[A case of IL-6 producing malignant mesothelioma of abdomen with thrombocytosis].

A 77-year-old man was admitted to our hospital with abdominal pain and ascites. He had an occupational history of working with asbestos. Abdominal CT showed multiple nodular lesions with enhancement by contrast medium in the cavity. Platelet counts, CRP and serum IL-6 level were increased. Biopsied materials obtained by laparoscopy showed oval cells with rich cytoplasm growing in an epithelial pattern. To clarify the characteristics of the cells, immunohistochemistry was performed. Calretinin and CK5/6 were positive, and CEA, S-100 protein, c-kit and CD34 were negative, result in confirmation of a diagnosis of malignant mesothelioma. Because IL-6, IL-6 receptor and VEGF were expressed markedly, the patient received chemotherapy for IL-6 suppression. During the treatment, thrombocytosis imploved satisfactorily.

Usefulness of a combined evaluation of the serum adiponectin level, HOMA-IR, and serum type IV collagen 7S level to predict the early stage of nonalcoholic steatohepatitis.

OBJECTIVE: Since nonalcoholic steatohepatitis (NASH) may progress to cirrhosis, it is important to differentiate NASH from simple steatosis, especially in its early stages. However, a liver biopsy cannot be performed in all patients with nonalcoholic fatty liver disease (NAFLD). We herein investigated whether serum biochemical markers are useful for predicting early-stage NASH. METHOD: Nineteen patients with simple steatosis and 66 patients with early-stage NASH (stage 1-2 in Brunt's criteria) were studied. The area under the receiver operating characteristic curve (AUC) was used to illustrate the diagnostic ability of serum biochemical parameters to distinguish between simple steatosis and early-stage NASH. RESULTS: The serum adiponectin level was found to be significantly lower with early-stage NASH group (3.6 mug/mL) than in the simple steatosis group (6.0 mug/mL) (P < 0.001). The AUC was high (0.765) in the early-stage NASH group, and it was also the highest among all other markers. The sensitivity of the serum adiponectin level in the diagnosis of early-stage NASH was 68%, which was higher than for any other factors, while its specificity was 79%. The corresponding sensitivity and specificity of HOMA-IR were 51% and 95%, respectively. For type IV collagen 7S, sensitivity was 41% and specificity 95%. The sensitivity of the combination of three markers was 94%, with a specificity of 74%. CONCLUSION: Approximately 90% of the patients with early-stage NASH can be predicted by a combined evaluation of the serum adiponectin level, HOMA-IR, and serum type IV collagen 7S level.

Effect of the inducer of interleukin-6 (ME3738) on rat liver treated with ethanol.

BACKGROUND: Recently, ME3738, a derivative of soyasapogenol B, was developed as an inducer of interleukin (IL)-6. It has been demonstrated that ME3738 is stimulate to produce IL-6 and that it protects against concanavalin A-induced liver failure. It has also been reported that IL-6 prevents alcoholic fatty liver in mice. These results suggest that ME3738 may prevent alcoholic liver injury. In the present study, we investigated whether ME3738 prevents fatty liver in ethanol-fed rats. METHODS: Twenty-four male rats were fed with liquid diets containing ethanol or carbohydrates for 8 weeks. Liquid diets were prepared with or without ME3738 (0.8 mg/mL). Liver sections were stained for histology and IL-6 expression. Fatty changes of liver were classified into 4 grades: 0, 1+, 2+, and 3+. Plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), triglyceride, total cholesterol, and IL-6 were measured, as was hepatic ATP content. RESULTS: The extent of fatty degeneration in ethanol-fed rats was significantly greater (p=0.023) than that in controls. Fatty changes in rats fed ethanol containing ME3738 decreased, but were not significantly different from those in rats fed ethanol. Immunohistochemical staining of IL-6 was observed in perivenular hepatocytes of all rats, with its intensity becoming stronger in the order of controls, controls containing ME3738, ethanol, and ethanol-containing ME3738-fed rats. Plasma levels of AST and ALT in rats fed ethanol were significantly higher than those in controls. In rats fed ethanol-containing ME3738, these levels decreased to those of control-fed rats, but were not significantly different from those in rats fed ethanol. Plasma IL-6 was not detected in any rats. Hepatic ATP content in rats fed ethanol was significantly (p<0.05) lower than that in control-fed rats; however, in rats fed ethanol-containing ME3738, it increased to that in control-fed rats. CONCLUSIONS: Oral administration of ME3738, inducer of IL-6 may prevent the development of fatty liver caused by chronic ethanol consumption.

