Triple-negative breast cancer (TNBC) is aggressive with limited treatments. Although the natural flavonoid Apigenin (API) shows anti-tumor potential, its mechanism in TNBC remains unclear. This study investigated API's role in inducing ferroptosis and inhibiting glycolysis to suppress TNBC. Molecular docking predicted API's binding to ferroptosis- and glycolysis-related proteins. In vitro, 4T1 and MDA-MB-231 TNBC cells were used to assess API's effects on viability, migration, invasion, and key metabolic markers (Fe2+, MDA, ROS, GSH, lactic acid, glucose, ATP, OCR, ECAR), and protein expression (GPX4, SLC7A11, TFR, FPN1, FTH1, FTL, FSP1, PKM2, GLUT1, GLUT4, HK2, LDHA) using RT-qPCR and Western blotting. In vivo, a 4T1 tumor xenograft model evaluated API's impact on tumor growth, protein expression, and toxicity (H&E staining). Molecular docking indicated good binding affinity of API with ferroptosis- and glycolysis-related proteins. In vitro, API inhibited the viability, migration, and invasion of TNBC cells. API significantly increased Fe2+, MDA, and ROS levels while decreasing reduced GSH levels, downregulated GPX4, SLC7A11, FPN1, FTH1, FTL, and FSP1 expression, and upregulated TFR expression, indicating ferroptosis induction. API also decreased lactic acid, ATP, and ECAR levels while increasing intracellular glucose and OCR levels, downregulating PKM2, GLUT1, GLUT4, HK2, and LDHA expression, demonstrating glycolysis inhibition. In vivo, API significantly inhibited tumor growth in the 4T1 xenograft model without obvious toxicity and regulated the expression of ferroptosis and glycolysis-related proteins. These findings establish API as a promising natural anti-TNBC drug by simultaneously targeting ferroptosis and aerobic glycolysis.

Abstract Helminths systemically suppress host immunity, yet whether they impose immune tolerance by rewiring host-associated microbial metabolism remains unclear. Here we show that Trichinella spiralis infection remodels intestinal tryptophan metabolism to generate an AhR-dependent regulatory immune state. T. spiralis infection enriched the commensal bacterium Ligilactobacillus murinus , which converted tryptophan into indole-3-lactic acid (ILA), a microbial metabolite that directly engaged the aryl hydrocarbon receptor. Antibiotic-mediated microbiota depletion abolished infection-induced ILA accumulation, AhR activation and Treg/Th17 rebalancing, whereas fecal microbiota transplantation from infected donors or supplementation with L. murinus or ILA restored these effects. Pharmacological blockade or genetic deletion of AhR eliminated the ability of T. spiralis , L. murinus and ILA to restrain LPS-induced cytokine-storm-like lung inflammation, establishing AhR as an essential host node in this circuit. Extending these findings to viral inflammatory disease, oral ILA improved survival and reduced pulmonary immunopathology in SARS-CoV-2-infected K18-hACE2 mice. Re-analysis of human COVID-19 metabolomic data further revealed reduced circulating ILA in severe disease. These findings define a helminth-remodeled microbial tryptophan metabolic pathway that promotes disease tolerance and identify the ILA–AhR axis as a candidate postbiotic strategy for limiting hyperinflammatory tissue injury.

Cancer cachexia is a multifactorial syndrome of progressive skeletal muscle wasting and functional decline that affects 50-80% of patients with advanced malignancies, frequently overlaps with sarcopenia, and contributes to 22-30% of cancer-related deaths. Effective therapies remain lacking, in part because the driving mechanisms are incompletely understood. Systemic inflammation-particularly interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α)-has long been considered central to muscle wasting, yet cytokine-targeted trials have shown limited efficacy, prompting investigation of additional pathways. Among these, endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) have emerged as candidates, and this review focuses specifically on the IRE1α/XBP1 branch. The rationale rests on three observations from recent preclinical studies: XBP1s activity is increased in cachectic muscle; XBP1s occupies regulatory regions of autophagy-lysosome and ubiquitin-proteasome genes, a direct transcriptional link to protein degradation that distinguishes it from the translation-attenuating PERK and folding-oriented ATF6 branches; and genetic or pharmacological suppression of IRE1α/XBP1 attenuates wasting in these models. We examine how tumor-derived signals activate IRE1α/XBP1 to upregulate both the autophagy-lysosome pathway (ALP) and ubiquitin-proteasome system (UPS); its crosstalk with inflammatory (JAK-STAT3, NF-κB) and metabolic (mitochondrial dysfunction, fatty acid metabolism) networks; the evidence across cancer models and clinical contexts; and the therapeutic potential of IRE1α inhibitors, XBP1-directed strategies, and nutritional approaches including arginine. We frame the ER stress-autophagy axis as a mechanistically plausible, potentially tractable therapeutic target that requires further cross-model and clinical validation.

