrad experiments (comprising assessments of hypothermia, analgesia, locomotion inhibition, and catalepsy). Furthermore, the conditioned place preference (CPP) experiments and withdrawal experiments were conducted to evaluate the rewarding effect and cannabinoid withdrawal potential of the substances in question., Results: The results demonstrated that all three drugs exhibited certain acute toxic effects and could potentially induce tetrad effects. The data were analyzed using non-linear regression, and the corresponding ED 50 values and 95% confidence intervals (CI) were obtained. The rank order of potency was determined to be CUMYL-PEGACLONE > 5F-EDMB-PICA > NM-2201. In the CPP experiments, it was demonstrated that 5F-EDMB-PICA significantly induced an increase in CPP score at a dose of 0.3 mg/kg, while NM-2201 caused an increase in CPP score and a significant aversion effect at a dose of 2 and 3 mg/kg, respectively. It is noteworthy that all three types of SCs were observed to produce a significant biphasic effect, indicating that CPP scores were biphasic for all compounds. Following the administration of the CB1 receptor antagonist rimonabant, a notable increase in head twitches and paw tremors was observed, indicating that these three SCs induce cannabinoid withdrawal through the mediation of CB1 receptors., Conclusions: The results of this study indicated that these SCs possess cannabinoid-specific pharmacological effects and abuse potential, which provides substantial experimental data to support the future regulation of these substances., Competing Interests: Declarations. Ethics approval and consent to participate: All procedures followed the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Conflict of interest: The authors declare no conflict of interest related to this work., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Fructose, as a natural and simple sugar, is not significantly harmful to the human body when consumed in moderation and can provide energy for the body. High-fructose diets have been linked to an increased risk of a range of metabolic disorders, including hypertriglyceridemia, hypertension, and diabetes mellitus. These conditions are known to be associated with an elevated risk of developing cardiometabolic diseases. Cardiomyocytes in mammals possess the capacity to proliferate from the moment of their birth. However, this capacity diminishes over time, and cardiac growth is ultimately achieved through cardiomyocyte (CM) hypertrophy. Prior studies have demonstrated that fructose metabolism is enhanced in the heart during pathological hypertrophy [1]. The consumption of foods containing high levels of fructose has been linked to an increase in the size of cardiomyocytes, which can lead to damage to the heart. The impact of high fructose on cardiomyocytes at the point of their initial capacity for proliferation has not been previously documented. In this experiment, our purpose was to explore the impact of high fructose in cardiomyocyte proliferation. To establish an apical resection model in neonatal mice, neonatal ICR mice were randomly divided into a sham-operated group (Sham + PBS), a sham-operated combined high-fructose group (Sham + fructose), an apical resection alone group (AR + PBS) and an apical resection combined with a high-fructose group (AR + fructose). Next, echocardiography was employed to assess the cardiac function of all mice. Masson staining was carried out to analyze cardiac fibrosis. Immunostaining was performed by extracting primary rat cardiomyocytes after the high-fructose intervention to see if proliferation-related markers (Ki67, PH3, Aurora-B) changed, qRT-PCR and immunofluorescence were used to determine changes in the expression profile of Notch1 in the neonatal heart. The results suggest that high fructose could inhibit cardiomyocyte proliferation in vivo and in vitro, The possible mechanism is that high fructose levels inhibit cardiomyocyte proliferation through suppression of Notch1 signaling pathway. In conclusion, high fructose levels inhibit the proliferation of cardiomyocytes via the Notch1 signaling pathway., Competing Interests: Declaration of competing interest The author declares that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.)

This study evaluates the efficacy of Bifidobacterium longum subsp. longum BL21 in mitigating symptoms of polycystic ovarian syndrome (PCOS) in DHT-induced PCOS model mice. It focuses on BL21's role in modulating metabolic dysregulation, inflammation, and neuroprotection via the gut-brain-ovary axis. Employing an 8-week treatment regimen, this research assessed the effects of BL21 on prenatal androgen-induced PCOS in ICR mice. Evaluations included body weight, glucose tolerance tests, serum analyses of BDNF, inflammatory markers, sex hormone levels, and 16S rRNA gene sequencing for gut microbiota diversity and composition. Twenty-four ICR mice with induced PCOS served as subjects to examine the probiotic's impact. Mice were administered a daily oral dose of 1 × 10 9 CFU of BL21 continuously for a total of 8 weeks. BL21 significantly enhanced sex hormone levels ( P < 0.05), particularly those of follicle-stimulating hormone (FSH) and estradiol (E2), indicating improved ovarian function and offering a novel PCOS treatment approach. The intervention notably curbed weight gain and improved glucose tolerance in PCOS mice ( P < 0.05). BL21 reduced inflammatory markers such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and lipopolysaccharides (LPS), while increasing IL-10, BDNF, FSH, and E2 levels ( P < 0.05 for all). It also enriched gut microbiota diversity, enhancing populations of Bifidobacterium and Lactobacillus . Correlation analyses underscored the positive shifts in microbiota linked to beneficial hormonal and inflammatory profiles. BL21 shows promise in alleviating PCOS symptoms through metabolic regulation, inflammation reduction, and neuroprotection, validating its potential in integrated therapeutic strategies.IMPORTANCEPolycystic ovarian syndrome (PCOS) is a prevalent endocrine disorder affecting women of reproductive age, characterized by metabolic irregularities, hormonal imbalances, and chronic inflammation. Existing treatments are often inadequate, addressing symptoms without targeting the underlying etiological factors. The investigation of Bifidobacterium longum subsp. longum BL21 as a probiotic intervention offers a novel approach by potentially regulating the gut-brain-ovary axis. This could lead to innovative therapeutic strategies that not only manage but also potentially reverse the multifaceted symptoms of PCOS, enhancing quality of life and reproductive health., Competing Interests: The authors declare no conflict of interest.

This study focuses on the preparation and characterization of platelet membrane biomimetic nanocarriers (P-PLGA NPs) and investigates their interactions with the transplacental barrier. Poly (lactic-co-glycolic acid) nanoparticles (PLGA NPs) were coated with platelet membrane (PLTM) to construct P-PLGA NPs. Additionally, fluorinated polyethylenimine (F-PEI) was grafted onto PLGA NPs to prepare F-PEI-PLGA NPs, which were compared with PLGA NPs. In vitro placental barrier cell models using human choriocarcinoma cells (BeWo b30) and in vivo pregnancy animal models using ICR mice were utilized to evaluate the transplacental barrier efficiency of the PLGA NPs with different surface modifications. The results demonstrated that all three types of nanoparticles could accumulate in the uterus and placenta. The transplacental barrier efficiency of F-PEI-PLGA NPs was found to be the highest at 4 h, while P-PLGA NPs exhibited the highest transplacental barrier efficiency at 12 and 24 h. Furthermore, there were no significant effects on the main organs, structure, and quantity of uterine spiral arteries, indicating the safety of the nanoparticles (NPs). P-PLGA NPs are expected to be a safe and effective nano-delivery system for perinatal drug delivery. This study provides insights into the transplacental barrier mechanism of NPs with different surface characteristics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Microplastics (MPs), plastic particles with a diameter of < 5 mm, are intentionally produced or formed by the breakdown of a variety of larger plastics. Polyethylene terephthalate (PET) is a common source of MPs and PET-MPs are prevalent in the environment. Owing to their persistence, PET-MPs can enter ecosystems, air, and food sources, posing significant health risks. This study aimed to investigate the toxicological effects and in vivo accumulation of PET-MPs smaller than 10 µm. To track their biodistribution, fluorescently labeled PET-MPs were prepared. Particle size and morphology were confirmed using physical and chemical characterization. Following the oral administration of PET-MPs in ICR (CD-1®) outbred mice, accumulation occurred predominantly in lungs, as confirmed by IVIS spectrum CT analysis and in vivo and ex vivo imaging. Toxicity assays revealed the development of granulomatous inflammation in the lungs at medium and high doses, indicating a concentration-dependent response. The recorded no-observed-adverse-effect levels were 1.75 mg/kg for males and 7 mg/kg for females. This study highlights the potential of PET-MPs to induce persistent inflammation in respiratory tissues and reveals the need for further research to support the regulatory standards and long-term health effects of MP exposure., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethical approval: This study was conducted in accordance with the Institutional Animal Care and Use Committee (IACUC), the Preclinical Center of the KMEDI hub, with an approval code of. KMEDI-23112701-00., (© 2025. The Author(s).)

We developed a PCR-free, cost-effective colorimetric assay using gold nanoparticles (AuNPs) to quantify 5-hydroxymethylcytosine (5hmC) without expensive MS instruments or antibodies. This method employs a photo-elutable 8-17 DNAzyme label specific to 5hmC sites, achieving accuracy comparable to ELISA when measuring 5hmC levels in various ICR mouse tissues.

Objectives: this study aimed to explore the effects of different intake levels (20-80%) of highland barley on the anti-fatigue capacity of ICR mice, focusing on energy metabolism, metabolite accumulation, oxidative stress, and changes in the gut microbiota., Methods: male ICR mice were assigned to five groups: control (normal diet) and four experimental groups with highland barley supplementation at 20%, 40%, 60%, and 80% of total dietary energy. Anti-fatigue performance was assessed by behavioral experiments (rotarod, running, and exhaustive swimming tests), biochemical markers, and gut microbiota analysis., Results: the results showed that moderate supplementation (20%) significantly enhanced exercise endurance and anti-fatigue capacity, as evidenced by increased liver glycogen (134.48%), muscle glycogen (87.75%), ATP content (92.07%), Na + -K + -ATPase activity (48.39%), and antioxidant enzyme activities (superoxide dismutase (103.31%), catalase (87.75%), glutathione peroxidase (81.14%). Post-exercise accumulation of blood lactate, quadriceps muscle lactate, serum urea nitrogen, and the oxidative stress marker malondialdehyde was significantly reduced, with differences of 31.52%, 21.83%, 21.72%, and 33.76%, respectively. Additionally, 20% supplementation promoted the growth of beneficial gut microbiota associated with anti-fatigue effects, including unclassified&#95;f&#95;Lachnospiraceae , g&#95;norank&#95;f&#95;Peptococcaceae , Lachnospiraceae NK4A136 , Colidextribacter , and Turicibacter . However, when intake reached 60% or more, anti-fatigue effects diminished, with decreased antioxidant enzyme activity, increased accumulation of metabolic waste, and a rise in potentially harmful microbiota ( Allobaculum , Desulfovibrio , and norank&#95;f&#95;norank&#95;o&#95;RF39 )., Conclusions: moderate highland barley supplementation (20% of total dietary energy) enhances anti-fatigue capacity, while excessive intake (≥60%) may have adverse effects.

