It is known that lipopolysaccharides (LPS) could lead to kidney injury and may play a role in intracavitary lithotripsy surgeries such as percutaneous nephrolithotomy (PCNL). Remote ischemic preconditioning (RIPC) has been demonstrated to have protective effects against kidney injury caused by various factors; however, its role in LPS-induced kidney injury remains unclear and its mechanisms require further exploration. Previous studies have shown that extracellular vesicles (EVs) induced by hypoxia mediated the renal protective effects of RIPC. In this study, we established a kidney injury model in rats by ligating the left ureter and injecting LPS into the renal pelvis, along with simultaneous removal of the right kidney (U-L model); EVs were isolated from the plasma of rats subjected to RIPC or sham treatment and administered to U-L model rats. Separately, EVs obtained from healthy human volunteers were applied to LPS-treated human renal tubular epithelial cells (HK-2). Results indicated that RIPC-EVs(R) reduced kidney injury caused by U-L by inhibiting apoptosis, inflammation, and oxidative stress. In vitro, RIPC-EVs(H) alleviate LPS-induced HK-2 cell injury by attenuating apoptosis and inflammation. Moreover, we identified 37 significantly upregulated differential proteins in the EVs sourced from human plasma after RIPC through 4D label-free proteomics, including Tenascin-C (TNC). Mechanistic studies revealed that RIPC-EVs(H) activated β-catenin, inhibited the p53 and NF-κB signaling pathways, and alleviated kidney injury. Clinical trial data showed that RIPC reduced the expression of kidney injury biomarkers in patients undergoing PCNL for kidney stones. Overall, the present study suggested that RIPC could mitigate kidney injury induced by LPS, possibly through the protective role of Tenascin-C carried by EVs.

The genetic architecture of polycystic ovary syndrome (PCOS) has not been fully elucidated. Translating genome-wide association study (GWAS) loci into functional effector genes represents a key challenge for elucidating disease mechanisms and advancing targeted therapeutic strategies. In this study, the FinnGen R12 dataset was utilized in conjunction with the Genotype-Tissue Expression Project (GTEx) v8 eQTL dataset to conduct cross-tissue transcriptome-wide association studies (TWAS). External validation was performed in two independent datasets, comprising a PCOS meta-analysis of Rotterdam- and NIH-criteria cohorts and UK Biobank summary statistics. To prioritize high-confidence candidate genes, we integrated a cross-tissue (UTMOST) TWAS and tissue-specific (FUSION) TWAS with a gene-based association test (MAGMA). Furthermore, causal inference analyses including Mendelian randomization (MR), colocalization analysis, summary data-based MR, and the heterogeneity in dependent instrument (HEIDI) test were carried out on candidate genes. GeneMANIA, gene-chemical-disease analysis, and phenome-wide association study (PheWAS) were further performed on candidate genes. Finally, we explored expression profiles of core candidate genes using transcriptomic datasets from the Gene Expression Omnibus (GEO) database. We prioritized 15 candidate genes potentially associated with PCOS. External analyses across independent datasets showed limited consistency, which may reflect differences in statistical power and cohort heterogeneity. MCM6 was prioritized by all three approaches, whereas the remaining 14 genes were supported by two approaches. Among the 15 candidate genes, six genes were demonstrated to have causal associations with PCOS risk. GeneMANIA network analysis further revealed that these candidate genes were involved in key biological functions, including DNA replication and regulation, DNA repair and recombination, protein-DNA complex, cell cycle regulation, nucleic acid-enzyme activities, DNA structure and chromosomal regions, reproductive system development, and sex differentiation. Gene-chemical-disease analysis suggested that the prioritized genes may mediate crosstalk between PCOS genetic risk and environmental/metabolic factors. PheWAS analysis highlighted the pleiotropic roles of PCOS candidate genes, which contributed to both PCOS susceptibility and a spectrum of PCOS-related traits including metabolic and cardiovascular phenotypes. Finally, transcriptomic data showed downregulated MCM6 expression in granulosa cells of PCOS patients. Our study established a multipronged pipeline for PCOS gene prioritization and provided novel insights into the genetic architecture of PCOS.

