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Tissue ischemia, mechanical or thermal trauma, infection and foreign body reaction predispose to adhesion formation. Adhesion formation resembles an inflammatory process. Sometimes, they appear as a bridge to wound healing, however, in other times it may cause small bowel obstruction. Methylene blue is known to inhibit the generation of oxygen radicals. Therefore, it may be used as an antioxidant reducing adhesion formation. Hamsters are animals used in surgical experiments, including pancreatic and hepatic surgeries. Objective. To study the effectiveness of using methylene blue in hamsters that undergo hepatectomy or pancreatectomy. Methods. Syrian golden hamsters were divided in 6 groups: Partial pancreatectomy with splenectomy (PPS), PPS and MB (PPS-MB), Partial hepatectomy (PH), PH and MB (PH-MB), Sham operation (SO), and Sham operation and MB (SO-MB). MB was administered intraperitoneally before closing abdominal wall. Side effects were observed. Animals were sacrificed 10 days later and adhesions were quantified. Results. Adhesions were present in all animals (100%) of PH group and 3 of 5 (60%) of PPS group. In PH-MB and SO groups, 1 of 5 hamsters (20%) in each group showed adhesions. No adhesions were found in PPS-MB and SO-MB treated groups. None of animals showed loss of weight. Conclusion. This study suggests that methylene blue administration showed to be effective to prevent adhesion formation in hepatopancreatic surgical procedures.

This chapter summarizes previous experimental data on pharmacological effects of Pyrroloquinoline quinone (PQQ) specifically with respect to acetaldehyde metabolism following ethanol loading to rats and the protective effects of PQQ against hepatotoxin-induced liver injury in rats and against hydrocortisone-induced cataract formation in chick embryos. Pharmacological application of PQQ to the animals was performed, particularly with respect to the acceleration of alcohol metabolism and prevention of experimental liver injury and cataract formation. Toxic effects of PQQ on the kidney were also demonstrated. PQQ was administered at a dose of 11.5 mg/kg body weight twice, at 1 hour and 10 minutes, prior to the ethanol administration. However, blood and liver acetaldehyde levels were much lower in the PQQ-treated rats than in untreated rats. The minimum effective dose of PQQ was determined by administration of a single dose of PQQ given 10 minutes prior to carbon tetrachloride administration.

We describe a 43-year-old woman with clinical features compatible with autoimmune cholangiopathy recently reported by Ben Ari et al. She was negative for anti-mitochondrial antibody, positive for high titer anti-nuclear antibody with homogeneous pattern, high levels of serum immunoglobulin G and nearly normal levels of serum immunoglobulin M for more than five years. In the early stages of the disease, the elevations of serum transaminase, alkaline phosphatase and gamma-glutamyl transpeptidase were well controlled by the administration of ursodeoxycholic acid. After five years of follow-up, she showed the second exacerbation of liver function tests, which then rapidly improved by prednisone administration. To analyze the antigen diversity recognized by T-cells in the liver, T-cell receptor repertoire was examined by immuno-histochemistry. The liver biopsy obtained in the early stage showed clonal accumulation of V beta 5.1-positive cells in portal areas, which was found in patients neither with primary biliary cirrhosis nor autoimmune hepatitis. In conclusion, these data suggest that T-cell response in autoimmune cholangiopathy is different from those two autoimmune liver diseases, which may imply a distinct entity of the disease.

