Study on the Microflora Structure in a <i>Litopenaeus vannamei</i>–<i>Sinonovacula constricta</i> Tandem-Culture Model Based on High-Throughput Sequencing under Different Culture Densities

In this study, we evaluated the intestinal contents of Pacific whiteleg shrimp (Litopenaeus vannamei), the visceral mass of razor clams (Sinonovacula constricta) and the water columns and the substrate sediments in different culture-density groups in a L. vannamei–S. constricta tandem-culture model by high-throughput sequencing of the 16S rRNA gene. The results show that the culture density affected the bacterial floral structure of the water columns, substrate sediment and razor-clam gut masses without making significant differences in the bacterial flora structure of the shrimp gut; the Shannon diversity indexes of the bacterial communities in the substrate sediment, shrimp gut and razor-clam gut masses were not significantly different among the density groups, and the Shannon diversity index of the bacterial communities in the water column was higher in the group with higher culture densities; at the phylum level, the dominant bacteria common to the shrimp guts, razor-clam visceral mass, water columns and substrate sediment were Proteobacteria and Bacteroidetes; Chloroflexi was the dominant bacterium specific to the substrate sediment; and Firmicutes was the dominant bacterium specific to the shrimp gut and razor-clam gut mass. We used national standards (GB 17378.4-2007, China) to evaluate the content of water-quality factors through the environmental factors and the genus-level correlation analysis of bacterial flora that follow: the dominant bacterium in the water column, uncultured_bacterium_f_Rhodobacteraceae, was negatively correlated with PO43−-P; the dominant bacteria in the substrate sediments, uncultured_bacterium_f_Anaerolineaceae and Woeseia, were significantly and negatively correlated with DO; and the dominant bacteria Lactococcus spp. in the razor-clam gut mass and the shrimp intestines were positively correlated with DO. These results show that culture density directly affects water-quality factors, which in turn affect the culture environment and the composition structure of the bacterial flora in a cultured organism

Luteolin Pretreatment Ameliorates Myocardial Ischemia/Reperfusion Injury by lncRNA-JPX/miR-146b Axis

Background. In the present study, we aimed to find out whether luteolin (Lut) pretreatment could ameliorate myocardial ischemia/reperfusion (I/R) injury by regulating the lncRNA just proximal to XIST (JPX)/microRNA-146b (miR-146b) axis. Methods. We established the models in vitro (HL-1 cells) and in vivo (C57BL/6J mice) to certify the protection mechanism of Lut pretreatment on myocardial I/R injury. Dual luciferase reporter gene assay was utilized for validating that JPX could bind to miR-146b. JPX and miR-146b expression levels were determined by RT-qPCR. Western blot was utilized to examine apoptosis-related protein expression levels, including cleaved caspase-9, caspase-9, cleaved caspase-3, caspase-3, Bcl-2, Bax, and BAG-1. Apoptosis was analyzed by Annexin V-APC/7-AAD dualstaining, Hoechst 33342 staining, as well as flow cytometry. Animal echocardiography was used to measure cardiac function (ejection fraction (EF) and fractional shortening (FS) indicators). Results. miR-146b was demonstrated to bind and recognize the JPX sequence site by dual luciferase reporter gene assay. The expression level of miR-146b was corroborated to be enhanced by H/R using RT-qPCR (P<0.001 vs. Con). Moreover, JPX could reduce the expression of miR-146b, whereas inhibiting JPX could reverse the alteration (P<0.001 vs. H/R, respectively). Western blot analysis demonstrated that Lut pretreatment increased BAG-1 expression level and Bcl-2/Bax ratio, but diminished the ratio of cleaved caspase 9/caspase 9 and cleaved caspase 3/caspase 3 (P<0.001 vs. H/R, respectively). Moreover, the cell apoptosis change trend, measured by Annexin V-APC/7-AAD dualstaining, Hoechst 33342 staining, along with flow cytometry, was consistent with that of apoptosis-related proteins. Furthermore, pretreatment with Lut improved cardiac function (EF and FS) (P<0.001 vs. I/R, respectively), as indicated in animal echocardiography. Conclusion. Our results demonstrated that in vitro and in vivo, Lut pretreatment inhibited apoptosis via the JPX/miR-146b axis, ultimately improving myocardial I/R injury

Iron Homeostasis Determines Fate of Human Pluripotent Stem Cells Via Glycerophospholipids-Epigenetic Circuit.

10.1002/stem.2967Stem Cells374489-50