(A) The survival rates of WT B6 and TLR9 mice were monitored for 6 d after CLP (15 mice per group, with data pooled from three experiments, each of which had similar statistical significance)

Dilutions of blood (B) or peritoneal lavage fluid (C) obtained from TLR9 or WT mice 12 h after CLP were cultured on BHI agar plates, and the number of bacterial colonies was counted (five mice per group). The colony count indicated is of 1 ml of blood and 1 ml of peritoneal fluid. (D) Serum cytokine levels were determined 6 and 12 h after CLP by using a cytometric bead array (five mice per group). Data shown are means of values ± SEM obtained from individual mice and are representative of at least two independent experiments. *, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Toll-like receptor 9 inhibition reduces mortality in polymicrobial sepsis"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1277-1283.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413026.</p><p></p

(A) 12 h after CLP or sham laparotomy, the number of peritoneal granulocytes in WT or TLR9 mice (three to five mice per group) was determined by flow cytometry

(B) WT or TLR9 mice received 500 μg anti-GR1 antibody or isotype control 24 and 2 h before CLP, and the survival rates were monitored for 6 d after CLP (15 mice per group, with data pooled from three experiments, each of which had similar statistical significance). (C) 2 × 10 CD45.1 splenic granulocytes were injected i.v. into WT or TLR9 (CD45.2) recipients (three to five mice per group). 12 h after transfer, the mice underwent CLP or sham laparotomy, and 12 h later the number of CD45.1 granulocytes were enumerated from the peritoneal cells by flow cytometry. (D) 12 h after CLP or sham laparotomy, the number of granulocytes (Ly6G) was determined by flow cytometry in the peritoneal fluid of WT mice pretreated with 10 Flt3L-expanded WT or TLR9 DCs (three to five mice per group). Treatment with anti-GR1 antibody achieved >97% depletion of granulocytes at the time of and 24 h after CLP (not depicted). Data shown are means of values ± SEM obtained from individual mice and are representative of at least two independent experiments. *, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Toll-like receptor 9 inhibition reduces mortality in polymicrobial sepsis"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1277-1283.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413026.</p><p></p

(A) 12 h after CLP or sham laparotomy, the number of conventional DCs (CD11cMHCII) were determined by flow cytometry in the spleen and peritoneal fluid of WT or TLR9 mice (three to five mice per group)

(B) The survival rates of WT mice receiving WT or TLR9 DCs were monitored for 6 d after CLP (23 mice per group, with data pooled from two experiments, each of which had similar statistical significance). (C) 10 Flt3L-expanded WT or TLR9 DCs (CD45.2) were injected i.v. into CD45.1 WT recipients (three to five mice per group). 12 h after transfer, the mice underwent CLP or sham laparotomy, and 12 h later the number of CD45.2 DCs were enumerated from the peritoneal cells by flow cytometry. (D) Dilutions of blood or peritoneal lavage fluid obtained from WT mice pretreated with WT or TLR9 DCs 12 h after CLP were cultured on BHI agar plates, and the number of bacterial colonies was counted (five mice per group). (E) Serum cytokine levels were determined 12 h after CLP by cytometric bead array (five mice per group). Data shown are means of values ± SEM obtained from individual mice and are representative of at least two independent experiments. *, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Toll-like receptor 9 inhibition reduces mortality in polymicrobial sepsis"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1277-1283.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413026.</p><p></p

12 h after CLP or sham laparotomy, the numbers of viable splenocytes (A) and infiltrating peritoneal lymphocytes (B) were counted on a hemocytometer (BrightLine; Hausser Scientific) after Trypan blue staining (three to five mice per group)

Peritoneal cells are expressed per milliliter of peritoneal fluid. (C) Peritoneal cells pooled from three to five mice 12 h after CLP were cultured without restimulation for 24 h, and the supernatant cytokine levels were determined with a cytometric bead array. Peritoneal cells from mice that underwent sham laparotomy secreted a minimal amount of cytokines (not depicted). Data shown are means of values ± SEM obtained from individual mice (except for cells pooled in C) and are representative of at least two independent experiments. *, P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "Toll-like receptor 9 inhibition reduces mortality in polymicrobial sepsis"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1277-1283.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413026.</p><p></p

(A) Immediately before CLP, WT mice received 100 μg iCpG or control oligodeoxynucleotide (ODN), and survival rates were monitored for 6 d

(B) 12 and 18 h after CLP, WT mice received 100 μg iCpG or control ODN, and survival rates were monitored for 6 d. Survival curves of mice treated with iCpG at 3 and 6 h were similar to the one shown for the 12-h iCpG group (not depicted). Data shown include 15 mice per group, pooled from three experiments, each of which had similar statistical significance.<p><b>Copyright information:</b></p><p>Taken from "Toll-like receptor 9 inhibition reduces mortality in polymicrobial sepsis"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1277-1283.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413026.</p><p></p

