Intrahepatic cholangiocarcinoma (ICC) is an insidious and aggressive malignancy with poor prognosis. Our previous research has suggested that miR-7-5p modulates the ICC cell phenotype by targeting MyD88; however, its downstream molecular mechanisms remain poorly elucidated. Considering that TGF-β signaling and aerobic glycolysis provide a favorable growth environment for tumors, this study aims to explored the relationship between the miR-7-5p/MyD88 axis and these metabolic characteristics. Bioinformatics methods were used to analyze the MyD88 expression in the GSE107943, and inferred its association with TGF-β signaling activity and glycolysis scores. Next, a series of experiments was conducted to evaluate the biological functions of MyD88 and miR-7-5p, with a TGF-β activator applied to elucidate potential mechanisms. A subcutaneous xenograft mouse model was used for in vivo validation. MyD88 expression was highly expressed in ICC samples, and its levels were positively correlated with TGF-β signaling activity and glycolysis scores. MyD88 knockdown attenuated the viability, migration, and glycolysis of ICC cells, thereby inhibiting tumor growth in vivo. Furthermore, MyD88 acted in a TGF-β-dependent manner, and TGF-β activation reversed the effects of MyD88 knockdown on malignant phenotype. Experiments also showed that MyD88 downregulation was caused by miR-7-5p, and then MyD88 overexpression reversed the suppressive effect of miR-7-5p on glycolysis. Collectively, miR-7-5p specifically targets MyD88, weakening glycolysis in ICCs by reducing TGF-β signaling activity, thus exerting an inhibitory effect on ICC.
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are considered as classic targeted drugs for EGFR-mutated non-small cell lung cancer (NSCLC), but induce EGFR-TKI-resistance and immunosuppression at advanced stage. The targeted inhibition of YES-associated proteins (YAP) combined with immunomodulation is expected to be a distinctive supplementary approach for EGFR-TKI-resistant NSCLC therapy. Herein an endoplasmic reticulum (ER)-targeting Zn/Cu-bi-single-atom nanoplatform (Zn/Cu-BSRGT) was prepared for EGFR-TKI resistance reversion, cascaded ER stress and immunoactivation. Specifically, after precisely targeting to the ER, Zn/Cu-BSRGT NPs provided simultaneous release of ·OH and 1O2 through the efficient chemodynamic therapy (CDT) and sonodynamic therapy (SDT), triggering intense ER stress. Meanwhile, the released YAP-siRNA interfered with the expression of YAP and the EGFR bypass signaling pathway, reversing the AXL-mediated resistance to EGFR-TKI. Furthermore, significant glucose consumption and ER stress triggered the immunogenic cell death (ICD) and systemic immune activation, and down-regulated the PERK-Nrf2 signaling pathway and multidrug resistance protein (MRP1). In summary, the combined application of single-atom-nanozyme catalytic technology and gene-targeted silencing technology successfully reversed EGFR-TKI resistance and promoted immunoactivation in NSCLC under ER-targeting assistance, providing support for the new strategic development of drug-resistant NSCLC.
Heat stress (HS) markedly impairs broiler growth, muscle function, and meat quality. In this study, broilers were subjected to a 2 × 2 factorial design with sodium selenite or yeast β-glucan selenium nanoparticles (yeast β-Glu-SeNPs) as the selenium source (0.3 mg/kg total selenium) under thermoneutral or HS conditions. We aimed to investigate the protective effects and underlying mechanisms of yeast β-Glu-SeNPs against HS-induced muscle damage. HS markedly impaired growth performance and induced systemic oxidative stress and inflammation, while also compromising meat quality and disrupting postmortem glycolysis, as evidenced by reduced glycogen availability and excessive lactate accumulation. Yeast β-Glu-SeNPs significantly improved growth performance, mitigated oxidative stress and inflammation, and restored meat quality in both breast and thigh muscles. Postmortem energy metabolism was preserved, as reflected by increased muscle glycogen and glycolytic potential, reduced lactate accumulation and glycolytic enzyme activities, and a stabilized pH decline. Meanwhile, skeletal muscle Se deposition, glutathione peroxidase activity, and key selenoprotein expression were markedly enhanced. Notably, HS promoted a phenotypic shift toward fast glycolytic muscle fibers, as evidenced by increased expression of MYHC2b and Fast-MyHC (P < 0.05), accompanied by reduced levels of MYHC1, MYHC2a, and Slow-MyHC (P < 0.05). This maladaptive transition was effectively reversed by yeast β-Glu-SeNPs, which favored oxidative fiber formation, characterized by the upregulation of MYHC1 and MYHC2a, along with the suppression of MYHC2b (P < 0.05). At the mitochondrial level, yeast β-Glu-SeNPs preserved ultrastructural integrity and enhanced mitochondrial function, as reflected by increased ATP content, elevated mtDNA copy number, and the upregulation of mitochondrial biogenesis-related genes, including AMPK, PGC-1α, NRF1, and TFAM (P < 0.05). Correlation analysis, molecular docking, and co-immunoprecipitation demonstrated that SelO interacts with AMPK, supporting a SelO-dependent AMPK/PGC-1α axis that drives mitochondrial biogenesis and oxidative fiber remodeling. Overall, yeast β-Glu-SeNPs mitigated HS-induced muscle metabolic dysfunction and meat quality deterioration via SelO-mediated mitochondrial and myofiber reprogramming.