The preparation of nanomaterials is limited by operator-induced effects, which can have significant implications for nanomaterial reproducibility and applicability. In this study, gold nanoparticles (AuNPs) were controllably synthesized using an automated technique and were subsequently combined with a surface-enhanced Raman spectroscopy (SERS) system to achieve the automated trace detection of antibiotics. Following the automated and controlled cyclochemical synthesis of the AuNPs, their optical properties, morphologies, and sizes were evaluated using absorption spectroscopy and atomic force microscopy. Subsequently, an automated cyclochemical synthesis-SERS system was developed and applied for the detection of three common aquaculture antibiotics. Using this system, the average particle size of the synthesized spherical AuNPs was controlled at 85 +/- 10 nm, and the synthesized AuNPs exhibited significant SERS enhancement (enhancement factor = 1.6 x 10(6)). The SERS limits of detection for enrofloxacin, sulfamethazine, and nitrofurantoin were determined to be 0.09, 0.01, and 0.57 mu g/L, respectively. The linear correlation coefficients of the serial concentrations of the three antibiotics with Raman characteristic peak intensities were >0.95, indicating a good trace detection capability. A mixed solution of these three antibiotics was also tested, providing clearly distinguishable characteristic peaks for the three target compounds. In a spiked recovery experiment of enrofloxacin in aquaculture water, the relative standard deviation was similar to 10 % after six parallel determinations, and the spiked recovery ranged from 64 % to 87 %. The results of this study demonstrate that combining the automatically controlled synthesis of nanomaterials with SERS has great potential for the in situ detection of antibiotic residues in cultured waters.

Aloin (ALO), an anthraquinone derived from Aloe vera, exhibits antitumor activity; however, its precise mechanisms of action remain unclear. In this study, in silico molecular docking analysis first revealed that Aloin (ALO) bound effectively to ferroptosis-related proteins (SLC7A11, GPX4, ACSL4, and TFR1). Subsequently, in vitro ALO treatment triggered ferroptosis hallmarks in human cervical cancer cell line Hela and mouse colon cancer cell line MC38 at both transcriptional and protein levels-downregulating SLC7A11/GPX4, upregulating ACSL4/TFR1, with Fe2+/ROS accumulation, GSH depletion, and ferroptosis-specific mitochondrial cristae loss. Furtherly, ALO inhibited cancer cell proliferation, migration, and invasion, effects that were reversed by the ferroptosis inhibitor Ferrostatin-1. Concurrently, ALO induced autophagy, as evidenced by increased levels of LC3, LaminB1, and ULK1, decreased levels of P62, and TEM-visualized autophagosomes. Notably, the autophagy inhibitor chloroquine reversed ALO-induced ferroptosis, NEDD8 downregulation, and NEDP1 upregulation, linking ALO-induced autophagy to NEDD8 de-NEDDylation. Genetic and pharmacological perturbation of the NEDD8 pathway confirmed this: NEDD8 inhibition enhanced, while NEDP1 knockdown attenuated, ALO-induced ferroptosis. Co-immunoprecipitation and laser-scanning confocal microscope confirmed a direct NEDD8-GPX4 interaction diminished by ALO, positioning GPX4 as a key effector. In in vivo study, ALO effectively inhibited cancer cell growth in a murine colon carcinoma MC38 xenograft models, while exhibiting no obvious toxicity or side effects in mice. Moreover, ALO exhibited the same regulatory effects and trends on ferroptosis-related proteins in vivo as those in vitro. In summary, this study reveals a novel mechanism that ALO-induced autophagy promotes NEDD8 de-NEDDylation, driving ferroptosis via the NEDD8-GPX4 axis to suppress cancer cell growth.

