Helicobacter pylori eradication therapy outcome according to clarithromycin susceptibility testing in Japan.

BACKGROUND: Helicobacter pylori (Hp) infection increases the risk of gastric cancer. Therefore, eradication is a global goal, which requires continuous monitoring of therapeutic regimens and effectiveness. Clarithromycin resistance is an important contributor to eradication failure, and metronidazole is recommended as second-line treatment in such cases. Here, we retrospectively evaluated the clarithromycin and metronidazole resistance rates and treatment effectiveness in patients with Hp using tailored therapies according to clarithromycin susceptibility testing. METHODS: Data on drug susceptibility were obtained for 5249 Japanese Hp patients between July 2005 and August 2018. Clarithromycin/metronidazole resistance rates were analyzed according to year, gender, and age with Fisher's exact test. The relationship between clarithromycin resistance and Hp therapy outcomes was assessed for 1300 patients. Treatment regimens included a clarithromycin- or metronidazole-containing 7-day triple therapy with one of several proton pump inhibitors and vonoprazan. RESULTS: Clarithromycin resistance increased annually and was higher in women and younger patients (<30 years). Rates of metronidazole resistance were stable but decreased with age. Hp treatment regimens using PPIs had eradication rates of 88% and 45% among clarithromycin-sensitive and clarithromycin-resistant cases, respectively, while regimens including vonoprazan had eradication rates of around 90% regardless of clarithromycin susceptibility. In particular, triple therapy with vonoprazan, amoxicillin, and metronidazole achieved 98% eradication. CONCLUSION: Clarithromycin-containing triple therapy even using vonoprazan did not achieve satisfactory eradication rates even in the clarithromycin-sensitive group. To avoid antibiotic misuse in population with low metronidazole resistance, 7-day vonoprazan, amoxicillin, and metronidazole triple therapy might be a strong candidate as a first-line eradication therapy.

Early-stage blocking of Notch signaling inhibits the depletion of goblet cells in dextran sodium sulfate-induced colitis in mice.

BACKGROUND: Goblet cells, which contribute to mucosal defense and repair in the intestinal epithelium, are depleted in human and rodent colitis. The Notch signal pathway regulates the differentiation of intestinal stem cells into epithelial cells and inhibits the differentiation of secretory lineages, including goblet cells. The aim of our study was to clarify whether the blocking of the Notch pathway at an early stage of colitis would preserve goblet cells and facilitate the healing process in dextran sulfate sodium (DSS)-induced colitis in mice. METHODS: DSS was orally administered to C57/BL6 mice for 7 days, and dibenzazepine (DBZ), a Notch pathway blocker, was administered for 5 consecutive days, beginning on the first day of DSS treatment. Colonic mucosal inflammation was evaluated clinically, biochemically, and histologically. The expression of the goblet cell-associated genes Math1 and MUC2 and proinflammatory cytokines was evaluated by real-time reverse-transcriptase-PCR, with the expression of Math1 and MUC2 also visualized by immunohistochemical examination. RESULTS: The administration of DBZ at 4 mumol/kg significantly reduced the severity of the colitis. Compared with the DSS only-treated intestine, the number of goblet cells was relatively sustained, and the expression of Math1 and MUC2 was also elevated in the DSS/DBZ-treated intestine. DBZ treatment suppressed the mRNA levels for interleukin-1beta and -6, and matrix metalloproteinases-3 and -9 in the DSS-treated intestine. CONCLUSIONS: Early-stage blocking of Notch signaling may ameliorate acute DSS colitis by preventing reduction in the number of goblet cells.

Adenoendocrine cell carcinoma of the gallbladder clinically mimicking squamous cell carcinoma.

We present the case of a 62-year-old Japanese man whose histological diagnosis was adenoendocrine cell carcinoma of the gallbladder at autopsy, but whose antemortem diagnosis was squamous cell carcinoma. The patient was admitted to hospital with complaints of occasional vomiting and abdominal pain. Abdominal computed tomography revealed a large tumor on the gallbladder involving the adjacent liver, colon, and duodenum, with multiple metastases in the greater omentum and paraportal lymph nodes. The serum level of squamous cell carcinoma antigen (SCCA) was high, whereas that of carbohydrate antigen (CA) 19-9, as well as that of carcinoembryonic antigen (CEA) was within the normal range. Due to these clinical features, we first suspected advanced squamous cell carcinoma of the gallbladder. After two cycles of gemcitabine monotherapy, the tumor had become enlarged and the regimen was changed to a combination of docetaxel and cisplatin. Though tumor regression was achieved and his serum SCCA level normalized after 3 months, the patient rejected additional chemotherapy and died 8 months after the diagnosis. The histopathological findings made by autopsy demonstrated the tumor to be an adenoendocrine cell carcinoma without squamous carcinoma cells. The case is interesting in that the clinical features were similar to those of squamous cell carcinoma of the gallbladder.

