Michiko Yamagata

High proportion of mutant K-ras gene in pancreatic juice of patients with pancreatic cystic lesions

BACKGROUND/AIMS It was recently reported that the quantitative analysis of mutant K-ras gene in pancreatic juice could be useful for the diagnosis of pancreatic cancer. This methodology was applied to patients with pancreatic cystic lesions. METHODS DNA was extracted from pancreatic juice collected at the time of endoscopy with injection of secretin. The ratio of the K-ras mutant allele to the wild-type allele was measured by two methods to detect and semiquantify mutant K-ras gene: polymerase chain reaction/preferential homoduplex formation assay and enriched polymerase chain reaction/enzyme linked mini-sequence assay. RESULTS A high frequency of K-ras mutation was detected (more than 2% of all K-ras genes) in six of 14 patients (43%) with pancreatic cysts. This frequency was similar to those detected in patients with pancreatic adenocarcinoma and in intraductal papillary neoplasms of the pancreas. In contrast, the frequency of mutation was low (less than 2%) in patients without either pancreatic cysts or pancreatic neoplasms. CONCLUSIONS K-rasgene mutation may be derived from duct cells in the pancreas with a high potential for tumorigenesis. Therefore careful follow up of patients with a pancreatic cyst is recommended if they are found to have a high level of the mutant gene in the pancreatic juice.

Fulminant B hepatitis in a surface antigen and hepatitis B DNA-negative patient with diffuse large B-cell lymphoma after CHOP chemotherapy plus rituximab

The reactivation of hepatitis B in patients who are hepatitis B carriers is a well-known and often fatal complication of chemotherapy [1]. The current recommendations are that patients who are positive for hepatitis B surface (HBs) antigen should receive antiviral prophylaxis prior to chemotherapy [2,3] or should be carefully monitored when they receive cytotoxic or immunosuppressive therapy [4]. For HBs antigen-negative patients with evidence of previous hepatitis B virus (HBV) infection, however, the data are still insufficient to provide information on the incidence of HBV reactivation [5]. We experienced an individual with resolved HBV infection (i.e. HBs antigen-negative, anti-hepatitis B core (HBc) antibody-positive), in whom fatal reactivation of hepatitis B developed following chemotherapy that included rituximab for lymphoma. A 54-year-old man was diagnosed as stage IV diffuse large B-cell lymphoma in July 2005. Prior to chemotherapy, he was negative for HBs antigen. He had neither a past history nor a family history of liver diseases. He received CHOP (cyclophosphamideþdoxorubicineþ vincristineþdexamethasone) plus rituximab (R-CHOP). We occasionally added etoposide, ifosfamide, and methotrexate to that regimen. Seven courses of combination chemotherapy were completed in November 2005, and complete remission was accomplished. During the treatment for lymphoma, liver dysfunction was not demonstrated. In October 2005, during the chemotherapy period, the titers of HBs antigen (CLIA), anti-HBc antibody (CLIA), and HBV DNA (PCR) were 50.05 IU/ml, 10.8 S/CO, and 52.6 log10 copies/ml, respectively. In December 2005, when the chemotherapy was completed, HBs antigen was still 50.05 IU/ml, the titer of anti-HBc antibody was 10.5 S/CO, and the HBV DNA level was increased to 3.3 log10 copies/ml. The genotype of HBV was type C. In February 2006, he developed malaise and loss of appetite. He was admitted to our hospital in March 2006, when serum levels of aspartate aminotransferase, alanine aminotransferase, and total bilirubin were elevated to 622 IU/l, 531 IU/l, and 2.1 mg/dl, respectively. The titers of HBs antigen, anti-HBc antibody, and HBV DNA were 250 IU/ml, 8.2 S/CO, and [mt]7.7 log10 copies/ml, respectively. Anti-HBs antibody and IgM antibody to HBc antigen yielded negative results. Exacerbation of HBV infection was diagnosed and we initiated lamivudine at a daily dose of 100 mg to prevent HBV proliferation on the 1st day of admission. On the 8th day of admission, plasma exchange was introduced since fulminant hepatic failure developed

Serum endostatin levels in patients with hepatocellular carcinoma.