Characterization of hepatocellular adenoma and carcinoma using microRNA profiling and targeted gene sequencing - Fig 3

<p>HCC had significantly higher mean expression of chromosome 19 miRNA cluster than HCA and normal liver adjacent to HCA in (A) miR-518b, (B) miR-520c-3p, (C) miR-515-5p, and (D) miR-517a. Horizontal bars represent mean value +/- standard deviation.</p

Characterization of hepatocellular adenoma and carcinoma using microRNA profiling and targeted gene sequencing

<div><p>Background</p><p>Hepatocellular adenomas (HCA) are benign liver tumors that may transform into hepatocellular carcinoma (HCC), but the molecular drivers of this transformation remain ill-defined. This study evaluates the molecular changes in HCA and HCC and in comparison to their adjacent non-neoplastic liver.</p><p>Methods</p><p>11 patients with HCA and 10 patients with HCC without underlying hepatitis or cirrhosis were included in this pilot study. Tumor and non-tumor liver tissues were selected for immunohistochemical staining, small RNA sequencing, and targeted gene sequencing. We compared microRNA expressions and mutations between HCA and HCC and non-neoplastic liver.</p><p>Results</p><p>HCA were classified as inflammatory (n = 6), steatotic (n = 4), or β-catenin activated (n = 1) subtypes. MicroRNA profile of all 3 HCA subtypes clustered between that of normal liver and HCC in principal component analysis. In both HCA and HCC, miR-200a, miR-429, and miR-490-3p were significantly downregulated compared to normal liver, whereas miR-452, miR-766, and miR-1180 were significantly upregulated. In addition, compared to HCA, HCC had significantly higher expression of members of the chromosome 19 miRNA cluster (C19MC), including miR-515-5p, miR-517a, miR-518b, and miR-520c-3p.</p><p>Conclusions</p><p>This study indicates that while there are significant differences in the molecular profile between HCA and HCC, several miRNAs are similarly deregulated in HCA and HCC compared to adjacent normal liver. These results may provide insights into the drivers of hepatocarcinogenesis and warrant further investigations.</p></div

Differential expressions of miRNAs in both HCC and HCA compared to their normal liver.

<p>Differential expressions of miRNAs in both HCC and HCA compared to their normal liver.</p

Cholangiocarcinoma: Correlation between Molecular Profiling and Imaging Phenotypes

<div><p>Purpose</p><p>To investigate associations between imaging features of cholangiocarcinoma by visual assessment and texture analysis, which quantifies heterogeneity in tumor enhancement patterns, with molecular profiles based on hypoxia markers.</p><p>Methods</p><p>The institutional review board approved this HIPAA-compliant retrospective study of CT images of intrahepatic cholangiocarcinoma, obtained before surgery. Immunostaining for hypoxia markers (EGFR, VEGF, CD24, P53, MDM2, MRP-1, HIF-1α, CA-IX, and GLUT1) was performed on pre-treatment liver biopsies. Quantitative imaging phenotypes were determined by texture analysis with gray level co-occurrence matrixes. The correlations between quantitative imaging phenotypes, qualitative imaging features (measured by radiographic inspection alone), and expression levels of the hypoxia markers from the 25 tumors were assessed.</p><p>Results</p><p>Twenty-five patients were included with a median age of 62 years (range: 54–84). The median tumor size was 10.2 cm (range: 4–14), 10 (40%) were single tumors, and 90% were moderately differentiated. Positive immunostaining was recorded for VEGF in 67% of the cases, EGFR in 75%, and CD24 in 55%. On multiple linear regression analysis, quantitative imaging phenotypes correlated significantly with EGFR and VEGF expression levels (R² = 0.4, <i>p</i><0.05 and R² = 0.2, <i>p</i><0.05, respectively), while a trend was demonstrated with CD24 expression (R² = 0.33, <i>p</i> = 0.1). Three qualitative imaging features correlated with VEGF and CD24 expression (P<0.05), however, none of the qualitative features correlated with the quantitative imaging phenotypes.</p><p>Conclusion</p><p>Quantitative imaging phenotypes, as defined by texture analysis, correlated with expression of specific markers of hypoxia, regardless of conventional imaging features.</p></div

Supervised clustering showing 90 differentially expressed microRNAs between HCC and normal liver and between HCA and normal liver tumors.

<p>Data are median centered (white), with the lowest and highest intensity values in blue and red, respectively.</p