Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses.

BACKGROUND: Diabetic foot ulcers (DFU) are among the fastest-growing diseases worldwide. Recent evidence has emphasized the critical role of microRNA (miRNA)-mRNA networks in various chronic wounds, including DFU. In this study, we aimed to clarify the miRNA-mRNA axes associated with the occurrence of DFU. METHODS: Expression profiles of miRNAs and mRNAs were extracted from the Gene Expression Omnibus. Differentially expressed genes and differentially expressed miRNAs were identified, and miRNA-mRNA regulatory axes were constructed through integrated bioinformatics analyses. We validated the miRNA-mRNA axes using quantitative real-time PCR (qPCR) and dual-luciferase reporter assays. We conducted an immune infiltration analysis and confirmed the bioinformatics results using immunofluorescence staining. Single-sample gene set enrichment analysis (ssGSEA) was used to analyze the metabolic mechanisms. RESULTS: miR-182-5p-CHL1/MITF and miR-338-3p-NOVA1 interactions were identified using in silico analysis. The qPCR results showed apparent dysregulation of these miRNA-mRNA axes in DFU. The dual-luciferase reporter assay confirmed that miR-182-5p targeted CHL1 and MITF, and miR-338-3p targeted NOVA1. We conducted an immune infiltration analysis and observed that key genes correlated with decreased infiltration of M1 macrophages and resting mast cells in DFU. Immunofluorescence staining verified the co-localization of CHL1 and tryptase, while MITF and CD68 showed weak positive correlations. Metabolic pathways related to these three genes were identified using ssGSEA. CONCLUSIONS: In summary, the miR-182-5p-CHL1/MITF and miR-338-3p-NOVA1 pathway interactions and decreased infiltration of M1 macrophages and resting mast cells may provide novel clues to the pathogenesis of DFU. TRIAL REGISTRATION: The clinical trial included in this study was registered in the Chinese Clinical Trial Registry ( ChiCTR2200066660 ) on December 13, 2022.

Identification of potential diagnostic biomarkers and immune infiltration features in diabetic foot ulcer by bioinformatics analysis and validation.

Diabetic foot ulcer (DFU) is the most serious and costly chronic complication that may lead to disability and even death in patients suffering from diabetes mellitus (DM). However, the clinical diagnosis and prognosis of DFU is inadequate. There is still a lack of effective biomarkers for its early diagnosis. We obtained the circRNA expression dataset GSE114248 and mRNA expression dataset GSE80178 from the GEO. R software was used to identify the differentially expressed circRNAs (DECs). The mRNAs associated with DFU were identified by a random forest algorithm and intersected with mRNAs predicted by circRNAs. Then, the circRNA-miRNA-mRNA network was established and the hub genes were screened using GO semantic similarity and were validated by the GSE199939 dataset. Meanwhile, the expression level of the biomarkers was verified by RT-PCR assays and immunohistochemistry. Finally, GSEA was conducted to determine differential immune cell infiltration and the immunological cells' relationships with hub genes. We identified three hub genes including KIAA1109, ENPP5, and NRP1 that might play an important role in DFU. ROC curve results also showed a good performance of these three genes in the validation dataset. Furthermore, RT-PCR assays and immunohistochemistry confirmed the results above. Immune infiltration analysis indicated that DFU had a significant increase in Neutrophils. Moreover, three hub genes were closely correlated with a variety of inflammatory cells. KIAA1109, ENPP5, and NRP1 are key hub genes of DFU. They might play an important role in the development of DFU and could be potential biomarkers in DFU.

Asymmetric correlation between experienced parental attachment and event-related potentials evoked in response to parental faces.

This study aims to explore the modulation effects of attachment relationships with parents on the neural correlates that are associated with parental faces. The event-related potentials elicited in 31 college students while viewing facial stimuli of their parents in two single oddball paradigms (father vs. unfamiliar male and mother vs. unfamiliar female) were measured. We found that enhanced P3a and P3b and attenuated N2b were elicited by parental faces; however, the N170 component failed to discriminate parental faces from unfamiliar faces. An experienced attachment relationship with the father was positively correlated to the P3a response associated with the father's face, whereas no correlation was found in the case of mothers. Further exploration in dipole source localization showed that, within the time window of the P300, distinctive brain regions were involved in the processing of parental faces; the father's face was located in the medial frontal gyrus, which might be involved in self effect, and the anterior cingulate gyrus was activated in response to the mother's face. This research is the first to demonstrate that neural mechanisms involved with parents can be modulated differentially by the qualities of the attachments to the parents. In addition, parental faces share a highly similar temporal pattern, but the origins of these neural responses are distinct, which could merit further investigation.