Abstract This study assessed the antimicrobial susceptibility of clinical bacterial isolates collected from Chinese medical institutions in 2025. Antimicrobial susceptibility testing was performed on isolates from 80 hospitals using disk diffusion or commercial automated systems in accordance with the standardized China Antimicrobial Surveillance Network (CHINET) surveillance protocol. Results were interpreted using the 2025 Clinical and Laboratory Standards Institute breakpoints. Between January and December 2025, 516,309 clinical isolates were included, comprising 28.5% Gram-positive and 71.5% Gram-negative bacteria. The detection rates of methicillin-resistant strains among Staphylococcus aureus , Staphylococcus epidermidis , and other coagulase-negative staphylococci were 28.4%, 80.8%, and 76.5%, respectively, with no vancomycin-resistant strains identified. Vancomycin resistance among Enterococcus faecium isolates was 8.4%, and resistance to most antimicrobial agents was higher than that among Enterococcus faecalis isolates. Among nonmeningeal Streptococcus pneumoniae isolates from pediatric and adult patients, the respective detection rates of penicillin-nonsusceptible strains, comprising penicillin-intermediate S. pneumoniae and penicillin-resistant S. pneumoniae , were low at 3.9% and 5.9%. Except for Klebsiella spp., which showed resistance rates of 20.1% to imipenem and 20.8% to meropenem, resistance among other Enterobacterales was generally below 15.0%. After excluding intrinsically resistant genera, Enterobacterales remained highly susceptible to tigecycline and colistin, with resistance rates of ≤ 6.8%. Resistance rates to imipenem and meropenem were 22.6% and 18.8%, respectively, in Pseudomonas aeruginosa, and 72.0% and 71.9%, respectively, in Acinetobacter baumannii . Antimicrobial resistance among clinical isolates from major regions of China remained a serious concern in 2025. These surveillance findings should inform rational antimicrobial selection and support efforts to limit the emergence and spread of resistance.
Abstract This study sought to elucidate the protective role of Roundabout 4 (ROBO4) in maintaining blood-brain barrier (BBB) integrity during influenza virus-induced barrier dysfunction. The functional role of ROBO4 was examined using both in vivo and in vitro influenza infection models. Specifically, mechanistic investigations focused on the viral-induced down-regulation of ROBO4 and its subsequent impact on the vascular endothelial growth factor (VEGF)-VEGF receptor 2 (VEGFR2)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/endothelial nitric oxide synthase (eNOS) signaling axis and key tight junction (TJ) proteins, including Claudin-1, Occludin, and Claudin-5. Additionally, lentiviral-mediated ROBO4 overexpression was utilized to validate its potential protective efficacy. The results showed that influenza virus infection significantly down-regulated ROBO4 expression, which in turn enhanced the interaction between VEGF and its receptor, VEGFR2. Consequently, this molecular event triggered the PI3K/AKT/eNOS signaling cascade, leading to the depletion of TJ proteins and subsequent impairment of BBB integrity. Notably, ROBO4 overexpression effectively inhibited the VEGF-VEGFR2 interaction, suppressed the activation of the PI3K/AKT/eNOS pathway, and restored the expression levels of Claudin-1 and Occludin, thereby preserving barrier function. Overall, by identifying ROBO4 as a critical regulator of BBB integrity, this study highlights its potential as a novel therapeutic target. Furthermore, these findings provide theoretical support for the development of preventative and interventional strategies to mitigate the neurological complications associated with influenza virus infections.