Trichinella spiralis (T. spiralis), a zoonotic nematode that causes severe myositis and systemic morbidity, sustains chronic muscle parasitism through evolutionary adaptations; however, this globally prevalent disease lacks targeted therapies to disrupt chronic infection. Although the heme transport protein HRG-1 has been characterized as an intervention target in free-living species (e.g., Caenorhabditis elegans) and hematophagous parasites (e.g., Haemonchus contortus), the molecular machinery governing heme acquisition in the nonhematophagous parasite T. spiralis remains uncharacterized, and no drugs targeting HRG-1 have been reported until now. Herein, we demonstrate that T. spiralis, a parasite that lacks the ability to synthesize heme autonomously, has evolved a sophisticated mechanism to scavenge and utilize heme from its host. By employing an aspartic protease to degrade host hemoglobin and myoglobin in the parasitic niche, T. spiralis is able to liberate heme for its own growth and survival. The structurally and functionally conserved Ts-HRG-1 protein plays a key role in transporting heme to the entire worm, particularly to functional organs, such as the cuticle and stichosome. More importantly, we discovered that the interaction between Ts-HRG-1 and Ts-ATP6V0C results in the formation of a functional complex that is essential for the parasite's heme acquisition. The intervention effect achieved by Ts-ATP6V0C RNAi or inhibiting the activity of Ts-ATP6V0C with bafilomycin A1 (BafA1) was consistent with Ts-HRG-1 RNAi, resulting in impaired heme uptake, developmental arrest and a reduced larval burden in mouse hosts. These findings enhance our understanding of the parasite's heme acquisition mechanism and identify the development of drugs that target proteins that interact with HRG-1 as a new direction in anthelminthic drug research.

Brucellosis is a highly contagious zoonotic disease characterized by a non-specific clinical presentation and complex disease progression and outcome. Hemophagocytic lymphohistiocytosis (HLH) is an abnormal immune response syndrome marked by potentially fatal cytokine storms. Brucella-associated HLH is exceedingly rare and associated with a high mortality rate. We report a case involving a 23-year-old male residing in a brucellosis-endemic region, with a documented history of exposure to cattle and sheep. He was admitted to the hospital presenting with fever and arthralgia. Laboratory tests indicated splenomegaly, pancytopenia, elevated serum aminotransferases and ferritin levels, as well as decreased fibrinogen levels. Blood and bone marrow cultures yielded negative results. The Brucella serum agglutination test returned a positive result (titer, 1:200). Bone marrow aspirate results revealed an increased number of hemophagocytes, and PET-CT scans demonstrated splenomegaly, suggesting hemophagocytic changes. Following a comprehensive exclusion of hematological malignancies and neoplastic diseases, the patient was diagnosed with probable Brucella infection complicated by secondary HLH. Standard anti-brucellosis therapy was initiated immediately upon hospital admission. Remarkably, significant clinical improvement was observed within 7 days of targeted antibiotic treatment, without the need for corticosteroid therapy. This case, when analyzed alongside a systematic review of 12 published HLH cases associated with brucellosis, underscores the importance of maintaining a heightened clinical suspicion for this life-threatening complication in endemic regions, which may facilitate earlier diagnosis and optimized antimicrobial management strategies.

Osteoarthritis (OA) is a degenerative joint disease that is a major cause of deformity, swelling, pain and even loss of function in the knee joints of the elderly. Pantothenic acid (PA) plays a protective role in many organs due to its antioxidant and anti-inflammatory properties. Herein, we aimed to assess the protective role of PA on osteoarthritis and investigate the underlying molecular mechanism. The levels of inflammatory factors (IL-1β and TNF-α) in knee tissues were measured by ELISA. The Safranin O-Fast Green staining was used to assess the severity of OA and the H&E staining was used to assess the degree of synovitis. In vitro, the levels of iron, MDA, GSH were measured by the detection kits. Western blotting was used to assess the levels of signaling-related proteins. Our results showed that PA significantly attenuated the degree of cartilage degeneration in the MIA-induced osteoarthritis model. PA also reduced the expression of IL-1β, TNF-α, MMP1 and MMP3. In vitro, PA effectively reduced the concentrations of MMP1 and MMP3 in IL-1β-stimulated chondrocytes. PA decreased the levels of Fe2+ and MDA, while increasing GSH production and GPX4 and SLC7A11 expression in IL-1β-induced chondrocytes. Meanwhile, we found that PA was able to inhibit the phosphorylation level of p65, IκB protein in chondrocytes, which effectively blocked the NF-κB signaling pathway. Furthermore, PA also increased the level of SIRT1, Nrf2, and HO-1 protein expression. In addition, the inhibition of PA on IL-1β-induced MMPs production and ferroptosis were inhibited by the SIRT1 inhibitor EX-527. In conclusion, PA inhibited chondrocyte ferroptosis and cartilage destruction in osteoarthritis. The mechanism was through activating SIRT1/Nrf2 signaling pathway.

