Breast cancer (BC) is the most common malignancy among women worldwide, accounting for 11.7
Circular RNAs (circRNAs) are a novel class of non-coding RNAs that function as competing endogenous RNAs (ceRNAs) by sharing microRNA (miRNA) response elements with mRNAs, thereby modulating gene expression in various biological processes, including ferroptosis. Accumulating evidence has implicated circRNAs in the progression of acute kidney injury (AKI); however, whether circRNAs are involved in the regulation of ferroptosis during AKI and the underlying molecular mechanisms remain unclear. A circRNA-miRNA-mRNA interaction network was initially built via bioinformatics. Quantitative real-time PCR (RT-qPCR), Western blot, and immunohistochemistry were used to measure circ_ATP5A1, hsa-miR-145-5p, and ZFP36 expression. Dual-luciferase reporter assays combined with RNA immunoprecipitation verified the targeting relationships among the three molecules. In LPS-treated HK-2 cells, cell viability, inflammatory responses, and ferroptosis were evaluated using CCK-8, lactate dehydrogenase (LDH) release assay, flow cytometry, ELISA, and commercial reagent kits. An LPS-induced AKI mouse model was also established to assess the protective effect of circ_ATP5A1 expression. A ceRNA regulatory network comprising circ_ATP5A1/hsa-miR-145-5p/ZFP36 was established through bioinformatics prediction and experimental validation. Experimental findings revealed that expression of circ_ATP5A1 and ZFP36 was downregulated in both LPS-challenged HK-2 cells and AKI mice, whereas hsa-miR-145-5p was upregulated. Mechanistic studies demonstrated that circ_ATP5A1 acts as a ceRNA by sponging hsa-miR-145-5p, thereby relieving the repressive effect of hsa-miR-145-5p on its downstream target ZFP36. Functional assays showed that overexpression of circ_ATP5A1 restored the viability of LPS-exposed HK-2 cells, reduced inflammatory cytokine secretion (IL-1β, IL-6, TNF-α), increased the expression of ferroptosis-related markers glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11), and decreased acyl-CoA synthetase long-chain family member 4 (ACSL4) expression. In vivo, circ_ATP5A1 overexpression significantly ameliorated renal dysfunction and tissue injury in AKI mice. The circ_ATP5A1/hsa-miR-145-5p/ZFP36 axis attenuates AKI by inhibiting ferroptosis, providing a theoretical basis for AKI treatment.
The early detection of endometriosis (EM), a significant cause of dysmenorrhea, is essential for effective clinical management. This study sought to determine the functional role of miR-5584-5p in EM and the mechanistic involvement of its target, FZD2, in disease progression. This case-control study enrolled 217 dysmenorrhea patients (107 EM cases and 110 controls). Ishikawa cells with modulated miR-5584-5p and FZD2 expression were used to assess cell proliferation, migration, and invasion. The miR-5584-5p/FZD2 interaction and Wnt/β-catenin transcriptional activity were validated by dual-luciferase and TOP/FOP flash reporter assays. Serum miR-5584-5p was significantly downregulated in EM patients, demonstrating high diagnostic accuracy (AUC = 0.903) and acting as an independent protective factor. In vitro, miR-5584-5p overexpression suppressed Ishikawa cell proliferation, migration, and invasion, concurrently inhibiting Wnt/β-catenin transcriptional activity and the epithelial-mesenchymal transition (EMT) axis. FZD2 was confirmed as a direct target of miR-5584-5p. Crucially, FZD2 overexpression partially reversed the inhibitory effects of miR-5584-5p on malignant cellular phenotypes and restored Wnt/β-catenin signaling. Serum miR-5584-5p serves as a valuable potential diagnostic biomarker for EM. Functionally, it attenuates endometrial epithelial cell aggressiveness by targeting FZD2 and suppressing the Wnt/β-catenin-EMT axis. Future studies utilizing primary cells and in vivo models are warranted to validate these preliminary mechanisms.