Reduced expression of low-density lipoprotein receptor in hepatocellular carcinoma with paraneoplastic hypercholesterolemia.

BACKGROUND AND AIM: Hepatocellular carcinoma (HCC) is frequently associated with paraneoplastic hypercholesterolemia. In familial hypercholesterolemia, genetic mutation of the low-density lipoprotein (LDL) receptor gene has been recognized as being a pathogenesis of the disease. We investigate the expression of a LDL receptor protein and gene abnormalities of a LDL receptor in HCC cells in cases with paraneoplastic hypercholesterolemia. METHODS: Eleven patients with HCC associated with paraneoplastic hypercholesterolemia and seven patients with HCC who did not have hypercholesterolemia were studied. Paraffin-embedded tissues were obtained at operative resection, autopsy, or biopsy. Immunohistochemistry was performed using a monoclonal antibody against human LDL receptors. Confocal laser-scanning microscopy was used to observe the FITC-labeled LDL receptor. DNA samples were extracted from paraffin-embedded tissues. Since a LDL receptor gene is located on chromosome 19p13.2, a microsatellite marker, D19S413, was used to analyze the chromosomes. RESULTS: Immunoreactive LDL receptors were observed all over the surface of non-tumorous hepatocytes. However, expression of the LDL receptor was significantly decreased in all HCC cells derived from the 11 patients with hypercholesterolemia. In contrast, the expression was retained in the HCC cells of all patients without hypercholesterolemia. In two patients with hypercholesterolemia, DNA analysis revealed a loss of heterozygosity on chromosome 19p13.2. CONCLUSION: We demonstrated reduced expression of the LDL receptor in HCC cases with paraneoplastic hypercholesterolemia. LDL receptor genes with genomic disorders may cause decreased expression of the LDL receptor protein, leading to feed-back failure of the cholesterol regulation system, as seen in familial hypercholesterolemia. This is the first report considering the mechanism behind the development of paraneoplastic hypercholesterolemia in HCC.

Mutation of the p16/CDKN2 gene and loss of heterozygosity in malignant mucosal melanoma and adenoid cystic carcinoma of the head and neck.

The purpose of this study was to investigate the molecular biological characteristics of malignant mucosal melanoma (MMM) and adenoid cystic carcinoma (ACC) of the head and neck. We analyzed the common genetic abnormalities that may help to identify the loci in the genes involved in the development of MMM and ACC of the head and neck by PCR-LOH on chromosomes 1p, 6q, 9p, 10q, 11q, 12q, 17p, and 19q. LOH was observed in 6 of 12 cases of MMM and in 12 of 15 cases of ACC informative for at least one of the loci analyzed. One distinct deleted region was identified at chromosome 9p21. In addition, to identify a possible involvement of p16/CDKN2 PCR-SSCP and auto-DNA sequence analysis were also performed to detect any mutation of the p16/CDKN2. Particularly, 2 missense mutations were detected in codon 225 and 226, both in MMM and ACC. There were mutational hot spots in the p16/CDKN2 gene. These results suggested that mutation of the p16/CDKN2 gene was a common factor in the development of human MMMs and ACCs, while this gene may be correlated with development and/or progression of a subtype and play a role in the oncogenesis of these cancers.

Hepatic expression of gamma-glutamyltranspeptidase in the human liver of patients with alcoholic liver disease.

