Er-Chen decoction alleviates spermatogenic dysfunction in obese mice by tuning the SIRT1/p53 axis.

A novel "cut and sew" procedure for the natural reconstitution of essential oils prior to biological assays.

Alzheimer's disease (AD), a major neurodegenerative disorder, lacks effective early diagnostic and therapeutic strategies. This study aimed to investigate the diagnostic utility of Long non-coding RNAs HLA Complex Group 18 ( HCG18) in AD and elucidate its molecular mechanisms in neuronal injury.Eighty-three AD patients and 83 healthy controls (HC) were enrolled. Serum samples were analyzed for HCG18 expression using qRT-PCR and cerebrospinal fluid (CSF) samples were analyzed for AD biomarkers by ELISA. Diagnostic performance was assessed using ROC analysis. Aβ1-42-treated HT22 cells (Immortalized murine hippocampal neuronal-like cell line) were employed to model neuronal injury, with HCG18 knockdown and miR-425-3p inhibition experiments conducted to validate functional interactions. HT22 cell apoptosis, oxidative stress markers (SOD, GSH-Px, MDA, ROS), and HCG18/miR-425-3p interactions were evaluated through flow cytometry, biochemical assays, and dual-luciferase reporter systems.Serum HCG18 levels were significantly elevated in AD patients compared to HC (P < 0.001), exhibiting strong diagnostic accuracy (AUC = 0.889). HCG18 expression correlated negatively with CSF Aβ1-42 (r=-0.709) and MMSE scores (r=-0.657), but positively with t-tau (r = 0.591) and p-tau181 (r = 0.582). In Aβ1-42-treated HT22 cells, HCG18 knockdown reduced apoptosis, suppressed ROS, and normalized oxidative stress markers. Mechanistically, HCG18 directly bound to and acted as a molecular sponge for miR-425-3p, sequestering its function; the downregulation of miR-425-3p mediated by a synthetic inhibitor reversed the protective effects of HCG18 silencing.HCG18 serves as a potential non-invasive biomarker for AD, exacerbating neuronal injury via sponging miR-425-3p to disrupt redox balance. Targeting the HCG18/miR-425-3p axis may offer new therapeutic strategies for AD.

Heart failure (HF) represents the terminal stage of many cardiovascular diseases. Doxorubicin (DOX) can induce HF through oxidative stress (OS), inflammation, and apoptosis. Ginkgetin (GK) has potential cardioprotective effects, but its underlying mechanisms remain unclear. This study investigated the protective effects of GK against DOX-induced HF and explored its mechanisms, focusing on mitochondrial function and related signaling pathways. In vivo and in vitro experimental models. HF was induced by DOX in mice and H9c2 cardiomyocytes. Cardiac function, myocardial injury, OS, inflammation, and apoptosis were assessed using echocardiography, biochemical assays, enzyme-linked immunosorbent assay, histopathology, immunofluorescence, and Western blot. Mitochondrial function was evaluated via transmission electron microscopy, RT-qPCR, and Seahorse analysis. Compound C was applied to verify the involvement of the adenosine monophosphate-activated protein kinase (AMPK)/Sirt1/nuclear factor-κB (NF-κB) pathway. GK markedly improved DOX-induced cardiac dysfunction and myocardial injury, reduced cardiac injury markers and inflammatory cytokines, and alleviated fibrosis, hypertrophy, apoptosis, and reactive oxygen species accumulation. GK restored superoxide dismutase activity, decreased malondialdehyde levels, increased glutathione and ATP, and preserved mitochondrial structure and respiratory function. GK upregulated AMPK and Sirt1, inhibited NF-κB activation, and regulated apoptosis-related proteins, whereas Compound C reversed these effects. GK protects against DOX-induced HF by activating AMPK/Sirt1 and inhibiting NF-κB signaling, thereby mitigating OS, inflammation, apoptosis, and mitochondrial dysfunction.

