Multiple myeloma (MM) is an incurable malignancy of bone marrow plasma cells. Tumor-associated macrophages (TAMs) are the predominant immune cells in the bone marrow microenvironment of MM and play important roles in MM. The effect of RBMS1 on MM has not yet been reported. This study aimed to investigate the function of RBMS1 in MM. Through the analysis of GSE datasets, we identified RBMS1 as a potential pathogenic factor in MM. We further investigated the effects of RBMS1 on the proliferation of MM cells (RPMI8226, MM1S, and KMS11) and its regulation of TAM polarization. An animal model was established by intravenous injection of MM cells into 6-week-old male NOG mice. Bioinformatics analyses, including RRA, WGCNA, and GO enrichment, screened for potential pathogenic genes in MM. Kaplan - Meier survival identified RBMS1 as a prognostic marker associated with poor outcomes in MM. Functionally, RBMS1 enhanced MM cell proliferation, colony formation, and cell cycle. Moreover, RBMS1 promoted M2 polarization of macrophages, as evidenced by elevated levels of M2 macrophage markers, as well as increased CCL2 secretion. Consistently, in a male NOD/Shi-scid IL-2 Rγnull mouse xenograft model, RBMS1 accelerated tumor growth and enhanced M2 macrophage polarization. Mechanistically, RBMS1 bound to the 3'UTR of PDPK1 mRNA, enhancing its stability and activating the pro-tumorigenic β-catenin signaling pathway, thereby promoting tumor growth. Collectively, this study is the first to report the functional role of RBMS1 in MM and highlights the importance of the RBMS1/PDPK1/β-catenin signaling axis in MM, providing new insights for basic research on MM.

Oxidative stress is one of the crucial factors associated with the pathogenesis of age-related macular degeneration (AMD), particularly the degeneration of retinal pigment epithelium (RPE) cells. In this study, we identified nepetin, a natural flavonoid compound, as a potential inhibitor of hydrogen peroxide (H2O2)-induced ARPE-19 cell death. Pretreatment of nepetin significantly reduced intracellular generation of reactive oxygen species (ROS). Quantitative proteomics was applied to explore the underlying molecular response, revealing that 77 proteins were up-regulated, and 198 proteins were down-regulated significantly after nepetin treatment. Gene ontology (GO) analysis and the protein-protein interaction (PPI) network analysis showed that heme oxygenase 1 (HO-1), Kelch-like ECH-associated protein 1 (KEAP1), Sequestosome 1 (SQSTM1)/p62, and glucose-regulated protein 78 (GRP78) were associated with nepetin-mediated antioxidative responses. Western blotting confirmed the altered expression of these key proteins, with HO-1, p62, and GRP78 being upregulated and KEAP1 being downregulated. Immunofluorescence further showed nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear translocation, suggesting the involvement of Nrf2-related antioxidant signaling in nepetin-treated ARPE-19 cells. Although direct causal interactions were not established, the proteomic and bioinformatic analyses provide correlative and suggestive evidence that nepetin modulates key proteins within the oxidative stress response network. Based on our previous research and the current study, nepetin exhibits both anti-inflammatory and antioxidative properties in RPE cells, and may have potential implications for the prophylaxis and treatment of AMD, particularly dry AMD.

