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G microparticles from CD14 positive cells were correlated with severity of liverinflammation in patients with NAFLD [34]. Title Loaded From File However, we believe that sCD14 is a very convenient tool for evaluation of liver inflammation grade when compared with microparticles. Several limitations of our study should be discussed. First, we did not conduct liver biopsies in the healthy control group for ethical reasons. Second, some patient selection bias may exist because liver biopsy may have been reserved for patients with NAFLD who were deemed likely to have NASH. Third, using liver biopsy as the `gold standard’ for assessing the accuracy of sCD14 has important limitations associated with sampling errors, as well as intra- and inter-observer variability, which are at least partly linked to the biopsy size [32]. Finally, serum sCD14 levels may increase in other conditions such as cholestasis, biliary atresia, and ischemia reperfusion injury [35?6]. However, these are extremely unusual conditions. In conclusion, we confirmed that serum sCD14 may be a useful and non-invasive biomarker for diagnosis of NASH and assessing liver inflammation in patients with NAFLD, who are at high risk of progressing to advanced liver fibrosis. Further research, including larger-scale clinical studies or combination of serum sCD14 and other non-invasive biomarkers of NASH such as CK18, areTable 3. Clinical and serological characteristics of NAFLD patients with mild and severe liver inflammation.Grade 0? liver inflammation Number (n) Age (years) Gender (male; female) Body mass index (kg/m2) Visceral fat area (cm2) Subcutaneous fat area (cm2) Fasting Blood Sugar (mg/dl) AST (IU/l) ALT (IU/l) C-reactive protein (mg/l) Nt discarded. All pelleteted cells were resuspended in 10 ml of 0.5xYPDA HOMA-IR sCD14 (ng/dl) 43 47.2613.2 23;20 27.965.3 140.7635.1 199.5644.9 105.2613.1 42.3614.1 45.5612.9 0.7360.46 3.4361.33 25.7610.Grade 2? liver inflammation 70 52.3612.9 43;27 29.965.9 149.8646.2 191.9648.1 110.2613.4 43.2614.3 57.1617.6 1.1860.98 3.5961.31 31.2611.P value*0.046 0.037 0.042 0.051 0.226 0.251 0.430 0.048 0.043 0.431 0.Numbers represent the mean 6 SD. Abbreviations: AST, aspartate aminotransferase; ALT, alanine aminotransferase; HOMA-IR, homeostasis model for the assessment of insulin resistance. P values correspond to the comparison between grade 0? liver inflammation and grade 2? liver inflammation in NAFLD patients using the Student’s t-test for continuous factors. doi:10.1371/journal.pone.0065211.tsCD14 and Liver Inflammation in NASHFigure 2. Serum sCD14 levels for diagnosis of the grade of liver inflammation. Receiver operating characteristic (ROC) curve and area under the ROC curve (AUROC) for discriminating between patients with severe (grade 2?) or mild (grade 0?) liver inflammation using serum sCD14 levels in 113 patients are shown. Serum sCD14 levels can diagnose severe liver inflammation in patients with NAFLD with moderate accuracy. doi:10.1371/journal.pone.0065211.gFigure 3. Lipopolysaccharide (LPS) increases sCD14 in vitro. sCD14 in cell culture medium from sham- and LPS-treated RAW264.7 cells was compared by (A) Western immunoblot analysis and (B) a sandwich enzyme-linked immunosorbent assay. LPS increased sCD14 in cell culture medium from RAW 264.7 cells. The immunoblot is representative of three independent experiments. Results are presented as means 6 SD. Statistical significance was determined using ANOVA with Scheffe’s multiple testing correction (*p value ,0.05). doi:10.1371/journal.pone.0065211.gAuthor ContributionsConceived an.G microparticles from CD14 positive cells were correlated with severity of liverinflammation in patients with NAFLD [34]. However, we believe that sCD14 is a very convenient tool for evaluation of liver inflammation grade when compared with microparticles. Several limitations of our study should be discussed. First, we did not conduct liver biopsies in the healthy control group for ethical reasons. Second, some patient selection bias may exist because liver biopsy may have been reserved for patients with NAFLD who were deemed likely to have NASH. Third, using liver biopsy as the `gold standard’ for assessing the accuracy of sCD14 has important limitations associated with sampling errors, as well as intra- and inter-observer variability, which are at least partly linked to the biopsy size [32]. Finally, serum sCD14 levels may increase in other conditions such as cholestasis, biliary atresia, and ischemia reperfusion injury [35?6]. However, these are extremely unusual conditions. In conclusion, we confirmed that serum sCD14 may be a useful and non-invasive biomarker for diagnosis of NASH and assessing liver inflammation in patients with NAFLD, who are at high risk of progressing to advanced liver fibrosis. Further research, including larger-scale clinical studies or combination of serum sCD14 and other non-invasive biomarkers of NASH such as CK18, areTable 3. Clinical and serological characteristics of NAFLD patients with mild and severe liver inflammation.Grade 0? liver inflammation Number (n) Age (years) Gender (male; female) Body mass index (kg/m2) Visceral fat area (cm2) Subcutaneous fat area (cm2) Fasting Blood Sugar (mg/dl) AST (IU/l) ALT (IU/l) C-reactive protein (mg/l) HOMA-IR sCD14 (ng/dl) 43 47.2613.2 23;20 27.965.3 140.7635.1 199.5644.9 105.2613.1 42.3614.1 45.5612.9 0.7360.46 3.4361.33 25.7610.Grade 2? liver inflammation 70 52.3612.9 43;27 29.965.9 149.8646.2 191.9648.1 110.2613.4 43.2614.3 57.1617.6 1.1860.98 3.5961.31 31.2611.P value*0.046 0.037 0.042 0.051 0.226 0.251 0.430 0.048 0.043 0.431 0.Numbers represent the mean 6 SD. Abbreviations: AST, aspartate aminotransferase; ALT, alanine aminotransferase; HOMA-IR, homeostasis model for the assessment of insulin resistance. P values correspond to the comparison between grade 0? liver inflammation and grade 2? liver inflammation in NAFLD patients using the Student’s t-test for continuous factors. doi:10.1371/journal.pone.0065211.tsCD14 and Liver Inflammation in NASHFigure 2. Serum sCD14 levels for diagnosis of the grade of liver inflammation. Receiver operating characteristic (ROC) curve and area under the ROC curve (AUROC) for discriminating between patients with severe (grade 2?) or mild (grade 0?) liver inflammation using serum sCD14 levels in 113 patients are shown. Serum sCD14 levels can diagnose severe liver inflammation in patients with NAFLD with moderate accuracy. doi:10.1371/journal.pone.0065211.gFigure 3. Lipopolysaccharide (LPS) increases sCD14 in vitro. sCD14 in cell culture medium from sham- and LPS-treated RAW264.7 cells was compared by (A) Western immunoblot analysis and (B) a sandwich enzyme-linked immunosorbent assay. LPS increased sCD14 in cell culture medium from RAW 264.7 cells. The immunoblot is representative of three independent experiments. Results are presented as means 6 SD. Statistical significance was determined using ANOVA with Scheffe’s multiple testing correction (*p value ,0.05). doi:10.1371/journal.pone.0065211.gAuthor ContributionsConceived an.

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Author: Proteasome inhibitor