Abstract Background Acute liver injury (ALI) is a severe hepatic condition with high mortality and few therapeutic options aside from liver transplantation. Lonicera japonica Flos (LJ), a traditional herbal medicine, has been reported to have anti-inflammatory and hepatoprotective activities; however, the underlying mechanisms remain insufficiently clarified. Methods In this study, an integrative approach was used to investigate the effects of LJ on liver injury by combining network pharmacology, molecular docking, and experimental validation. Computational analyses were used to predict the potential targets and enriched pathways of LJ-derived active compounds on ALI. These predictions were subsequently validated in lipopolysaccharide (LPS) induced ALI murine models and LPS-stimulated HepG2 cells through biochemical assays and histological staining with evaluation of HIF-1α/IL-1β signaling pathway-related molecules. Results Network pharmacology highlighted that the HIF-1 signaling pathway is closely related to the potential mechanism of LJ on ALI. In silico docking demonstrated that loganin and loganic acid, main components of LJ, have potentially strong binding affinities on HIF-1α. Based on these predicted results, LJ lowered serum AST and ALT, alleviated histopathological injury, and suppressed hepatic TNF-α, IL-6, and IL-1β expression in LPS-induced ALI mice. In HepG2 cells, LJ inhibited cleaved IL-1β and HIF-1α protein expression without cytotoxicity in response to LPS, thereby dampening multiple inflammatory cascades. Conclusion Overall, LJ exerts hepatoprotective effects against LPS-induced ALI by targeting several interconnected pathways. These findings support LJ as a potential therapeutic candidate for inflammatory liver disease and demonstrate the value of combining computational prediction with experimental confirmation to investigate traditional herbal medicines. Graphical Abstract

Cyrtorhinus lividipennis, a key natural enemy of the brown planthopper, Nilaparvata lugens, has been observed to tolerate short-term high-temperature exposure; however, the physiological and molecular mechanisms underlying this heat tolerance remain unclear, which may hinder its effective conservation and utilization. Here, we combined physiological and biochemical assays with transcriptome sequencing to elucidate the physiological and molecular mechanisms of heat tolerance in C. lividipennis following 1 h exposure to three temperatures: 26 °C (control), 33 °C (moderate heat stress), and 40 °C (severe heat stress). At 40 °C, sorbitol, trehalose, lipid, and glycogen contents increased significantly, whereas glycerol levels declined. Transcriptomic profiling revealed temperature-dependent DEGs enriched in starch and sucrose metabolism, galactose metabolism, glycerolipid metabolism, oxidative phosphorylation, and protein folding, sorting, and degradation, with pronounced temperature-dependent upregulation of heat shock protein (HSP) gene families. Together, these results demonstrate that C. lividipennis coordinates its heat stress response through soluble polyol accumulation, which is known to act as a compatible osmolytes that help stabilize proteins and membranes and mitigate thermal damage, energy metabolic reprogramming, and HSP-mediated proteostasis, thereby providing a theoretical basis for its conservation and utilization in sustainable paddy agroecosystems.

The glenoid labrum is a fibrocartilaginous structure essential for shoulder stability, yet its regeneration remains an unmet clinical challenge. Current surgical approaches restore initial joint stability but frequently fail to reestablish native biomechanics, leading to recurrence and early degenerative changes. In this study, we investigated the feasibility of fabricating a patient-specific, anatomically scaled glenoid labrum scaffold using digital modeling based on magnetic resonance imaging and 3D cryo(bio)printing of a gelatin methacryloyl (GelMA) hydrogel. Printing was performed in a temperature-controlled platform (22.5 °C, 15 °C, and −20 °C) to evaluate the influence of thermal conditions on structural fidelity and biological performance. Quantitative analyses showed that cryogenic deposition markedly improved printing precision, reducing filament spreading and enhancing geometric accuracy in both sharp-angle and grid-pattern evaluations. Biological assays indicated high viability of human mesenchymal stem cells under all temperature conditions, validating the cytocompatibility of the methodology. Morphological assessment by structured-light 3D scanning demonstrated that bioprinted patient-specific scaffold at −20 °C achieved the highest correspondence to the digital reference model. Overall, the integration of anatomical modeling with cryo(bio)printing proved to be an effective approach for producing anatomically faithful, patient-tailored scaffolds. This study presents the first demonstration of human glenoid labrum bioprinting and establishes a foundation for future translational research in fibrocartilaginous tissue regeneration.