— Immunolocalization of Type I, Type III and Type IV collagens, laminin and prolyl hydroxylase (PH), a key enzyme in collagen synthesis, was examined to clarify the fibrotic process in chronic, active liver disease. In piecemeal necrosis of chronic, active hepatitis (CAH) and active liver cirrhosis (LC), fat-storing cells (FSCs) and transitional cells (TSCs), containing abundant rough endoplasmic reticulum (RER), were increased in number and stained intensely for PH. Immunodeposits of extracellular matrix (ECM) components were found in the RER, Golgi apparatus (GA) and vesicles of these cells, especially in cases with marked inflammation. On the other hand, in the periportal areas of chronic, persistent hepatitis (CPH) or inactive LC, immunoreaction of ECM components was seldom found in the RER of FSCs and TSCs. In the portal tract, immunodeposits of ECM components were seldom found in the organelles of fibroblasts, although ECM was increased there. These findings indicate that FSCs and TSCs in piecemeal necrosis might play a role in the production of ECM components in the progression of fibrosis during the development of chronic active liver disease. In addition, ECM component production by FSCs and TSCs is associated with marked inflammation.

— Monospecific antibodies, directed against type IV collagen and laminin, were used to clarify the process of sinusoidal capillarization in rats after carbon tetrachloride (CCl4) intoxication by the direct immunoperoxidase method. After acute intoxication, both type IV collagen and laminin were increased in the area of hepatic necrosis, adjacent to the central veins; however, sinusoidal capillarization was not found. During chronic intoxication, deposition of laminin was co-distributed with that of type IV collagen, but deposition proceeded more slowly than that of the type IV collagen. Deposition of laminin was increased in the Disse space. Sinusoidal capillarization was noted as thick deposition of both antigens by light microscopy. Immunoelectron microscopy showed that both components were continuously present in the Disse space. Intracellularly, both antigens were found in the rough endoplasmic reticulum (RER) of fat-storing cells (FSC) and endothelial cells, and these cells showed morphological changes, becoming slender and flattened. In contrast, few immunoreactive products of the two components were observed in the hepatocytes. These findings suggest that type IV collagen and laminin are indispensable for the establishment of sinusoidal capillarization, and that FSC play an important role in the production of both components.

We measured the concentrations of acetone and isoprene in the exhaled breath from students to evaluate their lifestyles for the annual medical checkup at our university. 451 students were examined for both gases simultaneously by a gas analysis device, Biogas Acetone Analyzer (BAS-2000). The average concentrations of acetone and isoprene in the breath were 0.53±0.45 ppm and 0.065±0.058 ppm, respectively, and the frequency of the abnormal high levels by judging the cut-off values of 1.5 and 0.20 ppm were 3.5% and 2.7% of them, respectively. Acetone concentration in the breath was higher in students having no breakfast in the day of the medical checkup than those having breakfast. Furthermore, acetone concentration was significantly correlated inversely with body fat percentage in students who took the medical checkup in the morning and significantly higher in students with body mass index (BMI) under 25 than obese students with BMI over 25. On the other hand, isoprene concentration was higher in male students than female students, and also higher in smokers than nonsmokers. However, isoprene level was not affected by ingestion of breakfast in the day. Acetone and isoprene concentrations were correlated positively with each other, although the relationship was not affected by having breakfast. The results suggest that the measurement of acetone and isoprene concentrations in the breath is useful for evaluating lifestyles such as lack of breakfast, smoking and obesity in students.

Nutritional supplement foods containing antioxidant vitamins and minerals and fish oil (mainly docosahexaenoic acid, DHA, C22:6n-3), referred to as capsules, were administered to seven smokers every day for 34 days. Concentrations of antioxidant vitamins and minerals in serum, activity of superoxide dismutase in plasma and the concentration of 8-isoprostane (8-epi-prostaglandin F2 α) in the urine showed an increase or a tendency to increase after the end of administration. The frequency of subjects showing poor state of psychological health evidenced by a total score of 8 points or more on the General Health Questionnaire (30-item edition) scale was 42.9%, although there was a significant decrease to 14.3% upon completion of administration of the capsules. These biochemical and psychological changes were mostly returned to the basal level one month after the end of administration of the capsules. The results suggest that administration of antioxidant vitamins and minerals and fish oil to smokers resulted in an increase in antioxidant capacity. Effectiveness in alleviating psychosocial stress likely to be attributable to DHA was also observed.