Tributyl phosphate (TBP) is an emerging hazardous material that is ubiquitous in the environment. However, whether TBP affects the male reproductive system remains unknown. In this study, we exposed male ICR mice and mouse spermatocyte GC-2 cells to TBP to explore the male reproductive toxicity of TBP. The results revealed that TBP (50 and 100 mg·kg -1 ·day -1 ) decreased sperm count and motility, damaged testicular structure, and induced apoptosis in mice after 35 days of oral gavage. TBP (10, 50, and 200 μM) inhibited cell viability, induced apoptosis, and reduced the ratio of BCL2/BAX in GC-2 cells after 24 h of exposure. TBP (50 and 200 μM) triggered the buildup of reactive oxygen species and induced a reduction in the mitochondrial membrane potential within GC-2 cells. In addition, TBP led to gut microbiota dysbiosis and metabolic profile alterations and increased the abundance of Prevotellaceae, which are negatively associated with sperm motility. Furthermore, TBP caused inflammation, induced the infiltration of Th17 cells into the mouse small intestine, and stimulated IL-17A signaling in mouse testes. In conclusion, TBP induced male reproductive toxicity via the gut-testis axis, oxidative stress, and apoptosis, offering fresh perspectives on TBP's reproductive hazards in male mammals.

Ethnopharmacological Relevance: The bitter resins in hops (Humulus lupulus) modulate GABA receptors, leading to central nervous system suppression, which induces sedative effects and enhances sleep., Aim of the Study: This study intends to explore the sleep-enhancing properties of Hongcheon-hop extract, a hop native to Korea, by analyzing sleep structure and its mechanisms through EEG., Materials and Methods: A pentobarbital-induced sleep model was used, along with EEG analysis to study sleep architecture. The receptor binding of Hongcheon-hop extract was analyzed using GABA receptor antagonists., Results: The extract contained 7.44 mg/g of xanthohumol, 133.04 mg/g of α-acids including 13.74 mg/g of humulone and 84.52 mg/g of β-acids. Administration of the extracts resulted in a dose-dependent enhancement of sleep duration. Both low (100 mg/kg) and high doses (200 mg/kg) of the extract increased non-rapid eye movement (NREM) sleep by enhancing δ wave sleep, which represents deep sleep. The extract exerted its sleep-promoting effects by binding to the γ-aminobutyric acid (GABA) binding site on GABA A receptor. In an insomnia model, low dose of the extract was more effective than the same dose of GABA in enhancing sleep. Administration of the extract for 3 weeks increased the expression and protein levels of 5-HT 1A receptors in addition to GABA A receptor, and also increased the content of GABA in the brain. A mixture of xanthohumol (1.49 mg) and humulone (2.75 mg) within Hongcheon-hop extract (200 mg) exhibited a sleep-enhancing effect comparable to that of the extract., Conclusion: These findings suggest that Hongcheon-hop extract, which contains xanthohumol and humulone, improves sleep quality by increasing NREM sleep through the enhancement of δ-wave sleep via GABA A receptors. Hongcheon-hop shows potential as a natural therapeutic agent for treating insomnia and enhancing sleep., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Polygonatum (Huangjing) genus has been used as both food and medicine in China for 2000 years, which was regarded as a "Top-grade" herb in the Shennong Bencao Jing. The most commonly used species is the rhizome of Polygonatum cyrtonema Hua (PC) that is traditionally utilized to invigorate Qi, nourish Yin, moisten lung, and tonify spleen and kidney., Aim of the Study: Excessive alcohol consumption causes severe upper-gastrointestinal diseases, notably gastric mucosal damage characterized by hemorrhagic gastritis, which lacks safe and effective intervention. This study aims to investigate the gastroprotective effects of nine-steaming and nine-drying processed Polygonatum cyrtonema Hua (PPC) on alcohol-induced gastric mucosal damage in mice., Materials and Methods: PPC extract was chemically characterized by UPLC-QE-MS analysis. ICR mice were subjected to an ethanol-induced gastric lesion model and were orally administered PPC aqueous extract for 5 consecutive days. After treatment, gastric tissues were stained with hematoxylin and eosin (H&E), and the pro-inflammatory and oxidative stress factors were determined using ELISA and Multiplex assay, while the gene expressions of gastric tissues were detected by RNA-seq and Western blotting., Results: PPC reduced the alcohol concentration of liquor in vitro and protected against alcohol-induced gastric mucosal lesion in mice. Notably, PPC aqueous extract relieved alcohol-induced pro-inflammatory and oxidative stress factors, including interleukin 6 (IL-6), IL-8, keratinocyte-derived chemokine (KC), monocyte chemotactic protein-1 (MCP-1), superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA). RNA-sequencing analysis revealed that ethanol exposure activated mitogen-activated protein kinases (MAPKs), tumor necrosis factor (TNF), and IL-17 signaling pathways in gastric tissue, and these activated signaling pathways were inhibited by the PPC treatment. Consistently, Western blot data showed that PPC treatment suppressed the activation of extracellular signal-regulated kinases (ERK), p38, c-Jun N-terminal kinases (JNK), TNF-α and IL-17A pathways in gastric tissue., Conclusion: In conclusion, the aqueous extract of PPC exerted a gastroprotective effect against alcohol-induced gastric injury by alleviating inflammation and oxidative stress, potentially through the inhibition of the MAPKs, IL-17 and TNF-α pathways. These findings supported the future development of PPC as an effective intervention for alcohol-induced gastric damage., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Jingfang Granules (JF) is a modified herbal compound preparation that is empirically used in clinical practice for the treatment of allergic diseases. Nevertheless, the role of JF in allergic rhinitis (AR) has yet to be demonstrated, and its potential mechanisms of action remain to be fully evaluated., Aim of Study: The objective of this research is to examine the underlying mechanisms by which JF can be used to treat AR. This will be achieved through the use of an ovalbumin (OVA)/aluminum hydroxide AR model in mice., Materials and Methods: ICR mice were administered an intraperitoneal (i.p.) injection of OVA/aluminium hydroxide in order to permit the establishment of an AR model. Following the intragastric administration of JF to the mice, testing nose scratching and sneezing behavior in mice to determine modeling status, and stained transverse sections of the mouse nose using the Hematoxylin and Eosin (H&E) method were in vitro evaluated to assess the histological effects of JF on mice with AR. The regulatory network was subjected to proteomic and metabolomic investigation. The expression of serum cytokines as well as histamine (HIS) was detected using ELISA kits. Protein expression in nasal mucosal tissues was identified through the use of a Western blot., Results: JF demonstrated a notable reduction in nose-scratching and sneezing in AR mice. Concurrently, JF markedly reduced IgE, IL-4, IL-6, IL-13, TNF-α and HIS levels while elevating IFN-γ levels in the serum of AR mice. This was achieved by inhibiting the endoplasmic reticulum (ER) stress-related protein associated proteins including GADD and ATF4, p-eIF2α, p-IRE1α, XBP1s and p-PERK. Proteomics, metabolomics, Western blotting and Quantitative Real-time polymerase chain reaction (qPCR) results confirmed that JF inhibits the glycolysis/arginine biosynthesis pathway by suppressing the ER stress (ERs) signaling pathway, which in turn inhibits the inflammatory response., Conclusion: Findings from the present study indicate that JF is an efficacious treatment for OVA/aluminum hydroxide-induced nasal mucosal injury and inflammation in mice. Furthermore, the study demonstrated that JF exhibited anti-AR clinic pharmacological effects by modulating the ERs signaling pathway and inhibiting glycolysis as well as arginine biosynthesis., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Garlic (Allium sativum L.) is a widely consumed spice and condiment around the world, applied both as a food and as a traditional medicine, and is a natural strengthening agent for the body's circulatory and nervous systems. Diallyl trisulfide (DATS) is the major volatile organosulfur phytochemical found in garlic, with antithrombotic, anticoagulant, and antiplatelet activities as well as antioxidant, anti-infective, and other pharmacological effects. However, the safe dose and the underlying mechanisms of its toxic effects remain elusive., Aim of the Study: DATS, an important pharmacologically active compound found in garlic, has garnered attention for its ability to fight cancer, antioxidant, anti-infective, and cardioprotective. The aim of this study was to evaluate the safety of DATS and to elucidate the potential mechanisms of its toxicity., Materials and Methods: In this study, ICR mice were selected for acute and subacute toxicity experiments according to OECD guidelines. The toxicity profile of DATS was analyzed by computer prediction software. Also, key differential proteins in spleen and serum were analyzed by proteomics. The binding stability of DAST to differential proteins was analyzed by molecular docking. Additionally, the regulatory relationship between DATS and differential proteins was verified by Western blot and ELISA experiments., Results: The results showed that the LD 50 value of DATS in acute toxicity was 188.67 mg/kg. In subacute toxicity, water consumption and food intake were reduced in both male and female mice. In addition, the spleen and small intestinal organ coefficients were significantly elevated in male mice at the high dose of DATS; the blood biochemical indices ALB and TP were also significantly elevated. HE staining results showed significant damage to the spleen, liver, small intestine, and kidney of mice at high doses of DATS. Spleen and serum proteomics analyses showed that DATS significantly inhibited ZBP1 expression and upregulated TEC. ADMETlab 2.0 software predictions identified DATS as having potential genotoxicity, dermal sensitization, carcinogenicity, and respiratory and ocular toxicity. Docking results showed that the binding energies between DATS and TEC protein (PDB: 6F3F) and ZBP1 protein (PDB: 4KA4) were -3.7 kcal/mol, -2.4 kcal/mol, respectively. Western blot results showed that DATS-H significantly inhibited the expression of ZBP1 (only in male mice) and Bcl-2 proteins. ELISA results showed that DATS-H significantly increased the level of TEC protein both in male and female mice., Conclusions: Long-term administration of high-dose DATS may carry some risk of toxicity. Based on the amount of DATS in garlic, it is recommended that adults should not take more than 359 mg of DATS and 84.5 g of garlic per day. The mechanism of toxicity may be related to the fact that DATS significantly inhibits ZBP1 expression, upregulates TEC, and promotes apoptosis. This study provides valuable toxicological data for the effective evaluation of the long-term toxicity of DATS and offers an additional experimental basis for developing DATS as a healthy food or drug., Competing Interests: Declaration of competing interest This manuscript has not been published or presented elsewhere. We have read and understand your journal's policies and do not believe this manuscript violates any of them. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Calcium release from the endoplasmic reticulum via sperm-derived phospholipase C zeta is crucial for oocyte activation during fertilization. Chloroquine (CQ) inhibits the increase in cytoplasmic calcium. This study investigated the effects of CQ on fertilization and oocyte activation. Oocytes were collected from ICR mice, and in vitro fertilization and artificial oocyte activation with strontium ions (Sr 2+ ) and ethanol were performed in the presence of 50 µM CQ. Pronuclear formation was assessed via Hoechst33242 nuclear staining, cortical granule release was evaluated using lens culinaris agglutinin-fluorescein isothiocyanate staining, and cytosolic calcium levels were measured using fluorescence microscopy with Cal-520 AM. In the presence of CQ, no pronuclei were formed even 8 h after Sr 2+ -induced oocyte activation. Furthermore, cortical granule release in CQ-treated oocytes was significantly suppressed, although not completely inhibited, and no increase in cytosolic calcium was detected. CQ also inhibited pronuclear formation during ethanol-induced oocyte activation. In in vitro fertilization, although the fertilization rate was decreased in the CQ-treated group, in which CQ treatment was continuously applied during insemination, pronuclear formation and cortical granule release were observed. The decrease in the fertilization rate was likely attributable to reduced sperm motility and decreased penetration of the zona pellucida. The findings indicate that the oocyte activation pathways triggered by ethanol/Sr 2+ and sperm are distinguishable by CQ, and that CQ can be used as a selective inhibitor of oocyte activation induced by Sr 2+ or ethanol treatment.