Mutation of mitochondrial DNA in livers from patients with alcoholic hepatitis and nonalcoholic steatohepatitis.

BACKGROUND: In alcoholic hepatitis (Al-Hep) and nonalcoholic steatohepatitis (NASH), triglycerides accumulate in hepatocytes. We examined the hypothesis that mutations in mitochondrial DNA may take place by mitochondrial overwork, resulting in dysfunction of mitochondria. SUBJECTS AND METHODS: Subjects of this research were 8 cases each of Al-Hep, NASH, and fatty liver (FL). Total DNA was extracted from the biopsied liver samples. DNA fragments were amplified by PCR and DNA sequences determined in the control and coding regions of mitochondrion. RESULTS: When the numbers of mutations per 1,000 bases of mitochondrial DNA were compared between each group, no significant differences were found among D-loop, HV1, and HV2 mitochondrial DNA regions. However, there were significantly more mutations in ND1 and COII of Al-Hep and NASH than in FL, and mutations were comparatively at random. Neither a region in which mutations were focused nor differences among the groups were recognized. When details of the base mutation in a control region were investigated by group, the transition type of mutation between T:A<<->>C:G occurred in at least 70%. Also, a transition-type mutation was found mostly in a coding region, which was similar to the mutation pattern in the control region, except for the ND1 and COII regions where there were hardly any mutations. CONCLUSIONS: As gene mutations of mitochondrial DNA appeared frequently in Al-Hep, and also in NASH, mitochondrial dysfunction caused by mutation in mitochondrial DNA may be involved in the pathogenesis of both diseases.

Effect of chronic dietary ethanol consumption on colonic cancer in rats induced by 1,1-dimethylhydrazine.

BACKGROUND: The pathogenetic correlation between chronic alcohol consumption and development of colon cancer is not clear. The role of alcohol abuse in the carcinogenic action of 1,1-dimethylhydrazine (DMH), which induces tumors in the colon, was evaluated. METHODS: Twenty male rats were fed liquid diets containing ethanol or carbohydrates for 39 weeks. DMH (20 mg/kg body weight, once a week) was injected subcutaneously from the 5th to the 20th week. Pair feeding was stopped at 10:00 am and DMH was administered at 02:00 pm. Ethanol was not detected in the blood at the time of injection. Liquid diets were provided again at 05:00 pm until 10:00 am next day. The animals were killed at the end of the 39th week, and the colons were removed for examination for the number of aberrant crypt foci (ACF) by methylene blue staining. Tissue sections were stained for histology and cytochrome P4502E1 (CYP2E1) expression. RESULTS: The number of ACF in colons obtained from ethanol-fed rats with DMH was 24 (n=5, 4.4+/-2.5/rat), which was significantly (p<0.001) more than that of the other treated rats: only 3 (n=5, 0.6+/-0.5/rat) in the pair-fed control rats with DMH, and none in the ethanol-fed or control-fed rats without DMH. Cytochrome P4502E1 staining demonstrated marked expression in the colon mucosa from ethanol-fed rats, but not in the pair-fed control rats. CONCLUSIONS: The increased expression of CYP2E1 induced by chronic ethanol consumption promotes the development of DMH-induced colon cancer.

Myelodysplastic syndrome complicated with inflammatory intestinal ulcers: significance of trisomy 8.

Three cases of myelodysplastic syndrome (MDS) complicated with inflammatory intestinal ulcers all had cytogenetic abnormalities with trisomy 8. The first two patients were diagnosed with intestinal Behçets disease and were successfully treated with salazosulphapiridine, and the third patient died after leukemic transformation. We review the reported cases of MDS complicated with Behçets disease. Most of these cases are Japanese, having intestinal involvement as well as cytogenetic abnormalities with trisomy 8. We discuss the significance of trisomy 8 in intestinal involvement in MDS.

[Mutation of mitochondrial DNA in patients with non-alcoholic steatohepatitis].

When the numbers of mutations per 1,000 bases of mitochondrial DNA were compared among three groups of Al-Hep, NASH and FL, there were significantly more mutations in ND1 and COII of Al-Hep and NASH than in FL. When details of the base mutation were investigated by group, the transition type of mutation between T:A and C:G occurred in control and coding regions. In FLS and FLS-ob mice, mutations in the D-loop and coding regions of mitochondrial DNA were investigated by sequencing analysis. The average number of mutations per clone in each region was more than 5-fold higher in FLS-ob than in FLS. In addition, the mutations were mostly transition-type mutations. These data indicate that nitric oxide plays an important role in mitochondrial DNA mutations.

