Search Results (11,884)

Metallothioneins (MTs) and heat shock protein 70 (HSP70) are key regulators of cellular stress response and metal homeostasis and play important roles in tumor biology. The aim of this study was to examine their expression patterns and potential prognostic significance in different molecular subtypes of breast cancer (BC), with special emphasis on triple-negative breast cancer (TNBC) and the Luminal A subtype, compared with benign breast lesions (fibroadenomas). A total of 90 tissue samples were included, and the expression of MTs in the cytoplasm and nucleus and HSP70 in the nucleus of tumor cells was analyzed immunohistochemically and correlated with clinicopathological features and treatment outcomes. Distinct expression patterns of HSP70 and MTs were observed between malignant and benign samples, as well as among the analyzed molecular subtypes of BC, suggesting their involvement in cellular adaptive mechanisms associated with malignant transformation. TNBC was characterized by less favorable clinicopathological features compared to the Luminal A subtype, including higher histological grade, increased proliferative activity, and a higher incidence of recurrence and metastatic disease. Survival analyses confirmed a worse outcome for patients with TNBC, while HSP70 and MTs expression did not show independent prognostic value in multivariate models. In conclusion, although HSP70 and MTs play important biological roles in the cellular response to stress and tumor adaptation, their expression in this study does not represent an independent prognostic indicator of clinical outcome. Nevertheless, the observed expression patterns provide insight into the complex mechanisms of tumor adaptation and emphasize the need for integrative approaches in BC biomarker research. Full article

Backgrounds: Roegneria ciliaris is a perennial tetraploid wild relative of wheat that is widely distributed in China. It can be used both as a forage crop and ecological grass (the grasses specifically bred for ecological restoration) due to its strong stress tolerance, early green-up, vigorous seedling growth in spring, and great palatability. Methods: It is necessary to select and validate appropriate reference genes (RGs) for gene expression normalization by qRT-PCR in order to decipher the stress tolerance mechanism of this grass species. Therefore, eight candidate RGs were identified from transcriptome data of R. ciliaris ‘Liao sheng’ in response to drought stress. The expression stability of these RGs was evaluated by five algorithms (∆Ct, geNorm, NormFinder, Bestkeeper and ReFinder) using samples from different tissues and drought stress. Results: The results showed that MDH and RPL19 were the most stable RGs among all samples, while GAPDH and TUBA presented the lowest expression stability. These representative RGs were further used to normalize the expression level of the pyrroline-5-carboxylate synthase (P5CS) and protein phosphatase 2C (PP2C) genes in different tissues and under drought stress. The results of P5CS and PP2C expression were consistent with transcriptome data. Conclusion: Our study provided the first systematic evaluation of the most stable RG selection for qRT-PCR normalization in R. ciliaris, which will promote further research on its tissue-specific gene expression and mechanism of drought tolerance. Full article

The risk of developing some types of head and neck squamous cell carcinoma (HNSCC) is seven times higher in males, and such disparities may not be associated only with tobacco and alcohol consumption or HPV infection. Therefore, the endocrine microenvironment is considered another risk factor, as epidemiologic studies have unequivocally shown the protective effect of estrogen in women. This research was focused on progesterone receptors (PRs), the least-studied sex hormone receptors in HNSCC. Our study included fresh tissue samples from 95 primary tumors, 25 metastatic lymph nodes and 40 healthy oral mucosa. Gene expression of nuclear (PGR) and seven membrane PRs (PAQR5, PAQR6, PAQR7, PAQR8, PAQR9, PGRMC1 and PGRMC2) was analyzed by qRT-PCR and associated with clinicopathological characteristics. The results showed that, compared to control tissue, PGR was increased in metastatic lymph nodes, while PAQR5, PAQR7, PAQR8 and PAQR9 were decreased in primary tumors (all p < 0.05). The expression of almost all PRs was greater in older patients and showed moderate to strong positive mutual correlations in both tumors and controls. PARQ8 and PAQR9 were increased in females and pT4 tumors (all p < 0.05). Survival analysis showed that higher PAQR5 (hazard ratio (HR) 2.8, 95% confidence interval (CI) 1.19–6.57, p = 0.019) and PAQR7 (HR 2.0, 95% CI 1.01–3.81, p = 0.048) were associated with worse overall survival, but their independence was not proven in multivariate analysis. Although most PRs were reduced in primary tumors, an increased PAQR5 expression, also associated with tumor invasion markers, could likely mark a specific aggressive, advanced stage of primary tumors and potentially serve as a negative prognostic biomarker for HNSCC. Full article