Objectives: This study explored the antidepressant mechanisms of aerobic exercise in CUMS rats by analyzing urinary metabolomics (LC-MS and NMR), with the aim of providing both theoretical and practical support for exercise-based depression interventions. Methods: (1) Thirty-two Sprague-Dawley rats were acclimatized for one week and then randomly assigned to four groups (n = 8 per group): control (C), control + aerobic exercise group (E), CUMS model (D), and CUMS + exercise (DE). Groups D and DE were subjected to nine types of CUMS stimuli. Behavioral indicators were assessed weekly, and the successful establishment of the CUMS model was confirmed at week 3. Following successful modeling, rats in groups E and DE underwent four weeks of aerobic exercise training. Throughout this period, groups D and DE continued to receive CUMS exposure, while groups C and E were maintained under standard control conditions. (2) At the end of week 7, behavioral tests were repeated. Twelve-hour urine samples were collected for metabolomic analysis using liquid chromatography–mass spectrometry (LC-MS) and 1H-NMR spectroscopy. The following morning, rats were euthanized under anesthesia. Whole blood was collected from the abdominal aorta, and serum was separated for subsequent biochemical assays. Bioinformatics approaches were employed to identify potential targets and signaling pathways associated with the antidepressant effects of aerobic exercise. (3) For statistical analysis, one-way or two-way analysis of variance (ANOVA) was applied to behavioral, physiological, and biochemical data, whereas multivariate statistical analysis was used for metabolomic data. Results: (1) By week 3, body mass, sucrose preference, rearing frequency, and the number of grid crossings were significantly lower in groups D and DE than in groups C and E (p &lt; 0.05 or p &lt; 0.01). These findings confirmed the successful establishment of the depression model. At week 7, all behavioral indicators in group DE showed significant recovery relative to group D (p &lt; 0.05 or p &lt; 0.01). (2) Compared with group C, corticosterone and blood ammonia levels were significantly elevated in group D (p &lt; 0.01). In contrast, these levels were markedly reduced in group DE compared with group D (p &lt; 0.01). (3) LC-MS analysis identified 25 urinary metabolites associated with depression in group D relative to group C. Among these, 21 were significantly downregulated and 4 were upregulated (p &lt; 0.05 or p &lt; 0.01), involving seven metabolic pathways. Following aerobic exercise intervention, six of these depression-related metabolites in group DE showed significant recovery (p &lt; 0.05 or p &lt; 0.01), which were associated with two metabolic pathways. (4) Integrated analysis of LC-MS and 1H-NMR data revealed glutamine as a common differential metabolite, linked to three metabolic pathways. All metabolic pathways modulated by aerobic exercise were related to amino acid metabolism. (5) Bioinformatics analysis indicated that AKT1, MTOR, IL6, RAF1, and TNF were core targets through which aerobic exercise regulated urinary metabolism in CUMS rats. Conclusions: A four-week regimen of aerobic exercise significantly improved depressive-like behaviors and enhanced anti-fatigue capacity in CUMS rats. This exercise regimen promoted urinary metabolic remodeling, primarily through the modulation of amino acid metabolism. Furthermore, its antidepressant effect is likely mediated through the regulation of core tissue targets—including AKT1, mTOR, IL-6, RAF1, and TNF—thereby influencing key pathways such as PI3K-AKT, MAPK/ERK, and neuroinflammatory signaling.