To explore the protective effect and mechanism of loganin, a iridoid glycoside isolated from Corni Fructus, on carbon tetrachloride (CCl4)-induced acute liver injury (ALI) mice model and L-02 cells. ALI mice model was developed by an intraperitioneal injection of 0.3% CCl4. Thirty 8-week-old male C57BL/6 mice were randomly divided into 5 groups using a random number table, including control, model, loganin (40 and 80 mg/kg), and silybin (100 mg/kg) groups (n=6). After 6 consecutive days of intragastric administration, serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using an automatic biochemical analyzer. Hepatic pathological changes were observed by hematoxylineosin (HE) staining. Levels of interleukin (IL)-6 and tumor necrosis factor-α (TNF-α) in liver tissue were detected by enzyme-linked immunosorbent assay (ELISA). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were measured using biochemical kits. Mitochondrial ultrastructure and autophagosome formation were observed by transmission electron microscopy (TEM). Co-localization of mitophagy markers LC3 and TOMM20 were assessed by immunofluorescence (IF). Protein expressions of NOD-like receptor protein 3 (NLRP)3, LC3 II/LC3 I, p62, Beclin-1, Atg-7, Parkin, and PINK1 were determined by Western blot. In vitro, an injury model was established in L-02 hepatocytes stimulated with CCl4. Cell viability was assessed by CCK-8 assay, and related mechanisms were evaluated using the aforementioned methods. Besides, CCl4-stimulated L-02 cells were intervened with NLRP3 shRNA or Mdivi-1. Compared with the model group, pre-treatment with loganin (40 and 80 mg/kg) significantly reduced serum AST and ALT levels (P<0.01), alleviated pathological injuries such as swelling, necrosis, inflammatory infiltration, and lipid vacuolation in liver tissue. Loganin also markedly decreased the levels of IL-6 and TNF-α, increased SOD activity, and reduced MDA content in liver tissues (P<0.05 or P<0.01). Mechanistically, loganin up-regulated the ratios of LC3 II/LC3 I and expressions of Beclin-1, Atg-7, Parkin, and PINK1, while down-regulated p62 and NLRP3 protein expressions (P<0.05). In vitro experiments further confirmed that loganin attenuated CCl4-induced injury in L-02 cells by enhancing mitophagy and inhibiting NLRP3 inflammasome activation, which was reversed by the mitophagy inhibitor Mdivi-1. Loganin protected against CCl4-induced ALI both in vivo and in vitro by suppressing the NLRP3 inflammasome and enhancing mitophagy.
Metastasis contributes to treatment failure and poor prognosis of esophageal squamous cell carcinoma (ESCC) patients. The 5'-3' exoribonuclease 2 (XRN2) is related to the pathogenesis and progression of various malignancies through its roles in transcription termination and metastasis promotion, but its function in ESCC remains unclear. Bioinformatic analysis showed that XRN2 was significantly overexpressed in ESCC tissues and was identified as a risk factor for ESCC patients. The analysis of our own cohort confirmed that XRN2 expression was overexpressed and significantly correlated with cancer stage in patients (P = 0.0100). Gain- and loss- of function analyses revealed that XRN2 promoted ESCC cell growth, migration and invasion capabilities, as well as experimental lung colonization foci. Interestingly, we found that the RNA level of XRN2 was upregulated by the RNA-binding protein polypyrimidine tract binding protein 3 (PTBP3). Specifically, PTBP3 bound to CUUUC motifs of the 3'UTR of XRN2, prolonging the half-life of XRN2 RNA. PTBP3 was found to be significantly overexpressed in ESCC, where its expression level correlated with tumor stage (P = 0.0164) and tumor size (P = 0.0495), positioning it as a risk factor for ESCC patients. PTBP3 upregulation promoted ESCC cell growth, migration, and invasion in vitro and in vivo. Notably, knockdown of XRN2 reversed the tumor promotion effects induced by PTBP3 overexpression. Collectively, our data reveal a novel function of XRN2 in ESCC metastasis and the critical roles of the PTBP3/XRN2 axis in ESCC metastasis, highlighting its promise as a novel therapeutic target in ESCC.
