Ulcerative colitis (UC) is non-specific inflammatory bowel disease. UC development and progression were closely associated with epigenetic modifications. Nevertheless, the specific relationship between N6-methyladenosine (m6A) modification at RNA transcription levels and UC pathogenesis remains unclear. We established UC cell models and mouse models through dextran sulfate sodium (DSS) induction. The expression levels of METTL14 were analyzed via qRT-PCR and western blot. In vitro functional experiments evaluated the effects of METTL14 overexpression on the viability of DSS-induced NCM460 cells and ferroptosis markers. Use of the m6A methylation detection kit, MeRIP-qPCR, and RNA stability experiments confirmed the molecular mechanism controlled by METTL14. In vivo experiments with inflammatory mice models elucidated the interaction between METTL14 and GPX4. Findings from this study indicated a notable reduction in m6A methyltransferase METTL14 expression in DSS-induced NCM460 cells and DSS-induced mice models. METTL14 overexpression effectively suppressed ferroptosis in DSS-induced NCM460 cells. In addition, METTL14 enhanced GPX4 mRNA stability through mediating m6A modification, and the interplay between METTL14 and GPX4 through m6A modification introduced innovative therapeutic approaches for UC management.
Purpose: Emerging evidences have indicated a role of the complement system in the pathogenesis of diabetic nephropathy (DN). Thus, this study was conducted to explore the complement system-related key biomarkers for patients with DN.
Methods: DN microarray datasets were downloaded from the GEO database, followed by differentially expressed genes (DEGs) screening. Complement system-related genes (CSRGs) were searched from various databases. Weighted Gene Co-expression Network Analysis (WGCNA) was employed to screen the DN-related genes, then the differential CSRGs (DCSRGs) were identified, followed by protein-protein interaction (PPI) network construction. In addition, key biomarkers were acquired by two machine learning algorithms, then immune infiltration analysis, Gene Set Enrichment Analysis (GSEA), and potential drugs screening were conducted. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and western blotting were utilized to detect the ITGB2 expression. Then the cell viability, inflammatory factors, and the expression of epithelial-mesenchymal transition (EMT) and fibrosis markers were determined by using Cell Counting Kit-8 (CCK-8) assay, enzyme linked immunosorbent assay (ELISA), western blotting assays, respectively.
Results: In total, 1012 DEGs and 974 DN-related genes were screened, and intersection analysis of the three (DN-related genes, DEGs and CSRGs) yielded 13 intersection genes, which were considered as the DCSRGs. Subsequently, 2 key biomarkers were identified by machine learning, namely VWF and ITGB2. The VWF and ITGB2 were both enriched in the pathways of chemokine signaling pathway, CAMs, focal adhesion and natural killer cell-mediated cytotoxicity, and significantly correlated with the activated mast cells, resting NK cells, and macrophages. Also, VWF and ITGB2 were significantly related to the clinical features, including age, serum creatinine level, and GFR (MDRD). Besides, mRNA and protein expression levels of ITGB2 in HG-treated HK-2 cells were remarkably elevated. Moreover, the viability of HK-2 cells, expression of TNF-α, IL-6, IL-12, α-SMA, E-cadherin and vimentin in HK-2 cells changed by HG administration were reversed by ITGB2-silence.
Conclusion: Complement system-related gene ITGB2 was overexpressed in DN, and inhibition of ITGB2 attenuated EMT and inflammation in DN.
Background and purpose: Inflammatory bowel disease (IBD) is a chronic, non-specific inflammatory bowel disease caused by multiple causes. Lymphocytes migration is involved in the pathogenesis of IBD. The purpose of this study was to evaluate whether there were differences in blood lymphocytes levels between IBD patients in clinical remission and healthy people.
Patients and methods: A total of 94 Crohn's disease (CD) and 20 ulcerative colitis (UC) patients were included in this study. Ninety-four people who underwent physical examination in our hospital were randomly selected as controls. We analyzed whether there were differences in white blood cell count, neutrophil count, neutrophil percentage, lymphocyte count, lymphocyte percentage between CD patients, UC patients, and healthy people.
Results: There were significant differences in lymphocyte count (P < 0.001), lymphocyte percentage (P < 0.001), neutrophil count (P = 0.038), and neutrophil percentage (P < 0.001) between CD patients and normal people, but no statistically significant differences in sex (P = 0.216), age (P = 0.745), and white blood cell count (P = 0.757). UC patients had significant differences in white blood cell count (P = 0.005), lymphocyte count (P = 0.010), and neutrophil count (P = 0.023), but no difference in lymphocyte percentage (P = 0.968) and neutrophil percentage (P = 0.461).
Conclusions: The white blood cell count of CD patients was not significantly different from that of normal people, but the lymphocyte count and lymphocyte percentage were significantly different from that of healthy people. Similar results were not found in UC patients.
