Search Results (27,898)

Background/Objectives: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive and lethal forms of malignant neoplasm of the endocrine system, and osteopontin (OPN) has been shown to be aberrantly expressed in this tumor type. Among the five OPN splicing isoforms (OPN-SI), OPN-4 has been recently reported in several tumor types, including ATC, but its functional role(s) have not yet been elucidated. Methods: To characterize OPN-4 roles in ATC cells, OPN-4 was ectopically overexpressed in the c643 ATC cell line, generating the c643/OPN-4 cells. OPN-roles were evaluated by cell functional assays, including cell proliferation and viability, using Carboxyfluorescein Succinimidyl Ester (CFSE), crystal violet, and trypan blue assays. For migration, clonogenicity, cell cycle and apoptosis assays were used. For assessment, c643/OPN-4 cells were cultured in two-dimensional (2D) monolayers or three-dimensional (3D) spheroids with the latter being maintained in a bespoke microfluidic system. Results: OPN-4 overexpression led to a significant reduction in cell proliferation, viability, migration and clonogenicity. c643/OPN-4 cells displayed a significant accumulation in the G0/G1 phase and a decrease in the S phase of the cell cycle; however this did not affect cell death or the expression levels of other OPN-SI. In a spheroid model of c643/OPN-4 cells, no significant differences were found in spheroid size or viability when compared to those formed by control cells. Notably, OPN-4 overexpression enhanced the effects of sorafenib on cell viability under dynamic treatment conditions involving continuous perfusion. Conclusions: These early findings point to the fact that OPN-4 may reduce some aspects of tumor progression features in ATC cells and open new avenues for investigating OPN-4 as a biomarker of therapeutic response in personalized treatment strategies. Full article

Garrya flavescens S. Wats. (GF) has been traditionally used to treat gastrointestinal spasms, yet its bioactivity within the central nervous system remains unexplored. This study aimed to characterize the bioactive constituents of GF and evaluate its anti-inflammatory and metabolic regulatory effects in lipopolysaccharide-activated microglia. Phytochemical profiling using LC-HRMS and HPLC identified rutin as a primary bioactive component, present at an exceptionally high concentration (9309 μg/g). In BV-2 microglial and RAW 264.7 cells, GF treatment significantly suppressed the expression of pro-inflammatory cytokines and mediators in a dose-dependent manner. Mechanistic studies revealed that GF specifically modulated the ERK signaling pathway. Furthermore, Seahorse XF analysis demonstrated that GF restored mitochondrial homeostasis by reducing basal respiration and proton leak while significantly enhancing spare respiratory capacity. Finally, conditioned medium from GF-treated microglia improved the viability of N2A neuronal cells. These findings highlight GF as a potent botanical source with significant neuroprotective potential, offering a promising candidate for functional food or nutraceutical applications targeting neuroinflammatory disorders. Full article

Grape (Vitis vinifera L.) is an important fruit crop, but its production is severely threatened by ripe rot, a fungal disease caused by Colletotrichum gloeosporioides. However, V. pseudoreticulata ‘Dongan-1’ has been reported to have significant resistance to ripe rot. To investigate the molecular basis of this resistance, we employed RNA-Seq to profile transcriptome changes in the leaves and berry skins of ‘Dongan-1’ following infection. Gene Ontology (GO) enrichment analysis suggested that differentially expressed genes (DEGs) were mainly linked to stress response, cellular processes, and metabolic processes. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that DEGs in both tissues were predominantly enriched in the plant MAPK signaling pathway, peroxisome pathway, plant–pathogen interaction pathway, and plant hormone signal transduction pathway. Notably, VpCML41 was identified as a highly induced gene. Functional characterization through heterologous overexpression in Arabidopsis thaliana and transient expression in ‘Thompson Seedless’ grape leaves demonstrated that VpCML41 enhances resistance to C. gloeosporioides. This enhanced resistance involves the coordinated regulation of salicylic acid and jasmonic acid signaling cascades. Our findings provide valuable genetic resources for understanding ripe rot resistance and offer a foundation for developing resistant grape varieties. Full article