Background: To date, disease-modifying antirheumatic drugs (DMARDs) are widely used as the primary first-line treatment option for patients with rheumatoid arthritis (RA), and the curative effect of methotrexate (MTX) and leflunomide (LEF; MTX + LEF) is greater than that of single-agent MTX therapy, but the synergistic mechanism of MTX + LEF is unclear. Methods: First, we explored the mechanism of action of MTX + LEF in RA through network pharmacology and molecular docking. Venn diagram analysis revealed 97 overlapping gene targets of MTX + LEF-RA and STRING, along with Cytoscape plug-in MOCDE and cytoHubba; and GO enrichment analysis revealed that the functions of 97 synergistic targets were related to 123 molecular functions (MF), 63 cell components (CC), and 1,068 biological processes (BP). The Cytoscape plug-in ClueGO demonstrated that these targets were enriched in KEGG pathways of 52 terms, whereas 9 pivotal genes were mainly involved in the signaling pathways of estrogen, Ras, Rap1, PI3K-Akt, relaxin, TNF, AMPK, FoxO, prolactin, IL-17, and adherens junction. Finally, CETSA and DARTS validated the direct binding of MTX or LEF to the selected target proteins EGFR, PPARG, MMP9, and SRC in RAW264.7 cells. Results: We identified 292 MTX targets and 247 LEF targets from 7 databases. Furthermore, 2,814 potential targets of RA were identified by merging 1,925 targets from 7 databases and 999 differentially expressed genes (DEGs) between normal controls and patients with RA extracted from 5 GEO databases. Nine pivotal genes, ESR1, ALB, CASP3, EGFR, HSP90AA1, SRC, MMP9, PPARG, and IGF1, were identified. Molecular docking verified that both MTX and LEF strongly bind to most of the 9 pivotal proteins except ESR1 and IGF1. Conclusion: These results contribute to our understanding of the enhancement mechanism of MTX combined with LEF and provide a targeted basis for the clinical treatment of RA.

Aspirin (ASA) induces autophagic death of human tumor cells and autophagy changes the susceptibility of Candida albicans biofilm to antifungal agents. This study investigates whether ASA suppresses C. albicans biofilm by autophagy regulation and its combination effect with antifungals. Biofilm sensitivity to ASA alone and in combination with antifungals was evaluated using the checkerboard method, and drug interactions were assessed by the fractional inhibition concentration index (FICI) and ΔE models. The effects of ASA on mTOR signaling were examined by western blotting. Alkaline phosphatase activity, acridine orange stain assay, and autophagy-related gene expressions were examined to evaluate autophagic activity. Autophagosomes were observed by transmission electron microscopy. Reactive oxygen species (ROS) were detected by DCFH-DA. Mitochondrial membrane potential (MMP), malondialdehyde (MDA), and ATP levels were determined using commercial kits. ASA inhibited C. albicans biofilm in a concentration dependent manner and showed synergistic effects against biofilms when combined with amphotericin B or 5-fluorocytosine. ASA treatment induced oxidative stress, evidenced by increased ROS and MDA levels, alongside a reduction in ATP and MMP. ASA inhibited mTOR signaling and induced autophagy in C. albicans biofilms by increasing oxidative stress and mitochondrial dysfunction, contributing to biofilm inhibition. This study provides valuable insights into the potential of ASA as an adjunct therapy in combination with antifungal agents for managing C. albicans biofilm-related infections.