Alzheimer's disease (AD) is characterized by oxidative stress-mediated memory dysfunction and neuronal cell death. This study investigated the effects of an ethanol extract from Bauhinia coccinea (EEBC) on memory impairment and neuronal damage in a memory deficit mouse model. EEBC was administered to ICR mice at doses of 50, 100, or 200 mg/kg daily for 3 weeks. Cognitive impairment was induced via scopolamine (SCO) injection. Brain tissues were analyzed for acetylcholine (ACh) levels, acetylcholinesterase (AChE) activity, neuronal apoptosis, and antioxidant markers. Behavioral tests showed that SCO injection induced memory loss, whereas EEBC significantly ameliorated SCO-mediated memory impairment. EEBC regulated the cholinergic system by decreasing ACh levels and enhancing AChE activity. Nissl staining and immunohistochemistry for NeuN showed that EEBC exerted neuroprotective effects in SCO-injected mice brains. Moreover, EEBC significantly reduced the number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive apoptotic cells increased by SCO treatment. EEBC also reversed the SCO-induced changes in apoptosis-related protein expression in brain tissues. Furthermore, EEBC significantly reduced malondialdehyde levels and activated catalase in SCO-administered brains. Quantitative RNA sequencing showed involvement of lipid metabolism in EEBC memory function regulation. Thus, EEBC is a promising candidate for attenuating AD progression as it targets the cholinergic system and neuronal apoptosis., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Cadmium (Cd) is a toxic heavy metal with a high propensity to accumulate within the body, and Cd accumulation has been shown to cause organ damage. However, it is unclear whether Cd accumulation is a cause of impaired decidualization, which induces to spontaneous abortion (SA). In this study, we found that the decidual Cd concentration was increased in patients with SA and positively correlated with the occurrence of SA. The levels of two decidualization markers (prolactin, PRL and insulin-like growth factor binding protein 1, IGFBP1) were reduced in the decidua of all-cause SA patients. Using 8-week ICR female mice, we further established a uterus-specific Cd accumulation mouse model and verified that Cd-accumulating mice had increased numbers of absorbed fetuses and defective decidualization. Finally, using in vitro-cultured human ENdometrial stromal cells (hEnSCs), we found that Cd accumulation significantly inhibited decidualization; and moreover, Cd treatment downregulated the regulatory genes upstream of PRL and IGFBP1 such as PGR, ESR1, ESR2 and FOXO1. This study suggests that Cd accumulation could produce impaired decidualization by downregulating the upstream regulators of PRL and IGFBP1, thereby increasing the risk of SA. Our study offered new possibilities for the prevention and treatment of spontaneous abortion., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

In recent years, coxsackievirus A6 (CVA6) has surpassed enterovirus A71 to become the main pathogen causing severe Hand, Foot, and Mouth disease (HFMD) in China with a substantial disease burden. However, there is currently no commercial CVA6 vaccine. The D3a genotype of CVA6 is the predominant genotype in China. In this study, virus-like particles (VLPs) and mRNA vaccines based on the CVA6 sub-genotype D3a were successfully developed. The immunogenicity and protective effects of the VLP of CVA6 combined with Al(OH) 3 and CpG adjuvant indicated that VLP-induced neutralizing antibodies against three CVA6 sub-genotype (D2, D3a, and D3b) strains in Institute of Cancer Research (ICR) mice, and the combination of the two adjuvants enhanced cellular immunity. Passive immunization with serum from mice immunized with VLPs protected suckling mice against CVA6 lethal challenge in both antiserum transfer and maternal immunization experiments. The immunogenicity and protective effects of the mRNA vaccine of CVA6 indicate that it induces robust T-cell immunity. T-cell immunity was found to cross-protect against coxsackievirus A10 infection in mice. This is the first trial of a CVA6 mRNA vaccine worldwide and the first comparison of the immunogenicity and protective effects of VLP and mRNA vaccines based on D3a CVA6. The study provides a theoretical basis for the development of enteroviruses vaccines and the formulation of immunization strategies., (© 2025 Wiley Periodicals LLC.)

Polysaccharides have been suggested to ameliorate metabolic diseases. However, their differential colitis-mitigating effects in mouse models with different colony structures remain poorly understood. Therefore, this study investigated the effects of polysaccharides from Lycium barbarum (LBP), sunflower (SP), and yam (YP) on colitis in C57BL/6 J (B6) mice born via vaginal delivery (VD) and in both caesarean section (CS)- and VD-born Institute of Cancer Research (ICR) mice. LBP was mainly composed of glucose (30.2 %), galactose (27.5 %), and arabinose (26.9 %). The main components of SP and YP were galacturonic acid (75.8 %) and glucose (98.1 %), respectively. Interestingly, LBP effectively alleviated body weight loss, reduced inflammatory cytokine levels, and restored intestinal barrier function in all three mouse models. Moreover, LBP decreased the abundance of norank&#95;f&#95;&#95;norank&#95;o&#95;&#95;Clostridia&#95;UCG-014, Coriobacteriaceae&#95;UCG-002, and norank&#95;f&#95;Eubacterium&#95;coprostanoligenes&#95;group in B6 mice, and the abundance of these genera positively correlated with pro-inflammatory cytokine levels. LBP increased the abundance of Lactobacillus, which was positively correlated with the levels of the protective factor, IL-10, in CS-born ICR mice. Collectively, our study suggests the potential application of LBP in the treatment of ulcerative colitis. We also provide an alternative method for restoring intestinal homeostasis in CS-born offspring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Exposure to stress in early life can have a significant impact on individuals. However, the effects of early-life stress (ELS) on prosocial behavior remain unclear, as do the underlying mechanisms. In this study, ICR juvenile mice were subjected to juvenile chronic social defeat stress (jCSDS) between postnatal days 32 and 41, during which body weight changes were continuously monitored. The behaviors of adult mice were evaluated using the open field test (OFT), the social interaction test (SIT), and the prosocial choice task (PCT). ELISA was used to quantify serum levels of oxytocin, serotonin, and dopamine. The density of dendritic spines in the basolateral amygdala was evaluated by Golgi staining. Behavioral test results showed that jCSDS induced anxiety-like behavior and decreased prosocial selection tendency in mice. Additionally, exposure to jCSDS increased the serum levels of oxytocin, decreased those of serotonin, and increased the density of dendritic spines in the basolateral amygdala. Correlation analysis indicated that prosocial behavior was negatively correlated with serum oxytocin levels and dendritic spine density in the basolateral amygdala. These results suggested that jCSDS reduced prosocial behavior, possibly due to changes in serum oxytocin contents and adaptive changes in amygdaloid neurons., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Methylmercury (MeHg) is a potent neurotoxicant with neurodevelopmental toxicity that is widely ingested into the body through drinking water and food. MeHg crosses the placental barrier and accumulates in the brain of the fetus, affecting the growth and development of the central nervous system. Although it has been demonstrated that MeHg induces neuronal calcium overload in the rat cerebral cortex, the role of calcium overload in MeHg-induced neurodevelopmental toxicity remains unclear. Here, we used ICR-pregnant mice and their resulting offspring and administered the BAPTA-AM calcium antagonist to investigate the molecular mechanisms by which MeHg exposure during gestation and lactation affects neurodevelopment. We found that exposure to MeHg during gestation and lactation resulted in developmental arrest and neurobehavioral dysfunction in the offspring, with calcium overload, disturbed mitochondrial dynamics, and apoptosis. However, the calcium overload inhibitor BAPTA-AM rescued MeHg-induced neurodevelopmental damage, attenuated the onset of calcium overload, reduced mitochondrial kinetic disturbances and apoptosis. Meanwhile, the activation of the CaM/CaMKII/DRP1 signaling pathway induced by calcium overload was inhibited, and the interaction between DRP1 and BAX was attenuated, which alleviated apoptosis to a certain extent. In summary, our study suggests that MeHg-induced calcium overload may induce disturbed mitochondrial dynamics through activation of the CaM/CaMKII/DRP1 signaling pathway, resulting in neuronal apoptosis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.)