Abstract This study analysed bacterial infections and antimicrobial resistance (AMR) profiles of the top five pathogens in adult intensive care units (ICUs) and paediatric ICUs (PICUs) in China from 2017 to 2022, aiming to inform AMR surveillance and treatment guidelines. Antimicrobial susceptibility was assessed via automated systems or the Kirby-Bauer method, interpreted per Clinical and Laboratory Standards Institute breakpoints, and analysed using WHONET 5.6. 22,278 and 152,117 unduplicated clinical bacterial isolates were collected from 17 PICUs and 44 adult ICUs. The rate of Gram-negative bacteria was higher in adult ICUs than in PICUs (78.5% vs. 62.6%, P < 0.001). The top five clinical isolates in both departments were Staphylococcus aureus , Acinetobacter baumannii , Klebsiella pneumoniae , Pseudomonas aeruginosa , and Escherichia coli. From 2017 to 2022, E. coli , K. pneumoniae , and P. aeruginosa generally showed decreased resistance to most antimicrobial agents in adult ICUs, but increased resistance in PICUs. Resistance of A. baumannii and S. aureus to most antimicrobial agents decreased in both PICUs and adult ICUs. The resistance proportions of E. coli to ceftazidime-avibactam, ceftriaxone, and imipenem; K. pneumoniae to ceftazidime-avibactam and ceftriaxone; and P. aeruginosa to amikacin were higher in PICUs than in ICUs. For other detected antibacterial agents, all five pathogens showed higher resistance in adult ICUs than in PICUs ( P < 0.05). These data reveal a severe AMR burden in ICUs, with notable differences between PICUs and adult ICUs. Enhanced ICU-specific AMR surveillance, rigorous infection control, and optimised antibiotic stewardship are imperative to counter multidrug-resistant pathogens. Although escalating resistance trends remain a major concern in PICUs, significant reductions in resistance for several pathogens in adult ICUs likely reflect the impact of strengthened antimicrobial stewardship and infection control programs in China.
Microplastic pollution poses significant threats to marine ecosystems and human health, requiring efficient and standardized monitoring within a One Health framework. This study presents the development and evaluation of an artificial intelligence (AI)-driven image segmentation model for detecting microplastics in images collected along the coastline of Sousse, Tunisia, a region particularly vulnerable to high pollution. The dataset includes 1080 images of plastics categorized into classes: high-density polyethylene (HDPE), low density polyethylene (LDPE), polyamide (PA), polyethylene terephthalate (PET), polypropylene (PP), and polystyrene (PS), as well as an "other" category comprising non-plastic and miscellaneous beach-collected items, such as cigarette butts and dried Posidonia oceanica. To ensure reliability with AI analysis, all fragments were first identified, photographed under controlled lighting to capture morphological variability. Using advanced polygon segmentation, the model enables pixel-level annotation and outperforms traditional bounding box methods, especially for irregularly shaped particles. Background subtraction and adaptive thresholds enhance accuracy, reducing false positives. Model performance is evaluated using the Intersection-over-Union (IoU) metric, measuring overlap between predicted segmentations and manually annotated ground truth data. Additionally, it discusses the ecological relevance of identified microplastic types in the regional context, highlighting potential impacts on coastal biodiversity and environmental health. Findings contribute to the growing field of marine pollution monitoring by demonstrating the practical utility and challenges of AI-powered segmentation, supporting future efforts toward scalable, ecologically informed, and standardized microplastic detection methods.
As an obligate intracellular parasite, Toxoplasma gondii is a widely distributed zoonotic pathogen posing a substantial threat to both human and animal health. Interestingly, T. gondii contains only a single and divergent mitochondrion that supports its metabolism and rapid proliferation. Coenzyme Q (CoQ), a lipid-soluble electron carrier, is essential for mitochondrial electron transport, cellular energy metabolism, and redox homeostasis. CoQ biosynthesis involves a series of enzymatic reactions, and its key proteins have been identified and functionally characterized in various eukaryotes. However, the functions of CoQ biosynthetic proteins in T. gondii remain poorly understood. In this study, we identified TgCoq4 as a conserved component protein of the CoQ biosynthetic pathway in T. gondii, and confirmed that it displays oxidative decarboxylation activity. Genetic depletion of TgCoq4 leads to impaired parasite growth, accompanied by mitochondrial structural damage and functional deficiency. In addition, TurboID-based proximity labeling technology combined with mass spectrometry analysis confirmed that TgCoq4 is spatially associated with multiple CoQ biosynthetic proteins. Therefore, this study identifies TgCoq4 as a key component of CoQ biosynthesis and provides new insights into mitochondrial mechanism in T. gondii.
