FASLG as a Key Member of Necroptosis Participats in Acute Myocardial Infarction by Regulating Immune Infiltration.

Background: Acute myocardial infarction (AMI) is a major cause of human health risk. Necroptosis is a newly and recently reported mode of cell death, whose role in AMI has not been fully elucidated. This study aimed to search for necroptosis biomarkers associated with the occurrence of AMI and to explore their possible molecular mechanisms through bioinformatics analysis. Methods: The dataset GSE48060 was used to perform weighted gene co-expression network analysis (WGCNA) and differential analysis. Key modules, differential genes, and necroptosis-related genes (NRGs) were intersected to obtain candidate biomarkers. Groups were classified and differentially analyzed according to the expression of the key biomarker. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, gene set enrichment analysis (GSEA), and construction of protein-protein interaction (PPI) networks are performed on differentially expressed genes (DEGs). Finally, CIBERSORT was used to assess immune cell infiltration in AMI and the correlation of key biomarkers with immune cells. Immune cell infiltration analysis revealed the correlation between FASLG and multiple screened immune cells. Results: WGCNA determined that the MEsaddlebrown module was the most significantly associated with AMI. Intersecting it with DEGs as well as NRGs, we obtained two key genes, FASLG and IFNG. But only FASLG showed statistically significant differences between the AMI group and the normal control group. Further analysis suggested that the down-regulation of FASLG may exert its function through the regulation of the central genes CD247 and YES1. Furthermore, FASLG was positively correlated with T-cell CD4 memory activation and T-cell gamma delta, and negatively correlated with macrophage M0. Conclusion: In conclusion, FASLG and its regulatory genes CD247 and YES1 might be involved in the development of AMI by regulating immune cell infiltration. FASLG might be a potential biomarker for AMI and provides a new direction for the diagnosis of AMI.

Construction and rescue of infectious cDNA clone of pigeon-origin Newcastle disease virus strain JS/07/04/Pi / 病毒学报

Based on the complete genome sequence of pigeon-origin Newcastle disease virus strain JS/07/04/ Pi(genotype VIb), nine overlapped fragments covering its full-length genome were amplified by RT-PCR. The fragments were connected sequentially and then inserted into the transcription vector TVT7/R resulting in the TVT/071204 which contained the full genome of strain JS/07/04/Pi. The TVT/071204 was co-transfected with three helper plasmids pCI-NP, pCI-P and pCI-L into the BSR cells, and the transfected cells and culture supernatant were inoculated into 9-day-old SPF embryonated eggs 60 h post-transfection. The HA and HI tests were conducted following the death of embryonated eggs. The results showed that the allantoic fluids obtained were HA positive and the HA could be inhibited by anti-NDV serum which indicated that the strain JS/07/04/Pi was rescued successfully. The rescued virus rNDV/071204 showed similar growth kinetics to its parental virus in CEF. The successful recovery of this strain would contribute to the understanding of the host-specificity of pigeon-origin NDV and to the development of the novel vaccines against the NDV infection in pigeons.

Raloxifene inhibits bone loss and improves bone strength through an Opg-independent mechanism.

The osteoblast-derived paracrine factor osteoprotegerin (OPG) is considered to play a key role in inhibition of osteoclast formation and activity. Recently, raloxifene, a nonsteroidal benzothiophene, was found to exert anti-resorptive effects via modulating OPG expression in osteoblasts. To explore whether raloxifene regulates bone metabolism via an OPG-dependant pathway in vivo, we investigated the effects of raloxifene on bone loss in Opg-deficient mice. The results show that bone mineral density and bone strength are increased in mice deficient for Opg after treatment with raloxifene for 30 days. Histomorphometric analysis shows that raloxifene can increase bone trabecular area and decrease the number of osteoclasts in Opg (-/-) mice. Moreover, raloxifene reduces Rankl transcription and serum level of Rankl, which is dramatically increased in Opg knockout mice. These results suggest that raloxifene-induced inhibition of bone resorption may be independent of Opg pathway in mice.

Effect of 15-hydroxyeicosatetraenoic acid on calcium ion concentration of cerebral arterial smooth muscle cells in rats / 中华神经医学杂志

Objective To observe the effect of 15-hydroxyeicosatetraenoic acid (15-HETE) on the intracellular calcium ion ([Ca2+]i) concentration of cerebral arterial smooth muscle and discuss the source of ([Ca2+]i) mobilization induced by 15-HETE to reveal the mechanism underlying the vasoconstriction aroused by 15-HETE. Methods First, papain and collagenase were employed to isolate the cerebral artery smooth muscle cells (SMCs) from the rats. The cells were separated into experimental group (treated with 15-HETE) and control group. Confocal laser scanning microscope was used to investigate the ([Ca2+]i) signaling in cultured SMCs. Then, Ca2+ channel blockers and Ca2+ store depletion agent were added to identify the source of Ca2+ transients in SMCs of the 2 groups. Finally, internal carotid artery (ICA) rings were used to observe the effect of non-Ca2+ solution on 15-HETE-induced ICA vasoconstriction on both groups. Results Compared with the control group, 15-HETE treatment group enjoyed a significantly increased level of ([Ca2+]i) in cultured SMCs (P＜0.05). After the addition of Ca2+channel blockers (nifedipine, lanthanum ion) and calcium-free solution, 15-HETE treatment group still enjoyed a significantly increased level of ([Ca2+]i) in cultured SMCs as compared with the control group (P＜0.05), while after the addition of Ca2+ store depletion agent (caffeine), no obvious difference was noted between the 2 groups (P＞0.05). Non-Ca2+ solution had no effect on 15-HETE induced vasoconstriction.Conclusion 15-HETE may activate Ca2+ releasing from intracellular stores to rise. ([Ca2+]i) level in SMCs, which subsequently induce the constriction of cerebral arterial smooth muscle.