Experimental Biology and Medicine最新文献

Diabetic nephropathy (DN) remains a major complication of diabetes, significantly impacting renal function. Emerging evidence suggests that NAD metabolism plays a crucial role in DN pathogenesis. This study investigates the roles of NAD metabolism-related genes in DN and how there are associated with different disease subtypes. We analyzed gene expression data from DN-associated datasets (GSE30528 and GSE30529) to identify differences in NAD metabolism-related genes between normal and DN samples. We classified DN into subtypes based on NAD gene expression and evaluated NAD scores using ssGSEA. Immune cell infiltration and pathway analyses were assessed using ssGSEA, Microenvironment Cell Populations-counter (MCPcounter), and Gene Set Variation Analysis (GSVA). Key biomarker genes were identified using machine learning algorithms and validated across multiple datasets. We further explored the relationship between gene expression and kidney function using the Nephroseq V5 tool. Thirteen differentially expressed NAD metabolism-related genes were identified, with distinctive expression patterns observed between normal and DN samples. Two distinct NAD-related subtypes were classified, demonstrating significant differences in gene expression, immune cell infiltration, and pathway activities. Immune-related pathways and cellular processes exhibited varied enrichment between subtypes. Six key NAD metabolism-related genes (FMO3, ALDH1A3, FMO5, TKT, LBR, HPGD) were identified as potential biomarkers. Higher levels of FMO3, ALDH1A3, TKT, and LBR were linked to worse kidney function, while FMO5 and HPGD were associated with better kidney function. The study highlights the significant involvement of NAD metabolism-related genes in DN pathogenesis and their association with disease subtypes and renal function. The identified biomarkers could be targets for new treatments and provide insight for future DN research.

Amorphous silicon dioxide nanoparticles (SiO₂ NPs) are abundant within the earth's crust and can be released into the air through industrial and manufacturing activities. Such materials are often used in industrial processes, in pharmaceutical and in the cosmetic industries. Amorphous SiO₂ NPs are pulmonary toxicants; however, the mechanism of toxicity is uncertain. In the current study, toxicity of SiO₂ NPs was assessed using inhalation exposure in an in vivo system to study a possible mechanism of pulmonary injury. Golden Syrian Hamsters were divided into 4 groups: 1- room air control, 2- vehicle control, 3- low concentration (6 mg/m³) and 4- high concentration (12 mg/m³). Hamsters were treated for 4 h a day for 8 days. Bronchoalveolar Lavage Fluid (BALF) analysis found increases in total cell counts (p < 0.0001), neutrophils (p < 0.0001), lymphocytes (p < 0.001), eosinophils (p < 0.01), multinucleated macrophages (p < 0.01), total protein (p < 0.0001), alkaline phosphatase (p < 0.0001), and lactate dehydrogenase (p < 0.001) in the high concentration group. Histopathological analysis found an increase in air space, quantified by Mean Linear Intercept (p < 0.0001), and a significant increase in TUNEL positive cells (p < 0.001), in the high concentration group. SEM and TEM found structural alterations to the lung tissue including increase in the number holes in the alveolar walls and in apoptotic bodies within tissue. Caspase 3 (p < 0.05), and 8 (p < 0.05), were significantly increased along with cellular inflammation markers TNF-α (p < 0.05), and HSP70 (p < 0.05) in the high concentration group. Results of the study indicate exposure to SiO₂ NPs may induce extrinsic apoptotic pathway, leading to tissue damage and significant airspace enlargement.