Homocysteine (Hcy) is an independent risk factor for atherosclerosis (AS). Hcy induces the transformation of vascular smooth muscle cells (VSMCs) into foam cells, which play a crucial role in this process. However, the detailed mechanism is still unclear. To identify the key regulatory proteins during this process and clarify the possible mechanism of Hcy-induced foam cell formation in VSMCs, thereby providing theoretical support for the intervention of AS. VSMCs were allocated into two groups: a control cohort and a group exposed to Hcy to simulate an AS-like state. Quantitative proteomic profiling was performed using the label-free quantitative DIA (LFQ-DIA) approach to detect differentially expressed proteins between these groups. To explore functional implications, enrichment analyses involving Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were conducted. Protein-protein interaction networks were constructed using the STRING database to identify central interactors. Target proteins were subsequently validated through parallel reaction monitoring (PRM). Furthermore, histological analyses (hematoxylin and eosin (HE) staining, Oil Red O staining), biochemical assays of lipid content (total cholesterol (TC) and triglycerides (TG)), and Western blot analysis were utilized to confirm the role and mechanism of identified proteins in the context of Hcy-driven foam cell conversion. The results showed that proteomic analysis identified 4804 proteins in total, of which 4799 passed missing-value filtering and were retained for downstream quantitative analysis. A total of 54 proteins were identified as differentially expressed using thresholds of adjusted p-value < 0.05 and fold change > 1.5. Among them, 13 proteins were upregulated, while 41 were downregulated in response to Hcy treatment. For PRM validation, 20 candidate proteins were selected according to proteomic evidence, biological relevance, and technical feasibility. Among them, 16 proteins (COX7C, STX5, UBQLN2, DDX50, TBCB, GSR, PCNP, CDV3, PEBP1, PPIA, S100A6, EIF4E2, UBQLN1, ARMC1, NUDCD2, and H1-2) showed the same direction of fold-change values as in the LFQ-DIA dataset, thereby underscoring the reliability of the proteomic analysis. Data are available via ProteomeXchange with identifier PXD064315. Histological staining demonstrated enhanced lipid accumulation, and the protein expression of the contraction phenotype marker a-SMA decreased, while the protein expression of the synthesis phenotype marker OPN increased. This indicates that Hcy induces VSMCs to transform from a contraction phenotype to a synthesis phenotype, resulting in the formation of foam cells. The protein levels of COX7C and sterol regulatory element-binding proteins (SREBP1C and SREBP2) were elevated upon Hcy exposure. Overexpression of COX7C further augmented the expression of SREBP1C and SREBP2, exacerbated lipid accumulation, and promoted foam cell transformation in Hcy-treated VSMCs. On the other hand, knockdown of COX7C had the opposite effect. Overall, the results of the present study suggest that COX7C plays a crucial regulatory role in Hcy-induced transformation of VSMCs into foam cells. Its pathogenic role is likely mediated through the upregulation of SREBP1C and SREBP2, thereby promoting lipid accumulation. These findings provide new insights into AS pathogenesis and identify COX7C maybe a potential therapeutic target.

This study aims to elucidate the synergistic protective mechanism of the AMPK agonist 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) in ischemia-reperfusion injury -associated acute kidney injury (IRI-AKI). By establishing a hypoxia/reoxygenation (H/R) injury model using human proximal tubule cells (HK-2) and IRI-AKI rat model, and employing molecular techniques including qRT-PCR, western blotting, serum biochemical assays, renal tissue hematoxylin and eosin staining, immunofluorescence, and transmission electron microscopy (TEM), we demonstrated that AICAR activates AMPK, leading to the significant downregulation of TXNIP and NLRP3, blocks Caspase-1-dependent release of IL-1β and IL-18, and ultimately suppresses pyroptosis, thereby alleviating renal inflammatory injury. Furthermore, AICAR restored mitochondrial membrane potential and ATP levels in H/R-treated HK-2 cells, reduced reactive oxygen species production in renal tissues of IRI-AKI rats, and elevated levels of antioxidant enzymes. Concurrently, utilizing targeted metabolomics technology, we discovered that AICAR effectively restores the levels of multiple metabolites associated with glycolysis, the TCA cycle, the urea cycle, and tryptophan metabolism and alleviates lipid deposition in IRI-AKI. This confirms that AICAR alleviates IRI-AKI by activating AMPK to restore impaired cellular energy metabolism, improve mitochondrial function, and ameliorate oxidative stress. Notably, this study is the first to reveal that AICAR, via AMPK activation, synchronously regulates dual protective pathways: "pyroptosis inhibition" and "energy metabolism remodeling." This synergistic protective mechanism may represent the core advantage distinguishing AICAR from other potential therapeutic strategies, highlighting its substantial translational potential as a multi-mechanism synergistic therapeutic agent. Our findings provide an innovative dual-regulatory ("pyroptosis-energy metabolism") therapeutic strategy for the clinical prevention and treatment of IRI-AKI.