To investigate the potential roles of histone lactylation, N6-methyladenosine (m6A) modification, and the Janus kinase-signal transducer and activator of transcription 3 (JAK-STAT3) pathway in regulating the wound healing following corneal alkali burns. An in vivo corneal alkali burn model was established using C57BL/6J mice. Corneal inflammation, fibrosis, histone lactylation, pan-lactylation, m6A modification, and JAK-STAT3 pathway activation were assessed at indicated time points post-injury using slit-lamp examination, hematoxylin and eosin (H E) staining, RT‒qPCR, and western blotting. In vitro, the effects of histone lactylation and m6A modification on JAK-STAT3 signaling and inflammation were investigated in interleukin-1β (IL-1β)-stimulated keratocytes. The impact of histone lactylation on corneal wound repair was evaluated by employing the glycolysis inhibitors 2-deoxy-D-glucose (2-DG) and rotenone, which modulates glycolysis via mitochondrial complex I inhibition. Analysis of the GSE191228 dataset revealed that JAK-STAT pathway plays an important role in corneal wound healing. Gene Set Enrichment Analysis (GSEA) of proteomic data further demonstrated substantial upregulation of the IL-6-JAK-STAT pathway in alkali-burned corneas. Additionally, the analysis revealed a positive correlation between both hypoxia and glycolysis pathways and the corneal alkali burns condition. In the mouse model, corneal alkali burns activated JAK-STAT3 pathway, elevated methyltransferase-like 3 (METTL3) expression, and increased global m6A modification. Furthermore, histone H3 lysine 18 lactylation (H3K18la) levels were significantly increased, accompanied by characteristic changes in pan-lactylation expression patterns. At day 7 post-alkali burn, treatment with 2-DG alleviated corneal inflammation and fibrosis while decreasing the levels of H3K18la, METTL3, and pSTAT3. In vitro, knockdown of LDHa significantly attenuated IL-1β-stimulated upregulation of H3K18la, METTL3 and pSTAT3 in keratocytes. Consistently, treatment with 2-DG or rotenone decreased or increased the expression of H3K18la, METTL3, and pSTAT3, as well as inflammatory factor IL-6, respectively. Moreover, reducing METTL3 expression via siRNA-METTL3 transfection decreased pSTAT3 activation. In TGF-β-induced fibrotic response of keratocytes, 2DG suppressed the TGF-β-mediated elevation of H3K18la, METTL3 and αSMA. Additionally, LPS similarly elevated H3K18la levels in keratocytes. Hypoxia and glycolysis following corneal alkali burn initiate lactylation, subsequently activating m6A modification and the JAK-STAT3 signaling pathway. This lactylation-induced cascade critically regulates corneal wound healing and represents a promising therapeutic target.

AIMS/BACKGROUND:Allergic rhinitis (AR) is an upper respiratory disease that affects inflammation levels, nasal function, and mental health in patients. However, the effect of AR severity on these indicators remains obscure. This study aimed to explore the impacts of AR severity on levels of inflammatory factors, nasal function, anxiety and depression. METHODS:The clinical data of 188 patients with AR from January 2022 to January 2025 were collected and retrospectively analyzed. The patients were divided into mild group (n = 90) and moderate/severe group (n = 98) based on the severity of AR. Meanwhile, 79 healthy individuals matched in age, gender, and body mass index (BMI) with the AR patients were included in the control group. Nasal airway resistance (NAR) and nasal mucociliary clearance time (NMCT) were detected. Hospital Anxiety and Depression (HAD) scale was applied for the assessment of anxiety and depression. Serum level of C-reactive protein (CRP) was measured using an automatic biochemical analyzer. Serum procalcitonin (PCT) and nasal lavage fluid levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were measured using commercial assay kits. RESULTS:Compared with the control group, the CRP, PCT, IL-1β, TNF-α, NAR, NMCT, and HAD anxiety and depression scores in AR patients were significantly increased (both p < 0.05). Compared with the mild group, the moderate/severe group exhibited increased levels of inflammatory biomarkers, NAR, NMCT, and HAD anxiety and depression scores (p < 0.05). In the mild group, anxiety and depression were correlated with the NAR, CRP, PCT, IL-1β, and TNF-α (p < 0.05); NMCT was correlated with the depression (p < 0.05). In moderate/severe group, anxiety and depression were correlated with the NAR, NMCT, CRP, PCT, IL-1β, and TNF-α (p < 0.05). The correlation between anxiety and depression and nasal function and inflammatory factors in moderate/severe group were stronger than those in mild group. CONCLUSION:The anxiety/depression and inflammation levels in AR patients increase, while the nasal function decreases, with the deteriorating severity of the disease. Anxiety and depression are correlated with nasal function and inflammation levels, with a more prominent correlation detected in patients with moderate/severe AR than those with mild disease.