Pathogenic microbial contamination in broiler drinking water constitutes a critical factor in disease occurrence and transmission within large-scale farming operations. Effective drinking water quality management is essential for optimizing broiler health and production performance. Functional additives, such as hypochlorous acid water (HAW) and hydrogen-rich water (HRW), are often added to farm drinking systems. HAW inhibits bacterial growth, and HRW enhances antioxidant capacity in broilers. This study aimed to investigate the combined effects of these two functional additives and evaluate their impacts on bacteria and biofilm in the drinking water system, broiler production performance, antioxidant capacity, and intestinal environment. A total of 480 broilers were randomly divided into four groups (n = 120 each) and raised for 42 days with distinct aqueous solutions: Group A (tap water), Group B (HAW, 0.3 mg/L), Group C (HRW, 1200 ppb), and Group D (combined HAW and HRW, hydrogen-rich hypochlorous acid water [HRHAW], 0.3 mg/L + 1200 ppb). The results indicated that the sterilization rate of planktonic bacteria in drinking water exceeded 99.90 % after the HRHAW intervention, and the biofilm biomass decreased by 62.85 %. Compared with controls, HRHAW showed no significant impact on the feed-gain ratio but significantly improved breast meat tenderness (39.15 %) and antioxidant capacity (SOD: 6.52 %-27.54 %; GSH-PX: 8.54 %-50.97 %). Intestinal health was enhanced through oxidative stress mitigation and antibacterial effects. In summary, HRHAW successfully integrates the antioxidant benefits of HRW with the antimicrobial efficacy of HAW, significantly reducing the bacterial content in drinking water and enhancing the antioxidant capacity of broilers, thereby positively influencing broiler health.

Probiotics have been established to exert a positive impact on the treatment of various diseases. Indeed, these active microorganisms have garnered significant attention in recent years for their potential to prevent and treat illnesses. Their beneficial effects have been hypothesized to be linked to their released extracellular vesicles. These nanoscale structures, secreted during the growth and metabolism of probiotics, possess favorable biocompatibility and targeting properties, thereby promoting intercellular material transport and signaling. This article aimed to review the bioactive components and functions of these probiotics vesicles, highlighting their role in the treatment of various diseases and discussing their potential future applications. By exploring the mechanisms of probiotic extracellular vesicles in disease development, this review aimed to provide a theoretical reference for further research on their therapeutic potential. © 2025 Society of Chemical Industry.

Trichinellosis is a globally distributed zoonotic parasitic disease. The Trichinella infective larvae migrate through the intestine after ingestion and settle in muscles, thus intestinal mucosal immunity plays a vital role against early infection with Trichinella. In this study, a recombinant adenovirus vector expressing the cysteine protease inhibitor of Trichinella spiralis (rAd5TsCLP) was constructed and combined with the recombinant protein rTsCLP in a heterologous prime-boost regimen. The regimen elicits strong, specific, and neutralizing antibodies in BALB/c mice, significantly enhancing cellular immunity through Th1 (IFN-γ, TNF-α) and Th2 (IL-13, IL-4) cytokine production in the peripheral blood, spleen, and cervical lymph nodes, driven by the activation of CD4+ and CD8+ T-cells. Notably, immunization with rAd5TsCLP:rTsCLP elevated mucosal secretory IgA (sIgA) levels, boosted histamine concentrations, and increased goblet cell numbers in the intestinal epithelium. Vaccinated mice showed a significant 61.17% reduction in adult worms and a 58.22% reduction in muscle larvae after the T. spiralis challenge. The adenovirus vector-delivered TsCLP amplifies local mucosal immunity, eliciting a Th1/Th2 mixed immune response that facilitates the expulsion of T. spiralis. Our study provides a feasible and promising approach for Trichinella vaccines, further highlighting the potential of an adenovirus vector for anti-helminth vaccine development.

Benzylpenicilloic acid (BPNLA) is a byproduct of the natural degradation and enzymatic hydrolysis of penicillin. BPNLA primarily enters and accumulates in the human body through the consumption of animal products. Previous research has mainly focused on drug resistance and the resulting allergic reactions, but as the accumulation of the drug increases, toxicity also manifests. The liver and kidneys are the main organs for drug metabolism and excre- tion of many drugs, and are most prone to toxic effects. Therefore, this study aimed to investigate whether BPNLA causes hepatorenal toxicity. All C57BL/6 mice were randomly assigned to four groups and administered orally 2.925, 146.25, and 7312.5 μg/kg b.wt (body weight) of BPNLA, or an equivalent volume of 0.9% saline (control) for 35 days. The results showed that BPNLA could lead to a decrease in the organ coefficient of the liver, as well as struc- tural abnormalities in the liver and kidneys. Further research found that liver and kidney function markers, lipid peroxidation markers (MDA), and proinflammatory cytokines (TNF-α and IL-1β) significantly increased compared to the control group. Moreover, the levels of antioxidant markers (GSH, SOD, GPX) decreased in a dose-dependent manner. In summary, the results clearly demonstrated that even relatively low concentrations of BPNLA can cause liver and kidney damage, highlighting the need for concern regarding human exposure to BPNLA.