Cyanide is generally considered a cytotoxic molecule. However, recent studies have shown that mammalian cells - including endothelial cells - can produce cyanide from glycine via a lysosomal pathway. Studies in hepatocytes indicated that cyanide, when administered at low concentrations, or when generated from endogenous sources, exerts regulatory, rather than cytotoxic effects. Here we show that human umbilical vein endothelial cells produce detectable levels of cyanide (∼0.1 nmoles/mg protein/h), and this is enhanced by administration of glycine (1 mM). Glycine stimulates endothelial cell proliferation, migration and tube formation. Low concentrations of the cyanide releasing molecules amygdalin or mandelonitrile (100 µM) exert similar effects. On one hand, cyanide induces the upregulation of VEGF protein in endothelial cells, while on the other hand, VEGF stimulates the generation of cyanide by endothelial cells, suggesting a positive feedback. VEGF-stimulated endothelial cell ATP generation, proliferation and migration is inhibited by the cyanide scavenger hydroxycobalamin (10 µM) as well as by pharmacological agents that prevent lysosomal acidification and thus inhibit cyanide formation by the endothelial cells. In conclusion, cyanide, at low concentrations, generated by endothelial cells, acts as a proangiogenic mediator, via stimulation of the VEGF pathway and the maintenance of cellular bioenergetics.

Although diabetic nephropathy (DN) stands as a prominent complication in individuals with diabetes, the specific molecular mechanisms remain unclear. In this study, we focused on one newly discovered lncRNA, MCF2L-AS1, and its target gene, BCOR, in individuals with various levels of DN. Twenty-eight participants with different stages of DN (14 early stage and 14 late stage), 12 non-diabetic individuals, and 12 with T2DM without microvascular complications were selected. The qPCR was done, and one-way ANOVA assessed gene expression. ROC curves analysis and Spearman correlations between levels of expression and clinicopathological parameters were explored. The expression of MCF2L-AS1 decreased in the early and late DN groups compared to the type 2 diabetes (T2DM) (P = 0.01 and P = 0.03, respectively) and non-diabetic groups (P = 0.01 and P = 0.03, respectively). However, BCOR gene expression analysis revealed that there was no significant difference between the groups (P = 0.27). MCF2L-AS1 levels negatively correlated with microalbuminuria (P = 0.003, r = -0.41), but not with creatinine (Cr) (P = 0.058, r = -0.29). Moreover, there was no correlation between BCOR and microalbumin (P = 0.85, r = 0.02) and Cr (P = 0.49, r = 0.10). ROC curves underscored significant diagnostic accuracy for MCF2L-AS1 in distinguishing DN from people without kidney diseases (P < 0.05). This study introduces MCF2L-AS1 as a potential key player in the molecular landscape of DN, shedding light on its multifaceted interactions. The results provide a basis for further exploration and therapeutic interventions in the management of DN.

Human Immunodeficiency Virus (HIV) and Human Papillomavirus (HPV) co-infections are significantly prevalent, especially among African Americans (AA), a situation further compounded by the prevalence of tobacco smoking. Extracellular vesicles (EVs) are integral to the mechanisms of viral pathogenesis, as they are pivotal in the modulation of immune responses and the inflammatory process. This research study examines the varying concentrations of EVs, their associated biomarkers, and the cytokine/chemokine profiles present in plasma obtained from individuals infected with HIV and those coinfected with HIV and HPV, with particular emphasis on the ramifications of smoking behavior. Our findings revealed that HIV infection markedly elevates EV formation and modifies their protein composition, whereas HPV co-infection does not significantly augment EV levels but does influence the specific cytokine packaging. Notably, monocyte chemoattractant protein-1 (MCP-1 or CCL2) and Regulated upon Activation, Normal T cell Expressed and presumably Secreted (RANTES or CCL5) exhibited substantial enrichment in EVs derived from individuals coinfected with HIV and HPV, implying a potential role of EVs in immune modulation related to viral persistence. Importantly, smoking was found to affect EV characteristics, resulting in an increase in EV size and the packaging of inflammatory mediators, such as MCP-1 and interleukin-18 (IL-18), from plasma into EVs in HIV- and/or HIV+HPV-infected samples. This observation suggests that oxidative stress induced by smoking may intensify immune dysregulation through modifications in EV-mediated cytokine signaling pathways. Nevertheless, smoking did not exhibit a significant impact on the expression of EV marker proteins or the overall levels of EVs. These outcomes underscore the intricate interactions between HIV, HPV, and/or smoking in influencing the immune milieu via EVs. Further comprehensive understanding of the role of EVs in the context of these viral infections could yield valuable insights into potential biomarkers for disease progression and new therapeutic strategies.