Next-generation sequencing (NGS) is the primary method for SNP genotyping in a large-scale population. However, it still faces a challenge to balance the paradox between SNP genotyping accuracy and sequencing cost in populations characterized by high heterozygosity and low genomic linkage disequilibrium. Although sequencing depth and depth of coverage (DP) are critical parameters influencing SNP genotyping accuracy, comprehensive investigation on genotyping consistency of SNP among different sequencing depths and impact of DP on SNP genotyping accuracy of low-depth sequencing is of absence. In this study, we utilized Lueyang Black-boned chickens (n=11) to evaluate genotyping consistency of SNP at 5× and 10× sequencing depths via two strategies: (1) comparison of SNP genotyping results between 5× and 10×; (2) comparison of genotyping accuracies of 5× and 10× by employing Sanger-genotyping results of 162 SNP at the SLCO1B3 locus as the criteria. Furthermore, we investigated the effect of DP filtering on genotyping consistency between 5× and 10× by increasing DP from 0 to 5. We studied the association of genotyping results of NGS with GC contents and repeat sequences for each chromosome. The results show that the ratio of genotyping consistency is 78.07% between 5× and 10×. The ratio is 78.40% between 5× and Sanger sequencing, and increased to 87.17% for 10×. Almost all (99.18%) of inconsistent results happen in heterozygotes, of which 5×AB-10×AA/BB is mainly present in 14 microchromosomes with lengths less than 10 Mb. The 5×AB-10×AA/BB subset of inconsistent results is significantly associated with repeat sequences and GC contents. The ratio of consistent loci increases from 78.07% to 87.33% with the increasing of DP from 0 to 5. However, calling rates of SNP dramatically reduces from 100% to 35.07%. The ratio of missing loci decreases from 13.16% to 4.08%. DP filtering has no significant effect on the ratio of inconsistent loci. The results indicate that challenge of the NGS-based SNP genotyping approach focuses on genotyping of heterozygotes. Sequencing depth and sequence features of chromosomes affect genotyping accuracy. To physically increase sequencing depth improves the genotyping accuracy of SNP, whereas to algorithmically raise DP has a negligible effect. Increasing DP can significantly improve the genotyping consistency between low- and high-depth sequencing. The improving effect quickly decays with the increasing of DP, whereas calling rates of SNP dramatically reduce.
Mechanical forces are persistent and dynamic physical cues during embryonic development and reproduction. Through mechanosensitive ion channels (MSCs), these forces are converted into intracellular signals-primarily mediated by Ca2+ flux, that regulate cell migration, lineage specification, and morphogenesis. In parallel, mitochondria serve as central hubs of energy metabolism and signal integration, and their metabolic state, redox homeostasis, and dynamic remodeling critically influence early developmental competence. Increasing evidence suggests that MSC-mediated Ca2+ signaling not only activates canonical signaling pathways but also functionally couples to mitochondrial activity, establishing a regulatory axis that links mechanical inputs to metabolic responses. This review focuses on the "MSC-mitochondrial function- embryonic developmental fate" axis. We systematically summarize the molecular characteristics and activation mechanisms of MSCs and discuss their roles in embryonic development and reproductive processes. Furtherly, we examine how MSC-dependent Ca2+ signaling modulates mitochondrial metabolic reprogramming, reactive oxygen species (ROS) homeostasis, and fusion-fission dynamics, and consider the potential implications of this coupling in key developmental events including blastocoel formation, lineage specification, and organogenesis. We also discuss the current limitations and technical challenges in the field, together with future research directions and potential translational implications. By integrating mechanotransduction with mitochondrial metabolic regulation in the developmental context, this framework provides insights into how mechanical and metabolic signals coordinately shape embryonic cell fate and morphogenesis, and offers a theoretical basis for elucidating the mechanisms underlying developmental abnormalities and reproductive disorders, as well as exploring potential intervention strategies.