Abstract Antimicrobial resistance (AMR) in livestock production systems represents a major One Health concern due to the potential transfer of resistant bacteria and resistance genes along the food chain and into the environment. This study investigated the occurrence of multidrug-resistant bacteria and associated antibiotic resistance genes (ARGs) in a pig production farm in Portugal. Samples were collected from slurry, animal skin swabs, feed, soil, and processed meat across three production batches. Enterobacteriaceae, Staphylococcus, and Enterococcus isolates were recovered using selective media, identified by 16S rRNA gene sequencing, and assessed for antimicrobial susceptibility using the disk diffusion method. ARGs were detected by multiplex PCR. Thirty-two isolates were identified, including Escherichia coli, Enterobacter spp., Kosakonia cowanii, Staphylococcus haemolyticus, Staphylococcus borealis, Staphylococcus devriesei, Staphylococcus pasteuri, Enterococcus faecalis, and Enterococcus faecium. Tetracycline resistance was widespread across all genera, with frequencies of 56.3% in Enterobacteriaceae, 77.8% in Staphylococcus, and 66.7% in Enterococcus. Among E. coli, bla TEM variants and tet(A) were the most frequently detected resistance genes, alongside occasional sulfonamide (sul1, sul2) and AmpC (CIT) determinants. Staphylococcus isolates exhibited high rates of resistance to clindamycin, erythromycin, and chloramphenicol, supported by the presence of ermB, ermC, and blaZ. Enterococcus isolates carried pbp5 and erm genes. Overall, 76.6% of isolates showed Multiple Antibiotic Resistance Index values above 0.2, indicating high selective pressure within the farm environment. These findings demonstrate the circulation of multidrug-resistant bacteria and clinically relevant ARGs across the animal-environment interface, emphasizing the need for improved sanitation, responsible antibiotic use, and strengthened antimicrobial stewardship within pig production systems.
Abstract The convergence of resistance to last-resort antibiotics in Escherichia coli represents a growing global threat. Yunnan, located at the edge of the Qinghai—Tibet Plateau and characterized by complex topography, monsoon climate, and highly diverse ecological niches, provides a unique setting in which the emergence of resistance in commensal reservoirs remains incompletely understood. Here, we performed selective screening of 1000 intestinal samples from healthy individuals in Yunnan, China, and identified 7.2% (72/1000) of bla NDM-positive E. coli isolates. Among these, 6.9% (5/72) of isolates co-harbored additional resistance determinants, including tet(X4) (n = 3) or mcr-1 (n = 2). All co-harboring isolates exhibited multidrug-resistant phenotypes, and conjugation assays confirmed the transferability of these resistance genes. Phylogenetic analysis incorporating 2194 publicly available genomes from the NCBI database revealed that bla NDM was distributed across diverse genetic backgrounds, supporting horizontal gene transfer as the primary driver of dissemination. Genomic context analysis further demonstrated both conserved and diverse resistance gene environments associated with mobile genetic elements. Together, these findings reveal the emergence of transferable combinations of bla NDM with tet(X4) or mcr-1 in intestinal E. coli, highlighting the gut microbiota as a potential reservoir for the accumulation and spread of resistance to multiple last-resort antibiotics.
Abstract Ceftiofur resistance in canine clinical Escherichia coli is usually associated with extended-spectrum β-lactamases (ESBLs) or AmpC β-lactamases. However, some isolates display elevated ceftiofur minimum inhibitory concentrations (MICs) without carrying these known resistance genes. In this study, we identified a phenotype-genotype discordant canine clinical E. coli isolate named 231255, with a ceftiofur MIC of 8 μg/mL. Routine resistance gene screening detected only bla TEM-1 and a chromosomal bla EC variant, bla EC-1149, which could not adequately explain the elevated ceftiofur MIC. To investigate the underlying mechanism, a genomic library was constructed from genomic DNA of isolate 231255 and screened on ceftiofur-containing plates. Positive clones revealed one candidate determinant: an altered ftsI fragment encoding penicillin-binding protein 3 (PBP3) with a four-amino-acid YRIN insertion downstream of residue P333, previously identified in human E. coli. Previous studies have shown that this type of YRIN/YRIK insertion alone can reduce susceptibility to PBP3-targeting β-lactams, particularly aztreonam and ceftazidime. Functional validation showed that recombinant plasmids carrying the altered ftsI consistently increased the ceftiofur MIC to 4 μg/mL in different recipient backgrounds. These findings provide experimental evidence that ftsI/PBP3 alteration can elevate ceftiofur MIC in canine E. coli. Notably, the isolate belonged to ST410, and phylogenetic analysis indicated that it was not confined to a dog-associated background but instead clustered within a broader lineage shared across multiple sources, highlighting the need for potential dissemination of this mechanism and its associated resistant lineages at the human-companion animal interface.