Breast cancer (BC) is a global health problem. BC is a biologically heterogeneous disease in which novel immunotherapeutic strategies are needed, particularly in the metastatic setting. The NKG2D/NKG2D ligand (NKG2DL) axis is a key component of innate antitumor immunity and represents a potential therapeutic target, but its relevance in BC has not been fully characterized. We performed an in silico analysis of NKG2DL expression in BC cell lines, healthy breast tissue, and tumor samples using publicly available transcriptomic databases (DSMZCellDive, ShinyTHOR, GTEx, TCGA, Human Protein Atlas), complemented by survival analyses from TCGA and KMPlot and a narrative review of the literature. NKG2DL transcripts were consistently expressed in BC cell lines and tumor tissues, with higher expression observed in ductal histology, higher tumor stage, and basal molecular subtype. Survival analyses showed heterogeneous and generally weak associations between individual NKG2DLs and clinical outcomes. In silico proteomics data are scarce, but the narrative review showed that NKG2DLs are expressed by immunohistochemistry in tumor tissues but absent in surrounding healthy tissues. The literature review also revealed concomitant dysfunction of NKG2D+ effector cells due to multiple resistance mechanisms (including ligand shedding). We also review potential therapeutic approaches. Full article

In this research, Fe-P scaffolds were successfully fabricated by the powder metallurgy method for the first time, using NaCl as the space holder for bone tissue engineering applications, with apparent porosities of approximately 70%. The Fe₃P powder was successfully synthesized by the mechanochemical method under an argon atmosphere using an initial mixture of Fe and P powders. The XRD patterns show that Fe₃P was obtained after sintering the milled powders at 1000 °C. Fe, Fe₃P, and Fe-50 wt% Fe₃P composite scaffolds and bulk pellets were prepared by sintering the milled powder at 1000 °C. Furthermore, the mechanical properties (compression strength) and bioactivity of the Fe-P scaffolds were determined. According to the compression test results, the composite scaffold showed higher compressive strength, lower fracture strain, and higher elastic modulus than the Fe and Fe₃P scaffolds, indicating that adding Fe₃P to Fe improves the mechanical properties. Moreover, among the scaffolds prepared by sintering at 1000 °C, the Fe scaffold exhibited the highest corrosion rate compared to the Fe₃P and composite samples, while the corrosion resistance of the composite sample was 3 times higher than that of the Fe sample. The ICP analysis showed that the amount of Fe released from the bulk pellets during soaking in PBS solution after four weeks was 3220 μg/dL, 4003 μg/dL, and 4774 μg/dL for the composite, Fe₃P, and Fe samples, respectively. The composite sample showed the highest cell viability, while the Fe sample had the lowest. The compressive strength (12.62 MPa) and fracture strain (5.98%) of the porous sintered composite scaffold at 1000 °C were within the range of trabecular bone, while the compressive strength of the composite sample was 17 times higher than that of the Fe sample. Furthermore, the MTS test showed that all the samples had good viability, while the composite sample had the best cell viability. The scaffolds were not cytotoxic. It can be concluded that the mechanical and biological properties of the composite sample were superior to those of the Fe and Fe₃P samples and that it may be a promising candidate for bone tissue engineering applications, especially for trabecular bone replacement. Full article

This study examined effects of two moderate-intensity aerobic exercise prescriptions on inflammatory cytokines and oxidative stress biomarkers in middle-aged obese Hispanic females. Fifty-four subjects were randomly assigned to a lower-moderate intensity group (55% VO₂max, LT, n = 18), an upper-moderate intensity group (70% VO₂max, HT, n = 19), or a non-exercise control group (CON, n = 17). Blood samples collected before and after a 12-week intervention were analyzed for tumor necrosis factor-alpha (TNF-α), adiponectin, C-reactive protein (CRP), total antioxidant status (TAS), and 8-hydroxy-2′-deoxyguanosine (8-OHdG). Body fat percentage (%BF) and visceral adipose tissue (VAT) were assessed using dual-energy X-ray absorptiometry. TNF-α significantly decreased in both LT (p = 0.004) and HT (p < 0.001). Significant increases in adiponectin (p = 0.001) and reductions in CRP (p < 0.001) were observed within the HT, whereas these changes were not significant within the LT. TAS significantly increased in both exercise groups (p < 0.001), and 8-OHdG significantly decreased in the HT (p < 0.001) and LT (p = 0.002). Both LT and HT demonstrated significant reductions in %BF (p < 0.001) and VAT (p < 0.05), with no significant changes in CON. Results indicate that moderate-intensity aerobic exercise improves inflammatory and oxidative stress profiles when total exercise volume is matched, regardless of differences in exercise intensity within the moderate range. Although post-intervention differences between groups were not statistically significant, the observation that CRP, adiponectin, and 8-OHdG changed significantly only within HT suggests that exercise intensity may influence biomarker responsiveness and warrants further investigation. Full article

