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In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy

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World Journal of Gastroenterology 2007年第15期13卷 2160-2165页

作者： Martin Goetz Beena Memadathil stefan biesterfeld Constantin Schneider Sebastian Gregor Peter R Galle Markus F Neurath Ralf Kiesslich Medical Clinic Johannes Gutenberg-Universit(a|¨)t MainzGermany Department of Pathology Johannes Gutenberg-Universit(a|¨)t MainzGermany

AIM： To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS： A novel rigid confocal probe （diameter 7 mm） was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation, Light emission was detected at 505 to 750 nm. The field of view was 475 μm × 475 μm. Optical slice thickness was 7 μm with a lateral resolution of 0.7 μm. Subsurface serial images at different depths （surface to 250 μm） were generated in real time at 1024 × 1024 pixels （0.8 frames/s） by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation.RESULTS： The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice. Real time microscopic imaging with the confocal minimicroscopy probe was easy to achieve. The different staining protocols （fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin） each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging.CONCLUSION： Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures. The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients.

关键词： Confocal microscopy in vivo imaging Endomicroscopy Fluorescence

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VEGF-D expression correlates with colorectal cancer aggressiveness and is downregulated by cetuximab

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World Journal of Gastroenterology 2008年第26期14卷 4156-4167页

作者： Markus Moehler Christian Frings Annett Mueller Ines Gockel Carl C Schimanski stefan biesterfeld Institute of Pathology Johannes Gutenberg University Mainz 55101 Germany Peter R Galle Martin H Holtmann 1st Department of Medicine Johannes Gutenberg University Mainz 55101 Germany Department of General and Abdominal Surgery Johannes Gutenberg University Mainz 55101 Germany

AIM:To gain mechanistic insights into the role played by epidermal growth factor receptor (EGFR) in the regulation of vascular endothelial growth factors (VEGFs) in colorectal cancer (CRC). METHODS:The impact of high-level expression of the growth factor receptors EGFR and VEGF receptor (VEGFR)3 and the VEGFR3 ligands VEGF-C and VEGF-D on disease progression and prognosis in human CRC was investigated in 108 patients using immunohistochemistry. Furthermore, the expression of the lymphangiogenic factors in response to the modulation of EGFR signalling by the EGFR-targeted monoclonal antibody cetuximab was investigated at the mRNA and protein level in human SW480 and SW620 CRC cell lines and a mouse xenograft model. RESULTS: Human CRC specimens and cell lines displayed EGFR, VEGF-C and VEGF-D expression with varying intensities. VEGF-C expression was associated with histological grade. Strong expression of VEGF-D was significantly associated with lymph node metastases and linked to a trend for decreased survival in lymph node-positive patients. EGFR blockade with cetuximab resulted in a significant decrease of VEGF-D expression in vitro and in vivo. CONCLUSION:In conclusion, the expression of VEGF-D in colorectal tumours is significantly associated with lymphatic involvement in CRC patients and such expression might be blocked effectively by cetuximab.

关键词： Human colorectal cancer Lymphangiogenesis Vascular endothelial growth factor-C Vascular endothelial growth factor-D Epidermal growth factor receptor

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Coexpression of receptor-tyrosine-kinases in gastric adenocarcinoma-a rationale for a molecular targeting strategy?

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World Journal of Gastroenterology 2007年第26期13卷 3605-3609页

作者： Daniel Drescher Markus Moehler Ines Gockel Kirsten Frerichs Annett Müller Friedrich Dünschede Thomas Borschitz stefan biesterfeld Martin Holtmann Thomas Wehler Andreas Teufel Kerstin Herzer Thomas Fischer Martin R Berger Theodor Junginger Peter R Galle Carl C Schimanski Department of General and Abdominal Surgery Johannes Gutenberg University of Mainz MainzGermany First Department of Internal Medicine Johannes Gutenberg University of Mainz MainzGermany Institute of Pathology Johannes Gutenberg University of MainzMainzGermany Third Department of Internal Medicine Johannes Gutenberg University of Mainz MainzGermany Unit of toxicology and chemotherapy German Cancer Research CenterGermany

AIM： To define the （co-）expression pattern of target receptor-tyrosine-kinases （RTK） in human gastric adenocarcinoma. METHODS： The （co-）expression pattern of VEGFR1-3, PDGFR（α/β and EGFR1 was analyzed by RT-PCR in 51 human gastric adenocarcinomas. In addition, IHC staining was applied for confirmation of expression and analysis of RTK localisation. RESULTS： The majority of samples revealed a VEGFR1 （98%）, VEGFR2 （80%）, VEGFR3 （67%）, PDGFRα （82%） and PDGFRβ （82%） expression, whereas only 62% exhibited an EGFR1 expression. 78% of cancers expressed at least four out of six RTKs. While VEGFR1-3 and PDGFRα revealed a predominantly cytoplasmatic staining in tumor cells, accompanied by an additional nuclear staining for VEGFR3, EGFR1 was almost exclusively detected on the membrane of tumor cells. PDGFRβ was restricted to stromal pericytes, which also depicted a PDGFRα expression. CONCLUSION： Our results reveal a high rate ofreceptor-tyrosine-kinases coexpression in gastric adenocarcinoma and might therefore encourage an application of multiple-target RTK-inhibitors within a combination therapy.

关键词： VEGFR EGFR PDGFR Cancer Adeno-carcinoma Gastric Stomach

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