Background: Pulmonary tuberculosis (PTB) development is linked to dysregulated immune responses, with the innate immune cell barrier playing a crucial role in both disease initiation and progression. Methods: The acquisition of transcriptomic datasets pertaining to PTB was conducted using the GEO database. Modules associated with the innate immune cell barrier were identified using weighted gene co-expression network analysis. The intersection of the differentially expressed genes and the module genes yielded a set of overlapping genes. Biological pathways were characterized and candidate genes were screened through functional enrichment analyses and protein-protein interaction network modeling. Key genes were identified through the application of machine-learning algorithms and further confirmed via qPCR using clinical samples. Diagnostic models were developed and independently evaluated. Multiple analytical approaches were employed to explore the biological mechanisms and therapeutic potential. These included gene set enrichment analysis, immune cell infiltration profiling, regulatory network analysis, and molecular docking. Results: A total of 360 overlapping genes were identified, enriched in immune activation and inflammatory pathways. Nineteen candidates were screened, and five key genes-OAS1, STAT2, IFITM1, CCR7, and CD19-were prioritized through machine learning. qPCR validation confirmed consistent expression trends for all genes except CD19. These genes were enriched in lysosomal and ribosomal pathways, with aberrant neutrophil and monocyte infiltration strongly correlating with gene expression. Molecular docking identified peginterferon-alfa-2a and isotretinoin as potential therapeutic agents. Conclusion: This study identified five key genes and two potential drug candidates associated with innate immune dysregulation in PTB, providing new insights into disease mechanisms and supporting their potential value in PTB diagnosis and therapy.
Introduction: Asthma affects over 260 million individuals globally; however, its heterogeneous subtypes pose significant challenges to precision prevention. Although gut microbiome dysbiosis and mitophagy and ferroptosis dysfunction have been implicated in asthma pathogenesis, the potential causal interactions between these pathways remain unclear. Methods: We performed two-sample Mendelian randomization (MR) to assess the causal effects of 473 gut microbial taxa and 49 mitophagy-related genes on four distinct asthma subtypes. Mediation and reverse MR analyses were further employed to explore potential mitophagy-mediated pathways and assess feedback effects from asthma liability on the gut microbiota. For the mitophagy-related mediator genes, we further dissected the ferroptotic pathway-mediated downstream mechanisms and performed drug candidate enrichment and molecular docking. Results: MR analysis identified 20, 19, 13, and 15 gut bacterial taxa that were causally associated with non-allergic, allergic, childhood, and eosinophilic asthma, respectively. Four mitophagy-related genes TP53, MAP1LC3A, NBR1, and CSNK2B exhibited subtype-specific associations. Mediation analysis suggested that TP53-mediated mitophagy mediates 7.7% of the total effect of Microvirga against childhood asthma. Approximately 49% of the total protective effect of TP53 was partially offset through its down-regulation of PARP11-mediated ferroptosis. DSigDB-selected baicalin and theaflavin both stably docked to TP53 at 7.8 kcal mol(-1), which offers natural origin, low toxicity, and high affinity. Conclusion: Our findings support the existence of a gut-mitophagy-ferroptosis-lung axis in asthma pathogenesis. Notably, the Microvirga-TP53-PARP11axis demonstrates a quantifiable protective association with childhood asthma. These insights highlight a potential opportunity for precision, subtype-specific prevention strategies targeting this pathway in early life.
Tooth tissue loss hinders full regeneration. Small intestinal submucosa (SIS) coated with hydroxyapatite (HA) possesses biocompatibility, biodegradability, and osseointegration properties. This study aimed to investigate the effects of SIS/HA on tooth defect. Herein, we used SIS/HA for tooth tissue engineering. Bone marrow mesenchymal stem cell (BMSC)-derived exosomes were inserted into SIS/HA. Histological analysis was detected using HE staining and ALP staining. Gene expression was analyzed by qRT-PCR and Western blot. Osteogenesis was analyzed by alizarin red staining. Cellular mineralization was analyzed by ALP staining. Cell viability was detected by CCK-8 assay. Angiogenesis was analyzed by tube formation assay. We found that SIS/HA effectively showed high cell viability. SIS/HA loaded with BMSC-Exo enhanced the effects of SIS/HA and promoted angiogenesis of RAOECs. Moreover, SIS/HA loaded with BMSC-Exo enhanced the effects of SIS/HA and mediated osteogenesis in vitro. BMSC-Exo-loaded SIS/HA enhanced new bone regeneration in vivo. In summary, BMSC-Exo-coated SIS/HA mediates osseointegration after tooth defect.
