The maternal-to-zygotic transition (MZT) is essential for early embryonic development, comprising zygotic genome activation (ZGA) as well as the degradation of maternal RNAs and proteins, whereas our understanding of repressors' role in this process remains limited in mammals. In our study, we inhibited protein degradation at the 1-cell stage using the proteasome inhibitor MG132 and observed impaired pre-implantation development and defective ZGA. Proteomic analysis of MG132-treated embryos showed that significantly up-regulated proteins in abundance were enriched for RNA-binding proteins (RBPs). Further functional screening highlighted one critical factor, 4E-T (EIF4ENIF1), whose overexpression caused 2-cell stage arrest and failure of ZGA initiation. Moreover, the abnormal accumulation of 4E-T translationally repressed key factors of the 1-cell embryo, including ZBED3 and KDM4A. Mechanistically, the reprsssion is linked to direct interactions of 4E-T with eIF4E and eIF4E1B. These findings suggest that specific maternal RBPs may act as repressors that require precise degradation post-fertilization to relieve translational repression and ensure successful MZT.
Malignant melanoma has a strong propensity for early metastatic dissemination, yet conventional clinicopathological predictors such as Breslow thickness do not fully capture individual metastatic risk. The endothelial protein C receptor (EPCR) exerts context-dependent functions across malignancies, but its clinical significance in melanoma remains unclear. We retrospectively analyzed primary tumor specimens from 62 patients with invasive melanoma; 61 with complete outcome data formed the analytic cohort for metastasis analyses. EPCR expression in melanoma cells was evaluated by immunohistochemistry using semi-quantitative intensity and extent scores (0-3). Tumor-infiltrating lymphocytes were graded according to the Melanoma Institute Australia scoring system. Higher EPCR expression was associated with lower odds of documented lymph-node and distant metastatic involvement. In prespecified parsimonious multivariable logistic regression models adjusted for tumor thickness and ulceration, higher EPCR extent remained independently associated with reduced lymph-node involvement (adjusted odds ratio: 0.36, 95% confidence interval:, 0.17-0.70). EPCR expression showed a positive association with CD8+ tumor-infiltrating lymphocyte scores. Higher EPCR expression in primary melanoma was independently associated with reduced lymph-node involvement and with a CD8+-inflamed tumor microenvironment, suggesting EPCR may complement conventional pathological predictors of metastatic risk.
The economic viability of methanol-based biomanufacturing, particularly with green methanol as feedstock, is often limited by the low value of single-product processes. Here, we developed an integrated co-production strategy in the methylotrophic yeast Pichia pastoris (Komagataella phaffii) for the simultaneous conversion of methanol into three products: the sweetener erythritol, the industrial biocatalyst β-mannanase, and single-cell protein (SCP) biomass. This new strategy explores the inherent spatial and functional separation between the ER-Golgi secretory pathway for enzyme production and the cytosolic pathway for chemical synthesis, thereby reducing interference between the two pathways. The engineered co-production strain achieved β-mannanase and erythritol titers comparable to those of the corresponding β-mannanase- and erythritol-producing reference strains in both shake-flask and fed-batch fermentor cultures. Furthermore, transcriptomic analysis revealed distinct regulatory responses related to enzyme production and erythritol synthesis, supporting the limited cross-pathway interference between the two pathways. In addition, we showed that ultrafiltration enabled efficient downstream separation of the small-molecule erythritol from the secreted β-mannanase, with recovery efficiencies exceeding 89%. These results demonstrate a feasible strategy for methanol valorization into multiple value-added products and expand the potential of methylotrophic yeasts for integrated biomanufacturing.