Salmonella enterica serovar Typhimurium (STM) causes severe colitis, necessitating the development of effective drugs. Here, the dockings of limonin with the STM T3SS-1 virulence factor SopB or SopE2 showed strong binding activity in silico and was verified by CETSA and DARTS assays in vitro. Limonin inhibited the enzyme activities and expression of SopB and SopE2 in vitro. Furthermore, we found that limonin treatment significantly reduced the number of STM colony-forming units (CFUs) in infected HeLa and Raw264.7 cells, which resulted in a decrease in the rate of membrane ruffling mediated by SopB-regulated Arf6/Cyth2/Arf1-, RAC1-, and CDC42-driven Arp2/3-dependent actin polymerization and the SopE2-regulated CDC42/Arp2/3 pathway, and the confocal laser scanning microscopy analysis revealed that limonin treatment repressed the recruitment of the Salmonella-containing vacuole (SCV) biomarkers LC3, Rab7, GAL8 and NDP52. Furthermore, limonin treatment ameliorated STM-induced colitis by reducing the disease activity index (DAI), colon shortening, and MPO and EPO activities; mitigating the severity of S. Typhimurium-induced colitis damage; and influencing the levels of inflammatory factors (IL-1β, IL-6, IL-10, TNF-α and IFN-γ) while increasing the levels of colonic epithelial barrier and tight junction genes (Mucin 1, Mucin 2, Occludin, Claudin-3 and ZO-1). A gut microbiota analysis revealed that limonin treatment influenced α- and β-diversity of the flora and increased the counts of the beneficial bacteria Muribaculum and Faecalibaculum to regulate gut microbiota dysbiosis. Finally, colon SCFA measurements revealed that limonin treatment significantly increased acetate, butyrate, propionate and valerate concentrations. Thus, this study is an important reference for the anti-STM effects of limonin on induced colitis.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a positive-sense RNA virus with an unusually large genome of approximately 30 kb. It is highly transmissible and exhibits broad tissue tropism. The third most pathogenic of all known coronaviruses, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the clinical manifestation known as coronavirus disease 2019 (COVID-19), which has resulted in the loss of millions of lives on a global scale. This pandemic has prompted significant efforts to develop therapeutic strategies that target the virus and/or human proteins to control viral infection. These efforts include the testing of hundreds of potential drugs and thousands of patients in clinical trials. Although the global pandemic caused by the SARS-CoV-2 virus is approaching its end, the emergence of new variants and drug-resistant mutants highlights the need for additional oral antivirals. The appearance of variants and the declining effectiveness of booster shots are resulting in breakthrough infections, which continue to impose a significant burden on healthcare systems. Computer-aided drug design (CADD) has been widely utilized for predicting drug–target interactions and evaluating drug safety; it is regarded as an effective tool for identifying promising drug candidates to combat SARS-CoV-2. The CADD approach aids in the discovery of new drugs or the repurposing of United States Food and Drug Administration (FDA)-approved drugs, whose safety and side effects are already well established, thus making the process more viable. This review summarizes potential therapeutic agents that target SARS-CoV-2 or host proteins critical for viral pathogenesis, as identified using CADD approaches. Additionally, this study provides insights into the common in silico methods used in CADD and their current applications in the SARS-CoV-2 drug discovery process.