Introduction The indiscriminate use of pesticides poses a significant risk to human health and the environment. Plant-based biopesticides offer an alternative to insecticides for integration into insect pest management programs. Methods The current study assessed the toxicological effects of leaf extracts from indigenous weed plants, Chenopodium murale and Achyranthes aspera , in albino rats, Rattus norvegicus . The extract-mixed diet was fed in three different doses (100 ppm, 150 ppm, and 250 ppm) for 28 days, while the Cypermethrin insecticide was used as a reference insecticide at the same dose levels. Samples from liver and kidney tissues were collected for histopathological study, and the blood serum was obtained for biochemical assay. Results and discussion Histopathological analysis of cypermethrin revealed congestion in the central vein, hemorrhage in hepatic tissues, and necrosis of liver tissues, while kidney tissues showed necrosis of renal tubules, fibrosis, and swelling in Bowman’s capsule. Moreover, hemorrhage was attenuated by degenerated inflammatory cells, edema, and shrinkage, and the rupturing of glomeruli was also observed. Mortality was also recorded at 28th day. In contrast, no physical signs of toxicity and significant alteration in liver and kidney tissues were shown by both plant extracts. Acetylcholinesterase (AChE) and phosphatase enzymes also showed non-significance with plant extracts and significant results with Cypermethrin. Similarly, genotoxicity through the comet assay revealed no changes for either plant. Hematological parameters showed no significant change with plant extracts. Non-significant results revealed that both plant extracts had no difference when compared to the control for all parameters, which indicates that the weed plants are less toxic as compared to Cypermethrin in vertebrates. Conclusion These findings suggest that these weed plants have the potential to be used as biopesticides for future integrated pest management (IPM) programs.

The present study employed two complementary in vitro chemical assays, namely Ferric Reducing Antioxidant Power (FRAP) and ABTS⁺ radical scavenging, to evaluate and compare the antioxidant potential of methanolic extracts derived from five distinct Jasminum species. The electron-donating capacity of the extracts, mediated through redox-based mechanisms, as well as their ability to neutralize free radicals, was determined using spectrophotometric analysis. Results revealed significant interspecific variation in antioxidant activity. Based on absorbance values, FRAP measurements (µmol Fe²⁺/g), and antioxidant power indices, J. azoricum and J. nudiflorum exhibited the highest antioxidant potential, whereas J. sambac demonstrated the lowest activity across both assays. Specifically, the FRAP assay quantified the extracts’ capacity to reduce Fe³⁺ to Fe²⁺, while the ABTS assay measured their ability to scavenge ABTS⁺ radical cations. The observed variability is likely attributable to differences in phytochemical composition, particularly in phenolic and flavonoid content. Overall, this comparative evaluation underscores the promise of certain Jasminum species as potential natural sources of antioxidants for pharmaceutical and nutraceutical applications. KEY WORDS: Jasminum species, Spectrophotometric analysis, Antioxidant activity, ABTS assay, FRAP assay Bull. Chem. Soc. Ethiop. 2026, 40(3), 679-689. DOI: https://dx.doi.org/10.4314/bcse.v40i3.14

N-methyl-D-aspartate-type glutamate receptors (NMDARs) initiate the synaptic plasticity underlying learning and memory. In forebrain excitatory neurons, NMDARs are heteromeric tetramers composed of two GluN1 subunits and two glutamate ionotropic receptor NMDA type subunit 2A (GluN2A) or GluN2B subunits. At birth, NMDARs contain primarily GluN2B, but within weeks, GluN2A-containing receptors predominate the forebrain, comprising over 65% of total NMDARs in adulthood. This rapid subunit switch is essential for neonatal cognitive development, yet mechanisms driving it remain unclear. Particularly, while GluN2B levels remain relatively constant, GluN2A increases several 100-fold, despite its mRNA rising by only ~10-fold, strongly suggesting involvement of unknown posttranslational regulation. Here, we show that in the neonatal mouse forebrain, the linear ubiquitination axis, composed of the E3 ligase complex LUBAC and the deubiquitinase OTULIN, shifts transiently toward higher activity, with HOIP upregulated and OTULIN downregulated. In neonatal mice, experimentally reducing the axis activity by OTULIN overexpression causes persistent synaptic immaturity and adult cognitive deficits. Using proteomic and biochemical assays, we identified GluN2A as a key substrate: Linear ubiquitination at six lysines in the GluN2A C-terminus stabilizes the subunit and promotes its synaptic expression, whereas disrupting this modification destabilizes GluN2A by promoting lysosomal degradation. Consistently, overexpression of wild-type GluN2A rescues OTULIN-induced synaptic immaturity, whereas the ubiquitination-deficient GluN2A-6KR mutant fails to rescue and further exacerbates this defect. OTULIN overexpression selectively promotes GluN2A degradation, thereby delaying the GluN2B-to-GluN2A switch and synaptic maturation. These findings reveal a role for the linear ubiquitination axis in selectively stabilizing GluN2A, supporting rapid synaptic and cognitive development.

