Proteolytic enzymes in human pure pancreatic juice (PPJ), which was collected by cannulating the main pancreatic duct using endoscopy, were investigated by two-dimensional zymography (2-DZ). 2-DZ was carried out by combining isoelectric focusing (IEF) in the first dimension with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension, using gels containing casein or gelatin as a substrate for the proteolytic enzymes. After electrophoresis, the gels were incubated in Triton X-100 followed by incubation at 37 degrees C in Tris buffer (pH 8.5) containing CaCl2. By staining the gels with Coomassie Brilliant Blue (CBB R-250), proteolytic enzymes were detected as clear spots and zones against a blue background. Proteinase inhibitors, such as a cysteine proteinase inhibitor (E-64), a metalloproteinase inhibitor (EDTA), and a serine proteinase inhibitor (Pefabloc SC), were added to PPJ in order to determine the types of proteinases. In patients with pancreatic cancer, spots of molecular weight (Mr) 70,000 and isoelectric points (pI) 5.3-5.5 were clearly detected on the gels containing casein and gelatin, while these spots were not detected in the PPJ from healthy subjects. The proteolytic activities of these spots were strongly inhibited by EDTA and Pefabloc SC but not E-64. These results suggest that the spots of Mr 70,000 and pI 5.3-5.5 in PPJ of pancreatic cancer might be matrix metalloproteinase 2, which is a candidate for tumor-associated proteinase. 2-DZ proved to be a tool for analysis of proteolytic enzymes in PPJ and for the clinical diagnosis of pancreatic cancer.

BACKGROUND Little is known about genetic aberrations associated with development and progression of biliary tract carcinomas. METHODS To study chromosomal aberrations associated with development and progression of biliary tract carcinomas, the authors used comparative genomic hybridization to examine 50 such carcinomas. RESULTS Gains in part or in whole of chromosomes 1q, 8q, and 20q and losses of 5q, 8p, 9p, and 18q were detected frequently in early stage (T1/T2 classification) biliary tract carcinomas (≥ 40% of 19 early stage tumors), and they also were found in advanced stage (T3/T4 classification) tumors. In particular, loss of 9p was the most frequently observed aberration in both early stage (15 of 19; 78%) and advanced stage tumors (21 of 31; 68%). The frequencies of gains of 7p12‐p14 (P < 0.003), 7p21‐pter (P < 0.007), and 7q31 (P < 0.01) differed significantly between biliary tract carcinoma with and without distant metastasis. Also, gains of 5p and 19q13 and loss of 6q14‐q16 were more frequent in tumors with lymph node metastasis than in those without it (P < 0.02). CONCLUSIONS It is likely that loss of 9p is one of the genetic aberrations critical for the development of biliary tract carcinoma, whereas gains of 5p, 7p, 7q, and 19q and loss of 6q are considered later events associated with tumor progression and are thought to confer metastatic potential to biliary tract carcinomas. Cancer 2001;91:570–7. © 2001 American Cancer Society.

A 68‐year‐old man presented with melena in June 1993, and was diagnosed as having adenocarcinoma of the rectum with liver metastasis. He underwent anterior resection of the rectum and was given weekly chemotherapy. In October 1993, he developed jaundice due to severe stricture of the middle to lower common bile duct caused by metastatic spread to the lymph nodes around the pancreatic head. A 10mm wide, 51mm long WallstentTM (Schneider) was inserted into the lower end of the bile duct through the papilla, resulting in rapid normalization of the serum bilirubin level. Three months later, the Wallstent eroded through the duodenal wall, causing massive fatal hemorrhage. We, therefore, emphasize the necessity of being alert to the potential for such complications in cases involving placement of a self‐expanding metallic stent for malignant biliary stricture.

We studied the role of the increase in the calcium concentration in pure pancreatic juice of alcoholic noncalcified chronic pancreatitis. Pure pancreatic juice was obtained endoscopically. The pancreatic juice from patients with chronic pancreatitis was adjusted to pH 7.5; then the calcium concentration was adjusted to 0.4, 2.9, 5.4, or 10.4 mmol/L. Artificial precipitates were produced by incubation of the samples at 37 degrees C for 6 hours. Proteins in the artificial precipitates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the protein patterns were compared with the patterns of natural protein plugs from patients with chronic pancreatitis. The amount of the precipitate increased as the added calcium increased. The protein patterns of SDS-PAGE of the artificial precipitates were similar to those of protein plugs. Albumin, a-amylase, lipase, trypsinogen, and chymotrypsinogen were identified by immunoblotting both in the precipitate and in the protein plug. The increased calcium concentrations in pancreatic juice induced the formation of precipitates whose protein composition was similar to that of protein plugs. An increased calcium concentration in human pancreatic juice may play an important role in the pathogenesis of protein plugs.

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