Cancer-related pain and anxiety significantly negate the quality of life in patients. Oligodendrocyte precursor cells (OPCs) were reported to involve in engulf synapses and remodel neural circuits. This study aimed to elucidate the role of OPCs-mediated phagocytosis of GABAergic synapses in a mouse model of bone cancer pain. Male C3H mice were utilized to establish a model of bone cancer pain. In vivo fiber photometry was used to monitor the activity of GABAergic neurons, and chemogenetic techniques were applied to modulate neuronal excitation. The phagocytosis of GABAergic synapses by OPCs was visualized via immunofluorescence-based 3D reconstruction and immunoelectron microscopy. Interventions targeting lipocalin-2 (LCN2) and its receptor SLC22A17 were carried out with adeno-associated virus (AAV), siRNA, and pharmacological tools. On the 21st postoperative day, mice with bone cancer displayed significant pain and anxiety-like behaviors. Tumor-bearing mice exhibited a compensatory increase in calcium activity among GABAergic neurons within the anterior cingulate cortex (ACC). Compared with sham-operated mice, OPC phagocytosis of GABAergic synapses was higher in the tumor-bearing mice than controls. Concurrently, a pronounced upregulation of LCN2 expression was observed in the tumor group. Administration of LCN2-neutralizing antibodies or AAV-mediated intervention markedly alleviated pain and anxiety-related behaviors in mice with bone cancer. Moreover, the LCN2 receptor SLC22A17 expression was significantly increased. Targeted inhibition of SLC22A17 induced cytoskeletal remodeling and decreased their phagocytic capacity of OPCs. Collectively, LCN2/SLC22A17 signal was involved in OPCs-mediated phagocytosis of GABAergic synapses and contributed to cancer pain and anxiety development.

Corneal alkali burns rank among the most severe ocular injuries, characterized by a dynamically evolving pathology that requires distinct therapeutic strategies at different stages—a critical challenge that conventional static drug delivery systems fail to address. To overcome this limitation, we developed an in situ-forming dual-crosslinked hydrogel (AlgMA/GelMA) capable of time-sequential drug release for stage-specific pharmacotherapy. It was engineered to release dexamethasone phosphate (Dex-P) rapidly to control acute inflammation, followed by sustained release of glycyrrhizin (Gly) encapsulated in PLGA nanoparticles (NPs). This design not only circumvents the ocular side effects associated with long-term glucocorticoid use but also targets HMGB1 to suppress the pro-fibrotic inflammation–fibrosis axis. In vivo studies demonstrated that this sequential release system significantly accelerated corneal epithelial regeneration, restored corneal transparency, and suppressed key inflammatory pathways. Mechanistic investigations revealed that the coordinated action of Dex-P and Gly potently suppresses the HMGB1–NF-κB–IL-1β pathway, thereby effectively attenuating both inflammatory and fibrotic cascades. This study establishes a versatile, pathophysiology-driven material platform, proposing a new paradigm for treating multi-stage ocular diseases and tissue injuries.

BACKGROUND:Fluoroquinolones (FQs) have been associated with an increased risk of heart valve regurgitation, yet the cellular mechanisms underlying this association remain unclear. Although the effects of fluoroquinolones on cardiac valvular interstitial cells have been explored in experimental models, the specific impacts and mechanisms of levofloxacin on heart valve interstitial cells remain insufficiently understood. MATERIALS AND METHODS:Aortic interstitial cells (AICs), mitral interstitial cells (MICs), and tricuspid interstitial cells (TICs) were extracted through collagenase digestion. The isolated cells were divided into the intervention group (treated with 20 μg/mL levofloxacin) and control group. The expression of matrix metalloproteinases (MMPs), collagen, and elastin was quantified by Western blot (WB). The impact of levofloxacin on cell apoptosis was explored using flow cytometry. RESULTS:The expression level of MMPs, which contribute to collagen degradation, was upregulated in AICs, MICs, and TICs treated with levofloxacin. Elastin and collagen, the key components of the extracellular matrix (ECM) of cardiac valve cells, were decreased in AICs and TICs, whereas no significant changes were observed in MICs following levofloxacin treatment. Moreover, prolonged levofloxacin exposure significantly enhanced apoptosis in MICs, while a pro-apoptotic trend was observed in AICs and TICs. CONCLUSIONS:Levofloxacin promoted extracellular matrix remodeling by modulating the expression levels of MMP, collagen and elastin, and inducing apoptosis of valve interstitial cells, which may contribute to pathological remodeling of heart valves.