Cells must continuously adjust metabolic output to maintain homeostasis under changing environmental conditions, yet the mechanisms that enable rapid and reversible control of pathway activity remain largely unknown. The methylerythritol phosphate (MEP) pathway, of bacterial origin and conserved in plastid-bearing eukaryotes, including plants and apicomplexan parasites, produces isoprenoid precursors essential for growth and stress adaptation. Here, we identify methylerythritol cyclodiphosphate (MEcPP) as a dual-function metabolite that serves both as a biosynthetic intermediate and a direct modulator of enzyme activity. Genetic perturbations and high light stress revealed step-specific MEcPP accumulation independent of transcriptional regulation. Biochemical and protease-protection assays showed that MEcPP destabilizes and inhibits methylerythritol cytidylyltransferase (MCT) while modestly stabilizing hydroxymethylbutenyl diphosphate synthase (HDS). Molecular docking analyses indicate that MEcPP interacts directly with the MCT catalytic site, displacing the natural substrate and thereby attenuating enzyme activity, suggesting a competitive, feedback-like mechanism of metabolic control. These results define MEcPP as a metabolic feedback signal that translates stress-induced changes into targeted enzymatic control. This mechanism illustrates how pathway intermediates dynamically coordinate biosynthetic activity with environmental cues, representing a broadly conserved strategy for metabolite-driven control of cellular metabolism.

Gastric ulcer (GU) is primarily caused by Helicobacter pylori (H. pylori) infection, ethanol-induced oxidative stress, and excessive gastric acid secretion. This is a significant global health burden. Lifetime prevalence of peptic ulcer disease (including GU and duodenal ulcer) in the general population has been estimated to be about 5-10%, and incidence 0.1-0.3% per year. Kahweol (a coffee-derived diterpene) exhibits broad pharmacological effects. Although Kahweol has strong antioxidant, anti-inflammatory and cytoprotective effects, its gastroprotective potential against ethanol-induced GU has remained unexplored. This is the first study demonstrating the multifaceted gastroprotective effects of Kahweol using an integrated computational, in vitro, and in vivo approaches, including direct comparison with omeprazole and evidence of dual acid-suppressive and anti-H. pylori activity. Molecular docking and 100ns molecular dynamics simulations demonstrated stable Kahweol binding to H⁺/K⁺-ATPase, nuclear factor-kappa B, NOD-like receptor protein 3, and cyclooxygenase-2. Kahweol exhibits inhibition of inflammatory pathways and acid secretion, having the highest affinity for H+/K+-ATPase (-7.92kcal/mol). In vitro assays revealed that Kahweol has dose-dependent antibacterial activity against H. pylori clinical isolates. In vivo, Kahweol significantly decreased the ethanol-induced ulcer index in a dose-dependent manner. Kahweol pretreatment inhibited GU up to 80% with 10mg/kg dose, which was comparable to that of omeprazole (85%). Histopathological analysis in Kahweol-treated groups confirmed preserved mucosal architecture, reduced vacuolation, and restored epithelial boundaries, which were comparable to those of omeprazole. Biochemical assays of Kahweol-treated groups showed enhanced antioxidant defenses, with increased levels of reduced glutathione, glutathione S-transferase, and catalase, along with decreased levels of nitric oxide and malondialdehyde. Results from the enzyme-linked immunosorbent assay of the Kahweol-treated group showed that the pro-inflammatory cytokines interleukin-1 beta and tumor necrosis factor-alpha were suppressed, confirming anti-inflammatory effects. Quantitative real-time polymerase chain reaction verified that the expression of H⁺/K⁺-ATPase was downregulated by Kahweol, indicating the acid-inhibition effect of Kahweol. These findings suggest Kahweol is the potential natural agent for the prevention and treatment of ethanol-induced GU, highlighting the comprehensive gastroprotective effects of Kahweol through its antioxidant, anti-inflammatory, anti-secretory, and anti-H. pylori activity.

