Single-Nucleus RNA Sequencing Reveals Cardiac Macrophage Landscape in Hypoplastic Left Heart Syndrome

Background:Hypoplastic left heart syndrome(HLHS)is one of the most challenging congenital heart diseases in clinical treatment.In cardiac tissues,resident macrophages fulfill critical functions in maintaining a stable cardiac state and have strong regenerative capacity and organ specificity.However,the molecular mechanisms of macro-phages in HLHS remained unclear.Methods:Single-nucleus RNA sequencing(snRNA-seq)data of HLHS and healthy control(donors)samples obtained from the Gene Expression Omnibus(GEO)database were normalized and clustered using the Seurat package.The“FindMarkers”function was used to screen differentially expressed genes(DEGs)between the HLHS and donor groups and to analyze the functional enrichment of the set of genes of interest.Finally,cell-cell communication,pseudotime,and single-cell regulatory network inference and cluster-ing(SCENIC)analyses were used to study the mechanisms of macrophages in HLHS.Results:Based on the snRNA-seq data of HLHS and donors,we identified a total of 9 cell clusters,among which the proportion of macrophages was significantly less in the HLHS group than in the control group.Subdivision of macrophage subpopulations(Macrophages 1,2,and 3)showed that Macrophages 1 was mainly involved in nervous system development,angiogenesis,and apoptotic processes.In addition,analysis of communication between Macro-phages 1 and cardiomyocytes revealed that ligand-acceptor pairs such as GAS6/AXL,IL6,IGF1,THY1,and L1CAM were present only in the donor group.Finally,pesudotime and SCENIC analyses demonstrated that FOXO3 and ELF2 played a critical role for Macrophages 1 to maintain cardiac function in patients with HLHS.Conclusion:Our study improved the current understanding of the molecular mechanisms of macrophage devel-opment in HLHS,showing that manipulating the regulatory role of macrophages in the heart can be a novel treat-ment for HLHS.

Influence of low temperature on lethal time extension for different life stages of Cryptolestes ferrugineus (Stephens) with strong resistance to phosphine fumigation

Knowing the time extension degree of full mortality in phosphine fumigation at low temperature significantly contributes to successful insect pest control,especially for reducing fumigation failure and inhibiting resistance development.The comparison ofmortality and lethal time on eggs,larvae,pupae and adults of Cryptolestes ferrugineus(Stephens)was conducted,and the strain with 1043 times of resistance factor to phosphine was assayed during fumigation with 300 mL/m^3 of phosphine concentration at 18,23 and 28℃.The LT50 values to eggs,larvae,pupae and adults of C.ferrugineus at 18℃ were postponed by 2,2,1 and 5 d comparedwith that at 23℃,and 5,4,5 and 7 d compared with that at 28℃,respectively.The LT99 values to eggs,larvae,pupae and adults at 18℃ were 5,1,2 and 7 d longer than that at 23℃,and 6,5,5 and 10 d longer than that at 28℃,respectively.The lethal time of different life stages of the C.ferrugineus strain significantly increased with temperature decreasing.The exposure time of full mortality on pupae at 300 mL/m^3 of phosphine at 18℃ reached 38 d,which was 12 d longer than that of larvae.The order of tolerance of different life stages of C.ferrugineus exposed to 300 mL/m^3 phosphine from high to low was pupae,eggs,adults,larvae at tested temperatures.