TEA domain transcription factors are critical regulators of tissue development and regeneration in mammals, yet their roles in aquatic invertebrate regeneration remain poorly understood. Here, a full-length cDNA encoding a putative transcriptional enhanced associate domain protein 1 (TEAD1) ortholog in Apostichopus japonicus (AjTEAD1) was cloned and characterized. The open reading frame (ORF) of AjTEAD1 is 1344 bp, encoding a polypeptide of 447 amino acids with a conserved TEA domain (Asp⁴⁰–Leu¹¹¹) and a protein-binding domain (Gly²³¹–Asp⁴⁴⁶). Function analysis demonstrates that AjTEAD1 is essential for intestinal regeneration. AjTEAD1 expression was significantly upregulated during the regeneration process. Functional impairment of AjTEAD1 suppressed intestinal regeneration and attenuated cell proliferation. At the molecular level, we identified the cell cycle gene in A. japonicus (AjCyclin E), whose expression pattern coincided with that of AjTEAD1 and was downregulated following AjTEAD1 knockdown. Dual-luciferase reporter assays further confirmed that AjTEAD1 binds to specific sites in the AjCyclin E promoter and transcriptionally activates its expression. In summary, our study reveals that AjTEAD1 promotes cell proliferation and drives intestinal regeneration in A. japonicus by directly upregulating AjCyclin E transcription. These findings identify the TEAD–Cyclin E axis as a key regulator of echinoderm regeneration, shedding new light on the regenerative processes and cytological mechanisms in economically important species. Full article

Peripheral nerve injuries involving critical-sized gaps remain a major clinical challenge. Although autologous nerve grafting is considered the gold standard for peripheral nerve repair, its clinical application is limited by the availability of donor nerve tissue and the risk of donor-site morbidity, including sensory deficits and functional impairment. Therefore, nerve guidance conduits (NGCs) have emerged as a promising alternative when combined with bioactive modulation strategies. In this study, we evaluated bisvinyl sulfonemethyl (BVSM)-crosslinked gelatin conduits integrated with electrical stimulation (ES) at different frequencies (0, 2, 20, and 200 Hz) in a rat sciatic nerve defect model over a 4-week recovery period (n = 10 per group). Structural regeneration was assessed by morphometric analysis, electrophysiology, macrophage infiltration, CGRP immunoreactivity, retrograde Fluorogold tracing, quantitative PCR of growth factors and inflammatory cytokines, and behavioral testing. Among all stimulation paradigms, low-frequency ES at 2 Hz produced the most pronounced regenerative effects. The 2 Hz group demonstrated significantly greater axon number, axonal density, and regenerated nerve area compared with control and high-frequency groups (p < 0.05). Electrophysiological assessments revealed improved nerve conduction velocity, higher MAP amplitudes, and shorter latencies. Enhanced macrophage recruitment and elevated CGRP expression were observed, suggesting coordinated neuroimmune and neurochemical activation. Gene expression analysis indicated upregulation of neurotrophic factors and balanced inflammatory cytokine responses under low-frequency stimulation. In contrast, high-frequency stimulation (200 Hz) failed to enhance overall regeneration and showed reduced axonal metrics, suggesting possible overstimulation-associated suppression. Collectively, these findings demonstrate that BVSM-crosslinked conduits provide a stable and biocompatible regenerative scaffold, and that appropriately tuned low-frequency electrical stimulation (2 Hz) optimally enhances structural, molecular, and functional recovery. The integration of material engineering with bioelectrical modulation represents a promising strategy for next-generation bioelectronic interfaces in peripheral nerve repair. Full article