Astrocytic activation is critically involved in the development and maintenance of bone cancer pain (BCP). Recent studies have shown that astrocytes participate in synaptic remodeling through synaptic phagocytosis, whereas neuronal cell adhesion molecule (NRCAM) restricts the extension of perisynaptic astrocytic processes and exerts an anti-phagocytic effect. However, whether astrocyte-mediated synaptic phagocytosis contributes to BCP remains to be explored. Hence, this study investigated the role of astrocytic NRCAM deficiency in synaptic remodeling and central sensitization in BCP. A mouse model of BCP was established by intrafemoral inoculation of fibrosarcoma cells. Pain-related behaviors were evaluated by spontaneous pain assessment and mechanical withdrawal threshold testing. Bone destruction and tumor infiltration were examined by hematoxylin and eosin staining. Synaptic alterations and astrocytic activation were assessed by western blotting and immunofluorescence. Astrocyte-mediated synaptic engulfment was analyzed using immunofluorescence, three-dimensional reconstruction, magnetic-activated cell sorting, and Golgi-Cox staining. Moreover, fluorocitrate was used to inhibit the astrocytic activation. Astrocytic NRCAM was overexpressed via adeno-associated virus (AAV)-mediated gene delivery. Fibrosarcoma cell inoculation induced persistent spontaneous pain and mechanical hyperalgesia in C3H/HeN mice, accompanied by trabecular bone destruction and tumor infiltration. In the spinal cord, expression of the excitatory synaptic markers VGLUT1 and PSD95 was increased, whereas expression of the inhibitory synaptic markers VGAT and Gephyrin was decreased, together with marked astrocytic activation. Multiple complementary assays consistently showed that astrocytes engulfed both excitatory and inhibitory synapses, with a clear preference for GABAergic synapses. Pharmacological inhibition of astrocytic activation with fluorocitrate alleviated pain behaviors and improved synaptic remodeling. NRCAM expression was significantly downregulated in tumor-bearing mice. Restoration of astrocytic NRCAM expression by AAV markedly reduced astrocyte-mediated engulfment towards GABAergic synapses, had minimal effect on excitatory synapses, and significantly attenuated BCP. Reactive astrocytes preferentially phagocytose GABAergic synapses in BCP, thereby contributing to synaptic imbalance and central sensitization. This process is associated with downregulation of astrocytic NRCAM. Restoring astrocytic NRCAM alleviates BCP by suppressing excessive astrocyte-mediated phagocytosis of GABAergic synapses. These findings identify astrocytic NRCAM-dependent synaptic phagocytosis as an unrecognized mechanism underlying BCP and as a potential therapeutic target.

Deltex E3 ubiquitin ligase 3L (DTX3L) is a well-established ubiquitin ligase implicated in various cancers, but its role in nasopharyngeal carcinoma (NPC) progression remains elusive. In this study, we confirmed for the first time that DTX3L was highly expressed in C666-1 and NPC/HK1 NPC cells. DTX3L overexpression promoted NPC cell proliferation, invasion, and migration. Conversely, DTX3L knockdown suppressed these malignant phenotypes. Notably, DTX3L activated the β-catenin pathway, as evidenced by increased β-catenin nuclear translocation, increased transcriptional activity, and elevated expression of its downstream target c-Myc. Mechanistically, we identified an upstream regulatory axis in which the oncogenic long noncoding RNA H19 acted as a molecular sponge for miR-423-5p, thereby alleviating the miR-423-5p-mediated repression of DTX3L. Dual-luciferase reporter assays confirmed that miR-423-5p directly targeted both DTX3L and H19, supporting the presence of a competitive endogenous RNA (ceRNA) network. Furthermore, rescue experiments demonstrated that DTX3L knockdown largely abolished the proliferative advantage conferred by H19 overexpression. Collectively, the results of our study revealed that the H19/miR-423-5p/DTX3L axis is a novel ceRNA-driven mechanism that promotes NPC progression via activation of β-catenin signaling.