Renal fibrosis is a final pathway leading to end-stage renal disease, with cellular senescence contributing to fibrosis and inflammation. Magnesium ions (Mg2+) are implicated in DNA stabilization and epigenetic regulation. In this study, we hypothesized that Mg2+ ameliorates renal fibrosis in association with reduced DNA damage responses and injury-induced cellular senescence, along with altered histone H3K4 trimethylation. To test this, we used murine models of radiation-induced organ injury and renal ischemia-reperfusion injury (IRI), along with primary cultured mouse renal proximal tubular cells. Mice received intraperitoneal MgSO4 (600 mg/kg) before radiation or IRI, with repeated dosing (300 mg/kg) after IRI. Cultured cells were treated with 6.4 mM MgSO4. We demonstrated that Mg2+ provided protection against radiation injury and reduced radiation-induced DNA damage markers in renal cells both in vitro and in vivo. Furthermore, Mg2+ suppressed IRI-induced morphological alterations, DNA damage, and cellular senescence in the kidneys, while inhibiting renal inflammation and cGAS-STING pathway activation, along with attenuation of renal fibrosis in IRI model mice. Consistent with these findings, a reduction in the expression of pro-inflammatory cytokines and fibrosis-related genes was observed. Finally, Mg2+ was associated with decreased p16INK4a transcription and reduced H3K4 trimethylation levels at its promoter in primary renal tubular cells. Our findings suggest that Mg2+ alleviates renal DNA damage while protecting against inflammation and fibrosis with accompanying epigenetic modulation. Although clinically relevant pharmacological Mg2+ dosing and therapeutic applicability require further investigation, these insights may inform therapeutic strategies targeting fibrosis and senescence-related kidney disease.

Neuroblastoma (NB) is a common pediatric tumor that exhibits significant clinical heterogeneity and substantial prognostic variability. However, there is still a lack of effective prognostic tools. It has been found that the differentiation of myeloid cells in the tumor microenvironment (TME) has a profound influence on tumor progression and immune regulation in neuroblastoma. But the correlation between NB and myeloid cell differentiation-related genes (MCDRGs) is still unclear. This study integrated bulk and single-cell RNA sequencing data to identify key prognostic genes associated with myeloid cell differentiation, construct and validate a risk prediction model, and thoroughly investigate the molecular mechanisms through which these genes regulate the TME and influence NB prognosis. Using bioinformatic approaches, we retrieved NB-related transcriptomic data from public databases, identified differentially expressed myeloid cell differentiation-related genes (MCDRGs), and constructed a prognostic risk model using Cox regression and machine-learning algorithms. Functional enrichment analysis, TME characterization, immune checkpoint profiling, drug sensitivity analysis, and somatic mutation analysis were performed between the high-risk group (HRG) and low-risk group (LRG). Single-cell RNA sequencing data were used to dissect the cell type-specific expression patterns of prognostic genes, which were further experimentally validated via reverse transcription quantitative polymerase chain reaction (RT-qPCR). Five key prognostic genes (FAXDC2, GP1BB, TRIB1, ETV2, and H4C12) were identified from 187 candidate genes, and a robust risk prediction model was constructed. Significant differences were observed between the HRG and LRG in terms of TME scores, immune cell infiltration, and immune checkpoint expression. Ribosome biogenesis and cell cycle-related pathways were enriched in the HRG, which displayed higher sensitivity to entinostat and sapitinib. Single-cell analysis further highlighted fibroblasts and myeloid cells as key cell subsets in which prognostic genes exhibited dynamic expression patterns. RT-qPCR further validated these gene expression trends. The prognostic model based on MCDRGs effectively predicts survival outcomes in patients with NB. This model systematically reveals the comprehensive mechanisms by which MCDRGs influence prognosis through regulating the TME, cell-cell interactions, and therapeutic responses. These findings provide a new theoretical basis and potential targets for precise risk stratification and individualized treatment strategies in NB.

Non-small cell lung cancer (NSCLC) accounts for ~85% of lung cancers, with platinum-based chemotherapy as the main treatment. Ferroptosis has been implicated in cancer chemoresistance, yet the molecular mechanisms linking metabolic reprogramming to ferroptosis-mediated cisplatin resistance in NSCLC remain elusive. In this study, we found that phosphoenolpyruvate carboxykinase 2 (PCK2), a metabolic reprogramming enzyme, was significantly upregulated, and its overexpression enhanced ferroptosis resistance, thereby promoting cisplatin chemoresistance. Mechanistically, RGB-286638 free base (RGB) inhibits PCK2 expression by directly binding to its R454 site to activate ferroptosis in cisplatin-resistant cells and restores cisplatin sensitivity both in vitro and in vivo. Targeting PCK2 with RGB provides a promising strategy to overcome chemoresistance, facilitating improved clinical interventions for NSCLC.