Background: Timing the fixator removal is vital for a successful external fixation treatment. The purpose of this study was to determine the effectiveness of axial load-share ratio in vivo as a supplemental decision support tool for the safe removal of an Ilizarov circular external fixator.
Methods: This prospective observational study consists of 83 patients undergoing tibial or femoral lengthening with Ilizarov circular external fixation in our institution, from January 2011 to October 2019. In group I (38 patients), the external fixator was removed based on the surgeon's clinical experience and radiographs from January 2011 to June 2015. In group II (45 patients), from July 2015 to October 2019, the supplemental axial load-share (LS) ratio test was accomplished without the knowledge of the clinical results by another medical team. The test was performed by electronically measuring forces in the fixator rods and in a ground force plate. When the LS ratio < 10% was consistent with the conclusion (dense bone formation was achieved in the distraction zone) drawn from the corresponding routine radiographs by the treating surgeon, the external fixator was removed.
Results: There was no statistical significance in demographic data between the two groups (P > 0.05). In group I, 4 of the 38 patients suffered refracture (the refracture rate was 10.5%) after fixator removal, and bone union was finally achieved with further intervention by intramedullary nail. In group II, 36 patients terminated the external fixation after the first mechanical test, and another 9 patients terminated the external fixation at the subsequent test. None of the 45 patients in group II suffered refracture (the refracture rate was 0%). There was statistical significance in the refracture rate between the two groups (P < 0.05).
Conclusions: Adequate assessment of bone regenerate is crucial before removing an external fixator to prevent deformation or refracture. The axial load-share ratio in vivo is a practically quantitative method to supplement radiography and clinical experience for the assessment of regenerate healing, and the axial load-share ratio dropped below 10% is a safe limit for the Ilizarov circular external fixator removal.
About 5% of the population suffers from fibromyalgia (FM), a chronic multi-symptom pain illness whose pathophysiology is still unknown. We aimed to be the first to investigate the possible association of sera levels of miR-217 and miR-532 in patients with fibromyalgia and correlate their expression levels to different clinical and biochemical disease criteria. This study included 80 participants who splitted into two groups: 40 fibromyalgia sufferers (12 male and 28 female), and 40 healthy volunteers (10 male and 30 female) who served as the control group. Venous blood samples were collected from all subjects. The miR-217 and miR-532 serum expressions were detected using quantitative real-time PCR (qRT-PCR). According to our data, the fold changes of miR-217 and miR-532 in fibromyalgia patients were significantly lower than in controls, for miR-217 (median = 0.1359, IQR: 0.038-0.287, P < 0.001) and miR-532 [median = 0.2199, IQR: (0.114-0.421), P < 0.001]. In addition, there was a significant negative relationship between Aspartate transaminase (AST) and both miR-217 and miR-532 (r = - 0.480, P = P < 0.001 r = - 0.462, P = P < 0.001), respectively. Serum miR-217 and miR-532 could serve as potential diagnostic biomarkers for fibromyalgia.
Objectives: The purpose of this study was to determine the relationship between protein-coding RNA (messenger RNA, mRNA) and long noncoding RNA (lncRNA) expressed in vascular endothelial cells (VECs) in keloids by reanalyzing Gene Expression Omnibus (GEO) microarray chip data.
Materials and methods: The GSE121618 database and clinical information of these samples were downloaded and reanalyzed by the R language package. Expression differences in mRNA and lncRNA between keloids and normal skin were calculated. GO/KEGG enrichment analysis was conducted to determine the function of these genes, and an interaction network of lncRNAs-mRNAs was constructed. Magnetic Sorting of VECs and qRT-PCR were used to verify these bioinformatic results.
Results: The expression of three hundred and five mRNAs in the keloid group was significantly different from that in the normal group, and 98 lncRNAs were different, 21 of which were upregulated and 118 of which were downregulated. The hub relationship between the upregulated lncRNA‒mRNA interaction was lncRNA LINC01546-RASAL3/COL13A1, while the downregulated hub was lncRNA LOC101929787-PRKAA2/KRT71/SSTR1. qPCR verification result showed no obvious statistical differences.
Conclusions: Through the in-depth mining of keloid microarray data using bioinformatic methods, we speculated that VECs can affect the development and progression of keloids by epigenomic regulation via lncRNA‒mRNA interactions.
Background: Adherence to dietary guidelines is a fundamental aspect of diabetes management; however, it poses a significant challenge for patients with diabetes. Our research aims to assess the level of dietary compliance among individuals with type 2 diabetes (T2DM) and to identify the factors that influence their adherence to dietary advice.
Methods: This study was a cross-sectional survey. The patients with T2DM undergoing treatment at our hospital from March, 2023, to June, 2024 were included. Compliance with dietary recommendations was assessed using the validated dietary compliance scale for type 2 diabetes mellitus patients (DCS-T2DM). Spearman correlation and logistic regression analyses were conducted to evaluate the factors influencing dietary compliance in patients with T2DM.