Neonatal hypoxic-ischaemic encephalopathy (HIE) remains one of the major causes of neonatal death and long-term neurological disability. Due to its complex pathogenesis, there are still many challenges in its treatment. In our previous studies, we found that quercetin can alleviate neurological dysfunction after hypoxic-ischaemic brain injury (HIBI) in neonatal mice. As demonstrated through in vitro experiments, quercetin can inhibit the activation of the TLR4/MyD88/NF-κB signalling pathway and the inflammatory response in the microglial cell line BV2 after oxygen-glucose deprivation. However, the in-depth mechanism still needs to be further elucidated. In the present study, 7 day-old neonatal ICR mice or BV2 cells were treated with quercetin with or without the SIRT1 inhibitor EX527 via neurobehavioural, histopathological and molecular methods. In vivo experiments have shown that quercetin can significantly improve the performance of HI mice in behavioural tests, such as the Morris water maze, rotarod test and pole climbing test, and reduce HI insult-induced structural brain damage, cell apoptosis and hippocampal neuron loss. Quercetin also inhibited the immunofluorescence intensity of the microglial M1 marker CD16 + 32 and significantly downregulated the expression of the M1-related proteins iNOS, IL-1β and TNF-α. Moreover, quercetin increased the immunofluorescence intensity of the microglial M2 marker CD206 and significantly increased the expression of the M2-related proteins Arg-1 and IL-10. In addition, quercetin limits the nucleocytoplasmic translocation and release of microglial HMGB1 and further suppresses the activation of the downstream TLR4/MyD88/NF-κB signalling pathway. The above effects of quercetin are partially weakened by pretreatment with EX527. Similar results were found in in vitro experiments, and the mechanism further revealed that the rebalancing effect of quercetin on microglial polarization is achieved through the SIRT1-mediated reduction in HMGB1 acetylation levels. This study provides new and complementary insights into the neuroprotective effects of quercetin and a new direction for the treatment of neonatal HIE., Competing Interests: Declarations. Conflict of interest: The authors declare that they have no competing interests. Ethical approval: For research involving animal experiments, we declare that all experimental protocols were approved by the Ethics and Welfare Committee of the First Affiliated Hospital of Nanchang University (Ethics No.: CDYFY-IACUC-202310QR027)., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Background: Food-derived nucleic acids exhibit various biological activities and may act as nutrients. Oral ingestion of the nucleic acid fraction (NAF) of salmon milt extract hydrolysates enhances cognitive function in mice, although their active ingredients have not yet been identified, and detailed mechanisms of action are unknown., Objectives: To identify active ingredients enhancing cognitive function contained in the NAF and its possible underlying mechanism., Methods: Because the NAF is rich in trinucleotides, proliferative effects of all 64 types of trideoxyribonucleotides were examined in primary cultured neural stem cells (pcNSCs). The active trideoxyribonucleotide was administered intrahippocampally (5 μg) in 7-wk-old male Institute of Cancer Research (ICR) mice (n = 4-6) or orally (1 mg) 3 times a week for 2 wk in 6-wk-old male ICR mice (n = 5), followed by evaluating neurogenesis and cognitive function by immunohistochemical analysis and spatial recognition test, respectively. The mechanism of action was examined by proteomic analysis of trideoxyribonucleotide-treated pcNSCs using the software DAVID, followed by western blot analysis in pcNSCs and hippocampus., Results: Among all trideoxyribonucleotides, only trideoxycytidine diphosphate (CCC) significantly promoted NSC proliferation (P < 0.05), whereas exposure to putative metabolites of CCC did not affect the proliferation. Intrahippocampal or oral CCC administration in mice increased doublecortin-positive cells in hippocampus (P < 0.05) and enhanced spatial memory (P < 0.05). Proteomic analysis revealed significant alterations in expression of PI3K-Akt signaling-related proteins, including eEF1A2 (P < 0.05). Intrahippocampal CCC administration significantly increased the Akt phosphorylation (p-Akt/Akt) (P < 0.05), which was abolished with the PI3K-Akt inhibitor, LY294002 (P < 0.05). CCC exposure increased p-Akt/Akt (P < 0.05) in pcNSCs, whereas LY294002 or small interfering RNA for eef1a2 suppressed CCC-induced increase in p-Akt/Akt (P < 0.05) and cell proliferation (P < 0.05)., Conclusions: A simple food-derived structural molecule CCC promotes NSC proliferation through eEF1A2/PI3K-Akt signaling pathway, thereby enhancing neurogenesis., Competing Interests: Conflicts of interest The authors report no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

The significant benefits of physical activity are well-documented in academic literature, with growing evidence highlighting its positive effects (among others) on memory and cognitive function. Exercise, particularly aerobic activities, has been shown to mitigate neuroinflammatory processes, promote neuronal regeneration, facilitate recovery from cerebral trauma, and reduce the risk of neurodegenerative diseases. Among neurological conditions, traumatic brain injury (TBI) is the most common in individuals under 50, with 80-90 % of cases categorized as mild traumatic brain injury (mTBI). This study investigates the impact of exercise on visual and spatial memory deficits in mice following mTBI. ICR mice were subjected to a seventeen-day treadmill training protocol initiated at four different time intervals post-mTBI (2, 7, 13, and 30 days). A battery of specific behavioral tests was used to assess anxiety-like behaviors, motor skills, and visual and spatial memory. Our results indicate that running positively affected mTBI in both novel object recognition (p < 0.001) and Y-maze (p < 0.001) regardless of the running protocol's initiation time, demonstrating that aerobic exercise significantly alleviates cognitive deficits associated with mTBI. Importantly, mTBI did not appear to impact motor abilities or anxiety-like behaviors based on the assessment paradigms utilized. In conclusion, aerobic exercise effectively enhances visual and spatial memory post-mTBI, with promising results observed even when the running protocol is initiated up to one-month post-injury., Competing Interests: Declaration of competing interest We declare that our study was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Zika virus (ZIKV) is a mosquito-borne, positive-stranded RNA virus, ZIKV infection during pregnancy threatens pregnancy and fetal health, and it remains a global health threat, there are no clinically approved vaccines or antiviral drugs for the treatment of ZIKV infection. VV116 is an oral drug candidate of nucleoside analog against SARS-CoV-2 that has demonstrated a satisfactory safety and tolerability in healthy subjects. Our study shows for the first time that VV116 has potent activity against ZIKV. We verified that VV116 can inhibit the RNA and protein level of ZIKV in vitro. Importantly, treatment with VV116 significantly enhanced survival and provided protection in ZIKV-infected ICR mice. VV116 acted at the replication stage of viral infection cycle, and exhibited reasonable inhibition of ZIKV replicons. Collectively, the in vitro and in vivo anti-ZIKV activity of VV116 suggests that it is a promising anti-ZIKV drug candidate., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Parkinson's disease (PD) is a neurodegenerative disorder caused by dopamine (DA) neuronal dysfunction. Although DA agonists and N-methyl-D-aspartate receptor (NMDAR) antagonists are used to treat PD, chronic use causes severe side effects. Puerarin (PUE) is a natural bioactive compound that affects the DA system; however, its effect on PD-associated motor functions is unknown. Therefore, we investigated whether PUE treatment in a 6-hydroxydopamine (6-OHDA) PD mouse model affects motor dysfunction., Methods: Adult male ICR mice received unilateral 6-OHDA microinfusion into the right medial forebrain bundle. After a 2-week recovery period, PUE (20 or 50 mg/kg) or the vehicle (saline, VEH) was administered intraperitoneally once daily for 21 days. Motor dysfunction was assessed using the locomotion, gait cycle, and rotation tests. Local field potentials (LFPs) were measured in the substantia nigra compacta (SNc), striatum (STR), subthalamic nucleus (STN), and primary motor cortex., Results: 6-OHDA-lesioned PD mice showed increased gait cycle disturbance and unidirectional rotation. PUE treatment ameliorated the gait cycle disturbance, but not unidirectional rotation of PD mice. These effects differed with DA agonist treatment (which improved PD symptoms) and NMDAR antagonist treatment (which aggravated PD symptoms). Moreover, locomotion was increased only in NMDAR antagonist treatment. PUE treatment induced no changes in the attenuated LFP of the beta wave in the STR and STN, and SNc-STN delta-wave coherence was shown in PD animals., Conclusions: This study suggests that PUE is a beneficial co-therapeutic agent for alleviating gait cycle disturbance in PD symptoms., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s) under exclusive licence to Maj Institute of Pharmacology Polish Academy of Sciences.)