We systematically explored PEG alternatives by synthesizing a combinatorial library of nine copolymers composed of three hydrophilic monomers including oligo(ethylene glycol) methyl ether methacrylate and 2-(methylsulfinyl)ethyl methacrylate and three hydrophobic monomers. These copolymers were evaluated for their hydrophobicity by reversed-phase chromatography and LCST behavior, and for their biodistribution. Extended hydrophilic brushes enhanced blood retention and tumor accumulation. For copolymers with the longest brushes, hydrophobic side chains minimally influenced kidney accumulation, whereas kidney accumulation of copolymers with medium-length brushes was strongly influenced. Liver accumulation of the copolymers was hardly affected by the hydrophobic side chain, except for the copolymer with the longest brushes and the most hydrophobic side chains, which showed increased liver accumulation. This study highlights the critical role of hydrophilic brushes and hydrophobic side chains in modulating biodistribution of polymeric carriers and provides a foundation for rational design of drug delivery systems.
Lumbar intervertebral disc degeneration (IVDD) is characterized by abnormal innervation and neurogenic inflammation, contributing to chronic discogenic pain. Electroacupuncture (EA) alleviates IVDD-related pain, yet its underlying mechanisms remain incompletely understood. This study aimed to investigate the role of the axonal guidance factor Netrin-1 in EA-mediated inhibition of myelinated nerve fiber ingrowth into degenerative intervertebral discs and to elucidate the downstream signaling pathways involved. A rabbit axial compression IVDD model was established. Animals were divided into sham, model, EA, model+OV-Netrin-1, EA+sh-Netrin-1, EA+μ-opioid antagonist (β-FNA), and EA+δ-opioid antagonist (NTI) groups. Tarlov scoring, MRI, HE, AB-PAS, immunohistochemistry, TEM, Western blot, RT-qPCR, Co-IP, ELISA and flow cytometry were used to evaluate disc histology, nerve ingrowth, Netrin-1 expression in annulus fibrosus (AF) and dorsal root ganglion (DRG), endogenous opioid peptides, receptor interactions, cAMP/cGMP levels and intracellular calcium concentration. EA improved Tarlov scores, restored disc structure and reduced pathological nerve ingrowth in degenerative discs. EA downregulated Netrin-1 in AF but upregulated Netrin-1 in L2 DRG. DRG Netrin-1 overexpression mimicked EA's anti-neoinnervation effect, whereas Netrin-1 knockdown abolished EA efficacy. EA elevated spinal POMC/M-ENK; blocking μ/δ opioid receptors reversed EA-mediated Netrin-1 upregulation in DRG. Mechanistically, EA promoted UNC5B-DCC complex formation, reduced cAMP, increased cGMP and lowered intracellular Ca2+ via Netrin-1 receptors to inhibit myelinated axon sprouting. EA upregulates DRG Netrin-1 by activating endogenous opioid peptide signaling. Netrin-1 subsequently modulates UNC5B/DCC downstream cAMP-cGMP-calcium pathways to suppress abnormal myelinated nerve fiber ingrowth, thereby alleviating discogenic pain and delaying IVDD progression.
To elucidate the expression pattern, clinical prognostic significance, and distribution characteristics of R-spondin 3 (RSPO3) within the tumor microenvironment (TME) of breast cancer (BC), to observe its effects on the malignant biological behaviors of MCF-7 cells, and to explore the possible mechanisms underlying its function. Integrated mRNA expression analysis was performed across multiple cohorts using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases and validated by quantitative reverse transcription polymerase chain reaction (qRT PCR) in clinical samples. Immunohistochemistry (IHC) was used to detect RSPO3 protein expression in breast cancer tissue microarrays (TMAs), and the results were combined with survival data to assess prognostic value. Single‑cell transcriptomics data were interrogated to identify the primary cellular sources of RSPO3, followed by differential gene expression and pathway enrichment analyses in key subpopulations. Finally, in vitro functional assays were conducted to investigate the effects of RSPO3 on breast cancer cell proliferation, migration, and apoptosis. Specifically, a co‑culture system of human breast cancer‑associated fibroblasts (HBCFs) and MCF‑7 cells was employed to examine changes in RSPO3 expression and the proliferative and migratory capacities of breast cancer cells after co‑culture. The function of RSPO3 was verified by knocking down RSPO3 in the co‑culture system, and further validation was carried out through Western blotting, CHIR99021 activator rescue experiments, and tissue analysis. Compared to adjacent non-tumor tissues, RSPO3 mRNA expression was generally downregulated in BC tissues; however, RSPO3 protein levels were significantly higher in tumor tissues. High RSPO3 protein expression was an independent risk factor for reduced overall survival (OS). Single-cell analysis revealed that within the TME, RSPO3 was primarily expressed in fibroblasts, endothelial cells, and epithelial cells. Notably, among fibroblasts, RSPO3 expression was specifically enriched in the inflammatory cancer-associated fibroblast (iCAF) subpopulation. RSPO3⁺ iCAFs exhibited functional characteristics related to cell proliferation regulation, angiogenesis, Wnt signaling, and oxidative phosphorylation. In vitro experiments demonstrated that RSPO3 knockdown suppressed the proliferation, migration, and invasion of MCF-7 cells while promoting apoptosis.Furthermore, co-culture of HBCFs with MCF-7 cells significantly increased RSPO3 protein levels in MCF-7 cells, accompanied by markedly enhanced proliferation and migration abilities. Knockdown of RSPO3 significantly reversed these pro-tumorigenic effects. RSPO3 knockdown downregulated the expression of Axin2, Cyclin D1, and β-catenin. Treatment with the Wnt pathway activator CHIR99021 partially rescued the inhibition of invasion and migration induced by RSPO3 knockdown. Immunohistochemical staining of breast cancer tissues further confirmed the positive correlation between RSPO3 expression and β-catenin and Cyclin D1 expression. RSPO3 exhibited significant discrepancies between transcriptional and translational levels in breast cancer tissues. High RSPO3 protein expression indicated poor prognosis and served as an independent prognostic risk factor. Breast cancer-associated fibroblasts may promote tumor cell proliferation and migration by regulating RSPO3 expression and subsequently activating the Wnt/β-catenin pathway. RSPO3 represents a potential prognostic biomarker and therapeutic target.
Intervertebral disc (IVD) injury is a major cause of low-back pain and can lead to structural deficits and mechanical instability. When the IVD is compromised, neuromuscular compensation by paraspinal muscles, such as the multifidus (MF) and longissimus (ML), is critical for maintaining spine stability. However, it is unknown how IVD injury and its interaction with nociception affect neuromuscular control. This study assessed the effects of IVD injury and additional muscle-derived nociception on trunk motor control during locomotion in a rat model. IVD injury was induced via needle puncture at L4/L5. One week later, hypertonic saline was injected into the lumbar MF to induce nociception. Trunk and pelvic kinematics, bilateral EMG activity of MF and ML were recorded during treadmill locomotion at baseline, one week after IVD injury, and immediately following hypertonic saline injection. Trunk and pelvic kinematics and bilateral muscle activation patterns remained largely consistent across conditions. No significant changes were found in stride duration, pelvic, lumbar and spine angle changes, variability, or movement asymmetry. MF activation was bilaterally synchronized, whereas ML showed left-right alternating activation patterns. Following IVD injury, right MF mean activation and EMG variability increased significantly compared to baseline. When muscle-derived nociception was added in the unstable spine (IVD injury) condition, left MF minimum amplitude was significantly reduced, and instability-related increases in right MF mean activation and variability were attenuated, but not fully reversed. These findings suggest that single-level IVD injury, alone or in combination with muscle-derived nociception, elicits localized neuromuscular adaptations without disrupting the global locomotor patterns.
Community-based chest X-ray (CXR) screening has been widely performed to improve tuberculosis identification in elderly populations, but comparative evidence against mode of symptom-driven case finding remains scarce. We conducted three annual rounds of mass CXR screening among individuals aged over 60 years in Lanxi City, China, from 2021 to 2023. Tuberculosis cases identified through CXR screening were compared with concurrent, symptom-driven case presentations via routine surveillance. Restricted cubic spline and multivariable logistic regression analyses were used to examine factors associated with distinct detection pathways. At the individual level, a total of 831 tuberculosis cases were identified through two modes. CXR mass screening detected more patients with less typical symptoms, while symptom-driven detection was more effective in females (aOR = 0.59, 95% CI 0.39-0.88), bacteriologically confirmed cases (aOR = 0.42, 95% CI 0.27-0.63), and those aged greater than 80 (aOR = 0.50, 95% CI 0.30-0.82). Age showed a nonlinear association with mass screening yield, with peak effectiveness between 60 and 66 years while no significance was found for BMI. In population-level analysis, CXR mass screening outperformed symptom-driven detection for case identification in the first round but declined and reversed by the third round. Subgroup analyses revealed potential benefits among individuals with diabetes or prior tuberculosis history, particularly within two years post-diagnosis or treatment. Community-based CXR screening could effectively detect incremental tuberculosis cases with atypical or mild symptoms. The young-old would benefit from mass CXR screening, while more accessible services were needed for the very elderly. Elderly individuals with diabetes or prior tuberculosis history should be prioritized, with intensified monitoring within two years after diagnosis or treatment completion.