The objective of this work was to investigate the role of ZFP36 in mitral valve prolapse (MVP). Mitral valve and plasma were collected to assess the expression of ZFP36, transforming growth factor β (TGF‐β), collagen and elastin and apoptosis rates. Mitral valve interstitial cells (MICs) were transfected with ZFP36 plasmids to observe changes in the secretion of collagen, elastin and matrix metalloproteinases (MMPs) and apoptosis rates. MICs were exposed to TGF‐β1 to evaluate the changes in expression of collagen, elastin, MMPs and ZFP36 and apoptosis. Subsequently, after transfection with ZFP36 plasmid, exogenous TGF‐β1 was added to the MICs, and the secretion of collagen, elastin and MMPs and apoptosis rate were re‐evaluated. Finally, transcriptome and RNA immunoprecipitation (RIP) sequencing was conducted to identify downstream genes of TGF‐β1 that could bind to ZFP36. Patients with MVP showed elevated levels of TGF‐β1 in plasma and increased rates of apoptosis, along with higher expression of ZFP36, TGF‐β1, collagen and elastin in the prolapsed valve. Overexpression of ZFP36 in MICs did not significantly alter the secretion of collagen or elastin or apoptosis rates. TGF‐β1 promoted apoptosis of MICs, increased the secretion of collagen, elastin, MMP‐3,9, ZFP36 and reduced the expression of MMP‐1,2,13. Moreover, overexpression of ZFP36 inhibited the effects of TGF‐β1 on MICs. Co‐analysis of transcriptome and RIP sequencing identified three genes: CFAP184, GTP binding protein 6 (GBP6) and HERC6. Knockdown of GBP6 reduced the pro‐apoptotic effects of TGF‐β1 on MICs. ZFP36 exerts a protective role in MVP by inhibiting the effects of TGF‐β1 on MICs. Notably, ZFP36 can mitigate the pro‐apoptotic effects of TGF‐β1 on MICs through the GBP6 pathway.

Background Allergic rhinitis (AR) affects the nasal mucosa, causing significant quality of life impairments. Sublingual immunotherapy (SLIT) is effective in mitigating AR symptoms. Autophagy regulates immune responses and is implicated in AR. We aimed to investigate the impact of SLIT on autophagy-associated proteins and their relationship with Th1 and Th2 cytokines in AR patients. Methods Sixty AR children were recruited. They underwent standardized SLIT with Der f extract. Efficacy was evaluated using combined symptom and medication score (CSMS). Blood samples were analyzed for autophagy-related proteins (ATG5, p62, Beclin-1, LC3I, LC3II) and Th1/Th2 cytokines (IFN-γ, IL-4). Results After 3 years’ SLIT, ATG5, LC3I, LC3II, and Beclin-1 expressions decreased, while p62 increased. Th1 cytokine expression increased, and Th2 cytokine expression decreased. The effective group showed greater changes in protein expression compared to the ineffective group. A positive correlation was found between autophagy-associated protein expression and Th2 cytokines. Conclusion SLIT alters autophagy-related protein levels and reverses the Th1/Th2 balance in AR patients, suggesting autophagy-associated proteins as potential biomarkers for SLIT efficacy.