Purpose: This study aimed to investigate the therapeutic effects and underlying mechanisms of the Ginseng and Rhubarb Combination (GRC) on sepsis-induced persistent inflammation, immunosuppression, and catabolism syndrome (PICS). Methods: The mouse model of sepsis-induced PICS was established using cecal ligation and puncture (CLP). Mice were randomly assigned to five groups: Sham, Model, Model + GRC, Model + GRC + oleic acid (OA), and Model + GRC + celastrol (CELA). GRC was administered by oral gavage once daily for seven days, whereas the agonists were injected intraperitoneally for three days. Additionally, KN-93 (a CaMKII inhibitor; 5 mg/kg) was used to verify the inhibitory regulation of CaMKII signaling. Seventy-two hours after surgery, serum and tissue samples were collected to evaluate the therapeutic effects of GRC. Outcome measures included survival, body weight, splenic immune cell distribution, histopathology, inflammatory cytokine levels, and ferroptosis-related indicators. Targeted metabolomics and differential proteomic analysis (DIA) based proteomics were performed to identify differential molecules and pathways associated with GRC treatment. Results: Compared to the model group, GRC markedly improved the survival rate and mitigated body weight loss in septic PICS mice. GRC reduced splenic neutrophil and macrophage infiltration and increased the proportions of CD4⁺ and CD8⁺ T cells. Histopathological examination revealed attenuation of organ damage. Levels of inflammatory cytokines (IL-6, TNF-α, IL-4, IL-13) were significantly decreased, and metabolic disturbances were ameliorated. Biochemical assays showed that GRC reduced Fe²⁺ and malondialdehyde (MDA) levels while increasing glutathione peroxidase (GSH-PX) and GPX4, indicating inhibition of ferroptosis. Western blot (WB) analysis demonstrated that GRC downregulated the CaMKII/HO-1 pathway, and this regulatory effect was reversed by CaMKII and HO-1 agonists. Multi-omics analysis further revealed that GRC modulated metabolic and proteomic networks associated with immune regulation and ferroptosis, confirming that its therapeutic benefits were mediated through suppression of the CaMKII/HO-1/GPX4 axis. Conclusions: GRC exhibited significant anti-inflammatory, immunomodulatory, and organ-protective effects in the murine model of sepsis-induced PICS. These benefits were mediated through the suppression of ferroptosis via modulation of the CaMKII/HO-1 pathway, providing a novel experimental foundation and a potential therapeutic strategy for managing sepsis-associated PICS.

RegIIIα is an antibacterial protein primarily operating in the digestive tract to defend against bacterial infection through direct bactericidal activity. A previous study proposed that RegIIIα forms hexameric pores on the membrane of Gram-positive bacteria, leading to cell lysis. These RegIIIα hexamers can further assemble into filaments, diminishing RegIIIα activity. However, the high-resolution structure of RegIIIα assembly remains elusive, impeding the comprehension of the molecular mechanisms underlying RegIIIα function. In this study, we determined the cryo-electron microscopy (cryo-EM) structure of RegIIIα filaments formed in vitro at a resolution of 2.2 Å. Our structure reveals a similar subunit arrangement but a distinct subunit orientation compared to the previously reported low-resolution model of RegIIIα filaments. Through structural analysis and biochemical assays, we identified two essential interfaces for RegIIIα assembly, offered a potential explanation for the necessity of lipids in RegIIIα assembly, and elucidated the inhibitory mechanism of the pro-segment of RegIIIα. Collectively, our study presents the first near-atomic structure of filaments formed by C-tyle lectin containing proteins, providing structural insights into RegIIIα assembly that are closely related to its physiological functions and regulations.