The Diannan small-ear pig, an indigenous breed in Yunnan of China, has significant conservation and utilization value due to its unique economic traits. To investigate the population genetic diversity and structure of this breed, this study applied Illumina Porcine 50K SNP chip to detect genome-wide SNPs in 424 Diannan small-ear pigs. Using PLINK software, a total of 29,468 SNPs were detected. The results showed that the average polymorphism information content (PIC) was 0.21, the minor allele frequency (MAF) was 0.18, and the average expected heterozygosity (He) was 0.24, indicating a moderately low level of genetic diversity in the Diannan small-ear pigs population. Subsequently, the phylogenetic tree was constructed using MEGA X software, showing that the Diannan small-ear pigs could be divided into eight linages. Computed with Plink software, the distance matrix was built from pairwise identity by state (IBS) proportions. The results revealed that the average IBS proportion across all individual pairs was 0.23. Analysis of genetic relationships within the population with GCTA software revealed that most Diannan small-ear pigs exhibited moderate genetic proximity. Additionally, a total of 5,301 runs of homozygosity (ROH) segments were identified. 59.54% of those ROHs ranged from 0 to 5 Mb in length. The average inbreeding coefficient based on ROH was 0.006, indicating a low level of inbreeding in the Diannan small-ear pigs population. This study provides a worthwhile genetic foundation for the future conservation and utilization of Diannan small-ear pigs. These findings indicated that the Diannan small-ear pig conservation farm could introduce other more gene flows from other subpopulations to promote population genetic diversity based on preserving their unique and desirable traits, and integrate modern biotechnology for breeding organisms like genomic selection to promote its potential for the discovery and utilization of the merit phenotype-related genes, contributing to the long-term scientific protection and sustainable development and utilization of this valuable genetic resource.
With advances in high-throughput sequencing technologies, whole-genome sequencing-based mutation mapping (mapping-by-sequencing) has become an important approach in modern genetic studies. However, in undergraduate life science courses, students have relatively few opportunities to engage with authentic sequencing data analysis workflows, and systematic teaching resources related to sequencing data analysis remain limited, particularly in conventional genetics teaching. Based on the Galaxy platform and the MiModD tool, this study reorganized the mutation mapping analysis workflow for instructional purposes and developed a case that can be applied in functional genomics and bioinformatics courses and integrated into genetics teaching. Using the published whole-genome sequencing data from the Caenorhabditis elegans mutant ot266 as a case, students are guided through the complete analytical workflow, including sequencing data quality control, read alignment, variant calling, allele frequency-based mapping, and candidate mutation identification. This teaching design provides a feasible teaching case for undergraduate bioinformatics or functional genomics education and can serve as a useful supplement to genetics teaching.
Homo erectus remains have been found in Africa, Eurasia, and Southeast Asia, with a fossil record dating back to 2 million years, holding a significant place in human evolution. However, due to limited molecular evidence, its genetic characteristics, diversity, and potential connections to other archaic homonins and modern humans have long remained unresolved. To address these issues, a research team led by Qiaomei Fu from the Institute of Vertebrate Paleontology and Paleoanthropology of the Chinese Academy of Sciences, in collaboration with other archaeological institutions, successfully extracted ancient enamel proteins from six Middle Pleistocene H. erectus teeth (~0.4 Ma) from the Zhoukoudian, Hexian, and Sunjiadong sites. Further in-depth paleoproteomic analyses revealed the following breakthroughs. First, a quantitative sex determination pipeline was established based on male-specific amelogenin, Y isoform (AMELY), confirming that five of these specimens are male and one is female. Second, two genetically specific amino acid variants in ameloblastin (AMBN) were identified in all specimens from three sites. One is a newly discovered variant, AMBN-253G, which has not been found in any other archaic or modern human populations. It represents a molecular marker specific to the East Asian Middle Pleistocene H. erectus lineage, providing solid evidence that H. erectus specimens from Zhoukoudian, Hexian, and Sunjiadong sites belonged to the same evolutionary lineage. The other variant, AMBN-273V, has previously been identified in Denisovans. However, genetic analysis in this study reveals that this variant may have been introduced into Denisovans through populations related to these Middle Pleistocene H. erectus, and some of which subsequently contributed to certain modern human populations from Southeast Asia or Oceania. This study obtains lineage-specific molecular information from Homo erectus fossils for the first time, and reshapes our understanding of hominin evolution and the history of genetic admixture in East Asia.
