Mechanism of wound repair in diabetic rats using nanosilver-free alginate dressing.

OBJECTIVE: Nanosilver-alginate dressing can effectively promote the healing of diabetic wounds in rats. However, due to the potential toxicity of nanosilver, its widespread application in hard-to-heal wound healing is limited. In the present study, the role and potential mechanism of nanosilver-free alginate gel (NSFAG) in the healing process of diabetic wounds were explored. METHOD: A diabetic rat skin wound model was established, and wounds were treated with saline (NC group), nanosilver gel (NSG group) or nanosilver-free alginate gel (NSFAG group) for seven consecutive days. RESULTS: NSFAG significantly promoted wound healing and increased the content of protein and hydroxyproline in granulation tissues, and was superior to NSG (p<0.05). Immunohistochemical analyses revealed that the skin wound tissue structure of the NSFAG group was intact, and the number of skin appendages in the dermis layer was significantly higher compared with the NC group and the NSG group (p<0.05). Western blot analysis found that the protein expression of the epidermal stem cell marker molecules CK19 and CK14 as well the proliferation marker of keratinocytes Ki67 in the NSFAG group was significantly higher compared with the NC group or NSG group (p<0.05). Additionally, the proliferation marker of keratinocytes Ki67 in the NSFAG group was significantly higher compared with the NC or NSG group (p<0.05). Immunofluorescence staining analyses indicated that the CK19- and CK14-positive cells were mainly distributed around the epidermis and the newly formed appendages in the NSFAG group, and this result was not observed in the NC or NSG groups. CONCLUSION: The present findings demonstrate that NSFAG can effectively accelerate wound healing in diabetic rats by promoting epidermal stem cell proliferation and differentiation into skin cells, as well as formation of granulation tissue, suggesting that it can be a potential dressing for diabetic wounds.

Downregulation of hsa-miR-203 in peripheral blood and wound margin tissue by negative pressure wound therapy contributes to wound healing of diabetic foot ulcers.

Negative pressure wound therapy (NPWT) has been widely used in the treatment of chronic wounds, including diabetic foot ulcers (DFU) as the severe manifestation of diabetic foot. Hsa-miR-203 is proven to be correlated with the severity of DFU. To investigate whether NPWT influences hsa-miR-203 levels in persons with DFU, we detected hsa-miR-203 levels in peripheral plasma and wound margin tissue from the following patients: type 2 diabetic (T2D) patients with DFU (DFU group), T2D patients without DFU (NDFU group), patients with chronic skin ulcer and normal glucose tolerance (SUC group), and healthy volunteers with normal glucose tolerance (NC group). All patients in SUC group received NPWT. As contrast, some of patients in DFU group received NPWT (NPWT group) while others chose routine dressing therapy (non-NPWT group). In vitro experiments were also performed to determine influences of negative pressure on cell proliferation and migration of HaCaT cells (human keratinocytes). Results showed that before NPWT, levels of hsa-miR-203 in peripheral plasma (P-miR-203) and wound margin tissue (T-miR-203) of DFU group were obviously increased compared to SUC group while expression of P-miR-203 decreased in NDFU group compared with NC group. After NPWT, levels of P-miR-203 and T-miR-203 in DFU and SUC group were significantly lower than before. Changes of P-miR-203 and T-miR-203 after NPWT were positively correlated with 4-week ulcer healing rate in NPWT and SUC group. In vitro, negative pressure lowered the expression of hsa-miR-203, enhancing cell proliferation and migration in HaCaT cells via up-regulation of p63 protein. Meanwhile, the effects of negative pressure on cells were remarkable reduced by high-glucose intervention. Our study suggests that NPWT promotes DFU healing by reducing the expression of hsa-miR-203 in peripheral blood and wound tissue. The changes of hsa-miR-203 in peripheral blood and wound tissue may be related to the therapeutic effect of NPWT.

Expression of miR-210 in the peripheral blood of patients with newly diagnosed type 2 diabetes mellitus and its effect on the number and function of endothelial progenitor cells.