The activation of endoplasmic reticulum stress (ERS), specifically the PERK/eIF2α/CHOP signaling, is a recognized consequence of ischemic stroke. However, the roles and mechanisms of CRM-1 regulating ERS in stroke are poorly elucidated. A murine model of stroke was generated via transient middle cerebral artery occlusion (MCAO). The endpoints included TTC-derived infarct volume, H&E/TUNEL histopathology, p-PERK immunohistochemistry, and Western blot. Oxygen-glucose deprivation/reoxygenation (OGD/R) was employed in HT22 neurons. The CRM1-ALKBH5 interaction and subcellular distribution were assessed by co-immunoprecipitation (co-IP), cytoplasm-nucleus fractionation, and confocal microscopy. m6A regulation of BANF1 was examined using MeRIP-qPCR, RIP-qPCR, and a dual-luciferase reporter assay. Knockdown of CRM-1 reduced the infarct size, decreased ER stress activation, decreased apoptosis in vivo, and improved cell viability after OGD/R. CRM-1 was attached to ALKBH5 and promoted its export from the nucleus, which increased m6A modification and expression on BANF1. IGF2BP2 and YTHDF1, respectively, enhanced the stability and translation of BANF1 mRNA, thereby upregulating its expression. BANF1 overexpression restored PERK/eIF2α/CHOP activation and apoptosis in the CRM-1 knockdown context. CRM-1 exacerbated ischemic stroke injury by exporting ALKBH5 to upregulate BANF1 in an m6A-IGF2BP2/YTHDF1-dependent manner. This cascade subsequently activated the PERK/eIF2α/CHOP ERS pathway.
Immunotactoid glomerulopathy (ITG) is a rare glomerulonephritis characterized by the deposition of hollow-centered microtubular structures on electron microscopy (EM), and by IgG, C3, and restricted light chain deposition on immunofluorescence (IF) staining. It is often associated with hematological malignancies, including lymphoma and lymphocytic leukemia. A 71-year-old Japanese woman presented with increased proteinuria (urine protein-to-creatinine ratio, 4.32 g/gCr) and impaired kidney function (serum creatinine, 1.24 mg/dL). A kidney biopsy was performed, and routine frozen-IF revealed positivity for C3 and κ-light chain along the glomerular basement membrane, while IgG was negative. EM demonstrated electron-dense deposits composed predominantly of microtubules with hollow centers measuring 15-20 nm in diameter and arranged in parallel arrays, raising suspicion of ITG. Mass spectrometry and paraffin-IF following pronase K pretreatment detected IgG2 along the capillary walls, leading to the diagnosis of monoclonal ITG with masked IgG. In addition, the presence of paraproteins in serum and urine prompted bone marrow examination, which confirmed symptomatic multiple myeloma. With chemotherapy, the M-protein level decreased, proteinuria improved to less than 0.5 g/gCr, and renal function remained stable. We describe a rare case of monoclonal ITG with masked IgG, diagnosed by mass spectrometry and paraffin-IF, in a patient with symptomatic multiple myeloma. Early diagnosis and therapy for multiple myeloma stabilized renal function and reduced proteinuria. When a discrepancy between IF and EM findings is observed, especially when false-negative immunoglobulin staining is suspected, restaining with paraffin-IF or evaluation by mass spectrometry should be considered to ensure accurate diagnosis and appropriate treatment.
Parkinson's disease (PD) and related Parkinsonian conditions involve progressive disruption of mitochondrial homeostasis, redox balance, synaptic function, and proteostasis. Available therapies remain predominantly symptomatic, motivating the search for multitarget compounds acting across stress-sensitive pathways. BjPro-7a (pEDGPIPP), a proline-rich oligopeptide from Bothrops jararaca venom, exerts cytoprotective effects under oxidative stress, but its activity in an in vivo Parkinsonian-like model is unknown. Here we investigated whether BjPro-7a attenuates the behavioral and proteomic disturbances induced by 1-methyl-4-phenylpyridinium (MPP+) in zebrafish larvae. Larvae were exposed to 500 µM MPP+ and post-treated with 10 µM BjPro-7a, followed by assessment of basal locomotion, light/dark-evoked behavior, and label-free proteomic profiling. BjPro-7a markedly reversed the MPP+-induced hypolocomotor phenotype by increasing the total distance traveled, improving bout-related parameters, reducing inter-episode intervals, and restoring visually evoked responsiveness. Proteomic analysis revealed broad rescue-associated remodeling, more pronounced among proteins suppressed by MPP+ than among those elevated by intoxication. Functional enrichment and protein-protein interaction analyses indicated coordinated modulation of vesicle trafficking and synaptic organization, mitochondrial and bioenergetic function, redox homeostasis, and protein quality control. Representative rescue-associated proteins included VAMP2 and SNAP25A (synaptic), SDHA and UQCRFS1 (mitochondrial), G6PD and PRDX3 (redox), and PSMC5 and STIP1 (proteostasis). Together, these findings show that BjPro-7a attenuates MPP+-induced dysfunction and promotes systems-level proteomic remodeling consistent with rescue in zebrafish larvae. These data support BjPro-7a as a promising candidate for future therapeutic studies in Parkinsonian-like neurotoxicity.