Despite recent developments,the genetics and biology of Alzheimer's disease remain insufficiently characterized.As an important first step toward developing effective treatment strategies to slow or prevent Alzheimer's disease onset,the identification of relevant genetic markers is crucial.In the present study,we analyzed transcriptomic and multi-omic datasets across multiple cohorts(the Alzheimer's Disease Neuroimaging Initiative,Religious Orders Study and Rush Memory and Aging Project,Mount Sinai Brain Bank,and Mayo Clinic Alzheimer's Disease Genetics Studies)using gene set enrichment analysis,machine learning algorithms,and polygenic risk scoring to identify gene sets relevant to Alzheimer's disease risk and pathological features.For prioritized gene sets,we performed epigenome-wide association studies to assess DNA methylation patterns,and used multi-omic mediation analysis to characterize the causal gene regulatory networks.Overall,we identified several key gene sets relevant to Alzheimer's disease pathology—particularly,those related to immune system function and mitochondrial dysfunction.Upregulated pathways,including neutrophil degranulation and tumor necrosis factor-α signaling pathways,correlated strongly with aspects of neuroinflammation in Alzheimer's disease.By contrast,downregulated oxidative phosphorylation pathways further suggested mitochondrial dysfunction.Gene sets that contained mitochondrially located genes(e.g.,SGK1 and LRRK1)were identified as significantly contributing to neurodegeneration.Moreover,genes such as CXCL1,TGFB2,and DUSP1 were consistently implicated in all datasets,thus emphasizing their involvement in immune modulation and mitochondrial function.The multimodal investigation outlined in the current study represents useful steps toward comprehending the genetic architecture of Alzheimer's disease,including an expanded understanding of the spatial interactions of genes associated with disease susceptibility.Mitochondrial dysfunction and immune modulation were pathological pathways that converged on Alzheimer's disease and future treatment novel options.Using the frameworks provided in the current comprehensive study,we present opportunities to explore targeted treatment strategies that may alter immune systems and mitochondrial function to optimize treatment outcomes for individuals at increased risk of or living with Alzheimer's disease.

Vision impairment caused by cornea-related diseases seriously affects patients' quality of life, and the shortage of corneal donors prompts the search for more substitutes. The three-dimensional bioprinting technology has rapidly developed in recent years and provides new hope for corneal transplantation and regeneration. This review discusses the crosslinking methods of different bioinks, such as ion crosslinking, physical crosslinking, and photocrosslinking for bioprinting. We then summarized the characteristics of biomacromolecule-based bioinks, including decellularized extracellular matrix and natural polymer-based bioinks, such as alginate, gelatin, gelatin methacrylate, hyaluronic acid, and collagen, highlighting the respective disadvantages of single-component hydrogel inks and improvements (mechanical strength, biocompatibility, and light transmittance) for cornea bioprinting. We also focused on the assistant bioinks, including support baths and sacrificial inks, and explored their potential in high-fidelity bioprinting and the preparation of porous hydrogels. In addition, bioinks for corneal structure bionics, regenerative functions, and clinical applications are discussed. Finally, we discuss the evaluation of bioinks for creating functional corneal substitutes and look forward to the combination of bioprinting and other biofabrication methods. In summary, this review presents the latest advancements of corneal bioinks, discusses the potential preparation strategy, and challenges in the development and evaluation of new generation bioinks in the field of corneal regenerative medicine.

Perineurial network (PNN) is a special extracellular matrix structure in the central nervous system, and its alterations are associated with the pain hypersensitivity. Recent studies have suggested a potential interaction between abnormal activation of spinal microglia and PNN. This study investigates whether S-ketamine mitigates neuropathic pain via inhibiting degradation of PNNs by spinal microglia. C57BL/6 mice were utilized for CCI modeling to induce neuropathic pain. Subsequent to modeling, we assessed the expression changes of spinal microglia, PNN and inflammatory factors. Microglia colocalization with PNN was evaluated via 3D reconstruction to quantify spatial overlap. Minocycline was administered to target microglia. S-ketamine was subsequently administered to CCI mice, and its effects on pain behavior, microglial activation, and PNN were investigated. Microglia-PNN colocalization was evaluated via 3D reconstruction to quantify spatial overlap. CCI mice exhibited significant neuropathic pain, accompanied by increased microglia-mediated phagocytosis of PNN. Minocycline and S-ketamine treatment of CCI mice led to improved pain thresholds, suppression of neuroinflammation, and reduction in microglia-mediated phagocytosis of PNN. Increased microglial phagocytosis leading to PNNs degradation in the spinal dorsal horn plays a critical role in neuropathic pain pathogenesis. The analgesic effects of S-ketamine may be attributed to its modulation of this mechanism.