Hepatocyte growth factor (HGF) can promote the regeneration of injured organs, including HGF gene therapy by electroporation (EP) for liver injury. In this study, we investigated the effect of HGF on dextran sulfate sodium-induced colitis and tried to clarify the regenerative mechanisms of colonic epithelial cells and the signaling pathway involved. Colitis was induced by dextran sulfate sodium in mice, together with HGF gene transfer by EP. On day 10, the colitis was evaluated histologically and by Western blot analysis. The colonic epithelial cell line MCE301 was exposed to HGF protein, and its proliferation and activated signaling pathway were analyzed. In vivo, the histological score improved and the number of Ki-67-positive epithelial cells increased in the HGF-treated mice compared with the controls. Western blot analysis showed enhanced expression of phospho-Akt in the HGF-treated mice compared with the controls. In vitro, HGF stimulated the proliferation of MCE301 cells. There was enhanced phospho-Akt expression for more than 48 h after HGF stimulation, although phospho-ERK1/2 was enhanced for only 10 min. LY-294002 or Akt small interfering RNA suppressed cell proliferation induced by HGF. Thus HGF induces the proliferation of colonic epithelial cells via the phosphatidylinositol 3-kinase/Akt signaling pathway. HGF gene therapy can attenuate acute colitis via epithelial cell proliferation through the PI3K/Akt pathway. These data suggested that HGF gene therapy by EP may be effective for the regeneration and repair of injured epithelial cells in inflammatory bowel disease.

Objective A late evening snack improves the catabolic state in patients with advanced liver cirrhosis. We tested whether long-term (3 mo) late evening snacking that included a branched-chain amino acid (BCAA)–enriched nutrient mixture produces a better nutritional state and better quality of life than ordinary food in patients with hepatitis C virus–positive liver cirrhosis. Methods In a multicenter, randomized study, 48 patients with liver cirrhosis received late-evening supplementation with the BCAA-enriched nutrient mixture or ordinary food, such as a rice ball or bread, for 3 mo. During the study period, each patient was instructed on energy and protein intake. Blood biochemical data, nitrogen balance, respiratory quotient, and health-related quality of life (Short Form 36 questionnaire) were evaluated at baseline and at the end of the study. Results Total and late-evening energy intakes were similar in the two groups at 3 mo. Serum albumin level, nitrogen balance, and respiratory quotient were significantly improved by the BCAA mixture but not by ordinary food. The parameters of the Short Form 36 did not statistically significantly improve over 3 mo in either group. Conclusion Long-term oral supplementation with a BCAA mixture is better than ordinary food in a late evening snack at improving the serum albumin level and the energy metabolism in patients with cirrhosis.

We measured the concentration of carbon monoxide (CO) gas in exhaled breath and the level of stress in students to evaluate their lifestyle and mental health for the annual medical checkup at our University. 993 students were examined for CO by gas analysis device and 445 selected randomly from them for stress levels by a Stressometer. CO concentration in breath is markedly higher in the case of smokers and slightly higher in non-smokers who are exposed to passive smoking. Students with a tremor of the nervous system at rest (TNR) higher than 70 in a Stressometer were found in 13.0%, and in non-smoking female students there was a significant correlation between stress levels and CO concentration in the breath which is produced in the cells by stress-induced heme oxygenase-1. The results of the present study suggest that measuring CO gas may be a convenient way to evaluate lifestyle and stress levels in students.