Lupus nephritis is a key factor affecting the prognosis of childhood-onset systemic lupus erythematosus (SLE). Immunoglobulin A (IgA) participates in immune regulation through pathways such as mucosal immunity; however, its specific role in pediatric lupus nephritis remains unclear. This study aims to investigate the association between peripheral blood IgA levels and the development of lupus nephritis in children with newly diagnosed SLE. This single-center cross-sectional study consecutively enrolled 380 children with newly diagnosed SLE at Kunming Children's Hospital between January 2023 and December 2025. According to the International Classification criteria for pediatric lupus nephritis, using 24-h urinary protein excretion > 0.15 g/24 h as the primary indicator, patients were divided into the lupus nephritis group (n = 238, 62.6%) and the non-lupus nephritis group (n = 142, 37.4%). Binary logistic regression models were used to assess the association between IgA levels (as both a continuous variable and quartile groups) and lupus nephritis, with stepwise adjustment for demographic characteristics, complement levels, autoantibodies, C-reactive protein, and urinary protein. Potential nonlinear relationships were explored using restricted cubic splines, and subgroup analyses were performed according to sex, anti-dsDNA antibody status, anti-nucleosome antibody status, and age. Serum IgA levels in the lupus nephritis group were significantly lower than those in the non-lupus nephritis group (median: 1.48 vs. 1.87 g/L, P < 0.001). Univariate logistic regression analysis showed that each 1 g/L increase in IgA level was associated with a 40% reduction in the risk of lupus nephritis (OR = 0.60, 95% CI 0.47-0.76, P < 0.001). Quartile analysis of IgA levels revealed a consistent decreasing trend in the proportion of lupus nephritis as IgA levels increased from the lowest to the highest quartile (78.9%, 62.1%, 59.4%, and 50.0%, respectively). In multivariate models, after adjusting for age and sex, the OR was 0.59 (0.46-0.75); with further adjustment for C3 and C4 levels, the OR was 0.61 (0.47-0.79); after adjustment for autoantibodies, the OR was 0.61 (0.47-0.78); and in the fully adjusted model (adjusted for age, sex, C3, C4, CRP, urinary protein, and autoantibodies), the OR was 0.51 (0.36-0.72). All models showed P < 0.001. Restricted cubic spline analysis revealed no nonlinear relationship (P > 0.05). Subgroup analyses indicated that the protective effect of IgA remained consistent across subgroups of sex, anti-dsDNA antibody status, anti-nucleosome antibody status, and age (all P for interaction > 0.05), although a marginal interaction was observed in the low C4 subgroup (P = 0.025).   Among children with newly diagnosed systemic lupus erythematosus, peripheral blood IgA level is independently and negatively associated with the presence of lupus nephritis, and this association remains stable across different clinical subgroups. This finding suggests a potential protective role of IgA in the pathogenesis of pediatric lupus nephritis, providing new insights into its immune mechanisms and laying a foundation for future prospective studies. Key Points • Serum IgA is significantly lower in pediatric SLE patients with lupus nephritis. • Each 1 g/L increase in IgA level was associated with a 40% lower odds of having lupus nephritis • The negative association remains independent of complements and autoantibodies. • IgA shows a consistent protective effect across sex and autoantibody subgroups. • IgA may serve as a protective biomarker for renal involvement in childhood SLE.