OBJECTIVE: This study aims to investigate the correlation between the expression of miR-210 in peripheral blood and the number of peripheral endothelial progenitor cells (EPCs) in patients with type 2 diabetes mellitus (T2DM). We also determined the effect of miR-210 on EPC proliferation, adhesion, migration, tube formation, and apoptosis. METHODS: A total of 32 patients with newly diagnosed T2DM (T2DM group) and 32 control subjects with normal glucose tolerance (NC group) were included. Peripheral blood samples were collected from each subject. The miR-210 level was determined by quantitative real-time polymerase chain reaction (qRT-PCR), and the number of positive EPCs indicated by CD34, CD133, and KDR expressions was detected by flow cytometry. After isolation, culture, and identification by fluorescent staining, EPCs were divided into four groups: NC group, untransfected type 2 diabetic group, miR-210 inhibitor NC group, and miR-210 inhibitor group. The expression of miR-120 in each group was detected by qRT-PCR, and the changes in the proliferation, adhesion, migration, tube formation, and apoptosis of EPCs after transfection with a miR-210 inhibitor were observed. RESULTS: The expression level of miR-210 in the T2DM group (5.83 ± 1.26) was significantly higher than that in the NC group (1.18 ± 0.54) (t = 17.26, P < 0.001). The number of EPCs was significantly lower in the T2DM group (39.3 ± 12.6)/106 cells than that in the NC group (76.2 ± 10.7)/106 cells (t = 10.49, P < 0.001). Spearman's correlation analysis showed that the expression of miR-210 in the peripheral blood of patients with T2DM was negatively correlated with the number of EPCs (r = -0.558, P = 0.001). Multiple linear stepwise regression analysis showed that the peripheral blood level of miR-210 was an independent correlation factor that affected the number of EPCs (P < 0.001). After transfection with the miR-210 inhibitor, the proliferation, adhesion, tube formation, and migration levels of EPCs in miR-210 inhibitor group were higher than those in untransfected type 2 diabetic group and miR-210 inhibitor NC group, whereas the apoptosis rate was lower than that in these groups, and these results were statistically significant (P < 0.05). CONCLUSION: The increased expression of miR-210 in patients with T2DM may be related to the decreased number and function of EPCs in peripheral blood.

Decreased expression of miR-24 in peripheral plasma of type 2 diabetes mellitus patients associated with diabetic foot ulcer.

To examine the correlations of miR-24 expression in peripheral plasma with the onset of diabetic foot ulcer (DFU) and diabetic foot osteomyelitis (DFO) in type 2 diabetes mellitus (T2DM) patients and explore the clinical value of miR-24 as a potential biomarker for the diagnosis and treatment outcomes of DFU and DFO, a total of 60 newly diagnosed T2DM patients without DFU (T2DM group), 112 T2DM patients with DFU (DFU group), and 60 healthy controls (NC group) were included. DFU group were further divided into DFO group (n = 64) and non-DFO group (n = 48). MiR-24 levels were determined by quantitative real-time PCR, while clinical features and risk factors of DFU and DFO were explored. The expression level of miR-24 in T2DM and DFU group was significantly lower than in NC group (P < .05), and that in DFU group was significantly lower than in T2DM group (P < .01). Additionally, the level of miR-24 significantly decreased in DFO group compared to non-DFO group (P < .01). Moreover, it was negatively correlated with the amputation rate in DFU group (P = .043) and positively correlated with healing rate after 8 weeks (P = .036). The multivariate logistic regression analysis confirmed that a low expression of miR-24 was an independent risk factor for DFU and DFO. The ROC curve analysis indicated that the AUC of miR-24 for the diagnosis of DFU and DFO was 0.849 (95% CI, 0.618-0.879, P < .001) and 0.782 (95% CI, 0.595-0.813, P < .001). Thus, a decreased expression of miR-24 of T2DM patients was closely related to the occurrence, development and prognosis of DFU and DFO, suggesting the use of miR-24 as a potential biomarker for the prediction of DFU and DFO.

Metabolic Heterogeneity of Tumor Cells and its Impact on Colon Cancer Metastasis: Insights from Single-Cell and Bulk Transcriptome Analyses.