While immune complex-mediated glomerulonephritis causes severe kidney dysfunction in diseases like lupus nephritis, the signaling pathways involved are poorly understood. Here we tested the role of the Src-family kinases Hck, Fgr and Lyn, primarily expressed in innate immune cells, in immune complex-mediated glomerulonephritis. Hck, Fgr and Lyn expression increased in human lupus nephritis, as well as in mouse nephrotoxic nephritis triggered by sheep antiserum against mouse glomerular components (nephrotoxic serum). Nephrotoxic serum induced robust albuminuria, serum creatinine level increase, hematuria and ascites in wild-type but not in Hck-/-Fgr-/-Lyn-/- mice. The Hck-/-Fgr-/-Lyn-/- mutation also abrogated crescent formation and leukocyte infiltration. However, the glomerular IgG/complement deposition and the humoral immune response against sheep IgG were not affected. Transplantation of Hck-/-Fgr-/-Lyn-/- bone marrow cells transferred disease protection to wild-type recipients. Mixed bone marrow chimeric experiments revealed an indirect role for Hck, Fgr and Lyn in leukocyte recruitment. Src-family kinases mediated immune complex-induced activation of Syk, a recently proposed therapeutic target. Our results suggest that myeloid Src-family kinases are indispensable for nephrotoxic nephritis. Inhibitors targeting those kinases may provide therapeutic benefit in immune complex-mediated glomerulonephritis.

Zinc is an essential trace element that plays a significant role in DNA damage repair (DDR). Cells must maintain zinc homeostasis to effectively respond to DNA damage caused by various stressful stimuli. Members of the Zrt-, Irt-related protein (ZIP) family are recognized as key regulators of intracellular zinc levels. Dysfunction of ZIPs has been associated with numerous diseases, including cancer, cardiovascular diseases, and neurodegenerative disorders. In the present study, human bronchial epithelial BEAS-2B cells were used to investigate the response of ZIPs to bleomycin, a well-known antibiotic that induces DNA damage. Among the ZIPs that exhibited altered expression following bleomycin treatment, ZIP1 showed the most prompt and significant induction. Both knockdown and overexpression experiments demonstrated that ZIP1 plays a crucial role in the cellular response to DNA damage. Mechanistically, we found that the level of TNFα was upregulated in bleomycin-treated cells, and inhibiting this cytokine reduced the induction effect of ZIP1 caused by bleomycin. Additionally, we observed an increase in the accumulation of NF-κB in the cell nuclei, and inhibition of NF-κB also diminished the effect of bleomycin on ZIP1. Two NF-κB binding sites were identified in the upstream sequence of SLC39A1, the gene that encodes ZIP1. The application of bleomycin significantly enhanced the binding of NF-κB to these specific sites. Taken together, our study reveals that bleomycin treatment may activate the TNFα-NF-κB pathway, which in turn upregulates ZIP1, helping cells better cope with the DNA damage induced by this antibiotic.

Zinc finger protein 217 (Zfp217) mediates adipogenesis via an N6-methyladenosine (m[6]A)-dependent mechanism; however, its role in hepatic lipid metabolism is unexplored. Nonalcoholic fatty liver disease (NAFLD), characterized by hepatic triglyceride (TG) accumulation resulting from disrupted lipid homeostasis, lacks well-defined epigenetic regulatory mechanisms. Here, we report that global Zfp217 heterozygous knockout alleviates high-fat diet (HFD)-induced hepatic steatosis in mice, as evidenced by reduced liver weight, decreased hepatic and serum TG and total cholesterol (T-CHO) levels, and hepatic lipid deposition. Mechanistically, Zfp217 deficiency suppresses hepatic de novo lipogenesis (DNL) by down-regulating sterol regulatory element-binding transcription factor 1 (SREBF1), a master regulator of lipogenic gene expression. Zfp217 physically interacts with methyltransferase-like 3 (METTL3) to repress its expression, thereby reducing m[6]A modification of SREBF1 mRNA at a specific coding sequence (CDS) site. Loss of Zfp217 enhances METTL3-dependent m[6]A modification at a specific site of SREBF1 mRNA, which promotes YTH domain-containing family protein 2 (YTHDF2)-mediated degradation of SREBF1 transcripts. Conversely, our findings identify a novel Zfp217-METTL3-m[6]A-YTHDF2-SREBF1 regulatory axis that controls hepatic DNL and NAFLD progression, establishing Zfp217 as a potential therapeutic target for NAFLD.