Sesame (Sesamum indicum L.), is one of the earliest domesticated oilseed crops. It is valued for its exceptional oil content, bioactive compounds, and adaptability to diverse agroclimatic conditions. However, its production remains highly susceptible to water scarcity particularly terminal drought and intermittent moisture deficits that in turn severely compromise the yield and oil quality. Recent advances in genomics, transcriptomics, proteomics, metabolomics, epigenomics, and phenomics have placed sesame as an emerging model for multi-omics-driven stress research. These approaches have uncovered key regulators of drought (NAC, MYB, WRKY), protective proteins (late embryogenesis abundant proteins, heat shock proteins, antioxidant enzymes), osmolyte- and redox-related metabolites, and hormonal signalling modules such as PYL-SnRK2-ABF (ABA), LOX/AOS/OPR (jasmonate), and EIN/ERF (ethylene). Addressing these gaps will require investments in precision phenotyping, robust pan-genomic databases, functional validation using CRISPR/Cas9 tools, and global data-sharing networks. This review highlights the current advances in sesame drought research across omics platforms, critically evaluates their relevance to breeding programs, and offers the first comprehensive multi-omics perspective on moisture-stress adaptation in sesame. Additionally, KEGG pathway-guided multi-omics integration connects ABA signaling, phenylpropanoid metabolism, and antioxidant pathways to drought adaptation in sesame. By bridging mechanistic insights with applied strategies, it highlights pathways to accelerate the development of climate-resilient sesame cultivars.
In the World Health Organization (WHO) 2022 classification of chronic myelomonocytic leukemia (CMML), the cutoff for absolute monocyte values was lowered from 1.0 × G/L (2016) to 0.5 × G/L to incorporate cases formerly referred to as oligomonocytic CMML. The prevalence of this criterion in the general population and its potential association with demographic parameters and comorbidities is unclear. In order to answer these questions, we used the previously published LEAD (lung, heart, social, body) cohort (ClinicalTrials.gov; NCT01727518), which is a longitudinal, observational, population-based Austrian cohort aiming to investigate the impact of genetic, environmental, social, developmental, and ageing factors on respiratory health and associated comorbidities through life. The prevalence of the 2022 criterion was 11.976% in the general population, as compared to 0.747% of the 2016 criterion. In the descriptive analysis, individuals with the 2022 criterion as compared to those without this criterion were more often males, had a greater height and bodyweight, more frequently suffered from cardiovascular (CVD) and chronic obstructive pulmonary disease (COPD), and had higher C‑reactive protein (CRP) values. In the multivariable logistic regression, higher CRP, COPD, and CVD were significantly associated with the 2022 criterion. Our results show that in the general population, the 2022 CMML blood diagnostic criterion is 16 times more common than the 2016 criterion. The 2022 criterion is associated with CVD, COPD, and inflammation, which should be considered in the interpretation of this criterion in clinical practice. In der Klassifikation der Weltgesundheitsorganisation (WHO) von 2022 für myeloische Neoplasien wurde der Grenzwert der absoluten Monozytenzahl für die Diagnose der chronischen myelomonozytären Leukämie (CMML) von 1,0 G/l (2016) auf 0,5 G/l herabgesetzt, um die Fälle der früheren oligomonozytären CMML mit zu erfassen. Die Prävalenz dieses Kriteriums in der Allgemeinbevölkerung und seine potenzielle Assoziation mit demografischen Parametern und Komorbiditäten ist unklar. Um diese Fragen zu beantworten, haben wir die LEAD-Kohorte („lung, heart, social, body“; ClinicalTrials.gov; NCT01727518) herangezogen, die als eine longitudinale, populationsbasierte österreichische Beobachtungsstudie publiziert und konzipiert wurde mit dem Ziel, den Einfluss genetischer, umweltbezogener, sozialer, entwicklungsabhängiger und altersabhängiger Faktoren auf Lungengesundheit und assoziierte Komorbiditäten im Laufe des Lebens zu untersuchen. Die Prävalenz des Kriteriums von 2022 in der Allgemeinbevölkerung war 11,976 % verglichen mit 0,747 % des Kriteriums von 2016. In der deskriptiven Analyse war dieses neue Kriterium bei Männern, bei Menschen mit vermehrter Körpergröße und höherem Körpergewicht häufiger nachweisbar sowie bei Personen mit kardiovaskulären Erkrankungen (CVD), chronisch obstruktiver Lungenerkrankung (COPD) und erhöhtem Wert für C‑reaktives Protein (CRP). Auch in der multivariablen logistischen Regression waren höhere CRP-Werte, CVD und COPD mit diesem Kriterium assoziiert. Die Assoziation dieser Faktoren mit dem 2022 eingeführten Monozytengrenzwert bei CMML sollte bei der Interpretation dieses neuen Kriteriums berücksichtigt werden.