We calculated the effective energy of a resistant cornstarch (HB-450) by using 13CO2, H2, and CH4 breath tests to assess HB-450 digestion and fermentation in healthy female subjects. On the basis of the areas under the curves (AUCs) for the concentrations of 13CO2 and H2 following ingestion of 40 g of HB-450, 23.0±6.8 g was digested in the upper digestive tract and 3.6±2.1 g was fermented by intestinal flora. The effective energy of the 40 g dose was thus calculated as 2.5±0.8 kcal/g. These results indicate that 13CO2, H2, and CH4 breath tests are useful for measuring the effective energy of resistant cornstarch.

deoxyguanosine (8-OHdG) in the urine as a consequence of oral administration of antioxidant health foods including plant-based SOD, vitamin C and vitamin E to seven healthy subjects every day for 15days. Although there was a significan ti ncrease i nt heconcentration of vitamins C and E in serum, there was as ignificant decrease in SOD (extracellular type) activity and Mn-SOD (mitochondrial type) content and a narrower range of variation therein. In contrast, there was a tendenc yt oward an increase in the amount of 8-OHdG in the urine (observed in 6 of 7 subjects). We looked into the possibility that SOD activity was being inhibited by pycnogenol (water extract of the bark of the French maritime pine) as the main ingredient of the antioxidant health foods, and it became clear that SOD activity is included in pycnogenol. These results suggest that oral administration of antioxidant health foods containing SOD originating in plants has the effect of lowering the activity and content of SOD in the blood.

Background and Aim: Fas-associated phosphatase-1 (FAP-1) has been thought as an inhibitor in Fas-mediated apoptosis. Here, we investigated the role of FAP-1 in Fas-mediated apoptosis of human colon cancer cells. Method: The viability of four colon cancer cell lines treated with agonistic anti-Fas antibody was determined using WST-1 assay and cell death detection ELISA. pRc/CMV-FAP-1 was transfected to a FAP-1-negative, Fas-resistant colon cancer cell line SW480 by lipofection and the clones expressing FAP-1 protein were selected by limiting dilution. In the clones, expression of 550 genes was analyzed by cDNA microarrays. Protein expression of FAP-1 and molecules related to apoptosis was examined by western blot. Results: We obtained two FAP-1 overexpressed clones which were much more susceptible to Fas-mediated apoptosis than control cells. In the clones, caspase 8 and caspase 3 were fully activated by agonistic anti-Fas antibody treatment. Bcl-2 family proteins were not related to the high susceptibility of these clones, because caspase 9 was not activated. Transfection of FAP-1 did not suppress the survival actions of insulin-like growth factor (IGF-1) which enhanced survival signal through Akt phosphorylation. Upregulation in 21 genes and downregulation in 29 genes was revealed by cDNA arrays. We confirmed protein expression of p21 and phosphorylated p21 were much more enhanced in the clones than in control cells. Conclusions: Overexpression of FAP-1 enhanced susceptibility to Fas-mediated apoptosis in SW480 and upregulation of p21 may contribute to this phenomenon. Our results indicate a novel function of FAP-1 in Fas-mediated apoptosis of human colon cancer cells.

Aims: The aim of this study was to examine diurnal variation in the concentration of methane in the breath of methane producers and to clarify the mechanism of methane production in enterobacterial flora. Subjects and methods : The study was performed on 11 methane producers (students) with a methane concentration of 5 ppm or more in their breath. Methane in the breath and hydrogen concentrations were measured simultaneously using an exhalation gas analyzer, and diurnal variations were followed. Results : Four diurnal variation patterns were observed in 11 methane producers. Pattern A, methane varies at a higher concentration than hydrogen; pattern B, methane and hydrogen vary in parallel at almost the same level; pattern C, hydrogen varies at a higher concentration than methane; and pattern D, methane concentration varies at a low level of 5 ppm or less. In the case of patterns A and B, the concentrations of methane and hydrogen were in a positive correlation (p

b Depart- We examined whether or not changes occur in the quantity of carbon monoxide (CO) production as a consequence of the intake of ordinary meals. The study was performed by following diurnal changes in the concentration of CO in the breath over approximately 12 hr after each meal on 17 healthy subjects. CO is a product of the reaction of the enzyme heme oxygenase- 1 (HO-1) induced through oxidative stress. The aver- age value for diurnal variations in the concentration of CO in the breath was 1.9 ± ± ± ± ± 0.5 (0.8 to 3.3) ppm, and the maximum range of variation in individual cases came within the narrow range of 0.4 to 1.3 ppm. There were no significant differences in average values be- fore and after meals. The value for the total quantity of CO in the breath 12 hr sought on the basis of the area under the diurnal variation curve was 22.8 ppm (0.39 mmol). This figure was very close to the heme breakdown quantity of 0.35 mmol (theoretical value, corresponding to the CO production quantity) every 12 hr.

