Role of dysregulated macrophage subpopulation ratios and functional changes in the development of coronary atherosclerosis.

Coronary heart disease is one of the most significant risk factors affecting human health worldwide. Its pathogenesis is intricate, with atherosclerosis being widely regarded as the leading cause. Aberrant lipid metabolism in macrophages is recognized as one of the triggering factors in atherosclerosis development. To investigate the role of macrophages in the formation of coronary artery atherosclerosis, we utilized single-cell data from wild-type mice obtained from the aortic roots and ascending aortas after long-term high-fat diet feeding, as deposited in GSE131776. Seurat software was employed to refine the single-cell data in terms of scale and cell types, facilitating the identification of differentially expressed genes. Through the application of differential expression genes, we conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional enrichment analyses at 0, 8 and 16 weeks, aiming to uncover pathways with the most pronounced functional alterations as the high-fat diet progressed. The AddModuleScore function was employed to score the expression of these pathways across different cell types. Subsequently, macrophages were isolated and further subdivided into subtypes, followed by an investigation into intercellular communication within these subtypes. Subsequent to this, we induced THP-1 cells to generate foam cells, validating critical genes identified in prior studies. The results revealed that macrophages underwent the most substantial functional changes as the high-fat diet progressed. Furthermore, two clusters were identified as potentially playing pivotal roles in macrophage functional regulation during high-fat diet progression. Additionally, macrophage subtypes displayed intricate functionalities, with mutual functional counterbalances observed among these subtypes. The proportions of macrophage subtypes and the modulation of anti-inflammatory and pro-inflammatory functions played significant roles in the development of coronary artery atherosclerosis.

Sources and migration similarly determine nitrate concentrations: Integrating isotopic, landscape, and biological approaches.

Rapid land use change has significantly increased nitrate (NO3-) loading to rivers, leading to eutrophication, and posing water security problems. Determining the sources of NO3- to waters and the underlying influential factors is critical for effectively reducing pollution and better managing water resources. Here, we identified the sources and influencing mechanisms of NO3- in a mixed land-use watershed by integrating stable isotopes (δ15N-NO3- and δ18O-NO3-), molecular biology, water chemistry, and landscape metrics measurements. Weak transformation processes of NO3- were identified in the river, as evinced by water chemistry, isotopes, species compositions, and predicted microbial genes related to nitrogen metabolism. NO3- concentrations were primarily influenced by exogenous inputs (i.e., from soil nitrogen (NS), nitrogen fertilizer (NF), and manure & sewage (MS)). The proportions of NO3- sources seasonally varied. In the wet season, the source contributions followed the order of NS (38.6 %) > NF (31.4 %) > atmospheric deposition (ND, 16.2 %) > MS (13.8 %). In the dry season, the contributions were in the order of MS (39.2 %) > NS (29.2 %) > NF (29 %) > ND (2.6 %). Farmland and construction land were the original factors influencing the spatial distribution of NO3- in the wet and dry seasons, respectively, while slope, basin relief (HD), hypsometric integral (HI), and COHESION, HD were the primary indicators associated with NO3- transport in the wet and dry seasons, respectively. Additionally, spatial scale differences were observed for the effects of landscape structure on NO3- concentrations, with the greatest effect at the 1000-m buffer zone scale in the wet season and at the sub-basin scale in the dry season. This study overcomes the limitation of isotopes in identifying nitrate sources by combining multiple approaches and provides new research perspectives for the determination of nitrate sources and migration in other watersheds.

Coupled effects of landscape structures and water chemistry on bacterioplankton communities at multi-spatial scales.

Bacterioplankton communities in rivers are strongly influenced by the surrounding landscape, yet the relationships between land use and bacterioplankton communities at multi-spatial scales and the mechanisms that shape bacterioplankton communities remain unclear. Here, we collected surface water samples from 14 tributaries of the Yuan River, a secondary tributary of the Yangtze River, which has been heavily impacted by human activities. We characterized the bacterioplankton communities by high-throughput sequencing techniques, and managed to identify the mechanisms governing bacterioplankton community assembly. The results showed that, in general, both landscape compositions and landscape configurations had significant effects on bacterial communities, and the effects were greater at the buffer scale than at the sub-basin scale. Additionally, there was no distinct distance-decay pattern for the effects of landscape structures on bacterial communities from the near-distance (100 m) to the long-distance (1000 m) buffer zones, with the maximal effects occurring in the 1000 m circular buffer (wet season) and 500 m riparian buffer (dry season) zone, respectively. Land use influenced the bacterioplankton community both directly through exogenous inputs (mass effect) and indirectly by affecting water chemistry (species sorting). Variance partitioning analyses showed that the total explanations of bacterial community variations by water chemistry and the intersections of water chemistry and land use (56.2% in wet season and 50.4% in dry season) were higher than that of land use alone (6.1% in wet season and 25.4% in dry season). These suggest that mass effects and species sorting jointly shaped bacterial community assembly, but that the effects of species sorting outweighed those of mass effects. Nevertheless, more biotic and abiotic factors need to be considered to better understand the microbial assembly mechanisms in anthropogenically influenced riverine ecosystems.