BACKGROUND: Gamma-glutamyltranspeptidase (GGT) has been recognized as an enzyme that converts glutathione into cysteine, and it is localized predominantly within the liver. Serum GGT is clinically recognized as the most useful marker for diagnosis of alcoholic liver disease (ALD). METHODS: GGT localization within the liver was examined immunohistochemically using an anti-GGT antibody and was visualized by confocal laser scanning microscopy in ALD and normal livers. Double immunostaining for GGT and dipeptidylpeptidase-IV (DPP-IV) was carried out to evaluate GGT localization in greater detail. RESULTS: Expression of GGT protein and mRNA was studied with immunoblot analysis and in situ hybridization, respectively. Immunohistochemically, the expression of GGT in the normal liver was faintly demonstrated in the bile canaliculi of hepatocytes and in biliary epithelial cells. In ALD livers, GGT was clearly demonstrated at the same sites. Double immunostaining demonstrated that GGT and DPP-IV were colocalized in hepatocytes in the ALD liver. In situ hybridization clearly demonstrated GGT-mRNA within the cytoplasm of hepatocytes and biliary epithelial cells. Immunoblot analysis revealed that GGT protein expression was increased in the ALD livers compared with that seen in the normal livers. CONCLUSION: These findings indicate that GGT in control and alcoholic livers is synthesized in hepatocytes and biliary epithelial cells, and is localized within the bile canalicular membrane and the luminal membrane in those cells, respectively. In conclusion, GGT synthesis and protein expression are increased in ALD livers, leading to the elevation of serum levels of GGT that are commonly noted in patients with the disease.

Altered expression of alkaline phosphatase (ALP) in the liver of primary biliary cirrhosis (PBC) patients.

Serum alkaline phosphatase (ALP) is a representative marker of cholestasis, in diseases such as primary biliary cirrhosis (PBC). However, the hepatic localization of ALP in patients with cholestatic liver diseases has not been fully clarified. Accordingly, we studied the expression of ALP in the liver of PBC, chronic hepatitis C and controls. By immunohistochemistry, in the liver tissue of controls and chronic hepatitis C patients, ALP was found to be localized in the canalicular membrane of hepatocytes and the apical area of the cytoplasm of bile duct epithelial cells. In PBC, ALP was localized in both the canalicular and baso-lateral membranes of hepatocytes and in the whole cytoplasm of the remaining bile duct epithelial cells. The expression of ALP in liver tissues evaluated by Western blotting was increased to 3.6-fold in PBC compared with that in the controls and chronic hepatitis C patients, while the expression of mRNA of ALP evaluated by RT-PCR was increased to 7.0-fold in PBC compared with that in the controls and chronic hepatitis C patients. The present study is the first study to reveal altered localization and increased expression of ALP which may result in the elevation of serum ALP in PBC.

Rapid-onset primary biliary cirrhosis resembling drug-induced liver injury.

A 54-year-old woman was admitted to our hospital because of acute liver injury. Since she had a history of having used a diet product, drug-induced liver injury (DILI) was initially considered. However, the patient was subsequently diagnosed as suffering from primary biliary cirrhosis (PBC) based on the findings of liver histology and serum anti-mitochondrial antibody positivity. Overlap syndrome between PBC and autoimmune hepatitis (AIH) was also suspected, however, serum levels of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase became normal three months later, after treatment with combination therapy comprising ursodeoxycholic acid plus bezafibrate. We therefore concluded that the liver disease in this patient was actually PBC, but that it resembled overlap syndrome or DILI. In cases of PBC, a rapid onset, as frequently seen in the case of DILI, viral hepatitis or AIH, is not common. We herein report a rare case of PBC which resembled DILI.

Nonalcoholic steatohepatitis associated with psoriasis vulgaris.

A 24-year-old man was admitted to our hospital because of liver dysfunction. He had been diagnosed as having psoriasis vulgaris at 18 years of age. Physical examination demonstrated obesity, general erythema, and hepatomegaly. Laboratory data revealed elevated serum levels of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and glucose. A histological examination of the liver revealed macrovesicular fatty change and infiltration of inflammatory cells, including lymphocytes and polymorphonuclear cells, within the liver lobules. Pericentral fibrosis and pericellular fibrosis were also recognized. He was diagnosed as having nonalcoholic steatohepatitis (NASH), based on the fact that he had no habit of drinking alcohol, as well as psoriasis vulgaris and diabetes mellitus. We herein report a very rare case of NASH associated with psoriasis vulgaris.

Hepatocellular carcinoma in a patient with liver cirrhosis associated with negative serum HCV tests but positive liver tissue HCV RNA.

A 70-year-old woman was admitted to our hospital because of a liver tumor. Laboratory data revealed mild liver dysfunction. Neither serum anti-HCV antibody nor HCV-RNA was detected. Computed tomography revealed a tumor lesion measuring 2 cm in diameter within the liver. Histological examination of the tumor revealed moderately differentiated hepatocellular carcinoma while the non-tumorous liver tissue demonstrated liver cirrhosis. By the RT-PCR method, HCV-RNA was detected from the non-tumorous liver tissue. We herein report a very rare case of hepatocellular carcinoma in a patient with liver cirrhosis associated with negative serum HCV findings, but positive finding for liver tissue HCV RNA.