Background: Periodontal disease is a chronic inflammatory condition characterized by progressive destruction of the supporting tissues of the teeth. MicroRNAs (miRNAs) have emerged as key post-transcriptional regulators of immune and inflammatory responses; however, their coordinated expression patterns in periodontal disease remain incompletely understood. Objective: This study aimed to evaluate the expression of miR-141-3p, miR-146a-5p, and miR-148b-3p in periodontal disease and to explore their potential co-regulation in relation to periodontal inflammation. Methods: Gingival samples were collected from individuals with periodontal disease and periodontally healthy controls. miRNA expression levels were quantified by real-time quantitative PCR using TaqMan assays and normalized to miR-191-5p. Relative expression was calculated using the ΔΔCt method. Statistical analyses included group comparisons and correlation analyses to assess relationships among miRNA expression levels. Results: miR-146a-5p expression was significantly reduced in periodontal disease compared with healthy controls (p < 0.05), while miR-141-3p showed a consistent downward trend without reaching statistical significance. No significant differences were observed for miR-148b-3p expression between groups. Notably, strong positive correlations were identified among the three miRNAs, indicating coordinated expression patterns independent of disease stage or grade. Conclusions: The findings demonstrate altered expression of miR-146a-5p and coordinated regulation of miR-141-3p, miR-146a-5p, and miR-148b-3p in periodontal disease. These results support the concept that miRNAs act within regulatory networks rather than as isolated markers and contribute to a more nuanced understanding of miRNA-mediated regulation in periodontal inflammation. Full article

Culturing cells and micro-tissue samples in 3D bio-scaffolding structures is gaining popularity; however, precise control of tissue micro-environment in such systems remains challenging. We describe a family of new hybrid bio-scaffolds with 3D O₂-sensing ability, produced by simple means from readily available bio-scaffolding and O₂-sensing materials. Three different types of phosphorescent O₂-sensing materials—polymeric microparticles (MPs), supramolecular probe MitoXpress and nanoparticulate probes NanO2 and Nano-IR (NPs)—were integrated in Matrigel and agarose scaffolding materials and evaluated. Key working characteristics of such hybrid scaffolds, including heterogeneity, stability, cytotoxicity, optical signals and O₂-sensing properties, ease of fabrication and use, were compared. The results show superiority of the Matrigel hybrids with NanO2 and Nano-IR probes. Demonstration experiments were conducted with HCT116 cells and individual spheroids derived from these cells, culturing them in the Matrigel–NP hybrid scaffolds and monitoring oxygenation and local O₂ gradients on a time-resolved fluorescence plate reader and by phosphorescence lifetime imaging microscopy (PLIM). Full article

Medical image classification is a fundamental task in medical image analysis and underpins a wide range of clinical applications, including dermatological screening, retinal disease assessment, and malignant tissue detection. In recent years, diffusion models have demonstrated promising potential for medical image classification owing to their strong representation learning capability. However, existing diffusion-based classification methods often rely on oversimplified prior modeling strategies, which fail to adequately capture the intrinsic multi-scale semantic information and contextual dependencies inherent in medical images. As a result, the discriminative power and stability of feature representations are constrained in complex scenarios. In addition, fixed noise injection strategies neglect variations in sample-level prediction confidence, leading to uniform perturbations being imposed on samples with different levels of semantic reliability during the diffusion process, which in turn limits the model’s discriminative performance and generalization ability. To address these challenges, this paper proposes a Confidence-Guided Adaptive Diffusion Network (CGAD-Net) for medical image classification. Specifically, a hybrid prior modeling framework is introduced, consisting of a Hierarchical Pyramid Context Modeling (HPCM) module and an Intra-Scale Dilated Convolution Refinement (IDCR) module. These two components jointly enable the diffusion-based feature modeling process to effectively capture fine-grained structural details and global contextual semantic information. Furthermore, a Confidence-Guided Adaptive Noise Injection (CG-ANI) strategy is designed to dynamically regulate noise intensity during the diffusion process according to sample-level prediction confidence. Without altering the underlying discriminative objective, CG-ANI stabilizes model training and enhances robust representation learning for semantically ambiguous samples.Experimental results on multiple public medical image classification benchmarks, including HAM10000, APTOS2019, and Chaoyang, demonstrate that CGAD-Net achieves competitive performance in terms of classification accuracy, robustness, and training stability. These results validate the effectiveness and application potential of confidence-guided diffusion modeling for two-dimensional medical image classification tasks, and provide valuable insights for further research on diffusion models in the field of medical image analysis. Full article