Abstract The emergence of extended-spectrum β-lactamase-producing Escherichia coli (ESBL-Ec) in food animals poses a significant public health concern. While the prevalence and genetic traits of ESBL-Ec in commercial farms have been widely studied, data from backyard farms remain limited. Herein, we investigated the occurrence of ESBL-Ec among backyard animals across 12 villages in Shandong province, China. Notable variations in ESBL-Ec prevalence were observed among animal species, ranging from 78.7% in pigs, 61.5% in cattle, 50.0% in sheep, 28.6% in ducks, 14.1% in dogs, 11.5% in chickens, and 5.3% in cats. In addition, 74 ESBL-Ec were recovered from 180 flies captured in household backyards. Genomic analysis revealed substantial genetic diversity, with dominant sequence types (STs) including ST10, ST48, ST206, and ST744 identified across multiple host species. Bayesian clustering showed that all seven lineages were distributed across at least two sources, suggesting frequent inter-host transmission. Several isolates from different animal species exhibited high similarity (0–65 single nucleotide polymorphisms), indicating potential clonal spread. The bla CTX-M-14, bla CTX-M-65, and bla CTX-M-55 genes were predominant, accounting for 79.1% of detected bla CTX-M variants, and were predicted to be located on IncFIB, IncHI2, IncFII, and IncX1 plasmids. Notably, isolates from backyard animals shared STs and bla CTX-M genotypes with previously reported human and environmental isolates from the same area, suggesting potential transmission across hosts and environmental niches. These findings highlight backyard farming systems as potential reservoirs of ESBL-Ec and emphasize the need for improved antimicrobial stewardship and management practices to limit resistance spread.
Abstract Mycoplasma bovis is a globally significant pathogen in dairy cow associated with pneumonia, arthritis, mastitis, otitis, and reproductive failure with different age, resulting in substantial economic losses in the cattle industry. M. bovis relies on host-derived cholesterol for its membrane integrity and intracellular survival, as it cannot synthesize this lipid itself. However, the specific mechanisms by which M. bovis alters host cell cholesterol metabolism remain unclear. Simvastatin, a common cholesterol-lowering drug that inhibits HMG-CoA reductase, provides a tool to investigate this host–pathogen interaction. This study aimed to: (1) determine the effect of M. bovis infection on cholesterol metabolism in bovine mammary epithelial cells (bMECs); (2) assess the role of intracellular cholesterol in bacterial proliferation; (3) investigate whether cholesterol depletion activates anti-bacterial autophagy via the AMPK–mTORC1–TFEB pathway; and (4) evaluate the anti-M. bovis efficacy of simvastatin in both in vitro and in vivo models. Results showed that in bMECs, M. bovis infection dysregulated the expression of cholesterol metabolism genes. Depleting cholesterol (using MβCD or simvastatin) significantly reduced the intracellular M. bovis load. This reduction was accompanied by the activation of autophagy and the AMPK–mTORC1–TFEB signaling pathway. In a murine mastitis model, simvastatin treatment enhanced autophagy, reduced the M. bovis burden, and mitigated infection-induced pathology. Our findings demonstrate that simvastatin exerts anti-M. bovis effects by reducing host cholesterol, activating the AMPK–mTORC1–TFEB pathway, and enhancing autophagic clearance of the bacteria. This reveals a novel potential therapeutic strategy for combating M. bovis infections.
Abstract Zoonotic viruses, such as the Nipah virus (NiV), Hendra virus (HeV), Rift Valley fever virus (RVFV), and Crimean–Congo hemorrhagic fever virus (CCHFV), pose substantial threats to global public health. Early and precise detection of these pathogens is crucial for their effective prevention and control; however, there are limitations to field applications of conventional diagnostic approaches which rely on large equipment or complex operations. In this study, we developed a rapid one-tube visual detection assay for four viruses, respectively, by integrating reverse transcription recombinase polymerase amplification (RT-RPA) with CRISPR/Cas12a technology. The system incorporates highly specific CRISPR RNAs that target conserved regions of the nucleocapsid (N) genes from the 4 viruses under isothermal conditions at 39 °C. To simulate clinical conditions, we constructed recombinant vesicular stomatitis viruses expressing the N gene (rVSV-N) of NiV, HeV, RVFV, and CCHFV, respectively. Results indicated that the RT-RPA–CRISPR/Cas12a system could visually detect rVSV-N in lung samples from infected mice, with a detection limit of 1 copy/μL of viral RNA, within 40 min in a one-tube system at 39 °C isothermally. Our highly sensitive, specific, and visual one-tube detection system for NiV, HeV, RVFV, and CCHFV offers significant potential for early and accurate on-site detection of these viruses.