Keratin 6A (KRT6A) is an epithelial-specific type II keratin localized within cytoskeletal intermediate filaments and functions in cooperation with KRT16/17 to maintain epidermal homeostasis and tissue repair. Accumulating evidence highlights its multifaceted roles in cancer. Aberrant KRT6A expression promotes cell cycle progression, epithelial-mesenchymal transition, migration, and invasion, thereby driving tumor initiation and metastasis, although tumor-suppressive effects have been observed in specific contexts. Mechanistically, KRT6A regulates adhesion, cytoskeletal remodeling, and critical signaling pathways, thereby reshaping tumor immunity and metabolism to facilitate immune evasion and metabolic dysregulation. Elevated KRT6A expression is strongly associated with resistance to chemotherapy, targeted therapy, and radiotherapy. Therapeutic approaches targeting KRT6A include nucleic acid-based interventions, protein degradation strategies, inhibition of upstream regulatory pathways, and combinatorial regimens to overcome drug resistance. Clinically, KRT6A has emerged as both a diagnostic and prognostic biomarker, supporting treatment monitoring and enhancing predictive models for risk stratification and individualized outcome evaluation. Beyond oncology, mutations in KRT6A underlie pachyonychia congenita, and its dysregulation contributes to epidermal hyperproliferative disorders such as psoriasis. Overall, systematic elucidation of the structure-function-pathway-clinical axis of KRT6A offers new opportunities for precision medicine and supports its potential as a therapeutic target in cancer management.

ALI/ARDS are clinical syndromes with diverse etiological origins and are characterized by high mortality rates and a lack of specific therapeutic options. Garlic oil (GO) has been utilized in both culinary and medicinal applications for millennia. However, its complex chemical composition and inherent instability have limited further development and clinical application. We aimed to encapsulate GO within liposomes to increase its solubility and stability. The therapeutic efficacy of GO-loaded liposomes (GO-lips) against LPS-induced ALI was subsequently evaluated in vivo. A novel GO-lip formulation was developed, and its preparation process was optimized to ensure its stability and bioavailability. A murine model of LPS-induced ALI was established. The animals were randomly assigned to the normal control, LPS model, GO treatment, or GO-lip treatment group. Therapeutic outcomes were evaluated by lung tissue histopathology, inflammatory cytokine quantification and oxidative stress biomarker measurement. PCR and molecular dynamics simulations were used to verify the ALI treatment-related pathways influenced by GO-lips. We successfully developed GO-lips using a novel fabrication method. GO-lips demonstrated favorable physicochemical characteristics, with a mean particle diameter of 175 ± 3 nm, a PDI of 0.27 ± 0.02, and an encapsulation efficiency of 70.74 ± 2.11%. Compared with the LPS model group, the GO-lip treatment group exhibited significant protection against LPS-induced ALI. GO-lips demonstrated greater efficacy than free GO, as evidenced by the improved lung histopathology, reduced pulmonary edema, decreased inflammatory responses, and attenuated oxidative stress. PCR analysis demonstrated that GO-lips significantly protect mice primarily via Nrf2 pathway activation. These findings suggest that liposomal encapsulation of GO increases its anti-inflammatory and antioxidant activities, protecting against LPS-induced ALI. This research offers a novel clinical therapeutic approach for ALI and contributes to foundational knowledge supporting the development and utilization of GO-derived formulations.