Human umbilical cord blood (HUCB) contains stem/progenitor cells, which can differentiate into a variety of cell types. In this study, we investigated whether HUCB cells differentiate into hepatocytes in vitro and in vivo. We also examined whether CD34 could be the selection marker of stem cells for hepatocytes. HUCB cells were obtained from normal full-term deliveries, and CD34+/−cells were further separated. For in vitro study, HUCB cells were cultured for 4 wk, and expressions of liver-specific genes were examined. For the in vivo study, nonobese diabetic/severe combined immunodeficient mice were subjected to liver injury by a Fas ligand-carried adenoviral vector or only radiated. Mice were treated simultaneously with or without cell transplantation of HUCB, CD34+, or CD34−cells. After 4 wk, human-specific gene/protein expression was examined. In the in vitro study, human liver-specific genes were positive after 7 days of culture. The immunofluorescent study showed positive staining of α-fetoprotein, cytokeratin 19, and albumin in round-shaped cells. In the in vivo study, immunohistochemical analysis showed human albumin-positive, hepatocyte-specific antigen-positive cells in mouse livers of the Fas ligand/transplantation group. Fluorescence in situ hybridization analysis using the human Y chromosome also showed positive signals. However, no difference between transplanted cell types was detected. In contrast, immunopositive cells were not detected in the irradiated/transplantation group. The RT-PCR result also showed human hepatocyte-specific gene expressions only in the Fas ligand/transplantation group. HUCB cells differentiated into hepatocyte-like cells in the mouse liver, and liver injury was essential during this process. The differences between CD34+and CD34−cells were not observed in human hepatocyte-specific expression.

Background & Aims: Nutritional intervention with branched-chain amino acid (BCAA) is reported to increase serum albumin concentration in patients with decompensated cirrhosis. However, a definite conclusion on whether it can improve patients' survival has not yet been reached. The present study aimed to test possibilities of improving survival of patients with decompensated cirrhosis by using a BCAA preparation that is suitable for long-term oral administration. Methods: A multicenter, randomized, and nutrient intake-controlled trial on the comparative effects of BCAA orally administered at 12 g/day for 2 years versus diet therapy with defined daily food intake (1.0–1.4 g protein kg−1 day−1 including BCAA preparation and 25–35 kcal kg−1 day−1) was conducted in 646 patients with decompensated cirrhosis. The primary end point was a composite of death by any cause, development of liver cancer, rupture of esophageal varices, or progress of hepatic failure (event-free survival). The secondary end points were serum albumin concentration and health-related quality of life (QOL) measured by Short Form-36 questionnaire. Results: The incidence of events comprising the primary end point significantly decreased in the BCAA group as compared with the diet group (hazard ratio, 0.67; 95% confidence interval, 0.49–0.93; P = .015; median observation period, 445 days). Serum albumin concentration increased significantly in the BCAA group as compared with the diet group (P = .018). The "general health perception" domain in Short Form-36 measures was also improved (P = .003). Patients' adherence to the prescription was favorable. Conclusions: Oral supplementation with a BCAA preparation that can be administered for a long period improves event-free survival, serum albumin concentration, and QOL in patients with decompensated cirrhosis with an adequate daily food intake.