Background: Metabolic reprogramming plays a crucial role in the development of colorectal cancer (CRC), influencing tumor heterogeneity, the tumor microenvironment, and metastasis. While the interaction between metabolism and CRC is critical for developing personalized treatments, gaps remain in understanding how tumor cell metabolism affects prognosis. Our study introduces novel insights by integrating single-cell and bulk transcriptome analyses to explore the metabolic landscape within CRC cells and its mechanisms influencing disease progression. This approach allows us to uncover metabolic heterogeneity and identify specific metabolic genes impacting metastasis, which have not been thoroughly examined in previous studies. Methods: We sourced microarray and single-cell RNA sequencing datasets from the Gene Expression Omnibus (GEO) and bulk sequencing data for CRC from The Cancer Genome Atlas (TCGA). We employed Gene Set Variation Analysis (GSVA) to assess metabolic pathway activity, consensus clustering to identify CRC-specific transcriptome subtypes in bulkseq, and rigorous quality controls, including the exclusion of cells with high mitochondrial gene expression in scRNA seq. Advanced analyses such as AUCcell, infercnvCNV, Non-negative Matrix Factorization (NMF), and CytoTRACE were utilized to dissect the cellular landscape and evaluate pathway activities and tumor cell stemness. The hdWGCNA algorithm helped identify prognosis-related hub genes, integrating these findings using a random forest machine learning model. Results: Kaplan-Meier survival curves identified 21 significant metabolic pathways linked to prognosis, with consensus clustering defining three CRC subtypes (C3, C2, C1) based on metabolic activity, which correlated with distinct clinical outcomes. The metabolic activity of the 13 cell subpopulations, particularly the epithelial cell subpopulation with active metabolic levels, was evaluated using AUCcell in scRNA seq. To further analyze tumor cells using infercnv, NMF disaggregated these cells into 10 cellular subpopulations. Among these, the C2 subpopulation exhibited higher stemness and tended to have a poorer prognosis compared to C6 and C0. Conversely, the C8, C3, and C1 subpopulations demonstrated a higher level of the five metabolic pathways, and the C3 and C8 subpopulations tended to have a more favorable prognosis. hdWGCNA identified 20 modules, from which we selected modules primarily expressed in high metabolic tumor subgroups and highly correlated with clinical information, including blue and cyan. By applying variable downscaling of RF to a total of 50 hub genes, seven gene signatures were obtained. Furthermore, molecules that were validated to be protective in GEO were screened alongside related molecules, resulting in the identification of prognostically relevant molecules such as UQCRFS1 and GRSF1. Additionally, the expression of GRSF1 was examined in colon cancer cell lines using qPCR and phenotypically verified by in vitro experiments. Conclusion: Our findings emphasize that high activity in specific metabolic pathways, including pyruvate metabolism and the tricarboxylic acid cycle, correlates with improved colon cancer outcomes, presenting new avenues for metabolic-based therapies. The identification of hub genes like GRSF1 and UQCRFS1 and their link to favorable metabolic profiles offers novel insights into tumor neovascularization and metastasis, with significant clinical implications for targeting metabolic pathways in CRC therapy.

Proteomics changes after negative pressure wound therapy in diabetic foot ulcers.

Label­free quantitative mass spectrometry was used to analyze the differences in the granulation tissue protein expression profiles of patients with diabetic foot ulcers (DFUs) before and after negative­pressure wound therapy (NPWT) to understand how NPWT promotes the healing of diabetic foot wounds. A total of three patients with DFUs hospitalized for Wagner grade 3 were enrolled. The patients received NPWT for one week. The granulation tissue samples of the patients prior to and following NPWT for one week were collected. The protein expression profiles were analyzed with label­free quantitative mass spectrometry and the differentially expressed proteins (DEPs) in the DFU patients prior to and following NPWT for one week were identified. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were conducted to annotate the DEPs and DEP­associated signaling pathways. Western blotting and ELISA were performed to validate the results. By comparing the differences in the protein profiles of granulation tissue samples prior to and following NPWT for one week, 36 proteins with significant differences were identified (P<0.05); 33 of these proteins were upregulated and three proteins were downregulated. NPWT altered proteins mainly associated with antioxidation and detoxification, the cytoskeleton, regulation of the inflammatory response, complement and coagulation cascades and lipid metabolism. The functional validation of the DEPs demonstrated that the levels of cathepsin S in peripheral blood and granulation tissue were significantly lower than those prior to NPWT (P<0.05), while the levels of protein S isoform 1, inter α­trypsin inhibitor heavy chain H4 and peroxiredoxin­2 in peripheral blood and granulation tissue were significantly higher than those prior to NPWT (P<0.05). The present study identified multiple novel proteins altered by NPWT and laid a foundation for further studies investigating the mechanism of action of NPWT.

Two cases of autoimmune pancreatitis with diabetes and literature review.

OBJECTIVE: To investigate the clinical features of autoimmune pancreatitis (AIP) with diabetes as the first manifestation, improve our understanding of the disease and highlight the recognition of special types of diabetes. METHODS: A retrospective analysis was performed on 2 AIP patients diagnosed with diabetes at the First Affiliated Hospital of Anhui Medical University. RESULTS: Two elderly patients with new-onset diabetes mellitus were admitted to the hospital with weight loss and yellowing of the skin. Imaging showed pancreatic enlargement, bile duct dilatation, and cholestasis. Auxiliary examination followed by histopathology or experimental hormone therapy revealed elevated IgG4 levels, and the patients were eventually diagnosed with AIP. CONCLUSION: For elderly diabetic patients with atypical clinical characteristics, such as unexplained gastrointestinal symptoms or weight loss, IgG4 levels should be examined to rule out diabetes secondary to AIP.