Kcnk3, Ggta1, and Gpr84 are involved in hyperbaric oxygenation preconditioning protection on cerebral ischemia-reperfusion injury.

The present study aimed to explore the potential mechanism of the effect of hyperbaric oxygenation (HBO) preconditioning on cerebral ischemia and reperfusion injury (CIRI). GSE23160 dataset was used to identify differentially expressed genes (DEGs) from striatum between the middle cerebral artery occlusion (MCAO)/reperfusion and sham rats. The gene clusters with continuous increase and decrease were identified by soft clustering analysis in Mfuzz, and functional enrichment analysis of these genes was performed using clusterProfiler package. The intersection set of the genes with significantly altered expression at post-reperfusion 2, 8, and 24 h were screened in comparison to 0 h (sham group), and the expression of these genes was detected in the MCAO/reperfusion model and HBO preconditioning groups by real-time PCR (RT-PCR) and western blotting. A total of 41 upregulated DEGs, and 7 downregulated DEGs were detected, among which the expression of Gpr84 and Ggta1 was significantly upregulated at each reperfusion phase as compared to the sham group, while the expression of Kcnk3 was significantly downregulated except in the postreperfusion 8 h in the striatum group. RT-PCR and western blotting analyses showed that the expression of Ggta1, Gpr84, and Kcnk3 genes between the MCAO/reperfusion and sham rats were consistent with the bioinformatics analysis. In addition, the HBO preconditioning reduced the expression of Ggta1 and Gpr84 and increased the expression of Kcnk3 in MCAO/reperfusion rats. Kcnk3, Ggta1, and Gpr84 may play a major role in HBO-mediated protection of the brain against CIRI.

[The significance of erythrocyte fragment count to predicting hemolysis in roller pumps].

Hemolysis in blood pumps has been measured by various in vitro tests, in which normalized index of hemolysis (NIH) was established. As NIH is complicated and difficult to calculate, erythrocyte fragment count (EFC) is proposed in the present study to predict hemolysis in roller pumps. Five paired in vitro tests were conducted using the POLYSTAN pediatric pump (group A) and COBE pump (group B). Ten whole blood samples (400ml) were circled in the roller pump for 16 hours. Erythrocyte fragments count and plasma-free hemoglobin (FHb) were measured before pumping and every 2 hours through circulation after 4 hours pumping. The morphological changes of erythrocyte were observed by scanning electron microscope. Simple linear regression analysis showed the two groups' EFC and FHb levels increased linearly during a long period of pumping, and the coefficient of determination (R2) was 0.64. These data suggested that Erythrocyte fragments count could be used as an index in evaluating the in vitro hemolytic properties of blood pumps.

[The study of erythrocyte fragility and morphological changes caused by roller pump in vitro].

To observe the erythrocyte fragility and morphological changes of erythrocytes caused by roller pump. Ten tests were divided into two groups, Polystan pediatric pump group A (n = 5) and COBE pump group B (n = 5). Ten whole blood samples (each 400 ml) were circulated in the roller pump for 16 h. Erythrocyte fragility and free hemoglobin were measured before pumping and at every 2 hours during pumping. The possible morphological changes of erythrocytes caused by roller pump were observed by scanning electron microscope. The electron microscopic observation was made before pumping and at every 4 hours throughout pumping. Results showed that the erythrocyte fragility of two groups was not increased during a long period of pumping. The number of acanthocytes of two groups was 1.77/1.81% in the samples before pumping and 6.12/7.13, 9.18/8.73, 13.21/12.89, 16.53/17.21% at 4 h, 8 h, 12 h, and 16 h respectively. The free hemoglobin level of two groups was increased linearly during a long duration of pumping and the index of hemolysis of two groups was 0.296 mg/L/h and 0.3993 mg/L/h respectively. The result shows: 1. the erythrocyte fragility was not increased during a long period of pumping; 2. the erythrocyte membrane was injured or broken by roller pump directly; 3. the morphological changes of erythrocytes would be the basis of post operative hemolysis.