This multicenter study compared the effects of branched-chain amino acid granules (Livact((R)) Granules, LIV) and an enteral nutrient for chronic hepatic failure (Aminoleban((R)) EN, EN) on serum albumin in patients with decompensated liver cirrhosis. This study enrolled "patients with decompensated liver cirrhosis associated with hepatic encephalopathy who were suffering from hypoalbuminemia in spite of adequate food intake," a condition for which both drugs are indicated. Enrolled patients were randomized to the two groups according to the central registration method. This study continued for 24 weeks. Selected foods were supplied to each patient in principle so that caloric and protein intakes were standardized between the two groups. A total of 281 patients were enrolled. LIV was not inferior to EN concerning the primary efficacy endpoint changes in serum albumin.

Background/Aims : Human hepatoma cells have been reported to be resistant to Fas-mediated apoptosis. Sodium butyrate (SB) induced apoptosis of several cancer cells. We investigated the effects of SB on Fas-mediated apoptosis of hepatoma cells. Methods : In hepatoma cells (HuH-6, HuH-7, Hep-G2, and PLC/PRF/5), susceptibility to Fas-mediated apoptosis and Fas expression were assessed. Caspase-3 activation and cell cycle progression were evaluated in HuH-6. A cDNA microarray assay was performed to screen the changes in the expression of mRNAs. Results : Pretreatment with SB caused an enhancement of the sensitivity to anti-Fas-mediated cytotoxicity, though it did not increase the expression of Fas. The cDNA microarray assay revealed up-regulation of pro-apoptotic Bik, Bak, Bid and c-Jun N-terminal protein kinase-1, and down-regulation of anti-apoptotic Bag-1 and cellular Fas-associated death domain-like interleukin-1β-converting enzyme inhibitor protein. In some molecules, expression of the proteins was confirmed by Western blotting. An increase in truncated-Bid accompanying the reduction in Bid was also observed. Conclusions : SB enhances the susceptibility of hepatoma cells to anti-Fas-mediated cytotoxicity by altering the mRNA and protein expression and/or the activation status of proteins that could be involved in the Fas signaling pathway. SB may have an important role in the elimination of hepatoma cells.

Background/Aims : CD103, a mucosal integrin αEβ7, binds to E-cadherin expressed on hepatocytes and bile duct epithelium in the liver. Although CD103 + T cells are enriched in intestinal intraepithelial lymphocytes, the localization of those cells in the liver is unknown Methods : We investigated whether CD103 + cells are present in human livers, and how they are associated with the intrahepatic development of T cells by flow cytometry and immunohistochemistry. Results : Human livers contain significantly ( P + cells in CD4 + and CD8 + T cells (25.7±13.5 and 27.1±19.3%, respectively) than peripheral blood lymphocytes. Moreover, CD103 + cells in the liver, but not in peripheral blood, contained T cells with intermediate expression level of T cell receptor αβ. Those cells consist of mostly CD4 + and CD4 − CD8 − cells, and expressed low level of CD56 and interleukin-2 receptor beta chain in most of the population. These characteristics are distinct from natural killer T cells, which have been thought to be extrathymic T cells in human livers. Moreover, intrahepatic CD103 + cells expressed mRNA for recombination-activating gene-1, -2 and pre T cell receptor-alpha detected by reverse transcription-polymerase chain reaction Conclusions : CD103 + T cells are preferentially accumulated in human livers, and those T cells show characteristics of extrathymic T cells.

In Japan there are reportedly 2 million HCV carriers, half of whom may have been infected through blood transfusion.To fulfill the responsibility of our hospital to the community, we undertook screening for undiagnosed hepatitis C patients among 3, 266 patients who had received a blood transfusion in our hospital since its opening (or earlier at other hospitals) and November 1989 when HCV-antibody testing of blood products began, by initiating contact with them by mail. Among respondents, 429 individuals underwent HCV antibody testing. Of those tested, 377 were negative, 52 (12.1%) were positive, and 6 patients requiring treatment (as of April 2002) were discovered. We consider that this kind of effort to discover hepatitis C patients is important to improve their prognosis through early diagnosis.