An intermittent infusion during dialysis therapy across the membrane by backfiltration is expected to have a cleaning effect on the membrane in addition to stabilizing blood pressure. In this study, the effect of backfiltration infusion on the solute removal performance was investigated for both diafilters with polysulfone (PSf) and polymethylmethacrylate (PMMA) membrane in vitro experiment using bovine whole blood. To clarify the infusion method with a high cleaning effect, the infusion was performed under two conditions for dialysate flow: countercurrent and parallel flow to blood flow, aiming to induce backfiltration at different area in a diafilter. A scanning electron microscope (SEM) and a scanning probe microscope (SPM) were used for evaluation of the degree of fouling on the membrane after experiments. Although intermittent backfiltration did not show any effect on the removal efficiency of creatinine and inulin, it was slightly effective in maintaining protein permeability under certain conditions. SEM observations and SPM measurements revealed that protein adsorption to the membrane caused the macropore blocking, and the fouling was partly removed by intermittent infusion primarily in areas where backfiltration mainly occurs. In conclusion, backfiltraion had an effect on resolving fouling and recovering solute removal performance for both membrane materials; however, its effect was limited.
Hydatid cyst disease is a zoonotic infection that predominantly affects the liver and can lead to severe hepatic complications. Exosomes are important mediators of intercellular signaling and are capable of transferring bioactive molecules to target cells. This study aimed to investigate the biological effects of exosomes derived from hydatid cyst stem cells on human hepatocytes to better understand parasite-associated hepatic alterations. In this in vitro experimental study, stem cells were isolated from the germinal layer of hydatid cysts and cultured. Cell identity was confirmed by flow cytometry. Exosomes were isolated using sequential filtration and characterized by dynamic light scattering (DLS). Human hepatocytes were divided into control and exosome-treated groups. Cell viability was assessed using the MTT assay. Gene expression levels of apoptosis-related markers (Bax, Bcl-2, Caspase-3, and Caspase-7) were determined by qPCR, and protein levels were quantified using ELISA. Exosomes derived from hydatid cyst stem cells significantly reduced the viability of human hepatocytes (P < 0.0001). Moreover, exosome exposure was associated with increased expression and protein levels of pro-apoptotic markers, including Bax, Caspase-3, and Caspase-7, along with decreased expression of the anti-apoptotic marker Bcl-2. Hydatid cyst stem cell-derived exosomes promote apoptotic changes in human hepatocytes in vitro. These findings provide preliminary evidence suggesting a potential role for parasite-derived extracellular vesicles in hepatic cellular alterations observed during hydatid disease.
Stress granules are large cytoplasmic bodies formed in response to environmental insults by eukaryotic cells. Stress granule formation is key for post-stress recovery, and many diseases and infections are characterized by dysregulation of these membraneless organelles. How specific and non-specific macromolecular interactions drive the formation of stress granules and other large assemblies is an area of active research. Stress granules are comprised of dense, ~200 nm cores, and these are known to contain numerous RNAs and proteins. Now, we have discovered that more than half of the nucleic acid content of stress granule cores is circular, double-stranded DNA. We demonstrate cytologically that these extrachromosomal circular DNAs (eccDNAs) colocalize cytoplasmically with canonical stress granule marker proteins in HEK293T cells, and through CRISPR targeting in budding yeast, that they are required for stress granule formation upon stress. This discovery thus reveals a key function for eccDNA in the eukaryotic stress response.
