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. 2018 Jul 17;8:10814. doi: 10.1038/s41598-018-29144-9

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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pSPHK1 is overexpressed in biliary tract cancer with lymph node metastasis on immunohistochemistry. (a) Semi-quantitative analysis of the immunohistochemical pSPHK1 expression in gallbladder cancer tissues with (n = 10) or without lymph node metastasis (n = 10) was performed. A staining score was calculated by multiplying two scores of staining intensity (0: negative staining; 1: weak staining; 2: moderate staining; 3: strong staining) and extent (0: 0–25%; 1: 26–50%; 2: 51–75%; 3: 76% -) of biliary tract cancer cells staining positive. The score of 0 to 5 was considered pSPHK1-negative, and 6 to 12 pSPHK1-positive. The score was compared between gallbladder cancer tissues with (n = 10) or without lymph node metastasis (n = 10) using Fisher’s exact test. The test was two-sided, and P value < 0.05 was considered statistically significant. Scale bar = 200 µm. (b) The staining intensity was determined and scored as 0 (negative staining), 1 (weak staining), 2 (moderate staining) and 3 (strong staining). Scale bar = 200 µm. (c) The result of immunohistochemical staining of pSPHK1 for gallbladder cancer with or without lymph node metastasis is shown.