The hepatic low-density lipoprotein receptor (LDLR) mediates the clearance of circulating low-density lipoprotein (LDL) and is a key determinant of cholesterol homeostasis. However, how nutritional cues such as chronic dietary cholesterol regulate LDLR intracellular trafficking and protein turnover through specific molecular switches remains poorly understood. Dr. Alan Saltiel's team at the University of California San Diego discovered a cholesterol stress-response pathway mediated by Ral GTPases, revealing a new mechanism by which dietary cholesterol reshapes LDLR protein fate and thereby affects cholesterol homeostasis. The study found that chronic dietary cholesterol loading or sustained Ral activation reduces LDLR protein levels. Mechanistically, chronic cholesterol loading activates Ral GTPases by increasing RAS activity. Activated Ral, on the one hand, recruits the RalBP1-REPS1 endocytic complex to promote LDLR internalization and lysosomal trafficking while inhibiting LDLR recycling back to the cell surface. On the other hand, Ral activation promotes the processing and lysosomal localization of cathepsin A (CTSA) and reduces its extracellular secretion, thereby enhancing CTSA-mediated LDLR degradation. This process occurs independently of LDLR transcriptional regulation or PCSK9-mediated LDLR degradation. Inhibition of CTSA stabilizes LDLR protein and promotes LDL uptake and plasma cholesterol clearance. Human genetic analyses further indicate that key components of this pathway are associated with plasma lipid levels and cardiovascular disease risk. This study reveals a dietary cholesterol-activated Ral-CTSA-LDLR protein homeostasis pathway, providing a new mechanistic framework for understanding how nutritional stress disrupts cholesterol metabolism and identifying a potential therapeutic target distinct from existing treatment strategies for hypercholesterolemia and cardiovascular disease.
Yersinia pestis, the causative agent of plague, is a highly virulent pathogen that has caused three historical pandemics and currently persists in multiple active natural foci worldwide, posing a serious threat to human health and public safety. Distinct phylogenetic lineages of Y. pestis exhibit differences in virulence, among which the Microtus biovar within the 0.PE4 lineage is nearly avirulent to large mammals. Comparative genomics between low-virulence and highly virulent populations is therefore key to elucidating the evolutionary basis of virulence. However, comprehensive population-genomic analysis of the low-virulence lineage remains limited, particularly those integrating multiple types of genomic variations. In this study, we analyzed 169 low-virulence strains and 215 representative high-virulence strains collected from long-term national surveillance data in China and public databases. Phylogenetic and spatiotemporal analyses revealed geographic clustering within the low-virulence 0.PE4 lineage. The 0.PE4.3 subclade circulating in China and Mongolia was further subdivided into three regionally associated tertiary lineages, with two more finely resolved quaternary lineages identified specifically in Inner Mongolia. Comparative genomic analyses with representative high-virulence genomes identified mutations fixed in the most recent common ancestor (MRCA) of the low-virulence population and its subclades. Among these, 81 single nucleotide polymorphisms (SNPs), 19 insertion and deletions (Indels), and 5 large fragment losses are shared by all low-virulence strains, defining the genomic features of their MRCA. Together with selection pressure analysis, we identified five genes (ail, rovA, tssH, cdiA, and alr) under strong positive selection, which are involved in virulence, metabolism, and adaptation. Collectively, this study reconstructs the phylogenetic topology of the global low-virulence Y. pestis population and identifies key genomic variations that occurred during its evolutionary process, providing valuable insights for fine-scale tracing and identifying potential molecular targets for elucidating virulence mechanisms.