The long-term treatment outcomes for peri-implantitis (PI) are more unstable and unpredictable than periodontitis (PD). Histological evidence suggests that the epithelial barrier and immune system around the implant are weaker than those around natural teeth. In transcriptomic aspects, compared to periodontitis and PI, the specific molecules and cell types involved in the pathogenesis of PI have not been fully established. In the current study, gingival tissue samples were collected from patients, and single-cell RNA sequencing was used to analyze the heterogeneities of epithelial barriers, immune defenses, and related response molecules in soft tissues around natural teeth and implants. Our work supports that the disruption of the epithelial barrier around the implant was one of the causes of PI. The cell communication between epithelial cells and stromal cells was inhibited by the midkine (MDK)-nucleolin (NCL) pathway. The breakdown of the defense structure of connective tissue aggravated the progress of PI. In vitro evidence demonstrated MDK's functional role in stromal regeneration through overexpression experiments. These findings provide critical molecular insights into the pathogenesis of PI and offer a strong rationale for the development of early, targeted clinical interventions aimed at preserving epithelial integrity and restoring immune defense in peri-implant soft tissues.

Somatic mechanical stimulation (e.g., acupuncture) exerts systemic immunomodulatory effects, yet the cellular bridge translating peripheral physical force into visceral repair remains elusive. Here, employing a custom interpretable deep learning framework (CARSS) on single-cell RNA sequencing data, we identify CD34[+]PDGFRα[+] telocytes (CPTCs) as the primary mechanosensors in both fascia and colon during bacterial colitis. We show that somatic mechanotherapy triggers an AP-1/Hsp70-dependent transcriptional program in fascial CPTCs, inducing systemic Wnt elevation, which elicits a "transcriptional resonance" in colonic CPTCs, reprogramming their communication network from an inflammatory amplifier to a Wnt-driven regenerative hub. Mechanistically, this axis activates epithelial β-catenin/Myc signaling, suppressing apoptosis and restoring barrier integrity independent of immune cells. Our findings define a CPTC-Driven Mechano-Resonance Axis, where CPTCs serve as synchronized relay stations that convert somatic mechanical stimulation into systemic regenerative microenvironments.

Brucellosis is a zoonotic disease caused by Brucella spp. that poses a significant threat to human health. Copper resistance is a key determinant of the virulence of many intracellular pathogens, including those expressing the multicopper oxidase CueO. However, the role of CueO in Brucella remains poorly understood. In this study, bioinformatics tools were used to perform sequence homology comparison and conserved domain analysis of the CueO protein. Unmarked cueO deletion mutant and complemented strain of Brucella were constructed using homologous recombination technology. Subsequently, multiple experiments were carried out, including in vitro copper tolerance tests, RAW264.7 macrophage intracellular survival assays, in vivo virulence detection in mice, antibiotic susceptibility tests (AST), and so on. We found that CueO contains highly conserved domains across bacteria (motifs 1-10). Notably, the Brucella homolog contains only motifs 2, 3, 5, 6, and 10; it remains highly conserved within the genus. Despite this reduced domain composition, CueO is important for Brucella to resist copper toxicity. Notably, CueO contributes to intracellular survival in RAW264.7 macrophages and to the maintenance of copper homeostasis in Brucella. Deletion of cueO attenuated bacterial virulence in a mouse model. In addition, CueO influences bacterial susceptibility to doxycycline and levofloxacin. Overall, our results indicate that, despite lacking several typical domains, CueO retains critical functionality and links copper detoxification to intracellular survival, virulence, and antibiotic resistance. These findings enhance our understanding of CueO function and provide new insights into the pathogenesis of Brucella.