[Clinical features of NASH].

Nonalcoholic fatty liver disease (NAFLD), including simple steatosis and nonalcoholic steatohepatitis (NASH), is commonly associated with metabolic syndrome. Many of NASH patients do not have subjective symptoms. The NASH is often found by routine health checkups etc. It is difficult to distinguish the simple steatosis and the NASH by transaminase level. It is reported that 25% of the NASH patient progress to cirrhosis in a decade. However, there is no study by a lot of patients concerning the prognosis of the NASH, and further studies will be required in the future.

Effect of chronic dietary ethanol in the promotion of N-nitrosomethylbenzylamine-induced esophageal carcinogenesis in rats.

BACKGROUND: The pathogenetic correlation between chronic alcohol consumption and development of esophageal cancer is not clear. The role of alcohol abuse in the carcinogenic action of N-nitrosomethylbenzylamine, which induces tumors in the esophagus, has been evaluated. METHODS: Twenty male rats were fed liquid diets containing ethanol or carbohydrates for 30 weeks. N-nitrosomethylbenzylamine (0.1 mg/kg, twice a week) was injected i.p. from the 9th to 19th week. The pair feeding was stopped at 9.00 am and N-nitrosomethylbenzylamine was administered at 10.00 am. Ethanol was not detected in the blood at the time of injection. Liquid diets were provided again at 3 pm until 9 am next day. The animals were killed at the end of the 30th week. The esophagi were collected and examined for visible tumors. The tissue sections were stained for histology and CYP2E1expression. RESULTS: While 5-8 esophageal squamous polyps developed in all rats in the ethanol group, only one polyp each was formed in five out of the 10 rats in the control group. The size of the polyps was significantly larger in the ethanol group, when compared to the control group. Invasive squamous cell carcinoma was also observed in 50% of the animals in the ethanol group. Cytochrome P4502E1 (CYP2E1) staining demonstrated marked expression in the esophageal mucosa in the ethanol group, but not in the control group. CONCLUSIONS: The increased expression of CYP2E1 induced by chronic ethanol consumption promotes the development of N-nitrosomethylbenzylamine-induced esophageal tumorigenesis. However, the molecular mechanism of the increased production of esophageal tumors during alternative administration of N-nitrosomethylbenzylamine and ethanol is not clear.

Marked pneumoperitoneum 3 weeks after percutaneous endoscopic gastrostomy.

In November 2001, a 29-year-old woman was admitted to the hospital because of dysphagia due to an apallic state caused by cerebral anoxia. Nutritional support was maintained by nasogastric tube feeding for approximately 3 months. For improvement of the body state maintenance and quality of life, a percutaneous endoscopic gastrostomy (PEG) was performed. Three weeks after the PEG, the patient had a wound infection and abdominal distension appeared. Marked pneumoperitoneum was confirmed by radiological examination. No signs or symptoms of peritoneal inflammation developed. A gastrografin study showing that the PEG tube was in the stomach appropriately was checked, and it was noted to be firmly in place without extravasation of contrast. After suspension of the tube feeding and tube opening to decrease intragastric pressure, intravenous hyperalimentation was performed. The pneumoperitoneum resolved within 7 days. Forty days after the PEG, tube feeding was resumed successfully. No recurrence of pneumoperitoneum developed and the patient has remained stable until the present time. The etiology of this finding probably occurs by insufficient fixation of the PEG, causing leakage of air through the gastric wall which enters the free peritoneal space. We recommend that the external binder should be kept 1 cm away from the abdominal skin after the gastrostomy fistula has formed and matured, and periodic rotation of the tube to verify that the internal bumper is free and sufficiently fixed to the gastric wall. In the case of abdominal distension after PEG placement, a X-ray examination and computed tomography (CT) scan with contrast medium would be helpful to ascertain pneumoperitoneum.

Use of serum carbohydrate-deficient transferrin values to exclude alcoholic hepatitis from non-alcoholic steatohepatitis: a pilot study.