Nanogrooves provide instructive cues to cells in culture. Several nanofabrication techniques have been developed to create biomimetic substrates, advancing our understanding of cell adhesion. Their integration into nervous system models highlights the critical role of the extracellular matrix (ECM) in developing functional tissue constructs for in vitro platforms such as Brain-on-Chip (BoC) and Nervous System-on-Chip (NoC). This study presents a nanofabrication approach that integrates photolithography and microtransfer molding (μTM) to pattern nanogrooves using photocurable polymer NOA81 onto microelectrode array (MEA) plates. The resulting nanogrooves exhibited a pattern periodicity of 976 nm and a ridge width of 232 nm, as confirmed by scanning electron microscopy and atomic force microscopy. We assessed the biocompatibility and functional impact of these modified substrates using human induced pluripotent stem cell (hiPSC)-derived neuronal cultures. Neurons cultured on nanogroove-modified MEAs exhibited aligned neural processes due to the anisotropic surface features and expressed vivid spiking behavior and higher burst frequency compared to randomly cultured neuronal networks. In conclusion, the proposed fabrication technique integrates nanogrooves with commercial MEAs using a combination of microtransfer molding and photolithography, resulting in modified culture substrates that enhance spike activity and network organization, aiding in the development of more in vivo-like neural models. Full article

Clear cell renal cell carcinoma (ccRCC) is characterized by marked biological heterogeneity, which limits the prognostic accuracy of conventional clinicopathological models. Increasing attention has therefore focused on identification of biomarkers that can enhance risk stratification throughout all stages of the disease. Starting from the current state of the art, this narrative review summarizes and critically appraises the evidence published over the past decade regarding prognostic biomarkers in ccRCC. The analysis is structured into four overarching domains: (i) genomic biomarkers, covering somatic alterations and transcriptomic signatures; (ii) tissue-based biomarkers, including immunohistochemical surrogates and immune microenvironment features; (iii) circulating biomarkers, such as systemic inflammation parameters and indices; and (iv) integrated predictive models, represented by emerging multi-omic approaches. Going through the broad framework of potential prognostic biomarkers, emphasis is placed on their individual and integrative value in relation to classic clinical-pathological factors and survival parameters. At the tissue level, chromosome 3p-related alterations constitute a central molecular feature of ccRCC. Among these, BAP1 loss has emerged as one of the most consistently validated indicators of aggressive tumor behavior. Disruption of the SETD2/H3K36me3 axis and immune-related biomarkers, including PD-L1 expression, have demonstrated prognostic associations in selected settings, although with variable and context-dependent performance. In the circulating compartment, plasma KIM-1 has shown prognostic relevance following nephrectomy, while postoperative detection of circulating tumor DNA (ctDNA) may identify patients at increased risk of recurrence. However, limited analytical sensitivity and methodological heterogeneity currently restrict the broader clinical applicability of ctDNA-based strategies. Systemic inflammatory indices, such as the neutrophil-to-lymphocyte ratio, show reproducible associations with outcomes but largely reflect host inflammatory status rather than tumor-specific biology. However, no single biomarker currently supports routine prognostic implementation in ccRCC. Future progress will likely depend on integrative models combining genomic, tissue-based, immune, and circulating parameters with established clinical variables. Prospective validation and clear demonstration of incremental clinical utility will be essential before such strategies can meaningfully inform therapeutic decision-making. Full article

To improve the predictive accuracy of wind-field distributions during fires in high-rise buildings, this study targets the shortcomings of traditional prediction methods, including insufficient information fusion and dispersed feature representations under high-rise fire conditions. An efficient attention mechanism, termed Adaptive Channel and Multi-Scale Spatial Fusion Attention Mechanism (CSFAM), is proposed, which endows the model with enhanced adaptive focusing and multi-scale integration capabilities. CSFAM can account for environmental features across multiple dimensions to enable high-spatial-resolution wind-field reconstruction, thereby improving robustness and prediction accuracy in complex environments. To validate the effectiveness of CSFAM for predicting wind fields under high-rise-fire conditions, CFD-based scenario modeling was employed to generate a dataset of 1050 CFD-derived wind-field distributions across diverse inflow-wind and fire-source scenarios, partitioned into training, testing, and validation sets according to the fire-source size. When applying the CSFAM-enhanced multi-layer perceptron (MLP), the wind-field predictions achieved a mean squared error (MSE) of 0.0004, a mean absolute error (MAE) of 0.0141, and an R² of 0.9766, outperforming state-of-the-art methods. The results demonstrate that CSFAM plays a significant role in markedly improving wind-speed prediction accuracy during high-rise-building fires, and enhances the model’s ability to identify and express vortex-like and other key aerodynamic features generated by the fire, thereby improving the capture of the complex nonlinear aerodynamic structures induced by fire. Full article