Age-related macular degeneration (AMD) represents a leading cause of irreversible blindness among the older persons. Characterized by a complex pathogenesis and multiple risk factors, AMD poses substantial challenges for treatment and has emerged as a significant public health concern. The gut microbiota constitutes a vast and dynamically evolving ecosystem, with a healthy microbial community playing an essential role in maintaining host homeostasis through its involvement in digestion and immune defense. However, alterations in microbial composition or function can compromise intestinal barrier integrity, trigger systemic inflammation, and contribute to disease pathogenesis. Evidence now underscores the influence of gut microbiota on the development and progression of AMD. This review examines the mechanisms by which gut microbes may contribute to AMD pathogenesis and evaluates the therapeutic potential of interventions targeting the gut microbiome-including dietary modifications, Pharmacological and Biological Agents, probiotics, prebiotics, and fecal microbiota transplantation-for AMD management.

N6-methyladenosine (m6A) RNA methylation regulators have been implicated in colorectal cancer (CRC) progression. However, systematic evaluation using multiple machine learning approaches for prognostic prediction remains limited. This study aimed to develop and validate machine learning models for CRC prognosis based on m6A regulators and assess their potential for immunotherapy response prediction. We analyzed 1,047 CRC patients from TCGA and GEO databases (70% training, 30% validation). Twenty machine learning algorithms were systematically evaluated, with LASSO regression selecting optimal features from 27 m6A regulators. SHAP analysis provided model interpretability. Immune microenvironment characterization and immunotherapy response prediction were performed using established computational methods. LASSO regression selected eight m6A regulators (IGF2BP2, METTL3, HNRNPA2B1, METTL14, YTHDF2, VIRMA, FTO, ALKBH5) for model construction. Among 20 algorithms tested, Random Forest achieved optimal performance (training AUC = 0.895, validation AUC = 0.847). SHAP analysis identified IGF2BP2 (mean |SHAP| = 0.42) and METTL3 (mean |SHAP| = 0.36) as primary contributors to risk prediction. Risk stratification showed significant survival differences (HR = 2.41, 95% CI: 1.73-3.36, p < 0.001). Low-risk patients demonstrated enhanced immune infiltration with higher CD8⁺ T cells (17.8% vs. 10.2%, p < 0.001) and better predicted immunotherapy response rates (36.5% vs. 20.3%, p = 0.006). Our systematic machine learning analysis demonstrates that m6A regulators can effectively predict CRC prognosis and immunotherapy response. The eight-gene signature provides a practical tool for clinical risk assessment and treatment decision-making.

Even with the development of the Pfizer-BioNTech BNT162b2 vaccine, which provides protection against COVID-19 and demonstrates high efficacy in generating immune responses, the complexities of the dynamics linking pro- and anti-inflammatory cytokine profiles with anti-spike IgG production remain unclear. The study aims to elucidate these immune dynamics after vaccination. This prospective cohort research was done at the University of Diyala from January 2022 to January 2023, evaluating the immunological response to the Pfizer-BNT162b2 mRNA vaccine in 180 healthy students. Pro- and anti-inflammatory cytokines and anti-spike IgG antibodies were measured before vaccination, 1 month after the second dose, and 4 months after the second dose. Biomarkers were analyzed via ELISA and CRP assays. The study involved 180 healthy participants (80 males, 100 females; median age, 21 years; BMI, 25.7 kg/m²). After the first Pfizer-BNT162b2 vaccine dose, the level of anti-spike IgG increased by 330-fold, and the levels of pro- and anti-inflammatory markers, such as IL-1β, IL-10, and CRP, increased significantly. Four months after the second dose, anti-spike IgG levels were 136-fold above baseline. Significant correlations emerged between cytokine and IgG levels, with anti-spike IgG/IL-10 ratios elevated and sustained over the long term. Pfizer-BNT162b2 vaccine elicits a significant immune response associated with changes in pro-inflammatory cytokines, and the interaction between these cytokines and anti-spike IgG suggests a potential role for immune regulation in enhancing humoral immunity. Based on these findings, the IgG/IL-17 ratio may serve as a viable exploratory biomarker for assessing short- and medium-term vaccination efficacy.