Radiotherapy is a cornerstone treatment for unresectable lung cancer. However, radiation-induced bystander effects (RIBE) remain a major obstacle to its therapeutic efficacy. LINC01234, showing high expression in lung cancer RIBE, its role and mechanism remain unclear. Beas-2B cells were cultured in conditioned medium derived from A549 lung cancer cells exposed to 4.0 Gy X-ray irradiation to establish an in vitro RIBE model. Models with LINC01234 knockdown and MECOM overexpression were constructed. Mitochondrial alterations were examined by transmission electron microscopy. The expression of ferroptosis-related and MAPK pathway-associated factors was assessed to elucidate the relationship between LINC01234 and MECOM. Co-immunoprecipitation (Co-IP) was used to investigate interactions between MECOM and JNK, p38, and Erk1/2. RIBE reduced cell viability and promoted apoptosis in bystander cells. Knockdown of LINC01234 significantly alleviated RIBE-induced injury. Mechanistically, LINC01234 silencing suppressed Fe²⁺ accumulation, ROS generation, MDA levels, GSSG, and ACSL4 expression, while enhancing GSH, GPX4, and SLC7A11 expression. In parallel, phosphorylation of JNK, p38, and Erk1/2 was inhibited. The ferroptosis inhibitor ferrostatin-1 produced effects comparable to LINC01234 knockdown. Moreover, LINC01234 silencing downregulated MECOM expression. Overexpression of MECOM reversed the protective effects of LINC01234 knockdown, whereas co-silencing LINC01234 or treatment with the MAPK pathway inhibitor osmundacetone abrogated MECOM-induced effects. Co-IP confirmed that MECOM interacts with JNK, p38, and Erk1/2. Downregulation of LINC01234 suppresses ferroptosis in RIBE cells by inhibiting MECOM-mediated phosphorylation of p38/JNK/Erk1/2, thereby mitigating radiotherapy-associated side effects.
Some subtypes of head and neck squamous cell carcinoma (HNSCC) exhibit aggressive progression and poor prognosis, underscoring the need for novel therapeutic targets. While procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1) is implicated in tumour collagen remodelling, its functional role and regulatory mechanisms in HNSCC remain elusive. PLOD1 expression and clinical relevance were analysed using TCGA-HNSC data, patient tissues and cell lines. Functional impacts were assessed via in vitro assays (CCK-8, flow cytometry, Transwell) and in vivo xenograft models. Mechanistic insights were explored through co-immunoprecipitation, Western blotting, bioinformatics and pharmacological inhibition. PLOD1 was significantly upregulated in HNSCC tissues and correlated with adverse clinical outcomes. In vitro, PLOD1 overexpression potentiated proliferation, invasion and cell cycle progression while suppressing apoptosis; PLOD1 knockdown elicited opposing effects. PLOD1 activated the FAK/PI3K/AKT/mTOR pathway and directly interacted with prolyl 4-hydroxylase subunit alpha 2 (P4HA2). P4HA2 rescue reversed PLOD1 knockdown-mediated suppression of oncogenicity and pathway activation. The FAK inhibitor Y15 abrogated PLOD1-driven malignant phenotypes. In vivo, PLOD1 silencing inhibited tumour growth and reduced FAK/PI3K/AKT/mTOR phosphorylation. PLOD1 drives HNSCC progression by modulating P4HA2 and activating the FAK/PI3K/AKT/mTOR signalling cascade, positioning the PLOD1-P4HA2 axis as a promising prognostic biomarker and therapeutic target.
Microglia-mediated neuroinflammation is closely associated with the pathogenesis of epilepsy. Mammalian sterile-20-like kinase 4 (MST4) has been suggested a regulator of inflammation. However, the effect of MST4 on microglia neuroinflammation in epilepsy remains unclear. A pilocarpine-induced rat epilepsy model was constructed and a lipopolysaccharide (LPS)-stimulated microglia cell model was applied in the current research. Knockdown or overexpression of MST4 was established using lentivirus transfection. Electroencephalograph (EEG) was employed to measure brain activities of rats. The protein and mRNA expressions were detected using western blot and qRT-PCR, respectively. Immunofluorescent staining was conducted to detect the distribution of the proteins. TUNEL staining was performed to evaluate cell apoptosis. The protein interaction was evaluated with Co-IP assay. Our results showed that MST4 and nuclear PKM2 expressions were increased in epileptic rats compared to control and colocalized with microglia. MST4 overexpression inhibited microglia activation, the release of TNF-α and IL-1β, and improved neuronal apoptosis in epileptic rats. Furthermore, MST4 interacted with PKM2 and regulated PKM2 nuclear translocation. Inhibiting PKM2 nuclear translocation by TEPP-46 reversed the promoting effect of MST4 knockdown on microglia neuroinflammation. In summary, our study demonstrated that MST4 alleviated microglia-mediated neuroinflammation in epilepsy, and the mechanism of MST4-mediated anti-neuroinflammatory effects may be associated with the inhibition of PKM2 nuclear translocation.