Cancer pain, a frequent complication in patients with cancer, adversely affects quality of life and survival rates. Microglia promote nociceptive information transmission by modulating myelin integrity during pain perception. However, the specific mechanisms by which microglia regulate myelin in the context of cancer pain remain poorly understood. In this study, we developed a bone cancer pain model to examine the interactions among microglia, myelin, and oligodendrocyte precursor cells and their roles in cancer pain. Our study found that mice with bone cancer pain had oligodendrocyte differentiation defects and myelin loss, and that promoting myelination did not relieve pain. In addition, we observed that reactive microglia and inflammatory cytokines increased and microglia phagocytosed myelin in mice with bone cancer pain. Inhibition of microglia not only alleviated pain behaviors in mice with bone cancer but also mitigated myelin phagocytosis and the proliferation of oligodendrocyte precursor cells. Our study suggests that microglia-mediated myelin loss and oligodendrocyte precursor cell proliferation may be one of the pathological mechanisms underlying pain in mice with bone cancer.

BACKGROUND: Cervical cancer (CC) is caused by human papillomavirus (HPV) infection and has serious impacts on women's health. While some HPV vaccines are already available, there is still a need for deeper understanding of the pathogenesis of CC and better ways to diagnose and treat it. METHODS: qRT-PCR was used to determine the level of miR-141-5p expression in CC. A CCK-8 assay, colony formation experiment, and Transwell assay were used to determine the influence of miR-141-5p on biological characteristics. A dual luciferase assay was employed to examine the correlation between miR-141-5p and MRVI1. The expression of MRVI1 and proteins linked to epithelial-mesenchymal transition (EMT) were identified by western blot. Knockdown ofMRVI1 was performed to investigate its revertive effect on miR-141-5p down-regulation. A subcutaneous tumor-graft model was constructed to explore the impacts of down-regulating miR-141-5p on tumors in vivo, and the positivity of Ki-67 and MRVI1 in CC tissues was detected by immunohistochemistry. RESULTS: The level of miR-141-5p was markedly increased in CC. miR-141-5p knockdown reduced the viability, migration, and invasion of CC cells and hindered their EMT process. The opposite result was obtained when miR-141-5p was over-expressed. MRVI1 was screened as a target gene of miR-141-5p. MiR-141-5p negatively regulated MRVI1 expression, and silencing MRVI1 reversed the impact of miR-141-5p down-regulation on CC cells. Knockdown of miR-141-5p upregulatedMRVI1 expression in tumor tissues and inhibited CC formation in vivo. CONCLUSIONS: miR-141-5p was markedly increased in CC and regulated CC malignant progression by targeting and negatively regulating MRVI1. Additionally, knockdown of miR-141-5p inhibited CC progression in vivo.

BACKGROUND:miRNAs play a crucial role in the anti-inflammatory effects of inhaled corticosteroids (ICS) in asthma. Vitamin D can modulate the expression of several miRNAs and reduces asthma exacerbations, but its molecular interaction with ICS remains unclear. OBJECTIVE:We hypothesised that vitamin D influences long-term ICS response through miRNA regulation. METHODS:Baseline serum miRNAs were sequenced from 462 subjects in the Childhood Asthma Management Program (CAMP), with 187 randomised to ICS treatment included in this study. Linear regression assessed associations between miRNA expression and prebronchodilator forced expiratory volume in 1 s per cent predicted (FEV1%) change over 4 years, stratified by baseline vitamin D levels and tested in interaction models. Microarray analysis of lymphoblastoid B cells (lymphoblastoid cell lines (LCLs)) from 22 CAMP subjects treated with dexamethasone (DEX), vitamin D or sham identified differentially expressed genes (DEGs). An miRNA target gene network was constructed, clustered and annotated by enrichment analysis. Top miRNAs were evaluated for ICS response prediction. RESULTS:12 miRNAs were significantly associated with ICS-mediated FEV1% change in vitamin D insufficient subjects, and 11 miRNAs showed significant interaction with vitamin D (p≤0.05). Three miRNAs were approximately replicated in the Genetics of Asthma in Costa Rica Study. Microarray analysis identified 220 and 240 DEGs in DEX and vitamin D-treated LCLs, respectively. miRNAs hsa-miR-125a-5p, hsa-miR-181a-5p, hsa-miR-101-3p and hsa-miR-107 were enriched in haemopoiesis and leucocyte differentiation pathways (p≤0.05). Two miRNAs, hsa-miR-125a-5p and hsa-miR-181a-5p, predicted ICS response with an area under the receiver operating characteristic curve of 0.86 in the vitamin D insufficient group. CONCLUSIONS:Vitamin D may modulate ICS response through miRNAs involved in immune cell differentiation, which could serve as biomarkers for ICS response, particularly in vitamin D insufficient individuals.