Macrodactyly is a congenital abnormality characterized by overgrowth of single or multiple digits, frequently accompanied by pathological changes in bone, soft tissue, and nerve tissue, and is caused mostly by somatic variants. Previous studies have demonstrated that macrodactyly is predominantly caused by pathogenic variants in the PIK3CA gene, yet the underlying molecular mechanism remains unclear. In this study, we analyzed 13 patients clinically diagnosed with isolated macrodactyly. Through whole-exome and Sanger sequencing, we identified one pediatric patient carrying a novel mosaic de novo in-frame deletion mutation (c.307_312del/p.Glu103_Pro104del) in the PIK3CA gene, which was not detected in either parent. According to the American College of Medical Genetics and Genomics (ACMG) guidelines, we classified this mutation as a variant of uncertain significance (PM4+PM2_Supporting+PM6_Supporting). Protein structure prediction analysis indicated that this in-frame deletion altered the protein's local conformation and surface electrostatic potential. Furthermore, the wild-type and mutant expression vectors were constructed, including the well-established pathogenic mutation c.353G>A/p.Gly118Asp in PIK3CA gene as a control. We transfected these vectors into HEK293T cells for functional analyses, respectively. The results showed that although this mutation did not significantly affect PIK3CA mRNA or protein expression levels, it promoted the phosphorylation of downstream AKT protein at both Thr308 and Ser473. Therefore, the mutation may express the localized overgrowth phenotype by activating the PI3K/AKT signaling pathway. This study not only expands the genetic variant spectrum of macrodactyly, but also further clarifies the role of the PI3K/AKT pathway in the PIK3CA-related overgrowth.
Biological sex is a critical determinant influencing the incidence, progression, clinical presentation, and prognosis of human diseases. Beyond sex-specific tumors, men typically exhibit higher morbidity and mortality rates in nearly all tumors compared to women. However, the genetic mechanisms underlying the cancer sex bias are not definitive. Loss of Y chromosome (LOY), the most prevalent somatic mutation in healthy males, frequently occurs in individuals over 50 years of age. On one hand, LOY in peripheral blood immune cells is strongly associated with increased tumor susceptibility. On the other hand, LOY also frequently occurs within tumor cells themselves, leading to the loss of Y-chromosome-linked tumor suppressor genes and non-coding RNAs, thereby reshaping the tumor microenvironment through mechanisms such as metabolic reprogramming and altering tumor immunity. Strikingly, LOY is a "contagious" mutation, which means malignant cells with LOY cause the same mutation in tumor-infiltrating immune cells, and the synergistic interaction between two types of LOY cells accelerates tumor evolution. Given the widespread occurrence of LOY in tumors and its crucial role in malignant progression, LOY holds great promise as a biomarker for early diagnosis, classification, and staging, as well as a potential target for precision therapy. Here, we summarize the mechanisms by which LOY drives malignant progression in solid tumors from the perspectives of tumor-cell-intrinsic LOY, LOY in tumor-infiltrating immune cells, and their crosstalk. We also discuss the developmental opportunities and translational potential of LOY in precision oncology. This review aims to provide novel insights into the molecular basis of sex disparities in tumors and to identify potential molecular targets for precision prevention, diagnosis, and treatment.
Alzheimer’s Disease (AD) is a progressive neurodegenerative disorder characterized by complex and not yet fully understood pathological mechanisms. This study was designed to explore the molecular mechanisms of long non-coding RNA (lncRNA) FBXL19-AS1 in the pathological progression of AD. 110 AD patients and 110 healthy controls were recruited for this study. Relevant gene expression was measured using reverse transcription quantitative real-time PCR (RT-qPCR), and the diagnostic performance was assessed through receiver operating characteristic (ROC) curve analysis. AD cell models were established by treating SH-SY5Y and BV2 cells with amyloid β (Aβ)25–35. Cell proliferation was evaluated using the cell counting kit-8 (CCK-8) assay, cell apoptosis was analyzed by flow cytometry, and the secretion levels of inflammatory factors were determined via enzyme-linked immunosorbent assay (ELISA). A dual-luciferase reporter assay was performed to confirm the direct regulatory interactions. In AD patients, the FBXL19-AS1 and integrin subunit beta 3 (ITGB3) expression were significantly elevated, whereas microRNA-650 (miR-650) expression was markedly reduced. FBXL19-AS1 showed certain diagnostic potential for AD, and its expression was negatively correlated with Montreal Cognitive Assessment (MoCA) scores. Following treatment with Aβ25–35, SH-SY5Y cells exhibited reduced proliferative capacity and increased apoptosis, while BV2 cells showed elevated levels of inflammatory factors. Additionally, Aβ25–35 treatment led to upregulated expression of FBXL19-AS1 and ITGB3, along with downregulated expression of miR-650. Knockdown of FBXL19-AS1 effectively reversed these effects. However, inhibition of miR-650 partially attenuated the changes induced by FBXL19-AS1 knockdown. Furthermore, knockdown of FBXL19-AS1 markedly reduced ITGB3 expression, whereas concurrent inhibition of miR-650 partially reversed this downregulatory effect. FBXL19-AS1 appears to interact with and modulate miR-650, thereby affecting ITGB3 expression and potentially participating in the pathological progression of AD.
