Our previous study suggested that hypomethylation of perforin promoter of CD4 + T cells might be involved in the pathogenesis of autoimmune emphysema of rats. Whether transfer of this kind of cells hypomethylated in vitro into naive immunocompetent rats also results in emphysema is unknown yet. To test the hypothesis above, thirty Sprague Dawley (SD) rats were randomly divided into three groups: a model group (n = 10), a normal control group (n = 10) and a sham operation group (n = 10). In the model group, spleen-derived CD4 + T cells of normal rats were treated with 5-azacytidine (5-Aza), complete Freund's adjuvant and Phosphate Buffered Saline (PBS), then transferred into naive immunocompetent rats. The normal control group was injected with CD4 + T lymphocytes from spleens of normal rats and the same amount of adjuvant and PBS as above. In sham operation group, normal rats were injected intraperitoneally with complete Freund's adjuvant and PBS. Histopathological evaluations (mean linear Intercept (MLI) and mean alveolar numbers (MAN)), anti-endothelial cell antibodies (AECA) in serum and bronchoalveolar lavage fluid (BALF), lung vascular endothelial growth factor (VEGF)), the apoptotic index (AI) of alveolar septal cells and the methylation levels of perforin promoter of CD4 + T cells were investigated. The levels of the methylation above and MAN were lower in the model group than in the control and the sham operation group, while the AECA in serum and BALF, VEGF, MLI and the AI were greater (all p < 0.05). The methylation levels of perforin promoter were positively correlated with the MAN (r = 0.747, p < 0.05) and negatively correlated with AI, AECA, MLI, and VEGF (r was -0.789, -0.746, -0.743, -0.660, respectively, all p < 0.05). This study suggests that transfer of invitro CD4 + T cells with hypomethylation of perforin promoter into rats causes autoimmune emphysema, possibly by increasing expression of VEGF and promoting alveolar septal cell apoptosis.

Background. Stanford type A aortic dissection (TAAD) is one of the most life-threatening cardiovascular emergencies with high mortality and morbidity, and necroptosis is a newly identified type of programmed cell death and contributes to the pathogenesis of various cardiovascular diseases. However, the role of necroptosis in TAAD has not been elucidated. This study was aimed at determining the role of necroptosis in TAAD using bioinformatics analyses. Methods. The RNA sequencing dataset GSE153434 and the microarray dataset GSE52093 were obtained from Gene Expression Omnibus (GEO) database. Differentially expressed genes of necroptosis (NRDEGs) were identified based on differentially expressed genes (DEGs) and necroptosis gene set. Gene set enrichment analysis (GSEA) was applied to evaluate the gene enrichment signaling pathway in TAAD. The STRING database and Cytoscape software were used to establish and visualize protein-protein interaction (PPI) networks and identify the key functional modules of NRDEGs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of NRDEGs were also performed. Additionally, Spearman correlations were used to construct the necroptosis-related transcription factor-target genes regulatory network, immune infiltration patterns were analyzed using the ImmuCellAI algorithm, and the correlation between immune cell-type abundance and NRDEGs expression was investigated. The expression levels of NRDEGs and immune infiltration were additionally verified in the GSE52093 dataset. Results. We found that the necroptosis pathway was considerably enriched and activated in TAAD samples. Overall, 25 NRDEGs were identified including MLKL, RIPK1, and FADD, and among them, 18 were verified in the validation set. Moreover, GO and KEGG enrichment analyses found that NRDEGs were primarily involved in the tumor necrosis factor signaling pathway, nucleotide-binding oligomerization domain-like receptor signaling pathway, and interleukin-17 signaling pathway. The imbalance of Th17/Treg cells was identified in the TAAD samples. Furthermore, correlation analysis indicated that expression of NRDEGs was positively associated with proinflammatory immune-cell infiltrations and negatively associated with anti-inflammatory or regulatory immune-cell infiltrations. Conclusions. The present findings suggest that necroptosis phenomenon exists in TAAD and correlates with immune cell infiltration, which indicate necroptosis may promote the development of TAAD through activating immune infiltration and immune response. This study paves a new road to future investigation of the pathogenic mechanisms and therapeutic strategies for TAAD.