This study examined the renoprotective effect of Piribedil against cyclophosphamide (CP)-induced nephrotoxicity through modulation of adenosine monophosphate-activated protein kinase (AMPK)/sirtuin-1 (SIRT1), phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt), mitogen-activated protein kinases (MAPKs), and Toll-like receptor-4 (TLR4)/NOD-like receptor protein-3 (NLRP3) pathways, as well as renal injury markers kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL). Male rats were divided into four groups (n = 8). Controls received distilled water plus saline; the CP group received a single CP dose (200mg/kg, i.p.) on day 7; Piribedil groups received 15 or 40mg/kg/day for 10days with CP on day 7. Renal function, oxidative stress, inflammation, and injury markers (KIM-1 and NGAL) were assessed via biochemical assays, histopathology, immunohistochemistry, and quantitative real-time PCR (qRT-PCR). CP caused significant renal dysfunction, elevating blood urea nitrogen (BUN), serum creatinine (SCr), NGAL, and KIM-1, increasing oxidative stress (malondialdehyde [MDA], inducible nitric oxide synthase [iNOS]) and reducing nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and glutathione (GSH). CP also upregulated inflammatory mediators (interleukin-1β [IL-1β], interleukin-6 [IL-6], tumor necrosis factor-α [TNF-α], nuclear factor-κB p65 [NF-κB p65]) and enhanced TLR4, NLRP3, and MAPKs, while suppressing AMPK, SIRT1, and PI3K/Akt signaling. Piribedil reversed these changes, improving renal function, lowering oxidative and inflammatory markers, and normalizing BUN, SCr, KIM-1, and NGAL. Histology confirmed reduced renal damage. Piribedil effectively protects against CP-induced nephrotoxicity by modulating AMPK/SIRT1 and related oxidative and inflammatory pathways, supporting its potential use in drug-induced kidney injury.

ABSTRACTBacterial microcompartments (BMCs) are protein‐based organelles that spatially organise metabolic pathways in prokaryotes, playing critical roles in enhancing metabolic processes and microbe fitness. Notably, many bacterial species possess multiple types of BMCs. While recent studies have advanced our knowledge about the assembly and function of individual BMC types, the mechanisms governing the coexistence and interplay of distinct BMC families within a single bacterial cell remain poorly understood. Here, we engineered Salmonella enterica serovar Typhimurium LT2 to co‐express native 1,2‐propanediol utilisation (Pdu) BMCs and synthetic α‐carboxysomes (α‐CBs), providing a unique platform for dissecting their assembly dynamics and functional crosstalk. By exploiting super‐resolution fluorescence imaging, electron microscopy, biochemical and enzymatic assays, our studies demonstrate the formation of hybrid BMCs through the exchange of shell proteins between Pdu BMCs and α‐CBs, whereas cargo proteins exhibit only limited compatibility, highlighting the specificity of encapsulation mechanisms. Furthermore, the generated hybrid BMCs display altered mobility and enzymatic activities, revealing emergent properties arising from shell protein interchangeability. Our findings provide insights into the inherent structural plasticity and modular architecture of BMCs. More broadly, this study has implications for deciphering how bacterial cells modulate the construction and functions of diverse metabolic modules within a single cellular context and could inform the rational design and engineering of synthetic organelles and bio‐factories with tailored metabolic functions for biotechnological applications.

Mutations in the 3'-polypurine tract (3'PPT) of HIV-1 have been observed under pressure with two integrase strand transfer inhibitors, dolutegravir and cabotegravir. In the DOMONO randomized clinical trial, 3'PPT mutations emerged in a participant who experienced treatment failure under dolutegravir monotherapy. To understand the basis for this rare mutational pathway, we examined baseline viral sequences and identified the K156N natural polymorphism. Given the role of K156 in viral DNA binding, the potential relationship between K156N and 3'PPT mutations was further investigated. We assessed the impact of K156N on integrase using in silico modelling and biochemical assays with recombinant proteins. Infectivity, replicative capacity, and drug susceptibility of viruses carrying K156N, 3'PPT mutations, or both were measured. Viral evolution was assessed in cell culture. Structural models indicated that K156N altered viral DNA binding. K156N reduced strand transfer activity through decreased affinity for the LTR but increased 3'-processing. The K156N virus had normal infectivity, whereas the 3'PPT mutations decreased infectiousness sixfold and lowered maximal infectivity. K156N partially compensated for this defect, but maximal infectivity remained diminished. K156N also partially compensated for defects in replicative capacity imposed by 3'PPT mutations. K156N alone did not confer resistance against dolutegravir, nor did it increase the modest (2.5-fold) resistance conferred by the 3'PPT mutations. K156N alone promoted the spontaneous emergence of 3'PPT mutations distinct from those seen in DOMONO. These findings establish a direct functional relationship between natural variation in HIV-1 integrase and the emergence of 3'PPT mutations. People harbouring a virus with the K156N natural polymorphism may be predisposed to developing 3'PPT mutations upon failure with DTG. However, the clinical relevance of this association remains to be established.