Narihiro Ishiyama

Quantitative detection of hepatitis C virus RNA by multicyclic RT-PCR

Abstract We have developed a method to quantitate hepatitis C virus (HCV)-RNA from serum by using ‘multicylic’ RT-PCR. This method requires 100 μl of sera and synthetic HCV-RNA with a known copy number to be used as a standard. HCV RNA in serum are amplified by the RT-PCR technique and quantitated by comparing the darkness of the spot on the film with the standards after dot blot hybridization. Accuracy of this technique was comparable to the competitive RT PCR method (CRT-PCR) when compared using the same sample. Also, this technique uses one-tenth of the sample volume required in CRT-PCR with the additional advantages of low cost, faster assay time, and ability to run a larger number of samples. We analyzed the amount of HCV-RNA in 71 patients with chronic liver disease due to HCV subclassified into chronic inactive hepatitis (CIH), chronic active hepatitis (CAH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). Seventy-five percent of the patients contained 106 to 108 copies/ml of HCV-RNA. The number of samples which contained over 106 copies/ml in the four groups were CIH,5/8 (62.5%); CAH,15/20 (75.0%); LC,25/27 (92.6%); and HCC,14/16 (87.5%).

Detection and characterization of antibodies to bacterial heat-shock protein 60 in sera of patients with primary biliary cirrhosis

The enzyme‐linked immunosorbent assay (ELISA) with bacterial heat‐shock protein 60 (HSP60) purified from Yersinia enterocolitica (Ye) revealed that the antibodies directed against YeHSP60 existed in sera of patients with primary biliary cirrhosis (PBC). To characterize the epitope specificity of the antibodies in patients, the epitope mapping of HSP60 by means of the antibodies was performed. The results have suggested that the epitope recognized with anti‐HSP60 antibodies in PBC relates to the amino acid sequence of YeHSP60 molecule as follows: DLGQA‐KRVVINKDTTIIIDGVGDEAAIQGRLAQIRQQIEEATSDYDKEK.

Hepatocellular carcinoma containing sarcomatous lesions in a normal liver, accompanied by secondary Budd-Chiari syndrome