UFMylation is a conserved ubiquitin-like post-translational modification that controls protein stability and tissue homeostasis, while its role in amelogenesis remains largely uncharacterized. Here, we investigated the function of UFL1, the sole E3 ligase of the UFMylation pathway, in mammalian enamel development using K14-Cre-mediated epithelial-specific Ufl1 knockout mice. Ufl1 ablation caused severe amelogenesis imperfecta, with impaired enamel deposition, hypomineralization, and progressive tooth damage, accompanied by abnormal cervical loop development. Transcriptomic profiling revealed elevated endoplasmic reticulum stress and dysregulated expression of enamel mineralization genes, including significant downregulation of downstream targets of RUNX2, a master regulator of amelogenesis. We further demonstrated that UFL1 and DDRGK1 directly interacted with RUNX2, and UFL1-mediated UFMylation stabilized RUNX2 protein at the post-translational level. Taken together, this study identifies a novel UFMylation-RUNX2 regulatory axis essential for amelogenesis, providing new mechanistic insights into amelogenesis imperfecta and potential therapeutic targets for dental enamel defects.
The efficacy of cyclin-dependent kinase 4 and 6 inhibitors (CDK4/6i) in hormone receptor-positive/HER2-negative metastatic breast cancer is limited by resistance mechanisms, including mitogen-activated protein kinase pathway activation. We performed a phase I trial evaluating avutometinib, a dual RAF/MEK inhibitor (rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase), combined with abemaciclib and fulvestrant. Eligible patients had prior progression on CDK4/6i; prior fulvestrant was allowed. The primary endpoint was maximum tolerated dose (MTD). Planned dose levels were abemaciclib 50, 100, or 150 mg orally twice daily (BID), with avutometinib 2.4, 3.2, or 4.0 mg orally twice weekly (BIW; 3 weeks on/1 week off) plus fulvestrant 500 mg intramuscularly (IM) every 28 days (q28d). Secondary endpoints included safety, pharmacokinetics (PK), overall response rate (ORR), clinical benefit rate (CBR), and progression-free survival (PFS). Sixteen patients were treated (median age 61; 88% visceral disease; 63% prior selective estrogen receptor degrader). The MTD and recommended phase 2 dose (RP2D) were abemaciclib 100 mg BID, avutometinib 3.2 mg BIW, and fulvestrant 500 mg IM q28d. Common treatment-related adverse events (TRAEs) were creatine phosphokinase elevation (50%), neutropenia (50%), diarrhea (44%) and skin rash (44%). No grade 4-5 TRAEs occurred. ORR was 13% (2/15 patients with measurable disease), 24-week CBR 40% (6/15 patients with measurable disease), and median PFS 3.6 months (95% CI: 2.1-not reached). PK were consistent with prior reports. The regimen was well tolerated at the RP2D, with no new safety signals, and showed preliminary clinical activity. A phase II trial is ongoing.