Photosynthesis, the main energy source for life on Earth, confronts escalating challenges of high-light–high-temperature stress (HLHT). Our previous study identified a mutation in ATP synthase, F₁-α-C252Y, that significantly enhances the HLHT tolerance of Synechococcus elongatus PCC 7942 (Sye7942), although the underlying mechanism remains obscure. In this study, we found that this mutation led to elevated levels of the b subunit of F_o, F₁ subunits, and the ATP synthase within cells, without affecting ATP synthetic activity, indicating improved intracellular ATP synthesis activity. Additionally, the mutation altered the transcriptome of Sye7942, impacting the expression of genes involved in crucial processes, such as the electron transport chain, carbon fixation, and regulatory factors, which are crucial for cyanobacteria’s adaptation to stresses. Correspondingly, the mutant exhibited enhanced photosynthesis, accelerated growth, and increased glycogen under HLHT conditions, showing improved adaptation. The higher intracellular ATP synthesis activity, along with enhanced photosynthetic activity, suggests increased ATP production in the mutant under HLHT. Enhancing ATP production and remodeling the cellular transcriptome appear to be key strategies employed by the C252Y mutation for Sye7942 acclimating to HLHT. These findings provide valuable insights for enhancing photosynthetic efficiency and stress resilience in cyanobacteria and other photosynthetic organisms facing HLHT challenges. Full article

Background: Somatostatin receptors (SSTRs) are pivotal diagnostic and therapeutic targets in well-differentiated neuroendocrine neoplasms (NENs). SSTR-directed treatment strategies rely on sufficient SSTR2 expression. Thus, receptor loss during dedifferentiation limits therapeutic efficacy. Preclinical data suggest pharmacologic modulation of SSTRs’ expression. However, there are no robust data on the effect of NEN-specific systemic treatments on SSTRs’ expression and function. Methods: We systematically evaluated the effects of six systemic agents commonly used in NEN therapy—isplatin, etoposide, 5-fluorouracil (5-FU), streptozotocin (STZ), temozolomide (TMZ), and everolimus—on SSTR2 and SSTR5 expression, as well as on uptake of ⁶⁸Ga-DOTATOC, in BON-1 and QGP-1 cells, as well as the MS-18 cell line. Analyses included qRT-PCR, Western blotting, immunohistochemistry, and radiopeptide uptake assays. Results: Systemic agents modulated SSTR expression and radioligand uptake in a drug- and cell line-dependent manner. Etoposide consistently upregulated SSTR2 expression and significantly increased radioligand uptake across all three cell lines. TMZ enhanced SSTR2 expression and uptake in BON-1 cells, but reduced uptake in QGP-1 and MS-18 cells. In contrast, 5-FU, STZ, cisplatin, and everolimus showed heterogeneous, compound- and cell line-specific effects on SSTR2 expression and ⁶⁸Ga-DOTATOC uptake, including both up- and downregulation depending on the model. Conclusions: All agents under investigation affect SSTR expression in vitro, while etoposide is identified as the most consistent enhancer of SSTR2’s expression and function across cellular NEN models. Our findings highlight both the potential and the risks of systemic therapy-induced receptor modulation and therefore support further investigation of treatment sequencing strategies to optimize SSTR-targeted approaches. However, further studies are required to translate these observations to a clinical setting. Full article