Bone fracture induces an acute inflammatory response in the resident and peripheral monocyte/macrophage cells. Excessive amounts of proinflammatory cytokines can cause severe tissue damage and inhibit bone healing. The proinflammatory cytokine genes are mainly controlled by TLR4/NF-κB (Toll-like receptor 4/Nuclear factor κB). Thus, targeting the molecules in this signaling pathway to decrease the expression of proinflammatory cytokines is an effective strategy to inhibit the inflammatory response. Herein, we identified a naturally derived small molecule NDSM253 that specifically inhibited IKKα (Inhibitor of NF-κB kinase subunit-alpha), a critical component of TLR4/NF-κB signaling. Biochemically, NDMS253 decreased phosphorylation of IκB (Inhibitor of NF-κB), thereby increasing the binding of IκB-NF-κB and suppressing the proinflammatory cytokine gene expression. NDMS253 showed a much stronger inhibitory effect on proinflammatory cytokine gene expression than did the known IKK inhibitors, including ACHP (2-Amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)-3-pyridinecarbonitrile), IKK16, and Amlexanox. Administration of these IKK inhibitors in a mouse femoral fracture model showed that NDSM253 suppressed proinflammatory cytokine genes, thereby promoting bone healing, while the other three IKK inhibitors showed a weaker improvement of both bone healing and circulating proinflammatory cytokines. Collectively, our data suggested that NDSM253 might be an effective inhibitor of IKKα that could inhibit inflammatory cytokine action in bone injury.

IL-33 is a recently discovered member of interleukin-1 (IL-1) family. Binding to the complex of receptor ST2 and the associated signal transduction protein AcP receptor located on the cell surface, IL-33 is able to carry out the intracellular signal transduction and thus lead to a cascade effect. Acinetobacter baumannii, a non-fermentative and gram-negative genus Acinetobacter bacterium, often leads to human pneumonia and sepsis in clinical setting and has become a superbug in hospitals in China. In this study, a rat model with A. Baumannii pneumonia was constructed. Low and high dose of IL-33 were used to treat experimental animals for 24 h. The expression of IL-33 receptor ST2 gene in lungs and peripheral blood was detected by QRT-PCR. Expression of major proteins associated with apoptosis of neutrophils was detected by Western Blot. Cell apoptosis was detected using double staining flow cytometry. The results showed that IL-33 was able to elevate ST3 gene expression in pneumonia lung tissues and peripheral blood in rats infected by A. baumannii, inhibit the expression of caspase-3 and Bax, and in the meantime increase the expression of anti-apoptotic factor Bcl-2.

Objective To explore the changes in expression of endothelin-1 (ET-1) in overlap syndrome(OS) and the correlation to the sleeping respiratory index to presume the role and mechanism of ET-1 in OS. Methods The levels of ET-1 were detected by immuneohistochemistry in patients with chronic obstructive pulmonary disease(COPD) ,obstructive sleep apnea-hypopnea syndrome(OSAHS) and OS. Meanwhile,the sleeping respiratory index were measured to investigate the correlation between ET-1 and apnea hypopnea index(AHI),longest apnea time(LAT) ,Percentage of time when oxygen saturation lower than 90%(SLT90%)and lowest arterial O2 saturation (LSaO2). Results The ET-1 levels in OS group were higher than those in COPD group and OSAHS group. LSaO2 in OS group was significantly lower than that in COPD group and LAT,SLT90% in OS group were higher than those in COPD group. There were no differences in AHI and LAT between OS group and OSAHS group. SLT90% in OS group was higher than that in OSAHS, LSaO2 being siginificantly lower than that in OSAHS. Correlation analysis showed that ET-1 in COPD group had no correlation with AHI,LAT and SLT90% but negative correlation with LSaO2. ET-1 in OS group showed positive correlation with AHI,LAT and SLT90% but negative correlation with LSaO2. Conclusions The ET-1 participates in pathophysiology changes in OS. The level of ET-1 correlates with the deterioration of the process of the disease.