Colorectal cancer (CRC) remains a major therapeutic challenge due to toxicity, low tumor specificity, and multidrug resistance. Carvacrol, a natural monoterpenoid phenol with notable anticancer, anti-inflammatory, and antioxidant properties, has emerged as a promising compound. This study evaluated the anticancer efficacy and underlying molecular mechanisms of carvacrol in HT-29 CRC cells and CCD-18Co normal colon fibroblasts. Cell viability was assessed using CVDK-8, migration by wound healing assay, and apoptosis, reactive oxygen species (ROS) levels, and cell cycle distribution by flow cytometry. Expression levels of PDHA1, SOX2, CASP3, POLB, ABCC1, and E2F4 were quantified by qPCR. Bioinformatic analyses were performed using GEPIA2 for overall survival (OS) and disease-free survival (DFS) evaluation and STRING for protein-protein interaction (PPI) network analysis. Carvacrol exhibited dose-dependent cytotoxicity in HT-29 cells (IC50=237.3µM) and significantly inhibited cell migration, whereas its effects on CCD-18Co cells were limited (IC50=425.9µM). In HT-29 cells, carvacrol significantly induced apoptosis, promoted G0/G1 cell cycle arrest, reduced ROS levels, and upregulated CASP3 expression. In contrast, no significant apoptosis, cell cycle arrest, or CASP3 upregulation was observed in CCD-18Co cells, although PDHA1, SOX2, and POLB expression levels were reduced. Bioinformatic analyses revealed that elevated E2F4 expression was associated with poorer OS in CRC and identified a significant interaction sub-network among SOX2, ABCC1, and CASP3. In conclusion, the findings demonstrate that carvacrol effectively inhibits CRC growth while exerting limited effects on normal colon fibroblasts. These results support the further investigation of carvacrol as a potential complementary therapeutic agent for CRC.
As a pseudoprotease member of the rhomboid family, rhomboid domain containing 2 (RHBDD2) has been demonstrated to be associated with 5‑FU resistance. Our previous work found that RHBDD2 is also upregulated by cisplatin (DDP) treatment in esophageal squamous cell carcinoma (ESCC). However, the effect of RHBDD2 on DDP sensitivity and the underlying molecular mechanisms remain poorly understood. In this study, we demonstrate that RHBDD2 regulates the sensitivity of ESCC cells to DDP in vitro. In vivo, RHBDD2 knockdown significantly enhanced the antitumor effect of DDP in a subcutaneous xenograft model of ESCC. Enrichment analysis of RNA‑seq data from ESCC cells with RHBDD2 knockdown revealed that RHBDD2 is involved in regulating the biological process of ferroptosis. RHBDD2 inhibited ferroptosis by upregulating ferritin heavy chain 1 (FTH1), a major iron storage protein, and FTH1 knockdown reversed RHBDD2‑induced insensitivity of ESCC cells to DDP in vitro. Mechanistically, RHBDD2 promoted Wnt3a secretion, which in turn inhibited the phosphorylation and ubiquitination of β‑catenin, thereby enhancing β‑catenin nuclear localization and its transcriptional activity. Inhibition of β‑catenin‑mediated transcription by ICG‑001 reversed RHBDD2‑upregulated FTH1 expression and DDP insensitivity in vivo. Furthermore, we found that RHBDD2 mRNA is modified by N6‑methyladenosine (m6A), and its expression can be regulated by the m6A reader insulin‑like growth factor 2 mRNA‑binding protein 2 (IGF2BP2) in ESCC cells. Collectively, our results reveal a novel role for RHBDD2 in inhibiting ferroptosis and inducing DDP insensitivity by upregulating the Wnt3a/β‑catenin/FTH1 axis, which may provide a therapeutic target for improving the efficacy of DDP in ESCC.
Oxidized low-density lipoprotein (ox-LDL) and inflammation cause endothelial dysfunction in coronary heart disease (CHD). Although ribosomal protein S5 (RPS5) has been linked to cardiovascular disease, the mechanism by which it functions in the endothelium and its relationship to NF-κB signaling remain unknown. RPS5 was chosen as the hub gene for CHD after differential expression analysis was done on the GSE68506 dataset. The human coronary artery endothelial cells (HCAECs) injury model was established by ox-LDL. In vitro experiments were performed to evaluate the changes in RPS5 expression, apoptosis, inflammatory markers, vascular protective factors, cell proliferation, and Endothelial-to-Mesenchymal Transition (EndMT). Combining the dual luciferase reporter gene assay and the NF-κB nuclear translocation immunofluorescence assay, the effect of RPS5 on the NF-κB pathway was evaluated. Seven ribosomal genes (RPL11, RPL8, RPS13, RPS20, RPS3, RPS5, RPS8) were downregulated in coronary artery disease (CAD) samples in the GSE68506 dataset. In vitro, ox-LDL injury similarly reduced RPS5 expression in HCAECs. Functionally, RPS5 knockdown inhibits cell proliferation and enhances apoptosis and EndMT, while RPS5 overexpression has the opposite effect. Mechanistically, RPS5 overexpression diminished NF-κB nuclear accumulation, lowered κB-WT reporter activity, and reduced the phosphorylation levels of IKKα/β, IκBα, and p65. It also reduced pro-inflammatory mediators and decreased VEGF and HSP70. The pathway dependency was confirmed when LPS-induced NF-κB activation reversed these effects. Through the NF-κB signaling pathway, RPS5 plays a crucial role in controlling inflammation, apoptosis, EndMT, and vascular protection factors in injured HCAECs. Endothelial damage in CHD may be treatable by focusing on the RPS5-NF-κB axis.