Screening for Leishmania infection in dogs remains a major constraint in the control of dog-associated visceral leishmaniasis. This study evaluated the performance of four diagnostic assays—rK39, multi-antigen rapid diagnostic test strip (MRD), indirect fluorescent antibody test (IFAT), and quantitative polymerase chain reaction (qPCR)—using peripheral blood samples from asymptomatic dogs in endemic areas, with the aim of informing context-specific screening strategies. A total of 175 dogs were tested. Agreement between assays was assessed using Cohen’s and Fleiss’ kappa statistics, and paired differences were evaluated using McNemar’s test. Relative performance was further estimated using two-by-two tables and latent class analysis. The highest proportion of positives was detected by qPCR (30.9
Mycobacterium marinum is one of the main pathogens in aquaculture with direct or indirect impacts on human, animal, and environmental health, significantly slowing progress in productive and economic sectors. Microalgae are a potential source of new antimicrobial compounds; however, they are still little explored. In the present study, we propose the use of an alternative culture medium to produce bioactive compounds from Conticribra weissflogii against the important mycobacterial pathogen M. marinum. Complementary tools, 1H nuclear magnetic resonance and high-performance liquid chromatography, were employed to characterize the major compounds present in the bioactive extracts. The nonpolar extracts were rich in fatty acids and pigments, such as fucoxanthin, and exhibited potent antimycobacterial activity. The hexanic and ethyl acetate extracts from C. weissflogii were effective against M. marinum with a minimum inhibitory concentration of 0.1 mg·mL−1 and low cytotoxicity. This pathogen poses a threat to human society, affecting aquaculture production, leading to alarming economic losses, contaminating natural resources, and increasing the spread of zoonotic diseases with high morbidity and mortality. Addressing this issue through a One Health approach is necessary. In this context, the use of natural products with antimicrobial properties, such as the diatomaceous extract studied here, may be an effective solution.
Abstract The accelerating spread of extended-spectrum β-lactamase (ESBL)-mediated resistance threatens the clinical utility of β-lactam antibiotics across human, veterinary, and environmental health sectors, underscoring the need for reliable genomic frameworks within a One Health context. Public genomic repositories are central to antimicrobial resistance (AMR) research, yet their utility is constrained by annotation inconsistency, nomenclature ambiguity, and biased (non-random) data deposition. Here, we performed phylogeny-guided curation and variant-level fingerprinting of ESBL gene families using 2746 β-lactamase sequences retrieved from UniProt, GenBank, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Comprehensive Antibiotic Resistance Database (CARD), comprising bla CTX-M (n = 1316), bla TEM (n = 895), and bla SHV (n = 535). Conserved motif analysis, domain architecture assessment, and KEGG/CARD cross-validation revealed extensive annotation discordance: only 42.9% (1177/2746) of entries were correctly annotated, 3.8% (104/2746) required reclassification, and 53.3% (1465/2746) remained insufficiently resolved. Phylogenetic reconstruction enabled reclassification of cryptic variants and delineation of species-associated phylogroups consistent with lineage compatibility and horizontal gene transfer. Significant gene-host structuring was observed across species (χ2 = 479.64, df = 28, P = 1.04 × 10⁻83), with Escherichia coli enriched for bla CTX-M-15 (n = 124), Klebsiella spp. for bla SHV-11 (n = 105), and Salmonella spp. for bla TEM-1 (n = 52). These dominant allele patterns were independently supported by isolate-level genome surveillance metadata from NCBI Pathogen Detection (Create date ≤ 2022), confirming bla CTX-M-15 predominance in E. coli/Shigella and bla SHV-11 predominance in Klebsiella pneumoniae. Subvariant diversity (Shannon index) was highest in Acinetobacter spp. (H′ = 3.578), Enterobacter spp. (H′ = 3.385), and Proteus spp. (H′ = 3.251), whereas Vibrio spp. exhibited restricted diversity (H′ = 0.95). Morphological stratification confirmed the predominance of Gram-negative bacilli among ESBL-encoding organisms (99%; χ2 = 33.62, df = 2, P = 5.00 × 10−8). Analysis of database submissions spanning 1998–2022 demonstrated statistically significant temporal shifts in ESBL family representation across years (χ2 = 254.20, df = 36, P = 1.21 × 10⁻34), with a marked rise in bla CTX-M representation after 2016 period. Ecological attribution using curated metadata indicated that 85% (n = 2324) of ESBL entries were linked to species occurring at the human–animal interface, likely reflecting database deposition bias rather than true reservoir prevalence. Collectively, this study provides a high-confidence, variant-resolved ESBL fingerprint derived from curated public databases, highlights systematic annotation limitations, and supports database-aware interpretation of comparative AMR research within a One Health framework.
