Fluorescein as a topical fluorescent contrast agent for quantitative microendoscopic inspection of colorectal epithelium.

Fiber bundle microendoscopic imaging of colorectal tissue has shown promising results, for both qualitative and quantitative analysis. A quantitative image quality control and image feature extraction algorithm was previously designed for quantitative image feature analysis of proflavine-stained ex vivo colorectal tissue. We investigated fluorescein as an alternative topical stain. Images of ex vivo porcine, caprine, and human colorectal tissue were used to compare microendoscopic images of tissue topically stained with fluorescein and proflavine solutions. Fluorescein was shown to be comparable for automated crypt detection, with an average crypt detection sensitivity exceeding 90% using a combination of three contrast limit pairs.

Inhibition of fatty acid synthase attenuates CD44-associated signaling and reduces metastasis in colorectal cancer.

Fatty acid synthase (FASN) and ATP-citrate lyase, key enzymes of de novo lipogenesis, are significantly upregulated and activated in many cancers and portend poor prognosis. Even though the role of lipogenesis in providing proliferative and survival advantages to cancer cells has been described, the impact of aberrant activation of lipogenic enzymes on cancer progression remains unknown. In this study, we found that elevated expression of FASN is associated with advanced stages of colorectal cancer (CRC) and liver metastasis, suggesting that it may play a role in progression of CRC to metastatic disease. Targeted inhibition of lipogenic enzymes abolished expression of CD44, a transmembrane protein associated with metastases in several cancers including CRC. In addition, inhibition of lipogenic enzymes and reduced expression of CD44 attenuated the activation of MET, Akt, FAK, and paxillin, which are known to regulate adhesion, migration, and invasion. These changes were consistent with an observed decrease in migration and adhesion of CRC cells in functional assays and with reorganization of actin cytoskeleton upon FASN inhibition. Despite the modest effect of FASN inhibition on tumor growth in xenografts, attenuation of lipogenesis completely abolished establishment of hepatic metastasis and formation of secondary metastasis. Together, our findings suggest that targeting de novo lipogenesis may be a potential treatment strategy for advanced CRC.

Prefabrication of vascularized bone flap by demineralized bone matrix.

Demineralized bone matrix (DBM) has been reported to have osteoconductive and osteoinductive properties and has been clinically used as a bone graft alternative. In the present study we attempted to generate a vascularized bone flap by subcutaneous implantation of DBM with a vascular loop to provide blood supply in a rat model. Thirty male Sprague-Dawley rats were divided into two groups according to the presence or absence of blood supply. In the experimental group, the bone flap was created by application of 0.4 mL of DBM onto two pieces of gelatin sponge sheets between which a vascular loop was sandwiched. A prefabricated flap without a vascular loop served as the control. The flaps were biopsied at three different time intervals postoperatively (2, 4, and 6 weeks). The results showed that DBM induced subcutaneous bone formation in both of the groups. However, in the nonvascularized group, the amount of bony tissue had decreased at four postoperative weeks and continued to do so afterwards. In contrast, bone formation was active at four weeks in the vascularized group. Our study indicated that implantation of DBM can prefabricate a bone flap. Blood supply to the flap is considered a key factor of the success of this prefabrication.

Neuroma-in-continuity model in rabbits.

The present study presents a model for creation of neuroma-in-continuity in the rabbit, confirmed by histologic study, electrophysiology, and examination of molecular markers. Twelve New Zealand rabbits were used. The lateral fascicule of the peroneal nerve with 15-mm in length was resected. In the intervals from 4 to 6 weeks postoperatively, the histology showed the typical pathologic changes of neuroma by hematoxylin-eosin (HE), Luxol fast blue, and Van-Gieson staining. As compared with the healthy nerve, the motor nerve conduction velocity (MCV) and compound motor action potential (CMAP) were found significantly slowed and reduced. The expression of ciliary neurotrophic factor (CNTF) in nerve and calcitonin gene-related peptide (CGRP) mRNA in L7, S1 dorsal root ganglia were downgraded and upgraded, respectively, with formation of neuroma-in-continuity. These electrophysiologic and molecular markers' expression data can be used as reliable parameters for evaluation of management of the neuroma-in-continuity.