Circular_RNAs (circ_RNAs) are involved in the development and progression of human malignancies, including breast cancer. Novel circ_RNAs for breast cancer remain to be further determined. This study investigates the role and mechanism of a novel circ_RNA circ_0048766 in triple-negative breast cancer (TNBC) progression. Bioinformatics analysis was performed using the GSE165884 database to identify differentially expressed circ_RNAs. The expression of circ_0048766 was validated in various breast cancer cell lines through quantitative real-time PCR. Functional assays, including CCK-8, flow cytometry, colony formation, and Transwell assays, were conducted in BT-549 and MDA-MB-231 cells. The mechanisms involving methyltransferase-like 3 (METTL3), miR-329-3p, and C-X-C Motif Chemokine Receptor 4 (CXCR4) were examined through bioinformatics, dual-luciferase reporter assays, RNA immunoprecipitation, and RNA pull-down. In vivo studies were conducted using a nude mouse xenograft model to evaluate tumor growth and CXCR4 expression. Circ_0048766 was significantly upregulated in TNBC cell lines compared to normal breast epithelial cells. Knockdown of circ_0048766 resulted in reduced cell viability, colony formation, migration, and invasion while increasing apoptosis in BT-549 and MDA-MB-231 cells. Mechanistically, circ_0048766 was confirmed to function as a sponge for miR-329-3p. Moreover, miR-329-3p directly suppressed CXCR4 expression, and circ_0048766 regulated CXCR4 in a miR-329-3p-dependent manner. Additionally, circ_0048766 promoted epithelial-mesenchymal transition, as evidenced by its regulation of E-cadherin, N-cadherin, and Vimentin expression via miR-329-3p. MiR-329-3p inhibition or CXCR4 overexpression reversed the effects of circ_0048766 knockdown. In vivo, circ_0048766 knockdown significantly diminished tumor growth and weight, along with decreased CXCR4 levels. METTL3 was identified as an upstream regulator mediating m6A modification of circ_0048766. Functionally, METTL3 knockdown suppressed CXCR4 expression and cell proliferation, effects that were rescued by CXCR4 overexpression. Circ_0048766 is a novel oncogenic circ_RNA that promotes TNBC progression through the miR-329-3p/CXCR4 signaling axis, regulated by METTL3. This study highlights the potential of targeting the METTL3/circ_0048766/miR-329-3p/CXCR4 axis as a novel therapeutic strategy for TNBC treatment. Not applicable.
Depression involves dysregulation across distributed cortico-limbic circuits, and ketamine is notable for its rapid antidepressant effects. Although depression and ketamine treatment have been linked to altered brain network topology, how within-frequency and cross-frequency coupling are jointly reorganized at the brain-wide level remains unclear. Here, we developed a frequency-varying multilayer brain functional network (FMBFN) framework to analyze local field potential recordings from eight brain regions in male C57BL/6 mice. This framework integrates within- and cross-frequency coupling and extracts multi-scale network features to characterize brain network structure. Applying this approach in the chronic social defeat stress (CSDS) model, we found that CSDS was associated with frequency-specific hyperconnectivity and selective alterations in network integration during social interaction. Ketamine reversed social avoidance and induced the distinct reorganization of multilayer network topology, including region-specific nodal changes. Notably, the lateral habenula showed the response pattern opposite to that of the other recorded regions. As an exploratory cross-modal extension, we further examined gut microbial features and found that specific ketamine-associated microbial changes were linked to global network topology, suggesting candidate gut-brain association patterns. Together, these findings establish the FMBFN framework as a systems-level tool for characterizing brain-wide neural dynamics in psychiatric disorders and for linking network-level alterations to biological contexts.