Abstract Traditional Chinese medicine (TCM) plays a crucial role in maintaining the intestinal barrier, which is considered integral to overall health and is connected with the functions of the spleen and stomach and the balance of qi and blood. TCM principles, such as "separating the pure from the turbid," are related to gut flora equilibrium, with imbalances corresponding to the "struggle between good and evil." TCM supports the intestinal barrier through methods such as invigorating qi and strengthening the spleen. In this review, authoritative databases such as PubMed, Web of Science, and Google Scholar were systematically searched for studies on the protective effects of TCM and its constituents on the intestinal barrier from 2018 to 2025. The results show that TCM and its components have multiple advantages in maintaining intestinal barrier function. Classic formulations such as Qingchang Wenzhong and Huangqin decoction protect the intestines through various pathways, including regulating the gut microbiota and enhancing immune function. Single herbs such as Coptis chinensis (Huang Lian) and Scutellaria baicalensis (Huang Qin), whose properties include clearing heat and detoxifying and tonifying qi, directly or indirectly promote intestinal health. Additionally, active compounds in TCM, such as flavonoids and saponins, contribute to intestinal homeostasis through mechanisms such as anti-inflammatory and antioxidant effects. The multicomponent, multitarget characteristics of TCM provide comprehensive and long-lasting efficacy in intestinal barrier protection. This article summarizes the current research status of TCM in treating intestinal barrier injury and highlights future research directions, offering a theoretical foundation and research insights for further exploration of the role of TCM in safeguarding intestinal barrier function. Graphical Abstract
Abstract Silicon dioxide (SiO2) is widely used as an anti-caking agent in food and cosmetics. PANoptosis is a highly complex inflammatory programmed death pathway and plays a crucial role in disease and toxic injury. This study investigated the damage caused by 50 nm, 300 nm, and 1 μm SiO2 particles to the mice's colon and compared the toxicity differences among these particle sizes. The results showed that SiO2 exposure induced ciliary damage and reduced intracellular serine content by inhibiting spermidine/spermine N1-acetyltransferase 1/2 (SAT1/2), thereby affecting mitochondrial one-carbon metabolism. Meanwhile, downregulation of mitochondrial overlapping with the m-complex activity 1 (OMA1)/optic atrophy 1 (OPA1) led to mitochondrial damage and mitochondrial reactive oxygen species (mtROS) release, eventually activating RIPK1-PANoptosome-mediated PANoptosis. Among them, 300 nm had the strongest effect on mitochondrial damage, 50 nm on apoptosis, and 1 μm on amino acid metabolism, one-carbon metabolism, necroptosis, and pyroptosis. In conclusion, SiO2 exposure induced RIPK-mediated PANoptosis via the serine/mitochondrial/mtROS pathway. Graphical Abstract
Abstract Norovirus (NoV) is a leading cause of acute gastroenteritis worldwide. Irrigation water has been confirmed as a vector for transmitting NoV to crops. The study aimed to analyze the prevalence of NoV in irrigation water across China and assess its implications for food safety. Between December 2023 and April 2024, 200 irrigation water samples were collected from 27 cities. Viral concentration was performed using tangential flow filtration, followed by RNA extraction and RT-qPCR detection for NoV genogroups GI and GII. NoV was detected in 17 samples (8.50%, 17/200). Among the positive samples, GI was identified in 8 (4.00%), GII in 14 (7.00%), with 5 samples (2.50%) positive for both genogroups. The results indicated that the detection rate of NoV in irrigation water exhibited seasonal variation, with a significantly higher rate observed in winter compared to spring (p < 0.001). Irrigation water in China is a reservoir for NoV, with a significantly higher contamination level in winter. These findings highlight irrigation water as a potential risk factor for foodborne and waterborne NoV transmission and warrant focused attention in public health strategies.