Clear-cut evidence has linked defective autophagy to Alzheimer's disease (AD). Recent studies underscore a unique hurdle in AD neuronal autophagy: impaired retrograde axonal transport of autophagosomes, potent enough to induce autophagic stress and neurodegeneration. Nonetheless, pertinent therapy is unavailable. Here, a novel combinational therapy composed of siROCK2 and lithospermic acid B (LA) is introduced, tailored to dredge blocked axonal autophagy by multi-mitigating microtubule disruption, ATP depletion, oxidative stress, and autophagy initiation impediments in AD. Leveraging the recent discovery of multi-interactions between polyphenol LA and siRNA, ε-Poly-L-lysine, and anionic lipid nanovacuoles, LA and siROCK2 are successfully co-loaded into a fresh nano-drug delivery system, LIP@PL-LA/siRC, via a ratio-flexible and straightforward fabrication process. Further modification with the TPL peptide onto LIP@PL-LA/siRC creates a brain-neuron targeted, biocompatible, and pluripotent nanomedicine, named "Nano-dredger" (T-LIP@PL-LA/siRC). Nano-dredger efficiently accelerates axonal retrograde transport and lysosomal degradation of autophagosomes, thereby facilitating the clearance of neurotoxic proteins, improving neuronal complexity, and alleviating memory defects in 3×Tg-AD transgenic mice. This study provides a fresh and flexible polyphenol/siRNA co-delivery paradigm and furnishes conceptual proof that dredging axonal autophagy represents a promising AD therapeutic avenue., (© 2024 Wiley‐VCH GmbH.)

The increasing use of SARS-CoV-2 mRNA vaccines has raised concerns about their potential toxicological effects, necessitating further investigation to ensure their safety. To address this issue, we aimed to evaluate the toxicological effects of SARS-CoV-2 mRNA vaccine candidates formulated with four different types of lipid nanoparticles in ICR mice, focusing on repeated doses and administration routes. We conducted an extensive analysis in which mice received the mRNA vaccine candidates intramuscularly (50 μg/head) twice at 2-week intervals, followed by necropsy at 2 and 14 dpsi (days post-secondary injection). In addition, we performed a repeated dose toxicity test involving three, four, or five doses and compared the toxicological outcomes between intravenous and intramuscular routes. Our findings revealed that all vaccine candidates significantly induced SARS-CoV-2 spike protein-specific IgG and T cell responses. However, at 2 dpsi, there was a notable temporary decrease in lymphocyte and reticulocyte counts, anemia-related parameters, and significant increases in cardiac damage markers, troponin-I and NT-proBNP. Histopathological analysis revealed severe inflammation and necrosis at the injection site, decreased erythroid cells in bone marrow, cortical atrophy of the thymus, and increased spleen cellularity. While most toxicological changes observed at 2 dpsi had resolved by 14 dpsi, spleen enlargement and injection site damage persisted. Furthermore, repeated doses led to the accumulation of toxicity, and different administration routes resulted in distinct toxicological phenotypes. These findings highlight the potential toxicological risks associated with mRNA vaccines, emphasizing the necessity to carefully consider administration routes and dosage regimens in vaccine safety evaluations, particularly given the presence of bone marrow and immune organ toxicity, which, though eventually reversible, remains a serious concern., Competing Interests: Declarations. Conflict of interest: All authors declare that they have no conflicts of interest concerning the publication of this paper., (© 2024. The Author(s).)

Inorganic nanoparticles are promising materials for bone tissue engineering due to their chemical resemblance to the native bone structure. However, most studies are unable to capture the entirety of the defective environment, providing limited bone regenerative abilities. Hence, this study aims to develop a multifunctional nanoparticle to collectively control the defective bone niche, including immune, angiogenic, and osteogenic systems. The nanoparticles, self-assembled by biomimetic mineralization and tannic acid (TA)-mediated metal-polyphenol network (MPN), are released sustainably after the incorporation within a gelatin cryogel. The released nanoparticles display a reduction in M1 macrophages by means of reactive oxygen species (ROS) elimination. Consequently, osteoclast maturation is also reduced, which is observed by the minimal formation of multinucleated cells (0.4%). Furthermore, the proportion of M2 macrophages, osteogenic differentiation, and angiogenic potential are consistently increased by the effects of magnesium from the nanoparticles. This orchestrated control of multiple systems influences the in vivo vascularized bone regeneration in which 80% of the critical-sized bone defect is regenerated with new bones with mature lamellar structure and arteriole-scale micro-vessels. Altogether, this study emphasizes the importance of the coordinated modulation of immune, osteogenic, and angiogenic systems at the bone defect site for robust bone regeneration., (© 2024 Wiley‐VCH GmbH.)

Ethnopharmacological Relevance: Xanthoceras sorbifolium Bunge (X. sorbifolia), an oil crop native to northern China, is valued for both its edible and medicinal uses. It has various applications, including the production of edible and bioactive oils, and is used in traditional medicine for its antioxidant and anti-inflammatory properties. However, the toxicity of X. sorbifolia, particularly its widely used seed kernels, remains unclear., Aim of the Study: This study aimed to evaluate the acute toxicity and safety risks of X. sorbifolia seed kernels based on human-recommended doses by in vitro or in vivo experiments, and integrating network analysis., Materials and Methods: In this study, rats and mice were employed as model organisms to investigate the acute toxicity of X. sorbifolia seed kernels. The experiments included the Salmonella typhimurium reverse mutation test, red blood cell micronucleus test, spermatocyte chromosome aberration test in mice, and a 90-day exposure study in rats to assess the potential toxicity and safety risks of the seed kernels. Based on this, combined with The Comparative Toxicogenomics Database (CTD), the biological functions of the main active ingredients of X. sorbifolia were further explored through integrated network analysis, and the anti-inflammatory effect of X. sorbifolia was explored through cotton ball granuloma inflammation experiment., Results: During the experimental period, animals in all treatment groups demonstrated normal growth and development. Although some detection indicators showed significant differences in different treatment groups, the results were still within a reasonable range. In addition, by screening the CTD, 120 target genes with potential interactions of the main active ingredients in the kernel of X. sorbifolia were obtained for analysis, and it was found that these genes were involved in important biological processes such as response to oxidative stress, response to reactive oxygen species, and regulation of inflammatory response. The cotton ball granuloma inflammation experiment in rats also suggested that X. sorbifolia tended to inhibit the proliferation of granulomas, indicating that the kernel of X. sorbifolia has potential anti-inflammatory and antioxidant properties., Conclusion: The findings suggested that X. sorbifolia seed kernels were safe within the recommended dosage range. As a traditional Chinese medicine prescription, it has certain anti-inflammatory and antioxidant effects. This study provides valuable reference guidelines for the clinical application of X. sorbifolia seed kernels and encourages further research into its potential uses and safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Insomnia is a pervasive and prominent problem worldwide, afflicting approximately one-third of the population and profoundly affecting patients' quality of life. Efficient and safe sedative-hypnotic medications are required. Menyanthes trifoliata L. (Mt), a sleeping herb in China, is used as a hypnotic remedy in ethnomedicines; however, there are few studies on this herb., Aim of the Study: We systematically evaluated the potential of Mt as a sedative-hypnotic candidate., Materials and Methods: The chemical constituents of the Mt extract were analyzed by lLiquid chromatography with photodiode array detection and mass spectrometry (LC-PDA-MS). The sedative-hypnotic effects of Mt extract (0.5, 2, and 4 g/kg) were investigated using the pentobarbital-induced sleep test (PIST), the caffeine-induced insomnia model (CIIM), and the open field test (OFT). Furthermore, the effect of Mt on sleep architecture was ‌investigated using electroencephalography/electromyography (EEG/EMG). The safety of the Mt extract was evaluated using the maximum tolerated dose method., Results: Fifteen phenolic compounds were identified based on their UV absorption and MS fragmentation using LC-PDA-MS analysis. In the CIIM, PIST, and OFT, Mt extract exhibited a dose-dependent reduction in sleep latency, an extension of total sleep duration, and a decrease in locomotor activity. Moreover, it increased the duration of non-rapid eye movement (NREM) sleep and reduced wakefulness after one day's administration, according to EEG/EMG. Additionally, no signs of toxicity were observed at a dose of 30 g/kg (equivalent to 316.46 g/kg of crude drugs)., Conclusion: This study supports the potential medicinal use of Mt extract for sleep promotion., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Carthamus tinctorius L. (Safflower) was believed to have multiple benefits, including antioxidant effects, enhanced learning and memory, and improving neuronal injury. Safflower Yellow(SY) are the main active ingredients of Safflower, displays strong pharmacological potential treatment of Alzheimer's disease(AD). However, its effect on memory impairments remains insufficiently investigated., Aim of the Study: The study aims to investigate the effects of SY on cognitive functions in memory impairments model and to explore the mechanism of its action., Materials and Methods: We utilized the Morris Water Maze, Step-Through Test, Step-Down Test to assess the potential of SY in ameliorating learning and memory dysfunction caused by SCOP, NaNO 2 and ethanol in mice. Bioinformatic analysis and molecular biological approaches were used to study the related mechanisms of SY on anti-memory impairments., Results: The results of the Morris Water Test suggested that SY could shorten the escape latency and the time of the first crossing platform in the mice with memory acquisition and memory consolidation impairments, and increase the platform crossing times. The results of the Step-Though test and Step-Down test showed that the escape latency in the mice was prolonged and the number of errors was reduced after SY treatment. ELISA experiments indicated that SY decreased the AChE activities, increased the ChAT activities, and modulated oxidative stress markers (SOD, MDA, and GSH-PX) in scopolamine-induced mice. Western Blot and Nissl staining showed that SY could activated BDNF/TrkB/CREB signaling pathway and reduced neuronal damage., Conclusion: The findings present that SY can restore the function of the cholinergic system, inhibit oxidative stress, regulate the expression of upstream and downstream proteins in the CREB/BDNF/TrkB pathway, and alleviate brain tissue damage to improve memory impairment in mice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Fluoride, a common agricultural additive used to enhance plant resilience and pest control, poses toxicity risks when exposure surpasses safe thresholds, affecting ecosystems and human health. While its reproductive toxicity is recognized, the sex-specific and cross-generational effects remain underexplored. To address this gap, we employed an integrative approach combining transcriptomics (next-generation sequencing (NGS)), bioinformatic network analysis, gut microbiota sequencing, and in vivo functional assays. ICR mice (F0 generation), both male and female, were exposed to fluoride (100 mg/L in drinking water) for 35 days, continuing through gestation and offspring weaning. Our transcriptomic analysis revealed significant upregulation of autophagy (via the PI3K-AKT-mTOR pathway) and oxidative stress-induced mitochondrial dysfunction in gonadal tissue, with more pronounced effects observed in males. Further integrated analyses of transcriptomic and metabolomic data, supported by in vivo experiments, highlighted oxidative stress, mitochondrial dysfunction, and PI3K-AKT-mTOR pathway activation with stronger effects in males. The principal component analysis confirmed sex-specific transcriptome alterations, with males showing more substantial disruption. Additionally, 16S rRNA sequencing identified significant gut dysbiosis, particularly in males, with an increased Firmicutes/Bacteroidetes ratio and higher abundances of Oscillospirales and Anaerovoracaceae . Moreover, our study identified significant correlations between specific gut microbiota (e.g., Firmicutes , Proteobacteria ) and autophagy, oxidative stress, and mitochondrial dysfunction pathways, with notable sex-dependent differences. These findings suggest that gut microbiota may play a critical role in modulating fluoride-induced reproductive toxicity, particularly through their effects on oxidative stress and cellular homeostasis. The breakdown of the gut barrier and elevated serum/gonadal lipopolysaccharide (LPS) levels in fluoride-treated mice further established a link between gut dysbiosis and fluoride-induced reproductive toxicity. These findings underscore the importance of considering sex differences in xenobiotic-induced reproductive and developmental toxicity.