Immune rejection after liver transplantation remains a major challenge impacting the long-term survival of liver transplant recipients. Inhibiting macrophage M1 polarization exerts a positive effect on alleviating post-transplant immune rejection. Arctiin, an active component derived from traditional Chinese medicine, may suppress macrophage M1 polarization by inhibiting the release of pro-inflammatory cytokines. This study employed experimental techniques including ELISA, double-label flow cytometry, Western blot analysis, and cellular immunofluorescence to investigate the potential mechanism underlying the effect of arctiin on macrophage M1/M2 phenotypic switching in vitro. Results demonstrated that arctiin promoted the conversion of M0 macrophages to the M2 phenotype, significantly downregulated the levels of pro-inflammatory cytokines IL-6 and TNF-α, inhibited the expression of the M1 macrophage marker inducible nitric oxide synthase (iNOS), and reduced the proportion of M1-type macrophages. Furthermore, arctiin notably enhanced STAT6 phosphorylation and TRAF6 protein expression in M1 macrophages while suppressing STAT1 phosphorylation. However, these beneficial effects of arctiin were significantly reversed by a CB2R antagonist. Additional experiments showed that a CB2R agonist exerted similar effects to arctiin. Treatment with a TRAF6 inhibitor abrogated the inhibitory effect of arctiin or the CB2R agonist on macrophage M1 polarization, increased STAT1 phosphorylation, and decreased STAT6 phosphorylation. In conclusion, this study indicates that arctiin can activate CB2R and exert anti-inflammatory effects through the TRAF6-STAT1/6 pathway, thereby promoting the phenotypic switch of macrophages from M1 to M2.
Depression is a common psychiatric disorder during pregnancy and the postnatal period. Consequently, antidepressant treatment is primordial for the safety of the mother and the neonate. Clomipramine (CMI) is a tricyclic antidepressant that has been prescribed even when the evidence suggests potential adverse effects on the neurodevelopment of the offspring, considering that antidepressant drugs can be transferred through breast milk. Nevertheless, in some cases, when the benefits outweigh the risks, it is used to treat severe depression in pregnant women. In rodents, postnatal administration of CMI causes persistent behavioral and neurophysiological alterations in adulthood. By contrast, a rewarding experience, such as mating, improves motivational and copulatory behavior in rodents through neuroplasticity in brain structures involved in reproduction. In a previous work, we reported that postnatal exposure to CMI disrupts the motivational and copulatory components of female sexual behavior during a single copulatory test. Therefore, the purpose of this study was to examine the effects of postnatal CMI treatment on female sexual behavior and reproductive tissues, and to determine whether sexual experience serves as a modulatory factor that ameliorates its potential impact on sexual performance. Female pups were divided in two groups, CMI group (30 mg/Kg) and the control group (NaCl 0.9%). Each group received a daily subcutaneous injection with CMI or saline solution from the 8th to 21st postnatal days. Behavioral test and histological analysis were performed at 3 months of age. The results indicated that postnatal CMI administration disrupts receptive but not proceptive behaviors. Repeated sexual encounters with males partially reversed the receptivity impairment in CMI-treated females, as it occurs in control rats. Histological data showed that CMI reduces the population of primordial and primary follicles; however, no morphological modifications were detected in the uterine layers. In conclusion, the data show that even when sexual experience partially improved copulatory behavior in female rats exposed to CMI during the postnatal period, ovarian development was affected, which could compromise fertility.
Heavy alcohol consumption has multiple negative cognitive, psychological, and neurobiological consequences for people with epilepsy. However, the psychopharmacological interactions remain unclear with limited therapeutic interventions. In this study, we investigated the diverse impact of alcohol on experimental simulated pentylenetetrazol-induced seizures and alcohol-use disorder, and the effects of silymarin, a polyphenolic compound with neuroprotective properties. Following seven days of ethanol binge exposure (2 g/kg, oral gavage) in mice, maximal and sub-convulsive pentylenetetrazol-induced seizures were administered from days 8 to 14, alongside silymarin (50 and 100 mg/kg) or diazepam (3 mg/kg) oral administration. This study evaluated the interplay between ethanol and pentylenetetrazol-induced seizures, assessing behavioural comorbidities, dysregulation of the hypothalamic-pituitary-adrenal (HPA)-axis, neurochemical and neurotrophic alterations, oxidative stress, and neuroinflammation in the hippocampus, prefrontal-cortex, and striatum, which are involved in the disease. Ethanol increased seizure severity and frequency caused by pentylenetetrazol, and worsened anxiety-like and depressive behaviours, along with spatial working memory deficits linked to higher alcohol preference. These effects were reduced by silymarin. Ethanol also increased corticosterone release and reduced GABA-dependent glutamic acid decarboxylase activity, raising glutamate levels, while decreasing serotonin and brain-derived neurotrophic factor across the studied brain regions. Silymarin significantly reduced neuroinflammatory markers such as myeloperoxidase, TNF-α, IL-6, nitrite, and malondialdehyde, while enhancing IL-10 levels and antioxidant defenses, including catalase, superoxide dismutase and glutathione in the brain regions. These findings suggest alcoholism with alcohol-use disorders worsens epilepsy, notably involving neurochemical imbalance, neurotropic, HPA-axis upregulation, oxidative stress, and neuroinflammation, which were reversed by silymarin.