Abstract ADP ribosylation factor like 4c (Arl4c), a member of the small GTPase superfamily, has been implicated in tumor progression in various human cancers. However, its role in canine cancers remains largely unexplored. In this study, we demonstrated that Arl4c exhibits significant oncogenic activity in canine melanoma. Immunohistochemical analysis of canine clinical tumor samples indicated that Arl4c is highly expressed in canine melanoma. Functional assays in canine melanoma cell lines showed that Arl4c overexpression promotes cell proliferation and migration. Furthermore, a bioinformatics analysis identified a novel association between Arl4c and the nuclear factor kappa B (NF-κB) signaling pathway, subsequent experimental validation using Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) confirmed that Arl4c positively regulates NF-κB pathway activation, linking it to key tumorigenic mechanisms. These findings establish Arl4c as a pro-tumorigenic factor in canine melanoma and underscore its potential as both a diagnostic biomarker and a therapeutic target in veterinary oncology. Our study not only highlights Arl4c's role in canine tumorigenesis, but also reinforces the value canine models for informing novel treatment strategies for human cancers.
Abstract Sensitization of gold nanoparticle-based lateral flow assays (AuNPs-LFAs) will enable them to meet the requirements well across various applications. However, the sensitization of AuNPs-LFA commonly needs complicated operations, which will hinder their practical application. Herein, a novel LFA test strip integrated with a deceleration membrane (DM) was constructed, in which the DM was introduced between the test line and control line. The DM slows down the flow of sample solution in LFA test, thereby providing longer time for the binding between AuNPs-labeled antibody and target. Three types of DMs were prepared by modifying glass fiber membranes with polyvinyl alcohol (PGF), acrylic resin (AGF) and silicone resin (RGF), respectively. A common pesticide, carbendazim (CAR) was selected as a model target, and the analytical performance of the LFA integrated with three DMs (PGF-LFA, AGF-LFA, and RGF-LFA) in competitive format were evaluated. The results demonstrated that their visual detection limits for CAR were 50 ng/mL, 20 ng/mL, and 10 ng/mL, respectively. The limits of detection (LODs) for CAR were 0.38 ng/mL, 0.31 ng/mL, and 0.27 ng/mL using a portable reader, respectively. Compared with traditional AuNPs-LFA strip, their sensitivity had increased by 1.5-fold, 3.5-fold, and 7.5-fold, respectively. When applied to apple samples, the RGF-LFA for CAR achieved recovery rates of 96%–101.5%. In addition, the sensitization of RGF-LFA in sandwich format also was evaluated to be excellent by selecting SARS-CoV-2 N protein as model target, showcasing the versatility of this DM-integrated strategy.
Abstract Liver X receptors (LXRs) are members of the nuclear receptor superfamily, and LXR agonists have been recognized for their neuroprotective potential. This study aimed to investigate the regulatory effects and underlying molecular mechanisms of the LXR agonist LXR623 in modulating microglial polarization during infection with Japanese encephalitis virus (JEV). JEV infection significantly upregulated the expression of host LXRs. Treatment with LXR623 ameliorated JEV-induced morphological alterations and cellular damage in BV2 microglial cells. LXR623 also suppressed M1 macrophage polarization and promoted a shift toward the M2 phenotype, thereby attenuating the inflammatory response associated with JEV infection. These effects may be exerted through the pathway involving Toll-like receptor 4, nuclear factor-κB, and the NOD-like receptor family pyrin domain containing 3. The LXR inverse agonist SR9243 partially blocked the protective effect of LXR623. In JEV-infected mice, LXR623 administration significantly reduced mortality and alleviated clinical symptoms of encephalitis. These findings indicate that LXR activation modulates JEV-induced microglial polarization and confers neuroprotective effects in vivo, establishing a novel host-directed strategy with cross-species applicability for the prevention of Japanese encephalitis.
