Fluorescence-Guided Surgery of Liver Metastasis in Orthotopic Nude-Mouse Models

<div><p>We report here the development of fluorescence-guided surgery of liver metastasis. HT29 human colon cancer cells expressing green fluorescent protein (GFP) were initially injected in the spleen of nude mice. Three weeks later, established liver metastases were harvested and implanted on the left lobe of the liver in other nude mice in order to make an orthotopic liver metastasis model. Fourteen mice with a single liver metastasis were randomized into bright-light surgery (BLS) or fluorescence-guided surgery (FGS) groups. Seven mice were treated with BLS, seven were treated with FGS. Three weeks after implantation, the left lobe of the liver with a single metastasis was exposed through a median abdominal incision. BLS was performed under white light. FGS was performed using a hand-held portable fluorescence imaging system (Dino-Lite). Post-surgical residual tumor fluorescence was visualized with the OV100 Small Animal Imaging System. Residual tumor fluorescence after BLS was clearly visualized at high magnification with the OV100. In contrast, residual tumor fluorescence after FGS was not detected even at high magnification with the OV100. These results demonstrate the feasibility of FGS for liver metastasis.</p></div

Pre-operative and post-operative images from the orthotopic liver metastasis model treated with FGS.

<p><b>(A)—(C)</b> Upper panels show bright field images, and lower panels are images of tumor fluorescence obtained with the OV100. Residual tumor fluorescence could not be detected even at high magnification <b>(C). (D,F,H)</b> Pre-FGS tumor fluorescence was clearly visualized with the Dino-Lite imaging system. <b>(E,G,I)</b> Dino-Lite imaging showed no evidence of tumor after FGS. <b>(J-K)</b> Dino-Lite settings. <b>(J)</b> After exposing the left lobe of the liver, the mouse was put under the Dino-Lite. <b>(K)</b> Connection between the Dino-Lite and computer. Tumor fluorescence was imaged on the monitor during FGS. Magnifications are indicated above the columns.</p

Pre-operative and post-operative images from the orthotopic liver metastasis model treated with BLS.

<p><b>(A)—(C)</b> Upper panels show bright field images, and lower panels are images of tumor fluorescence obtained with the OV100. At low magnification, residual tumor fluorescence was marginally detected. <b>(B)</b> However, at high magnification, residual tumor fluorescence was clearly visualized (arrows) <b>(C)</b>. Arrowheads show residual tumor fluorescence in <b>B</b> and <b>C</b>. <b>(D)</b> Resected specimen. Magnifications are indicated above the columns.</p

Evaluation of tumor fluorescence at day 28 after surgery.

<p><b>(A)</b> Upper panel shows the bright field image, and lower panel shows the GFP tumor fluorescence image obtained with the OV100 at a magnification of 0.56. Laparotomy was performed at the 28^th postoperative day. Bright field image shows tumor in the resection site in the liver (arrows). Strong GFP fluorescence from the tumor is seen in the lower panel. Arrows show recurrent tumor in the resection site. Arrowheads show operative scar on the liver. <b>(B)</b> The GFP tumor fluorescence area was significantly larger in the BLS group compared to the FGS group, where only autofluorescence was detected. Error bars show SD. *<i>P</i><0.05.</p

Additional file 2: of Medullary carcinoma of the pancreas radiologically followed up as a cystic lesion for 9Â years: a case report and review of the literature

Immunohistochemical staining. The tumor was positive for cytokeratin (CK)-7 and CK-20, and focally positive for mucin (MUC) 5 AC and MUC6. The tumor was negative for MUC2 and caudal-type homeobox (CDX) 2. (PPTX 9523 kb

Additional file 1: of Medullary carcinoma of the pancreas radiologically followed up as a cystic lesion for 9Â years: a case report and review of the literature

Positron emission tomography (PET). The maximum standardized uptake value of the preoperative lesion was 6.8 (arrows). (PPTX 120 kb

Signal intensity distribution among MA (A), PLF (B), and CM (C) samples and among the groups (D).

<p>Signal intensity distribution among MA (A), PLF (B), and CM (C) samples and among the groups (D).</p

Relative expression levels of exosomal miRNAs in intraoperative peritoneal lavage fluid of T4 and T1–T3 gastric cancer patients.

<p>An asterisk indicates P < 0.05.</p

The number of detected miRNA species in 14 samples.

<p>The number of detected miRNA species in 14 samples.</p

Exosomal miRNA expression in malignant ascites (MA), peritoneal lavage fluid (PLF), and cell culture media (CM).

<p>Variation and commonality between the samples were examined. Evaluation of total exosomal RNA used a Bioanalyzer 2100. The electropherograms show the size distribution (nucleotides, nt) and fluorescence intensity (FU) of total exosomal RNA isolated from CM, MA, and PLF. The lowest spike (around 25 nt) in each sample is the marker of the Agilent RNA 6000 Pico kit.</p