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Objective To investigate the effect of exosomes derived from hepatocellular carcinoma cells on the polarization of tumor-associated macrophages (TAMs), and to reveal the novel mechanism of hepatocellular carcinoma formation. Methods Hepatocellular carcinoma cell-derived exosomes were isolated by ultracentrifugation, and the characteristics of exosomes were identified by transmission electron microscope (TEM), Dynamic Light Scattering (DLS), and Western blotting. The model of macrophage polarization was induced and verified by quantitative real-time PCR and Western blotting. The t -test was used for comparison of normally distributed continuous data between two groups. A one-way analysis of variance was used for comparison between multiple groups, and the LSD- t -test was used for further comparison between two groups. Results TEM showed that hepatocellular carcinoma cell-derived exosomes were round or oval vesicles, LDS showed that the exosomes had a particle size of 172.65±2.34 nm, and Western blotting showed highly positive expression of the biomarkers TSG101 and CD63 in exosomes. There was a significant increase in the expression of CD68 after the addition of 15 ng phorbol ester to induce human-derived mononuclear macrophages for 24 hours to achieve adherent growth (1.00±0.25 vs 6.67±0.98, t =11.20, P < 0.001). Western blotting showed that compared with the control group (L02 cell-derived exosomes), the hepatocellular carcinoma cell-derived exosomes (at low, middle, and high doses) induced M2 polarization of macrophages and increased the expression of the markers Arg-1 and CD163 (all P < 0.05). Conclusion Hepatocellular carcinoma cell-derived exosomes promote M2 polarization of TAMs.
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Objective To identify the circulating miRNAs which can be used to evaluate the diagnosis and prognosis of sepsis by microarray and quantitative real-time PCR,and to predict target genes of miR-519c-5p by bioinformatics analysis. Methods Three sepsis patients,3 septic shock patients and 3 normal controls were enrolled in this study. Plasma RNA was extracted,and was used for hybridized by miRCURY LNATMmicroRNA Array.The signals were scanned and used to conduct differential expression profilings and cluster analysis.Further-more,we performed qRT-PCR to confirm the expression of miRNAs chosen from microarray screening. We used the miRanda and Targetscan databases to predict target genes of the concerned miRNAs and used KEGG database to analyze the related pathways. Results Fifty-seven and 11 miRNAs were observed significantly upregulated in sepsis and septic shock patients,respectively(fold change≥2.0;P<0.05).qRT-PCR results showed that miR-519c-5p was significantly upregulated in patients with sepsis or patients with septic shock compared with the healthy normal controls(P<0.05).Twenty-nine target genes of miR-519c-5p were predicted by the bioinformatics analysis,and 7 potential target genes participate in the sepsis-related pathways.MiR-519c-5p might be a potential positive regulator for the critical cell cycle control gene of MAP2K4,contributing to the vascular endothelial cells apoptosis via MAPK signaling pathway. Conclusions We demonstrated that the plasma level of miR-519c-5p can be used for the diagnosis of sepsis and miR-519c-5p may be a potential therapeutic target for sepsis.
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Objectives To access the bacteriology in patients with sepsis due to biliary tract infection to provide a basis for empirical selection of proper antibiotic treatment.Methods This is a single-center retrospective study on 214 patients with biliary tract infection admitted from August 2014 to July 2016 to the surgical intensive care units (ICU) of The First Affiliated Hospital of Sun Yat-sen University.To study the demographic information,sequential organ failure assessment (SOFA),usage of antibiotics before ICU and duration of ICU were analyzed.Bile,peritoneal drainage and blood samples were collected.Results 47 septic shock patients and 25 septic patients due to biliary tract infection were enrolled in the trial.The two groups (the shock group vs.the sepsis group) had a significant difference in the duration of ICU stay [(6.4 ± 4.6) d vs.(2.3 ± 1.8) d,P ＜ 0.05].48 strains of pathogens were isolated from the bile samples.The major pathogens were Escherichia coli (E.coli) (n =23,47.9％),Enterococcus faecalis (n =8,16.7％) and Enterococcus faecium (n =2,4.2％).80 strains of pathogens were isolated from the peritoneal drainage culture samples.E.coli,pseudomonas aeruginosa,and Klebsiella pneumoniae ranked the top 3 species,accounting for 26.3％,11.3％ and 7.5％,respectively.The sensitivity of E.coli isolated from bile to amikacin,imipenem and panipenem were all over 90.0％.Conclusions E.coli was the principal gram-negative bacterium in biliary infection induced sepsis.Early administration of carbapenemes may reduce the occurrence of septic shock in these patients.