The loci of active HERV-Hs in colon tumor and adjacent normal tissues.

<p>*Active elements individually contributing to more than 10% of the transcripts in tumor or adjacent normal samples are highlighted in boldface and their transcript abundances are indicated in the ‘Abundance’ column (individual and total, respectively).</p><p>**HERV-HX is the colon cancer-related HERV-H element identified by us previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029950#pone.0029950-Liang2" target="_blank">[14]</a>. Inserts of PCR product clones were all HERV-HX fragments but not spliced sequences, in concordance with our previous finding that no spliced transcripts were produced from HERV-HX in colon tumor samples.</p><p>***The element located at 1q31.3 consists of 5′LTR and 3′LTR, with the entire protein coding region (<i>gag-pro-pol-env</i>) missing.</p><p>****Open reading frames (ORFs) were predicted by the online program ORF Finder and putative peptide sequences were subjected to Blastp search against the Non-redundant protein sequences at <a href="http://www.ncbi.nlm.nih.gov/" target="_blank">http://www.ncbi.nlm.nih.gov/</a>. Only predicted ORFs≥303 nt (peptide sequence ≥100 aa) and with Blastp matches are included. RT, Reverse transcriptase; CREB5, cAMP response element-binding protein 5; Gag, group-specific antigen.</p

The loci of active HERV-H elements in colon cancer cell lines.

<p>*Two adjacent HERV-H elements at 1p32.3 were combinedly active by making use of 5′ LTR of the first HERV-H and 3′ LTR of the second one (representative transcript sequence JK017392).</p><p>**The elements located at 16q24.1 and 19q13.31 are also actively transcribed in both tumor and adjacent normal colon tissues, while the one located at 20p12.1 is active in tumor tissue.</p><p>***Open reading frames (ORFs) were predicted by the online program ORF Finder and putative peptide sequences were subjected to Blastp search against the Non-redundant protein sequences at <a href="http://www.ncbi.nlm.nih.gov/" target="_blank">http://www.ncbi.nlm.nih.gov/</a>. Only predicted ORFs≥303 nt (peptide sequence ≥100 aa) and with Blastp matches are included. Gag, group-specific antigen; RT, Reverse transcriptase; CREB5, cAMP response element-binding protein 5.</p

OPN protein localization in the CRC and its liver metastasis visualized by immunohistochemistry. a

<p>In <i>IHC</i>, OPN proteins (brown positive stain) were localized in cytoplasm of CRC cells in primary lesions(x200). <b>b</b> Adjacent normal colorectal tissues were negative(x200). <b>c</b> Positive stain was found both in CRC cells and adjacent normal hepatocytes in liver metastatic tissues(x100). <b>d</b> As controls, OPN proteins stain in normal liver tissues without CRC metastasis was negative(x100).</p

OPN mRNA location in CRC and its liver metastasis with in situ mRNA hybridization. a,

<p>In <i>ISH</i>, the positive stain presents blue-purple under microscope. The OPN mRNA, which indicates a positive signal, was found in cytoplasm of CRC cells in primary lesions (×400). <b>b,</b> The stain in adjacent normal colorectal tissues was negative (×400). <b>c,</b> OPN mRNA was found in cytoplasm of CRC cells liver metastatic tissue. Adjacent normal liver tissues stains negative (×100).</p

Gap junctional intercellular communication (GJIC) with gap-FRAP in SW-480-pcDNA3.1(+) and SW-480-pcDNA3.1(+) OPN cells. a1,