Osteoarthritis (OA) is a prevalent whole-joint disease characterized by progressive cartilage degeneration, extracellular matrix (ECM) breakdown, synovial inflammation, and pathological changes in multiple joint tissues. Inflammatory signaling plays a central role in chondrocyte catabolic activation and cartilage matrix degradation. Sotetsuflavone (SF), a naturally occurring biflavonoid isolated from Cycas species and other medicinal plants, has been reported to possess anti-inflammatory and antioxidant properties; however, its protective effects and related molecular mechanisms in OA remain unclear. This study aimed to investigate whether SF attenuates OA-associated inflammatory and catabolic responses and to explore the functional involvement of phosphoinositide 3-kinase/protein kinase B/nuclear factor-κB (PI3K/Akt/NF-κB) signaling in its protective effects. An integrated strategy combining network pharmacology, molecular docking, and experimental validation was used. Potential SF-related targets and OA-associated genes were collected from public databases, and overlapping targets were analyzed by protein-protein interaction (PPI) network construction and functional enrichment analyses. Molecular docking was performed to evaluate the predicted binding modes between SF and selected pathway-related proteins. Primary mouse chondrocytes stimulated with interleukin-1β (IL-1β) were used to assess the effects of SF on inflammatory mediator production, ECM metabolism, and PI3K/Akt/NF-κB signaling. To further evaluate pathway involvement, a rescue experiment was performed using 740Y-P, a PI3K/Akt pathway activator. A destabilization of the medial meniscus (DMM)-induced mouse model was established to evaluate the protective effects of SF against OA-related structural and inflammatory changes in vivo. Network pharmacology analysis identified 68 overlapping targets between SF and OA-associated genes, with enrichment mainly involving inflammation-related and kinase-dependent pathways, including PI3K/Akt and NF-κB signaling. Molecular docking revealed favorable binding affinities of SF toward key targets. In vitro, SF reduced the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) and decreased the production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). SF also alleviated IL-1β-induced ECM catabolism, as shown by reduced matrix metalloproteinase-13 (MMP-13) and a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) levels and partial preservation of type II collagen (Col II) and aggrecan. Mechanistically, SF decreased PI3K/Akt phosphorylation, restored Inhibitor of nuclear factor-κB alpha (IκBα) expression, and reduced nuclear accumulation of NF-κB p65. Importantly, activation of PI3K/Akt signaling by 740Y-P partially reversed the inhibitory effects of SF on NF-κB activation and inflammatory mediator production, supporting the functional involvement of the PI3K/Akt/NF-κB axis. In vivo, SF administration alleviated cartilage destruction, reduced Osteoarthritis Research Society International (OARSI) scores, decreased MMP-13 expression, restored Col II expression, and lowered the expression of pro-inflammatory cytokines in joint tissues. These findings suggest that SF attenuates OA-associated inflammatory activation and cartilage matrix degradation in experimental models. The protective effects of SF are at least partly associated with suppression of the PI3K/Akt/NF-κB signaling axis. Further studies are warranted to clarify direct target engagement, pharmacokinetics, long-term safety, optimized delivery strategies, and therapeutic efficacy in more comprehensive OA models.
Osteoarthritis (OA) is a progressive and disabling joint disease driven by oxidative stress, chondrocyte senescence and extracellular matrix (ECM) degradation, yet lacks effective disease-modifying treatments. In this study, we identified miR-197-3p as a previously unrecognized, cartilage-protective miRNA significantly downregulated in both aged and osteoarthritic cartilage. Functional studies revealed that miR-197-3p restores ECM anabolism, suppresses senescence and directly targets G3BP1, a stress granule protein linked to redox imbalance and inflammatory signaling. To enable effective intra-articular delivery, we engineered a multifunctional microsphere platform (miR/PBNP@Gel) by co-encapsulating miR-197-3p and ultrasmall Prussian blue nanozymes (PBNPs) into GelMA hydrogel microspheres. This composite design synergistically enhances miRNA stability, facilitates cellular internalization and provides continuous reactive oxygen species (ROS) scavenging to protect mitochondrial function. miR/PBNP@Gel reversed mitochondrial dysfunction and senescence in OA chondrocytes, while promoting cartilage repair and joint function in vivo. Metabolomic profiling further revealed reprogramming of TCA cycle and antioxidant pathways. This work established miR-197-3p as a novel therapeutic regulator in OA and introduced a bioinstructive, injectable, and cell-free strategy that integrates miRNA therapy and redox modulation for disease modification and cartilage regeneration.