Coronary heart disease (CHD), a major cardiovascular disorder, is associated with T cell exhaustion (TEX) in its pathogenesis. This study aimed to identify TEX-related key genes in CHD and explore their potential mechanisms. The training set, validation set, and TEX-related genes (TEX-RGs) were downloaded from public databases. Differentially expressed genes (DEGs) were identified and intersected with TEX-RGs to obtain candidate genes. LASSO regression and Wilcoxon rank-sum test were then used to screen key genes. A nomogram for CHD prediction was constructed and validated via calibration curves and ROC curves. Finally, analyses related to functional enrichment, cell infiltration, T cell state score (TCSS) evaluation, regulatory network construction, drug prediction, and molecular docking were performed for key genes. Initially, the 48 candidate genes were identified, followed by the determination of two key genes (BST2 and XPC). A nomogram integrating BST2 and XPC showed high prediction performance. Functional enrichment analysis indicated their involvement in oxidative phosphorylation, spliceosome, and other processes. Notably, CHD patients exhibited altered infiltration of CD8+ effector memory T cells and chondrocytes, with imbalanced TCSS in Quiescence and Terminal Exhaustion T cell subsets. Regulatory network analysis identified transcription factors (e.g., GATA1, and SP1), microRNAs (e.g., hsa-miR-942-5p), and long non-coding RNAs (e.g., H19) associated with key genes. Potential drugs interacting with key genes (e.g., alvespimycin, MG-262) were predicted, though molecular docking revealed weak binding ability. Collectively, these findings provided valuable insights into the molecular mechanisms of CHD, offering a theoretical basis for understanding the disease and developing targeted therapeutic strategies.
This study aims to screen for and target DPP9 expression in patient-derived tumor xenografts (PDTX) of Undifferentiated Pleomorphic Sarcoma (UPS) to advance personalized therapeutic strategies. We found that DPP9 mRNA expression is significantly upregulated in UPS patients. Single-cell RNA sequencing (scRNA-seq) further confirmed elevated DPP9 levels within the UPS tumor microenvironment. Functional studies demonstrated that shRNA-mediated knockdown of DPP9 suppressed tumor growth in UPS mouse models. Mechanistically, DPP9 promotes UPS cell proliferation by inhibiting mitochondria-dependent ferroptosis. Specifically, DPP9 reduces intracellular reactive oxygen species (ROS) accumulation and confers resistance to ferroptosis. Conversely, DPP9 downregulation induces ferroptosis by enhancing ROS production and mitochondrial dysfunction. Moreover, N6-methyladenosine (m6A) methylation enhances DPP9 mRNA stability in UPS, contributing to its overexpression. DPP9 also activates the NRF2 signaling axis by suppressing KEAP1-mediated ubiquitination and subsequent degradation of NRF2. This stabilization of NRF2 underlies DPP9's ability to inhibit mitochondria-dependent ferroptosis. Collectively, our findings identify DPP9 as a critical regulator of ferroptosis resistance in UPS and suggest that targeting DPP9 may represent a promising therapeutic strategy-not only for UPS, but potentially for other malignancies driven by dysregulated ferroptosis.