Isoniazid (INH) is an antitubercular drug that exhibits high toxicity in dogs due to the absence of N-acetyltransferase activity in this species. Consequently, it has been implicated in both accidental and intentional poisonings in dogs. The aim of this study was to develop and validate an analytical method for the quantification of INH in canine liver samples and to apply it in the forensic investigation of seven suspected poisoning cases. The method, based on liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-QTOF-MS), enabled both accurate INH measurement and analysis of the molecular pattern of its metabolite formation. In addition, histopathological examination of the stomach, pancreas, liver, and brain was performed. Liver INH concentrations ranged from 11.822 to 30.484 μg/g and were associated with extensive necrotic lesions across all examined tissues. A strong signal for isonicotinic acid was observed in all samples, whereas the acetylated metabolite was negligible. The developed method allows precise quantification of INH in canine liver and facilitates identification of the characteristic molecular profile of its metabolites. Full article

Objective: Although Hu sheep are renowned for their high fecundity, the multi-tissue regulatory networks governing spermatogenesis, particularly within the hypothalamic–pituitary–testicular (HPT) axis, remain poorly understood. This study aimed to elucidate these mechanisms by performing a comparative proteomic analysis of the HPT axis in Hu sheep and three other breeds. Methods: We utilized data-independent acquisition (DIA) proteomics to analyze hypothalamic, pituitary, and testis tissues from 36 samples across four breeds. The experimental workflow included protein extraction, enzymatic digestion, LC-MS/MS, and subsequent bioinformatic analyses, complemented by histological examination. Results: Hu sheep exhibited accelerated testicular development and an earlier onset of spermatogenesis. Comprehensive proteomic profiling identified a total of 10,528 proteins, with 771 differentially expressed proteins (DEPs) detected in the testis. These testicular DEPs were significantly enriched in pathways related to spermatogenesis, the blood–testis barrier, and steroid hormone biosynthesis. Notably, the cAMP signaling pathway was consistently enriched across all three tissues, underscoring its pivotal role in regulating spermatogenesis. Protein–protein interaction (PPI) network analysis further highlighted hub proteins, such as MET, suggesting their potential involvement in somatic cell functions and the spermatogenic microenvironment. Key findings were validated by Western blot analysis. Conclusion: This study is the first multi-tissue proteomic investigation proposing a model in which the high reproductive performance of Hu sheep is potentially linked to the efficient, coordinated regulation of spermatogenesis-related proteins and signaling pathways—particularly in the testis. These findings offer novel insights into the molecular mechanisms of male reproduction in sheep and identify potential targets for future research and breeding applications. Full article

Background: Hypoxia is a key driver of cancer progression. However, its specific prognostic significance in gastric cancer (GC) remains insufficiently characterized. Methods: Single-sample gene set enrichment analysis (ssGSEA), weighted gene co-expression network analysis (WGCNA), univariate Cox regression, and least absolute shrinkage and selection operator (LASSO) regression were employed to identify a hypoxia-related prognostic signature. Subsequently, immune microenvironment profiling and single-cell RNA sequencing analyses were employed to further characterize the biological characteristics of the signature. In addition, quantitative real-time polymerase chain reaction (qPCR) was used to validate the expression levels of key hypoxia-associated genes in human GC tissues. Results: Elevated hypoxia levels were linked to worse survival outcomes in GC patients. Through integrated WGCNA, Cox, and LASSO analyses, a hypoxia-related prognostic signature (HYS) consisting of four genes—SPARC, AXL, NRP1, and VCAN—was established. Patients in the HYS-high group exhibited markedly poorer overall survival than their HYS-low counterparts [p = 0.000126, hazard ratio (HR) = 1.936]. Moreover, the HYS-high group exhibited increased infiltration of resting CD4⁺ memory T cells, monocytes, M2 macrophages, and resting mast cells, as well as elevated expression of immunosuppressive molecules, including PDCD1LG2 and HAVCR2. Single-cell RNA sequencing analysis revealed that the signature genes were predominantly expressed in cancer-associated fibroblasts. Consistently, qPCR analysis in five paired GC and para-carcinoma tissues confirmed higher expression of these genes in tumor samples (p < 0.01). Conclusions: Our findings indicate that hypoxia is a critical determinant of prognosis in GC and is closely associated with an immunosuppressive tumor microenvironment, highlighting its potential value as a prognostic biomarker and therapeutic target. Full article