BACKGROUND: Since no clinical or biochemical parameters allow an accurate diagnosis of non-alcoholic steatohepatitis (NASH), the diagnosis by exclusion of alcoholic hepatitis is necessary. However, it is difficult to get the accurate amount of alcohol consumed from the patients, especially from females. To differentiate between NASH and alcoholic hepatitis, we investigated whether serum biochemical markers of chronic alcohol abuse are useful or not. METHODS: Sera were obtained from 13 patients with NASH and 26 patients with alcoholic hepatitis. Diagnoses in these patients were confirmed histologically by needle biopsy of the liver. All patients with alcoholic hepatitis consumed more than 80 g of ethanol/day for more than 10 years. As markers of chronic alcohol abuse, serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), hyaluronate, mean corpuscular volume of red blood cells (MCV) and carbohydrate-deficient transferrin (CDT) were measured. RESULTS: Among alcohol markers, serum values of AST, AST/ALT ratio, GGT, CDT and MCV in patients with alcoholic hepatitis were significantly higher than those in patients with NASH, respectively. However, serum values of these markers, except for CDT, were overlapped in many cases of NASH and alcoholic hepatitis. Serum CDT values of all patients with NASH were lower than the cutoff value, 2.66%, and those of all patients with alcoholic hepatitis were higher than the cutoff value. CONCLUSION: The results of the present study suggest that serum CDT level could be used to differentiate between NASH and alcoholic hepatitis.

Expression of hyaluronic acid in N-nitrosodimethylamine induced hepatic fibrosis in rats.

Hyaluronic acid (HA) plays prominent role in the pathogenesis of liver fibrosis. The mechanism of increased serum and liver HA during hepatic fibrosis was studied in rats. Liver injury was induced by intraperitoneal injections of N-nitrosodimethylamine (NDMA) for 7 consecutive days. A group of animals were sacrificed on everyday during injection and also on days 14 and 21 after the start of NDMA administration. The alpha-smooth muscle actin (alpha-SMA) was stained as a marker for activated stellate cells. Liver HA was studied by histochemical methods and serum HA was monitored by HA binding protein assay. CD44 was stained immunohistochemically. After the start of NDMA administration, necrosis was initiated on day 3 and massive necrosis was observed on days 5 and 7. Fibrosis was developed on day 14 and early cirrhosis was present on day 21. Staining of alpha-SMA demonstrated activated stellate cells from day 3 onwards. Serum HA peaked on day 7 and reduced afterwards. Serial liver sections stained for HA revealed excessive accumulation of HA during NDMA administration. On days 14 and 21, alpha-SMA and HA staining was remarkable in fibrotic and cirrhotic areas. CD44 staining was negative except during necrosis. It is concluded that the early elevation of serum HA is due to the increased synthesis and simultaneous release from the necrotic liver. In latter stages the increase of both serum and liver HA is contributed by the increased synthesis by the activated stellate cells and reduced clearance by the impaired sinusoidal endothelial cells.

Correlation between adenosine triphosphate content and apoptosis in liver of rats treated with alcohol.

BACKGROUND: Chronic alcohol consumption depresses adenosine triphosphate (ATP) synthesis and induces mitochondrial DNA (Mt-DNA) deletion. ATP content in cells may play a critical role in inducing cell death, apoptosis, or necrosis. However, it is unknown which type of cell death occurs in alcoholic liver disease. In this study, the deletions of hepatic Mt-DNA, hepatic ATP content, and the number of single-stranded DNA (ss-DNA) of hepatocytes in rats treated with ethanol were determined to elucidate the relationship among Mt-DNA deletion, ATP synthesis, and/or hepatic apoptosis. METHODS: Sixteen male Wistar rats were fed with a liquid diet containing 36% ethanol (E group) or liquid diet without ethanol (C group) for 5 weeks. Hepatic ATP content was measured and the deletions of Mt-DNA encoding complexes I, IV, and V were determined in fresh liver tissue, and ss-DNA was stained in paraffin sections. RESULTS: Fatty change was observed in the E group, but not in the C group. Hepatic ATP content in the E group was 0.44 micromol/g of liver, which was significantly lower than that in the C group (0.84 micromol/g of liver). However, no deletion of Mt-DNA encoding complexes I, IV, and V was detected in either the E or the C group. ss-DNA staining was clearly observed in the nuclei of hepatocytes in both groups. The number of ss-DNA-positive hepatocytes in the E group was 5.6 +/- 1.8/10,000 hepatocytes, which was significantly less than that in the C group: 20.6 +/- 4.8/10,000 hepatocytes. There was a positive correlation between hepatic ATP contents and the number of ss-DNA-positive cells. CONCLUSIONS: The results suggest that mitochondrial function, at least in part ATP synthesis, was depressed before the damage of Mt-DNA by chronic ethanol consumption. Chronic ethanol consumption may not be responsible for the apoptosis of hepatocytes.