Dengue virus (DENV) infections persist as a significant global health threat despite decades of surveillance and control efforts. The disease may progress to severe dengue, marked by hemorrhage, plasma leakage, and vital organ impairment, contributing to substantial worldwide morbidity and mortality. The rapidly escalating DENV burden demands rapid and innovative diagnostic approaches that move beyond conventional detection methods. CRISPR-Cas-based technologies have emerged as a revolutionary approach, offering next-generation solutions for DENV diagnostics. This review outlines the recent advances in the use of CRISPR based technologies for robust and more sensitive detection of dengue virus nucleic acids, critically evaluating their advantages over conventional diagnostics, current limitations, and future prospects. The roles of Cas12 and Cas13 in DENV-RNA detection are discussed in detail. Additional key areas mentioned include field-deployable and portable CRISPR-Cas technologies, serotype-specific detection, hybrid and isothermal amplification-based approaches, and a combination of CRISPR with electrochemical sensing techniques and nanotechnology. Collectively, these advances highlight the potential of CRISPR-based diagnostics in evolving future strategies for rapid and effective dengue virus detection and control of infections.
PTEN-Induced Putative Kinase 1 (Pink1) is a key regulatory protein in mitochondrial autophagy: upon mitochondrial damage, Pink1 selectively binds to the mitochondrial outer membrane, thereby recruiting and phosphorylating Parkin. However, the mechanism by which the Pink1/Parkin signaling pathway functions in podocytes remains unclear, and this study aimed to investigate the role of this pathway in mitochondrial dysfunction associated with glomerular podocyte injury. For this purpose, flow cytometry was used to detect podocyte apoptosis rate; transmission electron microscopy was employed to observe the quantity and morphological changes of podocyte mitochondrial autophagosomes; and reverse transcription-polymerase chain reaction (RT-PCR) and western blot were performed to quantify the mRNA and protein expression levels of Pink1, Parkin, and LC3-II, respectively. The results showed that compared with the Control and Pink1 groups, the PAN group exhibited a significantly increased podocyte apoptosis rate; in the Pink1 group, mitochondria gradually became swollen and rounded, with disordered arrangement. These findings confirmed that PAN can induce podocyte injury and that this process is associated with the Pink1/Parkin pathway. In conclusion, the Pink1/Parkin signaling pathway plays a crucial role in mitochondrial dysfunction during glomerular podocyte injury, and these results provide a new perspective for the potential clinical application of the Pink1/Parkin signaling pathway in podocyte injury and future related research.
In recent years, the prevalence of diabetic nephropathy (DN) has been increasing year by year. Here, this experiment investigated the effects of PFKFB2 in DN and its molecular mechanisms of DN. DN mice were fed a high-fat diet for 12 weeks, and then injected with STZ. DN mice were transfected with negative or sh-DPP9 lentivirus using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). Human proximal tubular HK-2 cells stimulated with 20 mmol/L d-glucose. Analysis revealed a significant downregulation of PFKFB2 expression in DN patients. PFKFB2 was expression in renal cell of DN model using single-cell RNA sequencing. Sh-PFKFB2 aggravated DN in mice model. PFKFB2 up-regulation reduced oxidative stress and glycolysis in model of DN. The inhibition of PFKFB2 aggravated mitochondria-dependent ferroptosis in model of DN. Ferroptosis inhibitor reduced the effects of PFKFB2 down-regulation in mitochondria-dependent ferroptosis in model of DN. PFKFB2 suppressed HIF-1 alpha expression in model of DN by the inhibition of HIF-1 alpha ubiquitination. HIF-1 alpha inhibitor reduced the effects of PFKFB2 down-regulation in mitochondria-dependent ferroptosis in model of DN. In conclusion, PFKFB2 reduced oxidative stress and glycolysis of DN through the inhibition of HIF-1 alpha signaling pathway by the induction of Nrf2 ubiquitination, further elucidating the role of PFKFB2 regulated mitochondrial ROS-induced ferroptosis for DN. Targeting PFKFB2 is thus a potentially effective therapeutic strategy for DN.