Intracellular membrane fusion typically involves two distinct classes of GTPases. Dynamin-like GTPases mediate homotypic fusion between organelles, such as mitochondria and the endoplasmic reticulum (ER), while Rab GTPases facilitate fusion of transport vesicles with target membranes through vesicle tethering. Notably, these two classes of GTPases have not previously been implicated in the same fusion event. In this study, we demonstrate that Rab10 promotes ER membrane fusion driven by the dynamin-like GTPase atlastin (ATL). Rab10 interacted physically with ATL2, a human ATL predominantly expressed in non-neuronal cells, and co-localized with ATL2 throughout the ER, including at three-way junctions where fusion occurs. Fusion between ER microsomes isolated from HEK293T cells, in which ATL2 is the primary ATL isoform, was inhibited by affinity-purified anti-Rab10 antibodies, and was reduced in microsomes derived from Rab10 knockout cells. Moreover, co-reconstitution of Rab10 markedly enhanced fusion of ATL2-containing liposomes. Our findings reveal crosstalk between dynamin-like and Rab GTPases during ATL-mediated ER membrane fusion, uncovering a novel regulatory mechanism for organelle dynamics.

Early detection of obstructive sleep apnea (OSA) in adults with metabolic dysfunction associated steatotic liver disease (MASLD) remains a clinical challenge. Plasma proteomics provides a non-invasive tool for identifying individuals at elevated risk prior to symptom onset. We quantified 2911 plasma proteins in 17 375 OSA-free MASLD subjects from the UK Biobank (UKB). Participants were split into training and validation sets. Cox regression and Light Gradient Boosting Machine (LightGBM) with forward feature selection were used to identify and rank predictive proteins. Model performance was assessed by area under the receiver operating characteristic curve (AUC) in the validation set, and shapley additive explanations (SHAP) values were used to interpret feature contributions. Among 17 375 MASLD participants, 541 (3.1%) developed subsequent clinical diagnosis of OSA. Seven core proteins (FABP4, PON3, RTN4R, KIAA0319, CFC1, TFRC, CST3) were identified and combined into a protein risk score (ProRS). The ProRS stratified high- and low-risk individuals with C-index 0.727 and 10-year AUC 0.727, outperforming the clinical risk score (C-index 0.548). Integration with clinical factors further improved discrimination (C-index 0.765, 10-year AUC 0.765) and net benefit. The plasma protein-based ProRS predicts future OSA in MASLD and enhances risk stratification beyond clinical variables, supporting its potential for early, non-invasive screening and targeted intervention.

Heart failure (HF) resulting from chronic systolic overload is associated with increased burden on proteostasis and impaired proteasome function. Phosphorylation of proteasome subunit RPN6/PSMD11 at Ser14 (pS14-RPN6) mediates the activation of 26S proteasomes by PKA, but its significance in common forms of heart disease remains obscure. Hence, we investigated the impact of genetic blockade of pS14-RPN6 on cardiac remodeling and HF during systolic overload, a common condition occurring in hypertension and aortic stenosis. We detected marked increases in ubiquitin conjugates, along with elevated levels of RPN6 and pS14-RPN6 proteins in myocardial tissues from human patients with nonischemic HF. Similarly, myocardial pS14-Rpn6 was increased in wild-type mice 2 and 4 weeks after transverse aortic constriction (TAC). Compared with wild-type littermates, mice with genetic blockade of pS14-Rpn6 resulting from germline knock-in of Rpn6[S14A] (S14A mice) developed greater cardiac hypertrophy, fibrosis, left ventricular dysfunction, and lung congestion after TAC. Mechanistically, TAC induced comparable increases in 26S and 30S proteasomes at 2-week in both genotypes but, at 4-week post-TAC, the increases were remarkably attenuated in S14A mice. Meanwhile, TAC-induced increases in proteasome peptidase activities were significantly attenuated (2-week) or abolished (4-week) in S14A mice, accompanied by greater increases in total and K48-linked ubiquitin conjugates. Collectively, these findings demonstrate that pS14-RPN6 plays an essential role in upregulating myocardial proteasome activities and sustained upregulation of proteasome assembly, thereby assisting in maintaining proteostasis and protecting against maladaptive cardiac remodeling and HF during systolic overload.