Fluorescent biosensors offer a powerful tool for tracking and quantifying protein activity in living systems with high temporospatial resolution. However, the expression of genetically encoded fluorescent proteins can interfere with endogenous signaling pathways, potentially leading to developmental and physiological abnormalities. The EKAREV-NLS mouse model, which carries a FRET-based biosensor for monitoring extracellular signal-regulated kinase (ERK) activity, has been widely utilized both in vivo and in vitro across various cell types and organs. In this study, we report a significant defect in mammary epithelial development in EKAREV-NLS C57BL/6J female mice. Our findings reveal that these mice exhibit severely impaired mammary epithelial outgrowth, linked to systemic defects including disrupted estrous cycling, impaired ovarian follicle maturation, anovulation, and reduced reproductive fitness. Notably, estrogen supplementation was sufficient to enhance mammary epithelial growth in the EKAREV-NLS C57BL/6J females. Furthermore, outcrossing to the ICR genetic background fully restored normal mammary epithelial outgrowth, indicating that the observed phenotype is dependent on genetic background. We also confirmed the functional performance of the biosensor in hormone-supplemented and outcrossed tissues through time-lapse imaging of primary mammary epithelial cells. Our results underscore the critical need for thorough characterization of biosensor-carrying models before their application in specific research contexts. Additionally, this work highlights the influence of hormonal and genetic factors on mammary gland development and emphasizes the importance of careful consideration when selecting biosensor strains for mammary studies., Competing Interests: Declarations. Competing Interests: Z.S.K. is the Editor-in-Chief of the Journal of Mammary Gland Biology and Neoplasia. M.B. has no competing interest., (© 2025. The Author(s).)

Background: Sperm represent a heterogeneous population crucial for male reproductive success. Additionally, sperm undergo dynamic changes during maturation and capacitation. Despite these well-established processes, the complex nature of sperm heterogeneity and membrane dynamics remains elusive. The composition of phospholipids in the sperm membrane changes dynamically during maturation, with their release occurring during capacitation. This study aims to investigate the heterogeneity and dynamic changes in the sperm membrane during maturation and capacitation towards fertilization by visualizing these membrane dynamics., Methods: Sperm were collected from the cauda epididymis or testis of Institute of Cancer Research (ICR) male mice and stained with MemBright dye (commercial name: MemGlow™-560, MG-560), a fluorogenic live-cell membrane probe. Staining was performed either before, during, or after incubation for capacitation. In pre-staining experiments, sperm were stained with MG-560 before capacitation and then incubated to induce capacitation. Acrosome-reacted sperm were assessed after staining with peanut agglutinin FITC (PNA-Lectin FITC). Stained sperm were observed using fluorescence or confocal microscopy., Results: MG-560-stained sperm from the epididymis before capacitation showed four staining patterns: head-midpiece-tail (HMT), head-midpiece (HM), head (H), midpiece (M) positive, or totally negative, with ratios remaining unchanged during capacitation (30.5%, 29%, 11.3%, 3.7%, and 25.5%, respectively). In contrast, all testicular sperm were negative for staining. Pre-stained sperm exhibited an increased number of HM and M patterns over time, whereas the number of HMT-stained sperm decreased. Consistently, spontaneous acrosome-reacted sperm were detected predominantly in HM- or M-stained sperm. After in vitro fertilization (IVF) using pre-stained sperm, zona pellucida-attached sperm were mostly negative for staining. Finally, all sperm detected in the perivitelline space were only negative., Conclusions: Mature sperm membranes stained with MG-560 exhibited heterogeneous and dynamic changes during the capacitation and fertilization process. MG-560 staining identified sperm with the potential to undergo the acrosome reaction, and these MG-560-positive sperm eventually became negative as they penetrated the zona pellucida for fertilization. Thus, the MG-560 staining patterns likely reflect the physiological state and potential of the sperm. These findings provide new insights into sperm heterogeneity and dynamics, and this staining method may also prove useful for assessing sperm quality., (© 2025 The Author(s). Published by IMR Press.)

Humans may intake 0.02 mg/kg/day of short-chain chlorinated paraffins (SCCPs), and no study is available on mammalian ovarian damage caused by low-level SCCPs. In this study, four groups of 5-week-old female Institute of Cancer Research (ICR) mice were orally administered 0, 0.01, 0.1, and 1.0 mg/kg/day SCCPs for 21 consecutive days, and serum and ovaries were collected 20 h after the last SCCPs-administration. SCCPs at ≥0.1 mg/kg/day were found to reduce follicle counts at each stage, induce dose-dependent oxidative stress in mice, and lower serum E2 and ovarian anti-Müllerian hormone levels. The data indicated that cellular PANoptosis increased in the ovaries of all SCCP-treated mice. Furthermore, AIM2- and NLRP12-PANoptosome gene and protein levels were considerably elevated. Female germline stem cells (FGSCs) in the cortical portion of the ovary exhibited substantial damage in all SCCP groups, additionally, the expression of FGSC marker genes and major marker proteins was diminished in the ovaries. Oral administration of SCCPs with 0.01, 0.1, and 1.0 mg/kg/day to mice resulted in PANoptosis of the ovaries. Therefore, it was suggested that the oral administration of ≥0.1 mg/kg/day of SCCPs suppressed ovarian function, which may be attributed to the fact that SCCPs induced the generation of AIM2- and NLRP12-PANoptosome in ovary cells.

In humans, cerebral malaria is the most common cause of malaria-related mortality. Mouse C57BL/6 (B6) sub-strains are the major model system for experimental cerebral malaria (ECM) as they show similar pathophysiology to human cerebral malaria after infection with the rodent malaria parasite Plasmodium berghei ANKA. This model system has been used to analyze the molecular mechanisms of cerebral malaria. To develop new mouse models, we analyzed the ECM susceptibility of NOD/Shi (NOD) and NSY/Hos (NSY) strains established from the non-inbred ICR strain. Both NOD and NSY strains exhibited clinical symptoms and pathologies similar to ECM in C57BL/6J (B6J) mice and died within 11 days of infection. Thus, the NOD and NSY strains are susceptible to ECM and may be useful as new ECM models. The ECM susceptibility of both strains is suggested to be due to homozygosity for the cerebral malaria susceptibility allele of the ECM susceptible ICR strain. Although analyses using B6 sub-strains have proposed that complement component 5 (C5) plays an important role in ECM pathogenesis, we found that C5 was not essential as the ECM susceptible NOD strain is C5 deficient. Thus, results obtained from B6 sub-strains may not reflect the full picture of ECM in mice. Comparative analyses of multiple ECM models will contribute to a more accurate identification of the factors essential for ECM.