Background: Chronic stress is a major contributing factor to mood disorders, including depression and anxiety; however, the molecular mechanisms underlying individual differences in susceptibility to such disorders remain poorly understood. DNA methyltransferase 3a (Dnmt3a), a key epigenetic regulator, has been increasingly implicated in stress-related neurobiological adaptations. In this study, we employed a well-established mouse model of chronic social defeat stress (CSDS) to investigate the functional role of Dnmt3a in modulating individual susceptibility to social stress. Methods: Male C57BL/6J mice were exposed to chronic/submaximal social defeat stress (CSDS/SSDS). AAV vectors were used to achieve Dnmt3a overexpression or global and oligodendrocyte-specific knockdown in the nucleus accumbens (NAc). Behavioral tests, including social interaction, open field, and elevated zero maze, were conducted alongside Western blotting and immunofluorescence assays. Results: CSDS selectively increased Dnmt3a expression in NAc oligodendrocytes of stress-susceptible mice. Overexpression of Dnmt3a in the NAc enhanced susceptibility to stress, whereas its knockdown conferred resilience, without affecting baseline behaviors. Dnmt3a negatively regulated myelin basic protein (MBP) and dopamine D1 receptor expression. Stress-susceptible mice exhibited shortened myelinated segments and reduced D1 receptor levels, while D2 receptor expression remained unchanged. Conclusions: Dnmt3a in NAc oligodendrocytes modulates susceptibility to social stress through a Dnmt3a-MBP/D1 receptor-NAc pathway, highlighting a critical glia-neuron interaction. This mechanism extends our understanding of the neurobiological basis of stress-related disorders and positions Dnmt3a as a promising therapeutic target for developing precision interventions or biomarkers. Full article

Nosema ceranae is a common and highly contagious fungal pathogen that primarily infects the gut of adult honeybees, causing nosemosis. As a chronic disease of the digestive system, it poses a global threat to honeybee health and colony sustainability. This study aimed to investigate the inhibitory effects of different concentrations of Scutellaria baicalensis aqueous extract on N. ceranae in the intestines of infected Apis cerana through feeding experiments. In addition, the therapeutic efficacy of its major active component, baicalin, was evaluated, and its potential molecular mechanisms of action were explored. The results showed that, compared with the control group, administration of S. baicalensis aqueous extract at concentrations of 1 mg/mL, 5 mg/mL, and 10 mg/mL significantly reduced midgut spore loads (p < 0.05). Further experiments showed that a 0.5 mg/mL baicalin sucrose solution, prepared with 0.5% (v/v) DMSO as co-solvent, exhibited optimal solubility and significantly inhibited the proliferation of spores in the honeybee midgut. Transcriptomic analysis of A. cerana revealed varying numbers of significantly differentially expressed genes among the baicalin-treated (HG) group, the co-solvent control (DMSO) group, and the blank control (C) group. Four candidate DEGs associated with the effects of baicalin were further identified, namely LOC108003965, LOC108000905, LOC107996681, and CYP4G11. Gene Ontology enrichment analysis showed that, in the comparison between the HG group and the C group, these DEGs were significantly enriched in six functional categories: iron ion binding, phosphoric ester hydrolase activity, heme binding, tetrapyrrole binding, hydrolase activity (acting on ester bonds), and oxidoreductase activity (acting on paired donors, with incorporation or reduction of molecular oxygen). Collectively, these results demonstrate that S. baicalensis aqueous extract effectively inhibits the proliferation of N. ceranae within the host, and its active component, baicalin, exhibits a similar inhibitory effect. The present study proposes a novel strategy in which baicalin may enhance host endogenous chitinase-related activity to target and disrupt the spore wall, offering a new perspective for the prevention and control of honeybee nosemosis. Full article