To the Editor: We experienced a huge tumor in an otherwise-intact liver of a 53-year-old woman without any known predisposing causes. The tumor was a moderately differentiated hepatocellular carcinoma (HCC), intermingled with sarcomatous lesions. HCC in this case was complicated by secondary Budd-Chiari syndrome (BCS). It is quite unusual that such events occur simultaneously in a patient, although the precise pathoetiological link among them remains unclear. The patient was admitted to Kyorin University Hospital complaining of abdominal distension. She had received a blood transfusion during labor at the age of 25, but had had no episode of liver damage. There was no history of alcohol abuse, exposure to chemical carcinogens, or any chronic liver diseases. On physical examination, a hard liver was palpable. Longitudinal dilated veins were visible on the abdominal wall. Laboratory examinations showed an elevated lactate dehydrogenase (LDH) level of 884 IU/L and a decreased hemoglobin value of 8.5 g/dL. Liver function tests remained within the normal limit. Hepatitis virus infection, including hepatitis B, C, or other possible viruses, was negative. HCC tumor markers such as -fetoprotein were also negative. Computed tomography revealed a huge hepatic mass in the right lobe with a mixed appearance of solid and cystic components. The solid regions were stained with contrast medium until the delayed phase of the image. Duplex Doppler ultrasonography demonstrated reversed flat flows in the right and middle hepatic veins, which reflected dysfunction of the venous outlet into the inferior vena cava (IVC) possibly due to the compression of the mass, while a normal flow was confirmed in the left hepatic vein toward the IVC. Intrahepatic collateral vessels were also recognized communicating between the middle and left hepatic veins. These findings on ultrasonography were compatible with those of BCS. However, no membranous obstruction was found in the IVC or hepatic veins. Tumor cytology revealed malignant cells with round to spindle nuclei. All these clinical data led us to suspect the mass could be a hepatic sarcoma, and a right lobectomy was performed. The tumor occupied 23 × 22 × 12 cm of the resected hepatic lobe of 36 × 23 × 15 cm (3875 g) and contained large hemorrhagic necrosis foci. Pathologic examination revealed that the tumor was primarily a moderately differentiated HCC. However, lesions of sarcomatous appearance were also observed sporadically. These lesions were composed mainly of marked pleomorphic cells, including bizarre multi-nucleated giant cells, and partly of spindle-shaped cells lacking mutual adherence, resembling so-called pleomorphic sarcoma and spindle cell sarcoma, respectively. Noncancerous liver tissue showed no signs of chronic liver diseases except that mild dilatation of the central veins were noted in Zone 3. When multiple recurrences were found in the liver 19 months later, serum liver function tests and tumor markers were again within the normal limit, including LDH and hemoglobin. All these clinicopathologic findings indicate that the HCC in the present case was of unknown etiology. Although the development of HCC is often associated with chronic liver diseases, as represented by liver cirrhosis, it is unusual that HCC develops when liver tissues are intact. However, we cannot exclude the possibility that an unknown virus infection at the time of blood transfusion at the age of 25 was responsible for the carcinogenesis. A sarcomatous appearance has been observed in 9.4% of HCC cases in an autopsy study and in 1.8% of surgically resected cases. It is noteworthy that most of these cases have been shown to be related to at least one of a number of factors including chronic liver diseases, hepatitis virus infection, or sarcomatous transformation of HCC due to anticancer therapy, none of which, however, were confirmed in the present case. In comparison with ordinary HCC, this type of tumor tends to show certain characteristics such as low or negative levels of serum tumor markers, huge mass including hemorrhagic necrosis, and poor prognosis with intrahepatic metastasis, as were found in the present case. Another remarkable feature of this case is that HCC was complicated by the presence of secondary BCS. Although primary BCS could be a carcinogenic cause of HCC, in this case, BCS was considered to be the consequence of HCC. Collateral vessels disappeared in the remaining left lobe after surgery, which also supports this view. Although HCC is a possible though rare cause of BCS, occurring in only 0.6 to 3.2% of all BCS cases, atypical features in HCC in the present case, or HCC containing sarcomatous lesions, may be related to the pathogenesis of BCS.

Is Helicobacter pylori a causal agent in gastric carcinoma

The aim of this study is to clarify the relationship between Helicobacter pylori (H. pylori) infection and gastric carcinoma. 94 patients with gastric carcinoma and 111 patients with chronic gastritis were involved in this study. They were classified into 3 age groups: Group A (40 years and under), Group B (41-59), Group C (60 years and over). Serum samples were tested for H. pylori IgG antibodies by ELISA and for pepsinogen (PG) by RIA. The ratio of PG I/PG II was used as a marker for atrophic gastritis. Results were as follows. In the incidence of H. pylori antibodies, there was no significant differences between gastric carcinoma and chronic gastritis in any of the groups. In the quantity of H. pylori antibodies, there was no significant difference between gastric carcinoma and chronic gastritis in any of the groups. The ratio of PG I/PG II was significantly decreased in H. pylori positive cases when compared to H. pylori negative cases in each group with chronic gastritis and group A and B with gastric carcinoma. The ratio of PG I/PG II in gastric carcinoma was significantly lower than that of chronic gastritis. As conclusion, from the point of view of the prevalence of H. pylori, it is suggested that H. pylori is not a direct causal agent in the pathogenesis of gastric carcinoma.

A case of rheumatoid arthritis that developed autoimmune hepatitis associated with anti-Golgi complex antibody

Abstract A 50-year-old woman, who had been diagnosed as having rheumatoid arthritis (RA) 7 months earlier, was admitted to our hospital because of liver dysfunction. Her laboratory data and histological findings satisfied the criteria for autoimmune hepatitis (AIH) as revised by the International AIH Group. Laboratory examinations (indirect immunofluorescence and immunoblotting analyses) revealed that anti-Golgi complex antibodies (AGA) were positive in her serum. AGA are thought to be closely associated with AIH and/or liver dysfunction according to several reports.