In this paper, an oral liquid containing Magnolia officinalis polysaccharide was formulated and its hypoglycemic effects were investigated. An orthogonal test was conducted based on single-factor experiments to optimize the formulation guided by sensory evaluations. Its stability and safety were also assessed. The oral liquid was administered via gavage to STZ-induced diabetic mice at low (2.5 mL/kg), medium (5 mL/kg) and high (10 mL/kg) doses. Blood glucose levels were monitored weekly. After 8 weeks of treatment, the oral glucose tolerance test (OGTT) was performed and fasting insulin levels, lipid profiles, oxidative stress levels in serum and liver, and the activities of hexokinase (HK) and pyruvate kinase (PK) in the liver were measured. Results indicated that the optimal formulation contained 0.2% steviol glycosides, 0.2% ascorbic acid, a pH of 5.0, and 0.5% ginger juice, yielding a sensory evaluation score of 94 ± 0.58. The oral liquid remained stable at room temperature for 12 months and passed acute oral toxicity testing. After 8 weeks, diabetic mice exhibited a significant reduction in blood glucose levels (P < 0.01), enhanced glucose metabolism, increased fasting insulin levels, and decreased lipid levels. Additionally, antioxidant capacity improved, and HK and PK activities increased significantly in the diabetic mice. In conclusion, the Magnolia officinalis polysaccharide oral liquid demonstrated potential antidiabetic effects by promoting glucose utilization in peripheral tissues, increasing fasting insulin levels, and enhancing antioxidant capacity alongside HK and PK activities. This formulation could represent a promising hypoglycemic health product., Competing Interests: Declarations. Ethics Statement: All the experiments were conducted according to the Guidelines of Experimental Animal Administration austerely published by the State Committee of Science and Technology of People’s Republic of China. The study was reviewed and approved by the Laboratory Animal Ethics Committee of Hubei Minzu University(No.202039). Competing Interest: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Various wound dressings are under development to accelerate wound healing, and hydrogels in particular have the potential to create ideal conditions for wound healing. In this study, we investigated a novel hydrogel based on gelatin/oxidized carboxymethyl cellulose loaded with fucoidan derived from Ecklonia cava (ECF) for wound treatment. The mechanical stability and self-healing ability of the hydrogel were optimized, and its cytocompatibility was demonstrated against RAW 264.7 macrophages and human dermal fibroblasts (HDF). Furthermore, sustained drug release from the fabricated hydrogels depending on ECF concentration exhibited radical scavenging ability, induction of collagen production, promotion of cell migration, reduction of nitric oxide levels, and cytoprotective effects against oxidative stress in the ROS microenvironment, attributing these effects to the wound healing potential of bioactive ECF. In an in vivo experiments, GOC/F5 hydrogel was shown to significantly reduce wound area in a full-thickness ICR mouse model, and promoted re-epithelialization and collagen deposition to rapidly repair wounds. These results suggest that GOC/F5 hydrogels could be potentially used as an ideal dressing for wound healing., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The aim of this study is to investigate the differential impacts of high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) on neural circuit dynamics and neuronal firing in the hippocampal CA1 subregion (CA1) region and medial entorhinal cortex (MEC) of mice. Forty-two male ICR mice were randomized into control, HIIT, and MICT groups. Electrophysiological recordings were performed pre- and postintervention to assess neural circuit dynamics and neuronal firing patterns in the CA1-MEC pathway. Both exercise protocols increased local field potential (LFP) coherence, with MICT showing a more pronounced effect on δ and γ coherences ( P < 0.05). Both modalities reduced δ power spectral density (PSD) (HIIT, P < 0.05; MICT, P < 0.01) and elevated θ, β, and γ PSDs. Neuronal firing frequency improved in both CA1 and MEC following HIIT and MICT ( P < 0.05). HIIT enhanced firing regularity in CA1 ( P < 0.05), whereas MICT improved regularity in both regions ( P < 0.05). Both protocols reduced firing latency (HIIT, P < 0.05; MICT, P < 0.01) and enhanced burst firing ratio, interburst interval (IBI), burst duration (BD), and LFP phase locking ( P < 0.05 or P < 0.01). Notably, MICT significantly improved spatial working memory and novel recognition abilities, as evidenced by increased novel arm time, entries, and preference index ( P < 0.01). This study reveals that both HIIT and MICT positively impact neural processing and information integration in the CA1-MEC network of mice. Notably, MICT exhibits a more pronounced impact on neural functional connectivity and cognitive function compared with HIIT. These findings, coupled with the similarities in hippocampal electrophysiological characteristics between rodents and humans, suggest potential exercise-mediated neural plasticity and cognitive benefits in humans. NEW & NOTEWORTHY This study is the first to investigate HIIT and MICT's effects on neural activity in the mouse CA1-MEC circuit, demonstrating that exercise modulates processing, enhances integration, and boosts cognitive performance. Due to similar hippocampal electrophysiology in rodents and humans during movement and navigation, our findings suggest implications for human brain neural changes, advancing the understanding of neurophysiological mechanisms underlying exercise-cognition interactions and informing exercise recommendations for cognitive health.

Enterotoxigenic Escherichia coli (ETEC) is a major cause of diarrhea in humans and animals. The study aimed to evaluate the efficacy of bovine lactoferrin (bLf) as an adjuvant combined with AMP (N6) in the treatment of E. coli -induced bacterial enteritis. Firstly, 40 female ICR mice were randomly divided into four groups. The ETEC-A, ETEC-B, and ETEC-C groups were gavaged with 0.2 mL of ETEC K88 at 5 × 10 9 , 5 × 10 8 , and 5 × 10 7 CFU/mL for three consecutive days, respectively, the CK control group was given PBS. Based on the clinical symptoms and intestinal changes, the optimal model dose of ETEC K88 was determined to be 5 × 10 8 CFU/mL. Sixty female ICR mice were randomly divided into six groups: CK group (uninfected), NC group (infected and untreated), N6 treatment group (20 mg/kg), bLf treatment group (100 mg/kg), bLf + N6-A treatment group (10 mg/kg N6+100 mg/kg bLf), and bLf + N6-B group (20 mg/kg N6+100 mg/kg bLf). The clinical symptoms, intestinal morphology, inflammatory response and serum metabolites were monitored. The results showed that compared with the NC group, the bLf-N6-A and bLf-N6-B treatment groups had significant reductions in TNF-α and IL-6, significant increases in IL-10, and significant reductions in endotoxin and DAO in plasma ( p  < 0.05). Meanwhile, the bLf-N6-A and bLf-N6-B treatment groups significantly increased the expression of ZO-1, claudin-1 and occludin, increased the height of small intestinal mucosal villi and VH/CD after ETEC K88-induced intestinal injury ( p  < 0.05). The combination of bLf and N6 relieved enteritis by balancing intestinal mucosal immunity, improving intestinal morphology and barrier function. BLf combined with N6 can be used as an effective therapeutic strategy for the treatment of bacterial enteritis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Jellyfish stings can trigger abrupt heart failure via toxins, leading acute mortality rise. Proposed mechanisms involve oxidative stress and apoptosis, but evidence for effective treatments is lacking. To explore the concrete molecular mechanisms of jellyfish toxin-induced cardiotoxicity and to explore effective therapeutic approaches, we established tentacle extract (TE) of jellyfish Nemopilema nomurai induced cardiotoxicity models in vivo and in vitro based Intelligent Character Recognition (ICR) mice and H9C2 cells, respectively,.We assessed toxin-induced cardiac injury and screened antagonists from natural compounds to evaluate their antagonistic effects and explore their mechanisms of action. In vitro experiments showed that TE reduced the viability of H9C2 cells and induced a large number of cells apoptotic, accompanied by the elevation of reactive oxygen species (ROS), malondialdehyde (MDA) and the decrease of total superoxide dismutase (T-SOD), activated the phosphorylation level of mitogen-activated protein kinase (MAPK) nuclear transcription factors p38, extracellular regulated protein kinases (ERK) and c-Jun N-terminal kinase (JNK), and increased the transcription level of upstream cytokines interleukin-1β (IL-1β) and tumour necrosis factor-α (TNF-α), and OMT can significantly antagonize the above changes caused by TE; in vivo experiments demonstrated that TE could lead to the death of mice, as well as induce cardiac edema and rupture of myocardial fibers. In contrast, Oxymatrine (OMT) effectively counteracts the lethal effects of TE and reduces both cardiac edema and myocardial fiber rupture. In summary, OMT can antagonise TE-induced cardiac injury and lethal effects by inhibiting the activation of the MAPK pathway and reducing oxidative stress and apoptosis. As a natural compound, OMT offers a potential therapeutic strategy for jellyfish stings., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The primary adverse effect of chemotherapy drugs is the induction of spermatogenic disorders. Schisandra chinensis (Turcz) Baill. have been preliminarily shown to have a significant ameliorative in spermatogenic disorders. Nevertheless, there are relatively few studies on non-medicinal parts such as stems of S. chinensis, which have good therapeutic potential for side effects caused by cisplatin. Up to now, a total of 238 compounds have been isolated and identified from the vine stem of S. chinensi. The main components are lignans, polysaccharides, volatile oils, and organic acids. Therefore, the development of S. chinensis stem as a source of a novel chemotherapy drug protector is of great significance., Methods: In the present study, we explored the protective effect of the stem extract of S. chinensis (SCE) against cisplatin-induced spermatogenic disorders and its possible molecular mechanisms in vitro and in vivo. The network pharmacology was used to predict the targets that may act on spermatogenic disorders. Mice were injected intraperitoneally with cisplatin at 2 mg/kg for 7 days to induce spermatogenic disorders. SCE (75, 150, and 300 mg/kg) was administered to mice by gavage for 21 days. TM3 cells were treated with Schisandrol B (Sol B) (0.5, 1, and 2 µM) in the presence or absence of cisplatin (40 µM), and then cell viability, oxidative stress, apoptosis, and mitochondrial function were evaluated., Results: 16 constituents and target proteins were predicted to have possible effects on spermatogenic disorders by network pharmacology. Both in vivo and in vitro experiments showed a favorable protective effect of SCE against cisplatin-induced spermatogenic disorders. Sol B might as the major chemical component is responsibility for the protective effect. The mechanism may involve the regulation of Nrf2\HO-1 protein, PI3K\Akt, apoptosis, and MAPK signaling pathway to modulate 17βHSD, cycp450, Star and other crucial proteins in the testosterone synthesis pathway. Ultimately, this regulatory process aims to ameliorate the disruption of sex hormone secretion by cisplatin., Conclusion: These results indicated that the lignans in SCE could effectively improve the spermatogenesis barrier caused by cisplatin. These findings provided a theoretical basis for the development of non-medicinal parts of S. chinensis, and laid a foundation for improving spermatogenesis disorders caused by chemotherapy drugs., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Ticks vector a large number of significant pathogens, yet remain understudied due to the challenges in laboratory colonization. This study introduces innovative techniques for rearing Rhipicephalus sanguineus, focusing on in vivo tick feeding using ICR mice (Mus musculus) as a blood source. The research, conducted at the Walter Reed Army Institute of Research - Armed Forces Research Institute of Medical Sciences (WRAIR-AFRIMS), outlines the successful utilization of mice to support all stages of tick development. Ticks were retained on mice using Ethylene-Vinyl Acetate (EVA) foam capsules and cyclophosphamide was administered to the mice to prevent host immune response from interfering with tick feeding. These methods allowed the successful establishment and mass production of R. sanguineus tick colonies. The methods described herein hold promise for institutions seeking efficient tick production using a rodent model., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Bis(2-chloroethyl)2-chloroethylphosphphonate (B2CE2CEPP) was recently reported as a contaminant in underground water samples, however, information regarding its biotransformation and adverse effects is relatively rare. In this study, B2CE2CEPP and tris(2-chloroethyl) phosphate (TCEP; a typical organophosphate triester with a very similar structure as compared to B2CE2CEPP) were used as target contaminants to comparatively investigate their cytotoxicity, biotransformation, and alteration of lipid metabolism by use of an in vitro primary hepatocyte assay of ICR mice. This study aims to thoroughly investigate the cytotoxicity, biotransformation, and lipid metabolism effects of B2CE2CEPP and TCEP on primary hepatocytes of ICR mice, in order to reveal the potential health risks of these two organophosphorus compounds. The results showed that both B2CE2CEPP and TCEP were cytotoxic with LC50 values of 456 μM (B2CE2CEPP) and 1250 μM (TCEP), respectively. Both compounds underwent significant metabolic transformation follow exposure to primary mouse hepatocytes (PMHs), and a total of 4 potential metabolites were identified based on high-resolution orbitrap mass spectrometry technique. Exposure to TCEP and B2CE2CEPP led to lipid metabolism disorders in PMHs. TG (16:0/16:1/18:1) showed high sensitivity to B2CE2CEPP exposure and exhibited a significant upregulation trend, making it a potential biomarker significantly influenced by B2CE2CEPP exposure. Lipidomic analysis showed that B2CE2CEPP affected lipid metabolic pathways including thiamine metabolism, one carbon pool by folate, and folatesteroid hormone biosynthesis. The present study filled a knowledge gap in the current research on the biotransformation and adverse effects of B2CE2CEPP, and emphasized the importance on consistent monitoring of B2CE2CEPP and its structure-like compounds in various environmental samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