Background: Tumor hypoxia in non-small cell lung cancer (NSCLC) promotes malignant progression and treatment resistance by enhancing abnormal vasculature, invasiveness, and metastasis. However, the molecular mechanisms underlying hypoxia-driven tumor progression remain incompletely understood. Methods: In this study, patient samples, cell lines, single-cell transcriptomic data, and spatial transcriptomic data were comprehensively analyzed to investigate hypoxia-associated molecular alterations in NSCLC. Results: A global trend toward shortened 3’ untranslated regions (3’UTRs) was observed in hypoxic tumors. Analysis of hypoxia-related alternative polyadenylation (APA) events revealed preferential usage of proximal polyadenylation sites (poly(A) sites, PASs) in CARM1. Shortening of the CARM1 3’UTR was associated with hypoxia and may serve as a candidate biomarker. This APA event may reduce putative microRNA (miRNA) binding sites and contribute to increased CARM1 expression, while potentially influencing the expression of hypoxia-related genes such as SELENBP1. Drug sensitivity analysis further suggested that patients with shorter CARM1 3’UTRs may exhibit differential responses to cisplatin chemotherapy. Moreover, single-cell and spatial transcriptomic analyses demonstrated enhanced interactions between hypoxic tumor cells and fibroblasts, highlighting a potential role for APA in remodeling the hypoxic tumor microenvironment. Conclusions: Our findings identify hypoxia-related APA features and characterize hypoxia-associated alterations within the NSCLC tumor microenvironmen, providing new insights into the molecular landscape of hypoxia-associated tumor progression. Full article

Seed germination is a critical initial stage of the plant life cycle, regulated by signaling pathways such as phytohormones and reactive oxygen species (ROS). However, the low germination rate of immature grains is a key bottleneck limiting wheat speed breeding. This study used immature grains of the winter wheat cultivar Kenong 199 (KN199) collected 18 days post anthesis to establish an efficient germination protocol. By screening individual and combined treatments of hydrogen peroxide (H₂O₂, 1%), gibberellin (GA₃, 20 μM), and varying concentrations of abscisic acid (ABA) synthesis inhibitor sodium tungstate (Na₂WO₄), alongside transcriptome analysis, we identified the optimal reagent combination and gained preliminary insight into its molecular basis. The triple reagent combination of 0.5 mM Na₂WO₄ + 20 μM GA₃ + 1% H₂O₂ exhibited the highest germination rate of 80%, approximately sevenfold higher than single reagent treatments, with germination rate peaking after 4 days. Transcriptome profiling revealed that this combination modulated the expression of key genes related to dormancy release and germination, including upregulation of GA biosynthesis gene GA3ox2 and ABA catabolism gene TaCYP707A2, and downregulation of ABA biosynthesis and signaling genes (ABI5, TaNCED1, etc.). Additionally, genes associated with energy metabolism and transport pathways were enhanced. This optimized reagent combination significantly improves immature grain germination, shortens the breeding cycle, and provides a practical tool for achieving “five generations per year” speed breeding in winter wheat. Our findings contribute to seed biology by offering a chemical strategy to overcome dormancy in immature cereal grains. Full article

Alhagi camelorum is a dominant leguminous shrub distributed in the Taklamakan Desert, an area characterized by extreme drought and high soil salinization, which can complete its life cycle normally in salt-affected soils. However, the underlying molecular regulatory mechanism of its salt tolerance remains largely unclear. The AcABI5 gene was successfully cloned and characterized, and it encodes a typical nuclear-localized bZIP transcription factor. Functional characterization demonstrated that overexpression of AcABI5 markedly improved the salt stress tolerance of A. camelorum calli, whereas silencing of AcABI5 via virus-induced gene silencing (VIGS) rendered the plant more sensitive to salt stress. Further mechanistic investigations revealed that AcABI5 enhanced salt tolerance by regulating the expression of superoxide dismutase (SOD)- and peroxidase (POD)-related antioxidant genes. Compared with the wild type, AcABI5-overexpressing calli exhibited significantly increased SOD and POD activities and remarkably reduced malondialdehyde (MDA) content under salt treatment, whereas AcABI5-silenced lines exhibited the opposite physiological phenotypes. Furthermore, heterologous silencing of AcABI5 in Nicotiana benthamiana via virus-induced gene silencing (VIGS) produced comparable salt-sensitive phenotypes, similar to those observed in A. camelorum AcABI5-silenced lines. Collectively, these results provide insights into the molecular mechanism by which AcABI5 enhances salt tolerance in A. camelorum, and lay a solid theoretical foundation for the optimization of the A. camelorum genetic transformation system and the expansion of related salt-tolerant crop research. Full article