Detection of CTCs correlates with regional metastasis in inoperable SCCHN. Further follow-up is needed to evaluate the prognostic significance of CTC detection, in addition to clinical staging of lymph nodes, for regional or distant recurrence.
Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.
These results demonstrate that excessive keratinocyte proliferation and abnormal differentiation contribute to epidermal hyperplasia, while melanocytic proliferation is responsible for the pigmented lesions in SAK.
Autophagy is an essential intracellular self-degradation system to maintain the homeostatic balance between the synthesis, degradation and recycling of cellular proteins and organelles. Recently, it is reported that autophagy is associated with systemic lupus erythematosus, rheumatoid arthritis and idiopathic pulmonary fibrosis. The association with autophagy and scleroderma (SSc) is also suspected, however, the role of autophagy in the pathogenesis of tissue fibrosis is still unclarified, and the association of which phase of scleroderma is largely unknown. Therefore, we investigated the role of autophagy in the pathogenesis of SSc by using skin and lung samples of bleomycin (BLM)-induced SSc murine model and human SSc skin. BLM or phosphate-buffered saline (PBS) was injected into shaved back of C3H/HeJ mice for 2 or 4 weeks. Also, we used skin samples of edematous phase SSc and those of sclerotic phase SSc. We carried out haematoxylin and eosin (HE) stain for evaluation of skin sclerosis and immunofluorescent stain by anti microtubule-associated protein 1 light chain 3 (LC3) antibody, which is the most common autophagy marker. We evaluated the number of LC3positive dots in fibroblasts in the mice samples (skin and lung) and the human skin samples (both edematous and sclerotic phase). There was no clear difference between the number of LC3-positive dots of skin samples from PBS mice and those from BLM mice qualitatively. We are currently investigating the number of LC3-positive dots of mice lung tissues and human skin samples in more numbers.
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