Results: A total of 308 T2DM patients were included in our study. The results revealed that 46.10% of the participants had suboptimal dietary compliance. There were significant correlations between dietary compliance and several demographic and clinical factors, including age (r = 0.501), gender (r = 0.447), education level (r = 0.610), average monthly household per capita income (r = 0.627), and the duration, since T2DM diagnosis (r = 0.552), all of which were statistically significant (p < 0.05). Logistic regression identified age (OR = 1.705, 95%CI 1.262 ~ 1.987), gender (OR = 2.401, 95%CI 1.909 ~ 3.134), education level (OR = 3.083, 95%CI 2.434 ~ 3.957), average monthly household per capita income (OR = 3.721, 95%CI 2.553 ~ 4.405), and the time since T2DM diagnosis (OR = 2.470, 95%CI 1.755 ~ 3.262) as significant predictors of dietary compliance.
Conclusions: 46.10% of patients with T2DM exhibited suboptimal dietary adherence, with age, gender, education, income, and diabetes duration significantly predicting compliance. It is imperative for healthcare providers to devise individualized intervention strategies that incorporate these pivotal factors to enhance dietary adherence in patients with T2DM.
Background: Acute Osteofascial Compartment Syndrome (AOCS) stands as a critical surgical emergency, often secondary to various diseases. Its clinical manifestation arises from increased pressure within the fascial compartment, resulting in diminished tissue perfusion and consequential ischemic damage. Presently, clinical diagnostics lack effective biological markers, and patients face a grim prognosis, experiencing muscle contractures, necrosis, amputations, renal failure, and even mortality. The primary treatment, fasciotomy, poses infection risks and potential nerve damage. Hence, there is an urgent need for research elucidating AOCS's pathogenic mechanism and exploring novel treatments.
Methods: To address this, we established a rat model of AOCS, extracting toe flexor muscles from both experimental and control groups. Employing second-generation high-throughput sequencing, we obtained comprehensive mRNA, lncRNA, circRNA, and miRNA data. Comparative analysis of expression differences between AOCS and control groups, followed by in-depth examination, allowed us to unravel the intricacies of AOCS occurrence from a multi-omics perspective.
Results: Our research findings indicate that AOCS is an immune-mediated inflammatory disease, primarily involving immune cells, especially neutrophils. In addition, genes associated with ferroptosis, a form of regulated cell death, are found to be upregulated in the rat model, with non-coding RNAs playing a role in regulatory interactions.
Conclusions: These results suggest that neutrophils may undergo ferroptosis, thereby enhancing inflammation and immune responses in the fascial compartment, which promotes disease progression. Furthermore, these findings reveal the interactions between immune molecules and pathways in AOCS, which are significant for a deeper understanding of the pathogenesis of the disease and the development of targeted therapeutic strategies.
Background: Traumatic brain injury (TBI) is a major cause of disability and mortality among children and adults in developed countries. Transcription factor nuclear factor erythroid-derived 2-like 2 (Nrf2) has antioxidant, anti-inflammatory and neuroprotective effects and is closely related to TBI. Olfactory mucosa-mesenchymal stem cells (OM-MSCs) could promote neural regeneration. At present, the effects of OM-MSCs with overexpressed Nrf2 in brain diseases remain to be explored.
Methods: The OM-MSCs were prepared and transfected with Nrf2 overexpression plasmid. Those transfected cells were termed as OM-MSCs with Nrf2 overexpression (OM-MSCsNrf2) and co-cultured with rat pheochromocytoma cells PC12 or murine microglia BV2. The effects of OM-MSCsNrf2 on the survival and angiogenesis of PC12 cells were evaluated through cell counting kit-8 (CCK-8) and tube formation assay, and extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were calculated to reflect glycolysis. Immunofluorescence assay was applied to determine the effects of OM-MSCsNrf2 on microglial polarization, and the underlying molecular mechanisms were analyzed based on the quantification tests of RT-qPCR and immunoblotting.
Results: Co-culture of OM-MSCsNrf2 and PC12 cells increased the levels of anti-inflammatory cytokines and pro-angiogenesis factors, enhanced the cell survival and angiogenesis. Moreover, we also observed elevated phosphorylation of PI3K/AKT and suppressed BAX protein expression. Meanwhile, OM-MSCsNrf2 inhibited the levels of pro-inflammatory genes and affected the glycolysis in PC12 cells. In the co-cultured system of OM-MSCsNrf2 and BV2 cells, M2 microglial polarization was observed, and the levels of M2 microglia-relevant genes and the phosphorylation of STAT6/AMPKα/SMAD3 were elevated.
Conclusion: This study proved the effects of OM-MSCsNrf2 on modulating PC12 and BV2 cells in vitro, which, however, necessitates further in vivo validation.
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：