Endostatin is an angiogenic inhibitor, isolated from a murine hemangioendothelioma [1]. Endostatin, the 20-kDa C-terminal fragment of collagen XVIII [2], has been shown to prevent angiogenesis and drive large tumors into dormancy [3]. Thus, endostatin might become one of the therapeutic agents for angiogenesis-dependent cancers due to its antiangiogenic properties. Nevertheless, there have been no data on serum endostatin levels in patients with hepatocellular carcinoma (HCC), which is known to be hypervascular. The aim of the present study was to evaluate serum levels of endostatin in patients with HCC, in those with liver cirrhosis (LC) not accompanying HCC, and in healthy controls, to compare serum levels in patients with HCC before and after therapeutic procedures, and to study its activity in patients with HCC. In addition, we examined serum levels of vascular endothelial growth factor (VEGF), which is a specific endothelial cell mitogen [4], and their possible correlation with disease progression in patients with HCC. Clinical data were obtained from files at University of Tokyo Hospital, Department of Gastroenterology and HepatoBiliary-Pancreatic Surgery Division, Department of Surgery. Forty-two randomly-selected patients with HCC treated at the departments between 1995 and 1999 were included in this study, and serum levels of endostatin were measured before treatments. The diagnoses of HCC and background liver diseases were confirmed pathologically in all patients. HCC stages I, II, III, IVA, and IVB, based upon the classification of Liver Cancer Study Group of Japan [5], were seen in 6, 6,13, 9, and 8 patients, respectively. As for background liver, 40 patients were diagnosed as LC and 2 patients as normal liver (3 hepatitis B virus positive, 32 hepatitis C virus positive, and 7 both negative). Six tumors were graded as well-differentiated HCC, seventeen tumors as moderately-differentiated HCC, and nineteen tumors as poorly-differentiated HCC. Among these patients, serum levels of endostatin before and after therapeutic procedures were compared in seven patients treated by surgical operation, one patient treated by transhepatic arterial embolisation, and one patient treated by percutaneous microwave coagulation therapy. In addition, serum levels of endostatin in a panel of 10 patients with LC (1 hepatitis B virus positive, 7 hepatitis C virus positive, and 2 both negative) not accompanying HCC and 10 healthy controls were measured. For serum sample collection, blood samples without EDTANa were centrifuged after clot formation at room temperature, and then the serum was removed. Samples were stored at -20 °C until the measurement of endostatin and VEGF. For the measurement of serum levels of endostatin, a commercially available ELISA kit, whose minimum detectable dose was 1.95 ng/ml, was used (Accucyte human endostatin, Cytimmune Science, Inc., USA). For the measurement of serum levels of VEGF, an ELISA kit with antibodies which recognize VEGF (Quantikine human VEGF, R&D Systems, USA) was used. Its minimum detectable level was less than 9.0 pg/ml.

Washout of small stones in the bile duct by saline infusion using a side-holed balloon catheter in patients undergoing endoscopic papillary balloon dilation

BACKGROUND Complete bile duct clearance of stones should be achieved in patients managed with endoscopic papillary balloon dilation. However, complete retrieval of small stones or tiny fragments sometimes proves difficult using conventional devices. METHODS We attempted the removal of fine stone fragments by saline infusion using a specially designed retrieval balloon catheter with a blind tip and a side hole located proximal to the balloon in 14 patients. RESULTS Using this technique, bile duct clearance was achieved in a single attempt in 13 of 14 patients; 2 attempts were required in 1 patient. Cholecystitis occurred in 2 patients with gallstones in situ after the procedure. CONCLUSIONS Saline washout using a side-holed retrieval balloon catheter effectively cleans stones and stone fragments from the bile duct in patients treated with endoscopic papillary balloon dilation.

Periportal Stellate Cells in Subjects with Chronic Hepatitis C with a Varied Serum Alanine Aminotransferase Level

Recently it has been reported that HCV-related cirrhotic patients with persistently high serum levels of alanine aminotransferase (ALT) activity have higher risk of development of hepatocellular carcinoma (HCC) than those with persistently low levels of ALT activity [1]. Vitamin A has been demonstrated to have many biological functions in regulation of growth and differentiation of normal and cancer cells. Hepatic stellate cells (SCs) are the main storage site of vitamin A and regulate homeostasis of vitamin A. To analyze cellular and molecular mechanism in the relationship between persistent high serum level of ALT and HCC development from the viewpoint of vitamin A handling, this study was performed.

In vitro sonographic evaluation of common bile duct stones and fragments with a high-frequency microprobe

This study was conducted to evaluate the sonographic characteristics of common bile duct (CBD) stones and stone fragments scanned in vitro with a high‐frequency (20‐MHz) microprobe.