The effect of rabies virus infection on dendritic morphology and on the expression of the MAP2 protein in Purkinje cells in the cerebellum of mice was studied. ICR mice were inoculated with rabies virus, and six days later, the mice were sacrificed, the cerebellum was removed and processed for Golgi-Cox staining or MAP2 immunohistochemistry. Infection with rabies virus altered the dendritic pattern of Purkinje cells ranged from moderate changes to accentuated retraction in the dendritic tree of some Purkinje cells. The loss of dendritic branches in the samples of mice infected with RABV was also reflected in a decrease in intersections quantified using the Sholl technique, thus suggesting dendritic pathology. Immunoreactivity to MAP2 protein in the molecular layer of the cerebellum of control mice was mainly distributed in dendrites of Purkinje cells. Some somas were faintly stained. In infected mice immunoreactivity to MAP2 was intense in somas and dendrites of Purkinje cells and in some interneurons. These results are consistent with similar findings we previously reported for the cerebral cortex and spinal cord of rabies-infected mice. But they differ from studies in other pathologies where an association between dendritic pathology and loss of MAP2 immunoreactivity has been found. Our studies in rabies contribute to suggestion that MAP2 overexpression may also be associated with alterations in dendritic morphology. MAP2 protein contributes to maintaining cytoskeleton stability. However, in rabies, increased MAP2 expression here only determined by immunohistochemistry could destabilize the cytoskeleton of dendrites. Golgi staining is considered the gold standard for the study of dendritic morphology. Its association with changes in MAP2 expression appears to provide molecular support for the concept of dendritic pathology. These results contribute to the understanding of the effect of rabies virus infection on dendritic morphology. They therefore reinforce the idea that rabies not only has a dysfunctional effect on neurons, as some authors claim, but also affects their structure., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Conflict of interest: The authors declare that there are no conflicts of interest., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The highly valued oil of Rosa damascena Mill. (Rosaceae), widely used in high perfumery, cosmetics, and other spheres of human life, obliges us to know and study the safety profile of the product obtained from the water-steam distillation of fresh rose petals. The genotoxicity of the essential oil (EsO) has not been thoroughly studied despite its wide range of applications. That predetermined the object of this study-to evaluate, through classical cytogenetic methods, the possible cytotoxic/genotoxic activities of R. damascena Mill. EsO (EsORdm) in three different test systems: plant root meristem cells, mammalian bone marrow cells, and human lymphocyte cultures. The rose essential oil showed varying concentration- and time-dependent cytotoxic and genotoxic effects depending on the test system used, and it was established that the oil showed moderate cytotoxicity in lymphocyte cultures and non-high cytotoxicity in ICR mice but none in barley. Both barley and human lymphocytes showed a genotoxic effect with a dose-dependent increase in chromosomal aberrations (CAs) and a substantial rise in micronucleus (MN) frequency, while no genotoxicity was observed in bone marrow cells at the applied concentrations. Human lymphocytes exhibited the highest susceptibility to cytotoxic and genotoxic actions of the EsO. As a valuable plant-derived aromatic product with versatile uses in human life, R. damascena Mill. essential oil should be used in an appropriate concentration range tailored to cellular sensitivity.

Background: Sleep disorder is widespread and involves a variety of intricate factors in its development. Sleep deprivation is a manifestation of sleep disorder, can lead to energy metabolism disturbances, weakened immune system, and compromised body functions. In extreme situations, sleep deprivation can cause organ failure, presenting significant risks to human health., Purpose: This study aimed to investigate the efficacy and mechanisms of Astragalus Radix vesicles-like nanoparticles (AR-VLNs) in counteracting the deleterious effects of sleep deprivation., Methods: The ICR mice were divided into control, model, AR-VLNs high dose (equivalent to 20 g/kg crude drug), AR-VLNs low dose (equivalent to 10 g/kg crude drug), AR high dose (equivalent to 20 g/kg crude drug), and AR low dose (equivalent to 10 g/kg crude drug). The REM (rapid eye movement) sleep-deprivation model was established, and evaluations were conducted for motor function, antioxidant capacity, and energy metabolism indices. Moreover, CACO-2 cells damage was induced with lipopolysaccharide to evaluate the repairing ability of AR-VLNs on the intestinal cell mucosa by measuring permeability. Furthermore, metabolomics was employed to elucidate the mechanisms of AR-VLNs action., Results: AR-VLNs were demonstrated to enhance the motor efficiency and antioxidant capacity in REM sleep-deprived mice, while also minimized pathological damage and restored the integrity of the intestinal mucosal barrier. In vitro experiments indicated the anti-inflammatory effect of AR-VLNs against LPS-induced cell damage. Additionally, metabolomic analysis linked these effects with regulation of the amino acid metabolic pathways. Further confirmation from molecular biology experiments revealed that the protective effects of AR-VLNs against the deleterious effects of REM sleep deprivation were associated with the restoration of the intestinal mucosal barrier and the enhancement of amino acid metabolism., Conclusion: AR-VLNs administration effectively improved energy metabolism disorders in REM sleep deprived mice, by facilitating the repair of the intestinal mucosal barrier and regulating the amino acid metabolism., Competing Interests: Declarations. Ethics approval and consent to participate: All Institutional and National Guidelines for the care and use of animals were followed. All the animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Tsinghua University (23-JYY1). Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Objective: Ovarian tissue cryopreservation has become a promising alternative for fertility preservation in cancer patients, allowing ovarian tissue to be stored for future autotransplantation. Oxidative stress damage occurring during the cryopreservation process may impact tissue quality and function. This study aims to investigate the protective effects and potential mechanisms of Mitoquinone (MitoQ), a mitochondria-targeted derivative of the antioxidant ubiquinone, during the vitrification of ovarian tissue in mice., Methods: KGN cells were treated with various concentrations (0.1, 1, 10, and 50 μM) of MitoQ to determine the optimal concentration. Female ICR mice were divided into three groups: control, conventional vitrification, and MitoQ-supplemented vitrification. Ovarian samples were cryopreserved, thawed, and assessed for tissue morphology using Hematoxylin and Eosin (H&E) staining, and mitochondrial changes using immunofluorescence, transmission electron microscopy, and Western blot analysis. RNA sequencing (RNA-seq) was employed to explore potential protective mechanisms. Autotransplantation experiments were conducted, and the long-term effects of MitoQ on ovarian function were evaluated by counting follicle numbers through H&E staining and measuring serum estradiol and AMH levels using ELISA., Results: MitoQ at 1 μM was found to be the optimal concentration for maintaining follicular morphology after vitrification. It effectively reduced mitochondrial oxidative damage, preserved mitochondrial morphology, and regulated the expression of mitochondrial dynamics proteins (Drp1 and Mfn2). RNA-seq and Western blot analyses revealed that MitoQ inhibited the p38 MAPK pathway, thereby reducing apoptosis. Additionally, autotransplantation experiments showed that MitoQ treatment significantly increased follicle counts, estradiol (E2), and anti-Müllerian hormone (AMH) levels compared to conventional vitrification., Conclusions: MitoQ effectively mitigates vitrification-induced oxidative damage, maintains mitochondrial homeostasis, and preserves both follicular reserve and endocrine function. These findings suggest that MitoQ is a valuable adjunct in ovarian tissue cryopreservation and could significantly improve fertility preservation outcomes for cancer patients., Competing Interests: Declarations. Ethics approval and consent to participate: This study was approved by the Ethics Committee of the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital (Approval No. K2023097). All procedures followed the guidelines of the institution’s Animal Care and Use Committee and were in accordance with international standards for the ethical treatment of animals. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)