19-Hydroxyeicosatetranoic acid (19-HETE) is an oxylipin derived from arachidonic acid through the action of cytochrome P450. The S-enantiomer of 19-HETE (19(S)-HETE) has vasodilatory potential, acting through the prostacyclin receptor (IP). However, no research has explored whether 19(S)-HETE has the same relaxant potential in the airways and what intracellular signaling networks are leveraged. Relaxation of murine trachea and airway smooth muscle cells were measured following exposure to 19(S)-HETE by myography and traction force microscopy. cAMP was measured by ELISA and inhibitors for IP, protein kinase A (PKA), and exchange proteins directly activated by cAMP (EPAC1/2) were used to probe intracellular signaling networks. DNA synthesis was measured using EdU incorporation assay. Murine airways and ASM cells produce 19(S)-HETE. 19(S)-HETE promotes relaxation in a dose dependent manner and reduces DNA synthesis. Compared to isoproterenol, 19(S)-HETE is less potent but more efficacious, with a higher maximal relaxation. 19(S)-HETE stimulated the IP receptor, triggering the formation of cAMP and the activation of PKA and EPAC1/2 downstream. Inhibition of both PKA and EPAC1/2 blunted 19(S)-HETE induced relaxation, but only EPAC2 prevented 19(S)-HETE induced changes in myosin phosphorylation (pMLC2). Similarly, only inhibition of EPAC2 reversed the changes in DNA synthesis. 19(S)-HETE promotes ASM relaxation, through a mechanism dependent on the activation of IP receptor and downstream signaling through PKA and EPAC. However, the downstream events that lead to relaxation by PKA and EPAC are not the same, suggesting that more than reductions in pMLC2 are important in the action of 19(S)-HETE induced relaxation.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive impairment and pathological accumulation of amyloid-β and tau proteins. This study investigated the potential neuroprotective effects of a combined treatment consisting of artemisinin, N-acetylcysteine, resveratrol, and hesperidin in a streptozotocin (STZ)-induced intracerebroventricular (ICV) rat model of AD. Twenty 8-week-old rats were divided into four groups: control, SHAM, STZ-ICV, and STZ-ICV receiving oral administration of the compound combination for 30 days. Cognitive performance was evaluated using the Morris water maze and passive avoidance tests. Neurodegenerative and molecular changes were assessed through Western blot analysis of phosphorylated tau, amyloid-β-related markers, and apoptosis- and inflammation-associated proteins. Histological analyses included Nissl staining and immunofluorescence for amyloid deposition and caspase-3 expression. Results demonstrated that STZ-ICV administration induced significant cognitive impairment, neuronal loss, and increased amyloid-β and phosphorylated tau levels. Treatment with the combined compounds partially improved behavioral performance and was associated with reductions in amyloid-β deposition, tau phosphorylation, and caspase-3 expression, along with improved neuronal preservation in the hippocampus. These findings suggest that the combined administration of artemisinin, N-acetylcysteine, resveratrol, and hesperidin exerts multi-target neuroprotective effects in an experimental AD model, potentially through modulation of oxidative stress, neuroinflammation, and apoptotic pathways. However, further studies are required to evaluate pharmacokinetics, safety, and translational relevance before clinical application.