Macrophage polarization exerts a vital role in the progression of peri-implantitis. RNA methylation serves as an important regulator of macrophage polarization. However, reports on the role of N7-methylguanosine (m7G) regulators in peri-implantitis are scarce. The function of m7G regulators in peri-implantitis and macrophage polarization was investigated via in vivo and in vitro experiments. RNA sequencing (RNA-seq) and m7G methylated RNA immunoprecipitation sequencing (m7G MeRIP-seq), and molecular biology assays were performed to identify and validate downstream m7G-modified target genes. WDR4 was identified as the sole significantly upregulated m7G-related gene in peri-implantitis tissues. Overexpression of WDR4 promoted macrophage M1 polarization, which further inhibited osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). RNA-seq revealed 3001 differentially expressed genes (DEGs) in WDR4-overexpressing macrophages, including 1503 upregulated genes and 1498 downregulated genes. m7G MeRIP-seq identified 554 m7G differential peaks between the control and WDR4 overexpression group. Integrative analysis of RNA-seq and m7G-MeRIP-seq identified 33 genes with both increased expression and enhanced m7G methylation, among which ZBP1 was selected as a key downstream target. WDR4 interacted with ZBP1 mRNA, increased its m7G modification, and enhanced its mRNA stability. Silencing ZBP1 significantly attenuated WDR4-induced M1 macrophage polarization and restored the impaired osteogenic differentiation of BMSCs. WDR4 promotes M1 polarization of macrophages by enhancing the m7G modification of ZBP1, thereby inhibiting BMSC osteogenic differentiation and promoting peri-implantitis progression. Targeting the WDR4/m7G/ZBP1 axis may represent a potential therapeutic strategy for peri-implantitis.
Abstract When breeding apple cultivars for northern latitudes, early harvest and late flowering are desirable to manage the short growing season and ongoing climatic changes. Depending on the intended use, increased content of polyphenolic compound might be considered desirable (juice and cider) or undesirable (dessert). Thus, we phenotyped a total of 111, 189, and 88 apple genotypes for harvest date, flowering date, and peel concentration of procyanidin B2 (PRB2), respectively, across three Nordic locations in Sweden, Norway, and Finland. Using phased marker data from the 20 K apple Infinium ® SNP array we performed haplotype-based validation of previously described quantitative trait loci (QTL) intervals and genomic regions on Linkage Group (LG) 3 (harvest date), LG 9 and 12 (flowering), and LG16 (PRB2). We assessed the correlation between the traits and historical climate adaptation and identified haplotypes associated with increasing and decreasing effects for the three traits. These results are valuable for Marker Assisted Parent Selection in the Nordic breeding programs, as they support designing crosses to meet the demands of current and future climatic conditions, and fruit quality towards breeding of new dessert cultivars, or cultivars for juice or cider production.