Structural biomarkers determined by X-ray scattering of the tissues can complement conventional histopathology and facilitate a fast triage procedure of cancer biopsy samples. It has been shown previously that lipid reflexes can distinguish cancerous from benign samples, except for fibroadenomas. In the present study, we demonstrate that fibroadenoma samples can be recognized using X-ray scattering of collagen. Moreover, we show that modifications in collagen structure are manifested in the water reflexes. Examination of diffraction patterns from water using two-dimensional Fourier transformation and machine learning yields excellent classification metrics in both synchrotron images and laboratory diffractometer data. Full article

Mushroom-related intoxications pose a distinctive challenge for forensic medicine because early manifestations are non-specific, latency may be prolonged, and co-exposures can obscure the mechanism of death. This narrative review summarizes key toxic and psychoactive fungi and their principal compounds, spanning organ-toxic syndromes (amatoxins, orellanine) and functional neuropsychiatric intoxications—acute, predominantly functional effects causing impairment rather than organ failure (psilocybin/psilocin, ibotenic acid/muscimol). We propose an integrated diagnostic workflow combining exposure history, biochemical markers of organ injury, mycological assessment, and confirmatory toxicology. Particular emphasis is placed on postmortem interpretation: toxin instability and biotransformation, conjugation, matrix effects, postmortem redistribution (central vs. femoral blood), and postmortem fungal colonization that may alter analyte profiles or generate misleading metabolites. Because robust lethality thresholds are unavailable for most mushroom toxins, conclusions should rely on a multi-source synthesis of scene information, autopsy/histopathology, and time-dependent matrix selection (urine, gastric contents/vomitus, bile, and selected tissues; kidney for late orellanine confirmation). We review current screening and confirmatory methods—ELISA; LC-MS/MS, LC-HRMS/MS, GC-MS—and highlight pre-analytical requirements (rapid sampling, cold storage) to reduce false negatives. Finally, we discuss emerging directions such as point-of-care tests, portable mass spectrometry, and DNA barcoding for species identification. Full article

The study aims to investigate the skin microbiome composition of Yaks and the effects of different dietary nutrient levels on the skin microbiome diversity and metabolites. A total of 19 healthy Tianzhu White Yaks at two age stages (2.5 and 4.5 years old) were selected and fed either a high-energy diet (n = 9) or a low-energy diet (n = 10). After 90 days of feeding, skin microbiota and skin tissue metabolites were detected using 16S rRNA sequencing and LC-MS/MS untargeted metabolomics, respectively. The results showed: (1) the phyla Firmicutes, Actinobacteriota, Proteobacteria, and Bacteroidetes exhibited relatively high abundances in the skin of yaks, and the total abundance of these four phyla reached as high as 99.3%. Alpha diversity analysis indicated that the alpha diversity index of yak skin microbiota was significantly higher (p < 0.05) in the low-energy nutritional level group than in the high-energy nutritional level group in yaks of both 2.5 and 4.5 years of age. Principal coordinate analysis (PCoA) revealed a distinct separation of all skin microbiota samples into two clusters: the high-energy (H) and low-energy (L) groups. (2) A total of 114 differentially expressed metabolites were screened across both groups, significantly enriched (p < 0.05) in pathways including synaptic vesicle trafficking and glycerophospholipid metabolism; (3) Correlation analysis between microbiota and metabolites revealed significant positive correlations (p < 0.01) between Psychrobacter and choline, and between Corynebacterium and palmitic acid. In conclusion, A low-energy diet increases skin microbial diversity, which is beneficial for maintaining community stability; In contrast, a high-energy diet enriches bacterial genera such as Corynebacterium and Psychrobacter, enhancing functions related to antibacterial activity and barrier protection. Full article