Recombinant human serum albumin (rHA) is a promising alternative to human serum albumin (HSA) for managing ascites in cirrhotic patients. This phase Ib study aims to assess the safety, tolerability, and pharmacokinetics/pharmacodynamics (PK/PD) profiles of rHA in this population. This randomized, open-label, phase Ib trial was conducted between December 2019 and September 2020 at 3 medical centers in China. Patients with cirrhotic ascites were randomly assigned to receive rHA or HSA at 10 g/day, 20 g/day, or 30 g/day. Each group had 12 participants (nine receiving rHA and three receiving HSA as positive control). Treatment lasted up to 14 days or until serum albumin levels reached 35 g/L, followed by a 28-day follow-up. Adverse events monitored assessed safety and tolerability, while PK/PD was evaluated by tracking serum albumin levels and plasma colloid osmotic pressure (PCOP) before and after each dose (ClinicalTrials.gov No. NCT04701697). Thirty-six Chinese participants were enrolled, with 32 completing the study. The incidence of adverse events was similar between the rHA and HSA groups (44.4

The therapeutic potential of genistein (GEN) in rheumatoid arthritis (RA), particularly with regard to its role in exosome secretion and the inflammatory microenvironment through the Rab27/Mfge8/nSMase2 pathway.

L. monocytogenes is a significant foodborne pathogen. This study aims to explore the biodiversity and evolutionary characteristics of L. monocytogenes isolated from beef through pan-genome analysis, and to provide important reference value for its specific molecular detection. This study conducted an in-depth analysis of the virulence genes, antimicrobial resistance genes, and environmental resistance genes of 344 L. monocytogenes strains isolated from beef. Pan-genomic analysis revealed that L. monocytogenes from beef have open genomes, providing a solid genetic basis for adaptation to different environments. MLST analysis revealed that the most prevalent types of L. monocytogenes isolated from beef were ST9 and CC9. A total of 50 virulence genes were detected in these strains, with 26 virulence genes such as inlA, inlB, plcA, plcB, and prfA, present in all L. monocytogenes strains. The four most prevalent antibiotic resistance genes in L. monocytogenes were norB, lin, mprF, and FosX, indicating high resistance to fluoroquinolones, lincosamides, peptides, and phosphonic acid antibiotics. A total of 416 potential target genes were identified through pan-genomic screening, which were then further filtered using a hub gene selection method to mining novel target genes. Ultimately, 10 highly connected hub genes were selected: bglF_2, tilS, group_2105, group_2431, oleD, ndk, flgG, purB, pbpB, and fni. These genes play a crucial role in the pathogenesis of L. monocytogenes. The PCR results demonstrated the excellent specificity of the bglF_2 gene for L. monocytogenes. Moreover, in the artificial contamination experiment, the bglF_2 gene was able to effectively detect L. monocytogenes in beef samples. Therefore, the bglF_2 gene holds potential as a specific molecular target for the detection of L. monocytogenes strains in beef samples.

Salmonella enterica serovar Typhimurium (S. Typhimurium) poses a serious threat to human and animal health, and there is an urgent need to develop new therapeutic agents. In our in vivo study, ginsenoside Ro (Ro) reduced the mortality rate of S. Typhimurium-infected mice by effectively improving three key disease activity index (DAI) indicators. In particular, ginsenoside Ro inhibited S. Typhimurium-induced colitis by reversing colon length shortening; alleviating pathological damage to the colon; decreasing the levels of IL-1β, TNF-α, IFN-γ, and IL-6; and decreasing the activities of MPO and EPO, while increasing the levels of IL-10 as well as the colon epithelial barrier and tight junction-related genes (Mucin 1, Mucin 2, Occludin, Claudin-3, and ZO-1). Furthermore, ginsenoside Ro reduced CFUs in the liver, spleen, colon, and feces. In a mechanistic in vitro study, ginsenoside Ro reduced CFUs in HeLa and Raw264.7 cells, which was associated with ginsenoside Ro inhibition of the recruited S. Typhimurium-containing vacuole (SCV) biomarkers LC3, Rab7, GAL8, and NDP52. Molecular docking results revealed that the binding energies of ginsenoside Ro and SopB and ginsenoside Ro and SopE2 were as high as -11.3 and -9.7 kcal/mol, respectively, as verified by CETSA and DARTS assays. Moreover, ginsenoside Ro at 100 and 200 μM significantly decreased the enzyme activities and expression of SopB and SopE2. Finally, ginsenoside Ro inhibited the membrane ruffling caused by SopB-regulated Arf6/Cyth2/Arf1-, RAC1-, and CDC42-driven Arp2/3-dependent actin polymerization and the SopE2-regulated CDC42/Arp2/3 signaling pathway. In summary, our findings suggest that ginsenoside Ro is a potential lead compound for therapeutic use against S. Typhimurium infection, and these findings lay a foundation for its further development.