ObjectivesFerroptosis, a regulated form of cell death, has attracted significant attention in hearing loss research; however, the role of ferroptosis-related genes remains unclear. This study aimed to clarify diagnostic and therapeutic targeting of ferroptosis-related genes in hearing loss.MethodsDifferentially expressed genes related to hearing loss from the GEO database were intersected with ferroptosis-related genes. The Lasso and SVM-RFE models were applied to reduce the gene set, identifying model genes. Biological functions, pathways, and gene-drug associations related to these model genes were analyzed. Age-related hearing loss (ARHL) genes within the model genes were obtained from a genome-wide association study (GWAS) dataset. Further validation was conducted in HEI-OC1 cells and the cochleae of C57BL/6J mice, including auditory brainstem response (ABR) testing, qRT-PCR, Western blotting, Fe2+ detection, and immunofluorescence analysis.ResultsThe study identified 20 ferroptosis-related genes associated with hearing loss. Using Lasso and SVM-RFE models, a novel model was constructed, consisting of nine genes (SCD, ENPP2, PANX2, NEDD4, MEF2C, ABCC5, KLHDC3, CYP4F8 and IFNA2). Among these, MEF2C and NEDD4 were found to be associated with ARHL.ConclusionFerroptosis is a potential pathological mechanism in hearing loss research, and the nine ferroptosis-related genes identified provide promising targets for exploring new diagnostics and treatments for hearing loss. Notably, MEF2C and NEDD4 are associated with ARHL.

AIMS:Alzheimer's disease (AD) is a devastating neurodegenerative disorder where mitochondrial dysfunction is increasingly recognized as pivotal, yet its comprehensive molecular underpinnings remain incompletely understood. This study aimed to systematically identify and validate mitochondria-related biomarkers associated with AD risk and brain resilience, thus elucidating the molecular mechanisms underpinning mitochondrial dysfunction in AD. METHODS:We innovatively integrated a multi-omics approach, encompassing genomics, DNA methylation, RNA-sequencing, and miRNA profiles from the ROSMAP and ADNI cohorts (sample sizes ranging from 638 to 2090 per omic layer). Additionally, we applied 10 distinct machine learning methods to robustly identify and validate critical mitochondrial biomarkers relevant to AD progression. Subsequent validation was performed using a two-tiered approach: an in vivo AD mouse model to establish phenotypic relevance and an in vitro H2O2-induced oxidative stress model in HT22 cells to provide direct mechanistic validation. RESULTS:Our computational analyses identified key biomarkers such as hsa-miR-129-5p and SLC6A12 as pivotal regulators and highlighted the importance of the tricarboxylic acid (TCA) cycle. Experimentally, our AD mouse model exhibited significant cognitive deficits and brain remodeling, linked to a specific transcriptomic signature. Our in vitro model functionally recapitulated mitochondrial dysfunction and oxidative stress. Crucially, a cross-model analysis revealed a core signature of seven genes (including APOE, CDKN1A, and CLOCK) consistently dysregulated in both the cognitively impaired mouse brain and in neuronal cells subjected to direct oxidative insult. This provides powerful functional evidence linking our computationally derived targets, such as mitochondrial-epistatic genes (CLOCK), to AD-relevant pathology. CONCLUSION:These functionally validated findings provide deeper insights into the complex mitochondrial regulatory mechanisms involved in AD pathogenesis, offering robust biomarkers and novel potential avenues for developing targeted therapeutic strategies to address this challenging neurodegenerative disease.