BACKGROUND:Colorectal cancer (CRC) remains a major global health burden, and genetic factors such as vitamin D receptor (VDR) polymorphisms have been implicated in its pathogenesis. However, the translational relevance of these variants in clinical risk stratification remains unclear. METHODS:We conducted a comprehensive meta-analysis of case-control studies assessing the association between four common VDR single-nucleotide polymorphisms (Fok1, Apa1, Bsm1, and Taq1) and CRC risk, integrating data from PubMed, Embase, Google Scholar, and other sources through 2024. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated under multiple genetic models. Heterogeneity, publication bias, and sensitivity analyses were performed. Statistical power was evaluated using G*Power 3.1. RESULTS:Across 24 datasets from diverse ethnic populations, no significant associations were observed for any of the four VDR variants in allelic, dominant, recessive, or overdominant models. Statistical power exceeded 0.99 for all variants, indicating that the null results were unlikely due to sample size limitations. CONCLUSION:This study provides robust evidence that these common VDR polymorphisms are not clinically functional as biomarkers for CRC susceptibility. Eliminating these variants from biomarker panels can help redirect resources toward more promising genetic or molecular predictors. These findings also reinforce the need for integrative studies exploring gene-environment interactions, particularly vitamin D status, diet, and lifestyle, to clarify the role of vitamin D pathways in CRC prevention and treatment.
BACKGROUND:Colorectal cancer (CRC) is ranked among the most prevalent digestive system malignancies worldwide. Its progression is closely associated with angiogenesis, which not only supplies nutrients and oxygen to tumors but also facilitates metastasis. Dysregulation of lncRNA MANCR (MANCR) in multiple cancers influences tumorigenesis and development. However, the function and molecular mechanism of MANCR in CRC angiogenesis remain unclear. METHODS:This study constructed a MANCR/miR-20a-5p/GCNT4 axis using TCGA, starBase, and TargetScan Human databases. Expression of MANCR, microRNA-20a-5p, and GCNT4 in CRC was analyzed using the TCGA database, with validation performed in cell lines. In vitro functional assays (CCK-8, colony formation, transwell migration/invasion, tube formation, and Western blotting) were conducted to evaluate the impact of MANCR on CRC cell proliferation, migration, invasion, and angiogenesis. Bioinformatics analysis was used to analyze the Pearson correlation between MANCR and microRNA-20a-5p, and between microRNA-20a-5p and GCNT4. Functional interactions of MANCR and microRNA-20a-5p, and ceRNA regulatory mechanisms were verified via dual-luciferase reporter assays, RNA pull-down, and RNA immunoprecipitation, with further exploration of GCNT4's role in angiogenesis regulation conducted. RESULTS:Significant downregulation of MANCR expression was observed in CRC tissues and cell lines. Overexpression of MANCR robustly inhibited CRC cell proliferation, migration, invasion, and angiogenesis. Significant negative correlations were observed between MANCR and microRNA-20a-5p, and between microRNA-20a-5p and GCNT4. MANCR sponged microRNA-20a-5p to downregulate its expression, while microRNA-20a-5p repressed GCNT4 expression by binding to the 3'UTR of GCNT4 mRNA. Overexpression of microRNA-20a-5p reversed MANCR-mediated suppression of angiogenesis. CONCLUSION:LncRNA MANCR sponges microRNA-20a-5p, relieves the suppression of GCNT4 expression caused by microRNA-20a-5p, and elevates GCNT4 expression, ultimately inhibiting tumor growth and angiogenesis in CRC.
The recent trend of translational research has introduced a paradigm shift of our understanding of the applicability of bench top research. Translational research funnels in the outcomes of various techniques used in basic research and utilizes them for the benefit of patients in clinical research. Using the example of multiple myeloma, the aim of this article is to shed light on how our accumulated knowledge in different areas of basic research and application of cutting-edge technologies is leveraged in diagnosis, disease gradation, and prognosis. Patients suffering from this incurable disease benefit from the outcome of this research and experience significantly extended progress-free survival, which was beyond imagination even a few decades ago. The disease is currently managed by well-established and continuously evolving treatment options for patients, thus keeping their hope alive for a permanent cure.
