Upregulation of microRNA-451a improves endothelial cell function in atherosclerosis by direct targeting of macrophage migration inhibitory factor

This study aims to detect the expression of selected circulating microRNAs(miRNA),including miRNA-451a,miRNA-486-5p and miR-10b-5p,and their prospective roles as biomarkers in patients with atherosclerosis.For this purpose,levels of miRNAs were detected by real-time quantitative polymerase chain reaction(RT-qPCR)in case(N=30)and healthy control(N=30)groups.Receiver operating characteristic(ROC)curve analysis was carried out to evaluate the diagnostic ability of miRNAs.The correlations of miR-451a with lipid parameters were evaluated using Pearson’s correlation coefficients.HUVEC tested by ox-LDL was used as a cellular model of atherosclerosis.Cell viability and apoptosis were detected by methyl thiazolyl tetrazolium(MTT)and flow cytometry(FC)assays.Luciferase assay was used to determine the miRNA binding site on target genes.The results showed that miRNA-451a expression level was significantly lower in patients than controls.ROC curve analysis showed that the areas under the curve(AUC)of plasma miRNA-451a was 0.90.The miRNA-451a expression level exhibited significant negative correlations with cholesterol(TC),triglyceride(TG),and low-density lipoprotein(LDL).Besides,miRNA-451a specifically binds to the 3’UTR of macrophage migration inhibitory factor(MIF)and mediated the cell proliferation and apoptosis of HUVECs exposed to ox-LDL.Furthermore,overexpression of miRNA-451a promoted the proliferation and alleviated apoptosis of HUVECs exposed to ox-LDL,while this result was attenuated by macrophage migration inhibitory factor(MIF)overexpression.Therefore,miRNA-451a could be considered as a potential biomarker for atherosclerosis and miRNA-451a might contribute to regulating atherosclerosis through targeting MIF.

MIC17A is a novel diagnostic marker for feline toxoplasmosis

Toxoplasma gondii is a widespread parasitic pathogen that infect humans and all warm-blooded animals,causing abortion and stillbirth in pregnant women and animals,as well as life threatening toxoplasmosis in immune compromised individuals.Felines are the only defnitive hosts of Toxoplasma and oocysts shed by infected felines are the major source of infection for humans and other animals.Given the critical role of felines for T.gondii transmission,control of feline toxoplasmosis has signifcant impacts on reducing the overall prevalence of animal and human toxoplasmosis.However,reliable diagnosis of feline toxoplasmosis is still challenging.In this study,we found that the putative micronemal protein 17A(MIC17A)that was abundantly expressed in Toxoplasma merozoites is a good diagnostic marker for serological diagnosis of Toxoplasma infection in felines.T.gondii encodes four paralogs of MIC17A in total and the expression of three of them is drastically upregulated in merozoites than in tachyzoites.In contrast,when proteins like GRA1 and MIC3 that are more abundantly expressed in tachyzoites than in merozoites were used as diagnostic antigens to test feline toxoplasmosis,they reacted with Toxoplasma specifc IgG antibodies poorly.Taken together,these results suggest that merozoite antigens are better suited for the diagnosis of feline toxoplasmosis than antigens that are highly expressed at tachyzoite or bradyzoite stages.

Isolation of Sturgeon Peripheral Blood Lymphocytes and the Optimal Proliferation Response Conditions

In this study, sterlet （ Acipenser ruthenus ） was chosen as the model species of sturgeon, different solutions were used to isolate the sturgeon peripheral blood lymphocytes and study their optimal proliferate response condition. Phytohemagglutinin （PHA）, concanavalin A （ConA）and lipopolysaccharide （LPS） were used as lymphocyte proliferation mitogen, respectively. According to L 25 （5^6） five-factor five-level orthogonal experimental design, the conditions for sturgeons proliferation response of peripheral blood lymphocytes were optimized using enhanced cell counting Kit-8 （enhanced CCK-8 or WST-8）. Five factors were selected to explore the optimal response conditions, including culture time, culture temperature, cell concentration, fetal bovine serum （FBS） concentration, and mitogen concentration. The results showed that 70% Percoll （1.092 g/ml） used as the sturgeon lymphocyte separation solution had the best separating effect. The optimal proliferation conditions were as follows： 3.625×10 6 initial cells, 20 μg/ml PHA or 50 μg/ml ConA or 10 μg/ml LPS as mitogen, 10%-20% FBS, the temperature at 20-25 ℃, and the culture time of 2 d.

Analysis of Solvency of A-Share Listed Companies in China’s Real Estate Industry

The real estate industry is a capital-intensive industry and capital has become a particular concern for real estate enterprises.For a long time,China’s real estate enterprises rely on high-leverage development and carry out high-debt and high-risk operations.The solvency of real estate enterprises has been the focus of stakeholders’attention.In August 2020,China’s regulatory authorities introduced new financing regulations for real estate enterprises.They set up“three red lines,”which brought real estate enterprises’solvency into focus once again.This article takes A-share listed companies in China’s real estate industry as an example,analyzes and evaluates its debt solvency,and gives suggestions based on new policies and regulations,hoping to provide specific references to the enterpriser’s manager and external decision-makers.