Long non-coding RNAs (lncRNAs) function as pivotal regulators in gastric cancer (GC) pathogenesis via competitive endogenous RNA (ceRNA) networks. However, the precise role of GUSBP11 in GC progression remains inadequately characterized. This investigation aimed to delineate the biological function of GUSBP11 in GC and to ascertain whether it regulates malignant phenotypes via miR-29c-3p. GUSBP11, miR-29c-3p, and THBS2 expression levels were quantified in tissues and cells using RT-qPCR. Prognostic value of GUSBP11 in GC was assessed via Kaplan-Meier curve and Cox analysis. In vitro, AGS and HGC-27 cell lines were employed for loss- and gain-function assays. Cellular viability was measured via CCK-8, while migration and invasion capacities were examined using Transwell chambers. Epithelial-mesenchymal transition (EMT) markers were detected by RT-qPCR. Mechanistic interplay was validated through dual-luciferase reporter assays and miRNA inhibitor rescue experiments. GUSBP11 exhibited upregulation in GC tissues and cell lines, correlating significantly with lower 5-year overall survival. Functional assays demonstrated that GUSBP11 overexpression enhanced viability, migration, and invasion, whereas its knockdown elicited the opposite phenotype. Mechanistically, GUSBP11 operated as a ceRNA by sponging miR-29c-3p, thereby relieving suppression of downstream effector THBS2. Co-transfection with a miR-29c-3p inhibitor effectively reversed the tumor-suppressive consequences of GUSBP11 silencing, restoring THBS2 expression and EMT programming. The GUSBP11/miR-29c-3p axis exerts a pro-oncogenic influence on GC progression. This regulation offer fresh insights into GC pathophysiology and identifies prospective biomarkers for therapeutic intervention.
Postmenopausal osteoporosis (PMOP) is a prevalent female bone disease. It can progress to fractures at advanced stages and raise disability risks. This study evaluated the value of EPB41L4A-AS1 in PMOP diagnostic and fracture risk prediction, and explored its mechanism of regulating osteogenic differentiation in an in vitro ADSC model. A total of 118 healthy controls and 146 PMOP patients (75 non-fracture and 71 fracture) were enrolled. Serum levels of EPB41L4A-AS1, miR-302d-3p, and CUL3 were detected. ROC curves and logistic regression were performed to assess its clinic value. The regulatory mechanisms of EPB41L4A-AS1, miR-302d-3p and CUL3 were investigated using adipose-derived mesenchymal stem cells (ADSCs) models and a series of cellular experiments. Serum EPB41L4A-AS1 were downregulated in PMOP patients and further decreased in the fracture subgroup (P < 0.001). EPB41L4A-AS1 exhibited high diagnostic performance for PMOP (AUC = 0.818). Its expression was strongly correlated with key bone metabolism indicators. Low EPB41L4A-AS1 expression was an independent risk factor for PMOP (OR = 0.164) and fractures occurrence (OR = 0.208). During osteogenic induction of ADSCs, silencing EPB41L4A-AS1 significantly reduced osteogenic markers expression and mineralized nodule formation, and disrupted cellular function and bone microenvironment homeostasis. Mechanically, EPB41L4A-AS1 targeted miR-302d-3p, which further regulated CUL3. Rescue experiments confirmed that the EPB41L4A-AS1/miR-302d-3p/CUL3 axis modulated ADSC osteogenic differentiation and bone microenvironment homeostasis. EPB41L4A-AS1 serves as a marker for diagnosis and fracture risk prediction in PMOP patients, and in vitro evidence suggests it regulates ADSC osteogenic differentiation via miR-302d-3p/CUL3, providing an in vitro mechanistic proof-of-concept.
Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease, and renal oxidative stress (OS)–mediated tubular injury plays a critical role in its progression. Resveratrol (Res) possesses established antioxidative properties, yet its mechanistic involvement in ferroptosis-ferritinophagy signaling remains unclear. This study investigated the protective effects of Res on renal tubular injury in DKD and explored its regulatory role in the ferroptosis–ferritinophagy pathway. A DKD mouse model was established using unilateral nephrectomy followed by streptozotocin injection. A total of 42 male C57BL/6 N mice were used in this study. Six mice served as the blank control group, and the remaining 36 underwent model induction. The 30 successfully modeled mice were randomly divided into 5 groups (n = 6 per group): the DKD group, the positive control group (perindopril 0.48 mg/kg/d), and the resveratrol (Res) low-, medium-, and high-dose groups (1.0, 2.5, 5.0 mg/kg/d, administered by gavage for 8 weeks). Renal function, oxidative stress markers, and renal tubular injury markers were assessed. Hematoxylin and eosin (H E) staining, Masson staining, 4-hydroxynonenal (4-HNE) immunofluorescence, and Western blotting (for ferroptosis/ferritinophagy-related proteins) were performed. Compared with the Black group, DKD group mice exhibited marked OS, tubular injury, and activation of the ferroptosis–ferritinophagy axis. Res treatment significantly reduced fasting blood glucose, SCr, BUN, 24-hour urinary protein, MDA, Fe²⁺, β2-MG, RBP4, and NGAL levels, while elevating T-SOD and GSH-Px (P < 0.05). Res also downregulated Nrf2, SLC7A11, GPX4, LC3II/LC3I, and FTH1, alongside increased expression of p62 and NCOA4, demonstrating a clear dose-dependent effect. Resveratrol markedly attenuates renal oxidative stress and tubular injury in DKD. These findings suggest that its renoprotective effects may be mediated through inhibition of the ferroptosis–ferritinophagy pathway, highlighting Res as a promising therapeutic candidate for DKD.
Esophageal cancer (ESCA) is highly prevalent globally, but the role of RNA-binding proteins (RBPs) in it remains unclear. ESCA datasets were analyzed to identify differentially expressed genes (DEGs). Candidates were derived by intersecting DEGs with RBP-related genes and validated via machine learning, expression analysis, and receiver operating characteristic (ROC) curves. Prognostic value, functional enrichment, immune microenvironment, regulatory networks, drug prediction, and Eca-109 cell validation were assessed. Four biomarkers (CDC20, COL7A1, DNMT3B, UBE2T) were confirmed. Low UBE2T/DNMT3B correlated with better survival. In addition, CDC20/UBE2T were linked to DNA replication; COL7A1/DNMT3B to olfactory transduction. Immune analysis showed 15 cell types differed significantly, with most chemokines negatively correlating with biomarkers. COL7A1, DNMT3B, and UBE2T were associated with transcription factor TET1. The four biomarkers corresponded to 79, 26, 67, and 23 targeted drugs, respectively, and all were highly expressed in Eca-109 cells, advancing insights into RBP-related mechanisms in ESCA.
Colorectal cancer (CRC) is a prevalent malignant neoplasm characterized by high incidence and mortality rates. Currently, the role of microRNAs (miRNAs) in CRC is increasingly recognized. In this study, we aim to investigate the prognostic value of miR-363-5p and its associated molecular mechanisms. A total of 156 paired samples were collected from patients with CRC. The levels of miR-363-5p in the samples were detected by RT-qPCR. KM curves and multivariable Cox regression models evaluated the prognostic value of miR-363-5p. Flow cytometry, CCK-8 assay and Transwell assay examined the effects of miR-363-5p on cellular malignant phenotypes. Dual-luciferase reporter assays validated the targeting relationship. The expression level of miR-363-5p was declined in CRC, and patients with TNM stage III + IV exhibited lower miR-363-5p expression compared to those with TNM stage I + II. Furthermore, individuals with low miR-363-5p expression demonstrated shorter overall survival than those with high expression, and miR-363-5p was confirmed as an independent protective factor for CRC. In vitro, miR-363-5p suppressed cellular malignant phenotypes, primarily manifested by miR-363-5p mimic promoting apoptosis while inhibiting proliferation, migration and invasion. Mechanistically, we identified MEIS3 as a direct target of miR-363-5p, and rescue experiments confirmed that MEIS3 overexpression attenuated the tumor-suppressive effects of miR-363-5p. Downregulated miR-363-5p were involved in CRC progression and associated with poor patient prognosis. Mechanistically, upregulated miR-363-5p suppressed the malignant cellular phenotype by negatively regulating MEIS3, thereby inhibiting the progression of CRC.