<p>uniform fluorescence intensity before photobleaching in SW-480-pcDNA3.1(+). <b>a2,</b> weakened fluorescence intensity after photobleaching in SW-480-pcDNA3.1(+). <b>a3,</b> fluorescence redistribution after 10minutes in SW-480-pcDNA3.1(+). <b>b1,</b> uniform fluorescence intensity before photobleaching SW-480-pcDNA3.1(+) OPN cells. <b>b2,</b> weakened fluorescence intensity after photobleaching SW-480-pcDNA3.1(+) OPN cells. <b>b3,</b> faint fluorescence redistribution after 10minutes SW-480-pcDNA3.1(+) OPN cells. <b>c,</b> The percentage of fluorescence redistribution after photobleaching (FRAP%) in SW-480-pcDNA3.1(+) and SW-480-pcDNA3.1(+) OPN cells<b>.</b> After 5 minutes of photobleaching, the percentage of fluorescence redistribution after photobleaching (FRAP%) 24.65±4.08% in SW-480-pcDNA3.1(+) -OPN compared 44.74±6.23% in SW-480-pcDNA3.1(+)(<i>t</i> = 17.07, <i>P</i><0.001), and after 10 minutes, the FRAP% was still be inhibited at 25.98±4.48% in SW-480-pcDNA3.1(+) -OPN while it was redistributed to 64.92%±5.39% in the SW-480-pcDNA3.1(+) cells (<i>t</i> = 35.16, <i>P</i><0.001).</p

Expression of integrin αv and CD44v6 in normal hepatocytes with Immunohistochemistry. a,

<p>In <i>IHC</i>, integrin αv proteins (brown positive stain) were localized in cytoplasm of hepatocytes (×400). <b>b,</b> CD44v6 proteins (brown positive stain) were localized in cytoplasm of hepatocytes (×400).</p

Characterization of the active HERV-H elements in colon cancer cell lines and their spliced transcripts.

<p><b>A.</b> Schematic of the six commonly deleted regions in the active HERV-H elements in colon cancer cell lines. <b>B.</b> Schematics of the two extraordinarily short HERV-H elements and their transcripts. Pair-wise alignments for each HERV-H element were performed with the HERV-H consensus constructed by Jern P, <i>et al</i>. The shortened alignment results were shown to indicate the missing regions precisely. Color density represents the extent of homology with the HERV-H consensus. Gray areas represent deleted regions in the HERV-H elements as compared with the HERV-H consensus. Spliced transcripts are shown above the alignment results accordingly. Thick bars represent exons, and lines represent introns. Regions of LTRs, pre-gag, <i>gag</i>, <i>pro</i>, <i>pol</i> and <i>env</i> are labeled below. <b>C.</b> Schematics of the two combinedly active HERV-H elements located at 1p32.3 in HT29.</p

Feasibility and Accuracy of Sentinel Lymph Node Biopsy in Clinically Node-Positive Breast Cancer after Neoadjuvant Chemotherapy: A Meta-Analysis

<div><p>Sentinel lymph node biopsy (SLNB) has replaced conventional axillary lymph node dissection (ALND) in axillary node-negative breast cancer patients. However, the use of SLNB remains controversial in patients after neoadjuvant chemotherapy (NAC). The aim of this review is to evaluate the feasibility and accuracy of SLNB after NAC in clinically node-positive patients. Systematic searches were performed in the PubMed, Embase, and Cochrane Library databases from 1993 to December 2013 for studies on node-positive breast cancer patients who underwent SLNB after NAC followed by ALND. Of 436 identified studies, 15 were included in this review, with a total of 2,471 patients. The pooled identification rate (IR) of SLNB was 89% [95% confidence interval (CI) 85–93%], and the false negative rate (FNR) of SLNB was 14% (95% CI 10–17%). The heterogeneity of FNR was analyzed by meta-regression, and the results revealed that immunohistochemistry (IHC) staining may represent an independent factor (<i>P</i> = 0.04). FNR was lower in the IHC combined with hematoxylin and eosin (H&E) staining subgroup than in the H&E staining alone subgroup, with values of 8.7% versus 16.0%, respectively (<i>P</i> = 0.001). Thus, SLNB was feasible after NAC in node-positive breast cancer patients. In addition, the IR of SLNB was respectable, although the FNR of SLNB was poor and requires further improvement. These findings indicate that IHC may improve the accuracy of SLNB.</p></div

Meta-analysis of the IR.

<p>(A) Forest plot of the IR. The width of the horizontal line represents the 95% CI of individual studies. The vertical dotted line represents the overall expected IR. The combined estimate of IR was 89% (95% CI:85–93%, I² = 88.0%). (B) Funnel plot to assess publication bias effect on the IR. Each dot represents a separate study. The funnel plot revealed no apparent evidence of publication bias.</p