Hyperglycemia is a key factor in the development of diabetic complications, including the processes of atherosclerosis. Receptor-interacting protein 3 (RIP3), a mediator of necroptosis, is implicated in atherosclerosis development. Additionally, hydrogen sulfide (H2S) protects the vascular endothelium against hyperglycemia-induced injury and attenuates atherosclerosis. On the basis of these findings, the present study aimed to confirm the hypothesis that necroptosis mediates high glucose (HG)-induced injury in human umbilical vein endothelial cells (HUVECs), and that the inhibition of necroptosis contributes to the protective effect of exogenous H2S against this injury. The results revealed that exposure of HUVECs to 40 mM HG markedly enhanced the expression level of RIP3, along with multiple injuries, including a decrease in cell viability, an increase in the number of apoptotic cells, an increase in the expression level of cleaved caspase-3, generation of reactive oxygen species (ROS), as well as dissipation of the mitochondrial membrane potential (MMP). Treatment of the cells with sodium hydrogen sulfide (NaHS; a donor of H2S) prior to exposure to HG significantly attenuated the increased RIP3 expression and the aforementioned injuries by HG. Notably, treatment of cells with necrostatin-1 (Nec-1), an inhibitor of necroptosis, prior to exposure to HG ameliorated the HG-induced injuries, leading to a decrease in ROS generation and a loss of MMP. However, pre-treatment of the cells with Nec-1 enhanced the HG-induced increase in the expression levels of cleaved caspases-3 and -9. By contrast, pre-treatment with Z-VAD-FMK, a pan-caspase inhibitor, promoted the increased expression of RIP3 by HG. Taken together, the findings of the present study have demonstrated, to the best of our knowledge for the first time, that exogenous H2S protects HUVECs against HG-induced injury through inhibiting necroptosis. The present study has also provided novel evidence that there is a negative interaction between necroptosis and apoptosis in the HG-treated HUVECs.

Background: Children of mothers with gestational diabetes mellitus (GDM) are more prone to acquire type 2 diabetes and obesity as adults. Due to this link, early intervention strategies that alter the gut microbiome may benefit the mother and kid long-term. This work uses metagenomic and transcriptome sequencing to investigate how probiotics affect gut microbiota dysbiosis and inflammation in GDM. Methods: GDM and control metagenomic sequencing data were obtained from the SRA database. This metagenomic data helped us understand gut microbiota abundance and function. KEGG detected and extracted functional pathway genes. Transcriptome sequencing data evaluated GDM-related gene expression. Finally, GDM animal models were given probiotics orally to evaluate inflammatory response, regulatory immune cell fractions, and leptin protein levels. Results: GDM patients had more Fusobacteria and Firmicutes, while healthy people had more Bacteroidetes. Gut microbiota composition may affect GDM by altering the L-aspartate and L-asparagine super pathways. Mannan degradation and the super pathway of L-aspartate and L-asparagine synthesis enhanced in GDM mice with leptin protein overexpression. Oral probiotics prevent GDM by lowering leptin. Oral probiotics increased Treg, Tfr, and Breg cells, which decreased TNF-α and IL-6 and increased TGF-β and IL-10, preventing inflammation and preserving mouse pregnancy. Conclusion: Dysbiosis of the gut microbiota may increase leptin expression and cause GDM. Oral probiotics enhance Treg, Tfr, and Breg cells, which limit the inflammatory response and assist mice in sustaining normal pregnancy. Thus, oral probiotics may prevent GDM, enabling targeted gut microbiota modulation and maternal and fetal health.

In recent years hydrogen sulfide (H₂S) has demonstrated vasculoprotective effects against cell death, which suggests its promising therapeutic potential for numerous types of disease. Additionally, a protective effect of exogenous H₂S in HG‑induced injuries in HUVECs was demonstrated, suggesting a potential protective effect for diabetic vascular complications. The present study aimed to investigate the mechanism accounting for the cytoprotective role of exogenous H2S against high glucose [HG (40 mM glucose)]‑induced injury and inflammation in human umbilical vein endothelial cells (HUVECs). HUVECs were exposed to HG for 24 h to establish an in vitro model of HG‑induced cytotoxicity. The cells were pretreated with sodium hydrosulfide (NaHS), a donor of H₂S, or inhibitors of necroptosis and p38 MAPK prior to the exposure to HG. Cell viability, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP), IL‑1β, IL‑6, IL‑8, TNF‑α, phosphorylated‑(p)38 and receptor‑interacting protein 3 (RIP3) expression levels were detected using the indicated methods, including Cell Counting Kit 8, fluorescence detection, western blotting, immunofluorescence assay and ELISAs. The results demonstrated that necroptosis and the p38 MAPK signaling pathway mediated HG‑induced injury and inflammation. Notably, NaHS was discovered to significantly ameliorate p38 MAPK/necroptosis‑mediated injury and inflammation in response to HG, as evidenced by an increase in cell viability, a decrease in ROS generation and loss of MMP, as well as the reduction in the secretion of proinflammatory cytokines. In addition, the upregulated expression of RIP3 induced by HG was repressed by treatment with SB203580, while the HG‑induced upregulation of p‑p38 expression levels were significantly downregulated following the treatment of Nec‑1 and RIP3‑siRNA. In conclusion, the findings of the present study indicated that NaHS may protect HUVECs against HG‑induced injury and inflammation by inhibiting necroptosis via the p38 MAPK signaling pathway, which may represent a promising drug for the therapy of diabetic vascular complications.

Background and Objectives: The conventional diagnostic process for cardiovascular autonomic neuropathy (CAN) is often time-consuming and lacks standardization. The Cardiovascular Autonomic Nervous Function Multi-Parameter Evaluation System is the first intelligent device designed for CAN diagnosis, featuring wireless wearable modules and human‒computer interaction. This study aims to evaluate the accuracy and eficiency of the novel device. Methods: A two-part, paired-design multicenter study involving a total of 200 subjects (122 with diabetes) from three centers was conducted. Cardiovascular autonomic reflex tests (CARTs) and 5-min heart rate variability (HRV) analyses were performed via the study device. Concurrent manual measurements with physician diagnoses and nonstandardized conventional methods served as controls in Part I and Part II, respectively. Methods: In Part I, the study device diagnosed 19.3% (29/150) of the subjects with CAN, demonstrating 96.6% sensitivity and 100% specificity. There was excellent agreement with the physician's CAN diagnosis for CARTs (Cohen's kappa of 0.979, P < 0.001) and with the control device for HRV parameters (intraclass correlations [ICCs] > 0.9). Part II showed weak to moderate intermethod correlations with the nonstandardized conventional method (ICCs = 0.193‒0.632). Repeated tests revealed high reproducibility (coefficients of variation = 3%‒20%). The investigational device required only one examiner to perform a standardized assessment and saved 33 minutes compared with manual methods. Conclusions: The proposed system provides eficient and standardized testing with excellent accuracy and reproducibility for CAN assessment. This novel device will facilitate the evaluation of therapeutic efficacy and promote streamlined clinical workflows.