To clarify the clinicopathological significance of promoter hypermethylation of tumor suppressor and tumor-related genes in biliary tract carcinomas, we examined the promoter methylation status of multiple genes in primary biliary tract carcinomas. These consisted of carcinomas of the bile duct, gallbladder, and duodenal ampulla. Surgical specimens were obtained from a total of 37 patients with biliary tract carcinoma. The cohort consisted of 23 patients with bile duct carcinoma, 9 patients with gallbladder carcinoma, and 5 patients with ampullary carcinoma. The methylation status of CHFR, DAP-kinase, E-cadherin, hMLH1, p16, RASSF1A, and RUNX3 was examined by methylation-specific polymerase chain reaction (MSP). The correlation between methylation status and clinicopathological characteristics was then assessed. The methylation frequencies of CHFR, DAP-kinase, E-cadherin, hMLH1, p16, RASSF1A, and RUNX3 genes were 16.2%, 21.4%, 27.0%, 8.1%, 24.3%, 27.0%, and 56.8%, respectively, in primary biliary tract carcinomas. The number of methylated genes per sample was 2.17 ± ± ± ±0.28 (average ± ± ± ±SD) in bile duct carcinomas, 1.80 ± ± ± ±0.97 in ampullary carcinomas, and 0.89 ± ± ± ±0.35 in gallbladder carcinomas, with a statistically significant difference between bile duct carcinomas and gallbladder carcinomas (P = = = =0.02). As for clinicopathological significance, patients with a methylated RUNX3 promoter were significantly older than those with unmethylated RUNX3 (P = = = =0.01), and DAP-kinase methylation was more frequent in poorly differentiated tumors than in well to moderately differentiated ones (P = = = =0.04). The overall survival rate was significantly lower in patients with methylated DAP-kinase (P = = = =0.009) or RUNX3 (P = = = =0.034) compared to those with unmethylated genes. Furthermore, DAP-kinase methylation-positive status was independently associated with poor survival in multivariate analyses (hazard ratio = = = =8.71, P = = = =0.024). A significant proportion of primary biliary tract carcinomas exhibited promoter hypermethylation of tumor suppressor and tumor-related genes, although bile duct carcinomas are more prone to being affected by promoter methylation than are gallbladder carcinomas. Hypermethylation of DAP-kinase appears to be a significant prognostic factor in primary biliary tract carcinomas. iliary tract carcinoma is a disease with a poor prognosis. The 5-year survival rate is less than 25% for intra-and extrahepatic bile duct carcinoma, and 32% to 61% for gallbladder carcinoma, even after radical resection of the tumor.1-3) There is no effective therapy for biliary tract carcinomas except surgical resection. Moreover, the molecular-biological mechanisms of the development of biliary tract carcinomas are less well understood than those of carcinomas of the colon, stomach, and liver.DNA methylation is an important epigenetic mechanism for suppressing gene activity by changing the chromatin structure. 4,5) It has become clear that aberrant DNA methylation of promoter region CpG isl...

Early, distant and/or skip metastasis of squamous-cell thoracic esophageal cancer frequently occurs in the right recurrent nerve node (recR). However, the specific lymphatic route without a nodal relay, such as the submucosal ascending route, was not known for the recR afferent. Using 20 donated cadavers, macroscopic, and histological observations were performed on the recR and its surrounding lymphatics, especially afferent routes from the esophagus to the recR. Most afferent vessels of the recR originated from the right paratracheal node. However, the recR often (12/20) received a major submucosal lymphatic drainage route ascending along the thoracic esophagus. The submucosal vessel came out of the esophagus and ran in a longitudinal connective tissue mass along the right tracheo-esophageal groove. A direct drainage route was often (13/20) seen from the recR to the venous system. Moreover, because of the specific histology, collaterals seemed to be present around the recR. In the regional nodes of the intrathoracic esophagus, the recR histology was characterized by the high proportion of lymphocyte accumulating areas or the cortex. From the midthoracic level, metastatic cancer cells seemed to reach the recR via esophageal submucosal vessels in the early stage. Large lymphocyte accumulating areas of the recR suggested higher filtration capacity than other distal nodes. However, the collateral of the recR and its direct drainage to the venous system suggested that the recR involvement often corresponds to a systemic disease.

A regional lymphatic system is composed of the first, second, third and even fourth or much more intercalated nodes along the lymptatic route from the periphery to the venous angle or the thoracic duct. The third or fourth node is usually termed the last-intercalated node or end node along the route. Similarly, one of the supraclavicular nodes is known to correspond to the end node along the thoracic duct. It is generally called 'Virchow's node', in which the famous 'Virchow's metastasis' of advanced gastric cancer occurs. The histology of this node has not been investigated, although region-specific differences in histology are evident in human lymph nodes. We found macroscopically the end node in five of 30 donated cadavers. Serial sections were prepared for these five nodes and sections stained with hematoxylin and eosin. Histological investigation revealed that, on the inferior or distal side of the end node, the thoracic duct divided into three to 10 collateral ducts and these ducts surrounded the node. The node communicated with the thoracic duct and its collaterals at multiple sites in two to three hilus-like portions, as well as along the subcapsular sinus. Thus, the end node was aligned parallel to the thoracic duct. Moreover, the superficial and deep cortex areas of the end node were fragmented to make an island-like arrangement, which may cause the short-cut intranodal shunt. Consequenly, the filtration function of most of Virchow's node seemed to be quite limited.

Gaps and fragmentation of the superficial lymph node cortex are considered to provide intranodal shunt flow between the afferent and efferent vessels. Using serial sections of 205 nodes obtained from 27 donated cadavers more than 70 years of age, we examined the histological architecture of the abdominal and pelvic nodes in elderly Japanese. Secondary follicles were rare in the specimens. Cortex gaps were, to a greater or lesser degree, found in all nodes. We classified these nodes into three types according to how often the gap occurred. Type 1 nodes, with a relatively complete shield for the afferent lymph, were most frequently found in gastric nodes, whereas type 3 nodes, with numerous gaps, were often observed in the colic, para-aortic and pelvic nodes. The type 3 nodes showed a specific architecture characterized by a fragmented superficial cortex, three-dimensionally assembled cords and a common sinus between them. Primary follicles were located in the assembled cord structures as well as at the superficial cortex. Irrespective of the type, B and T lymphocyte areas were intermingled in the cortex-like areas. The present results reveal region-specific histological heterogeneity in aged human visceral nodes. Due to increased surface areas, the type 3 architecture seemed to accelerate systemic immunity rather than act as a local barrier in the para-aortic and pelvic nodes, which are located centrally along the lymphatic drainage routes. However, thick trabeculae often seemed to develop in the type 3 sinus to decrease nodal function with aging.

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