An IgG-type anti-IgA antibody was detected in 3 of 22 patients who developed blood transfusionrelated side effects in our hospital in 1998. These patients were females, and developed blood transfusion-related side effects during chemotherapy for malignant diseases. None of the patients had IgA deficiency. With regard to the subclass and allotype of the anti-IgA antibody, IgA1 and IgA2m (1) were common among the 3 patients. IgA2m (2) was present in 1 patient. A washed blood preparation (washed red blood cells, washed platelets) was transfused to 1 of the patients. In the remaining 2, a standard non-washed blood preparation was administered. The anti-IgA antibody titer markedly decreased in the patient treated with the washed preparation. In the remaining 2 patients treated with the non-washed blood preparation, in contrast, no decrease in anti-IgA antibody titer was seen. No side effects of blood transfusion occurred in the patient with a marked decrease in anti-IgA antibody titer, whereas in 1 of 2 patients without reduction of anti-IgA antibody titer, 3 adverse events related to blood transfusion were observed. Administration of washed blood preparation to patients with anti-IgA antibody may be required to prevent complications of blood transfusion.

OBJECTIVE: We compared the habitual food intake and plasma fatty acid composition in cirrhotic patients living in two different regions in Japan, Okayama and Toyama, and evaluated the effects of dietary polyunsaturated fatty acid and α-tocopherol intake on serum alanine aminotransferase (ALT) activity. METHOD: A quantitative food-frequency questionnaire method was used. RESULTS: The significantly higher intake of fish in the patients living in Toyama resulted in higher plasma levels of docosahexaenoic acid and lower levels of arachidonic acid. Serum ALT activity correlated negatively with plasma arachidonic acid (r = −0.456, P < 0.05) and α-tocopherol (r = −0.505, P < 0.05) levels. Dietary intakes of vitamin E and polyunsaturated fatty acids (mg/g) correlated negatively with serum ALT (r = −0.377, P < 0.05). Dietary intake of linoleic acid and the ratio of polyunsaturated to saturated fatty acid in dietary fat correlated significantly with serum ALT (r = 0.604, P < 0.01, and r = 0.622, P < 0.01, respectively). The amount of vegetable intake correlated with intake of vitamin E and polyunsaturated fatty acid (r = 0.527, P < 0.02). CONCLUSIONS: These findings suggest that habitual food intake affects the plasma fatty acid profile and that elevated serum ALT may be related to arachidonic acid deficiency and vulnerability to lipid peroxidation in cirrhotic patients with hepatitis B and C viruses.

It has been reported that H. pylori-containing cagE was associated with duodenal ulcer. The aims of the present study were to clarify the association between the cagE gene and clinical outcome and to analyze the relationship between the cagE gene and two other virulence factors—cagA and vacA—in two areas in Japan (Fukui and Okinawa) where the prevalence of duodenal ulcer and gastric cancer risk are quite different. Eighty of 81 isolates possessed the cagE gene, and all isolates possessed the cagA gene. The vacA genotype s1c/m1 was a major genotype in both areas in Japan. There was no significant association between cagE, cagA status, or vacA genotype and clinical outcome. Phylogenetic analysis of the cagE gene indicated that most Japanese isolates formed a different cluster from strains isolated in the West with an association with the vacA genotype. In conclusion, the strains with cagE, cagA, and the s1c/m1 genotype of vacA are predominant in Japan regardless of clinical outcome and construct a different phylogenetic cluster from those in the West.

Background and Aims: Little is known about whether Helicobacter pylori infection alters fatty acid metabolism in gastric mucosal cells. By using cultured rat gastric mucosal cells (RGM-1), we investigated the effect of H. pylori broth culture filtrates on this point. Furthermore, our study aimed to find out whether n-6 long chain polyunsaturated fatty acids from linoleic acid are formed in RGM-1 cells. Methods: Rat gastric mucosal cells were incubated with 10, 20 and 40 μg/mL of linoleic acid or medium alone. Phosphatidylcholine content extracted from whole RGM-1 cells was quantitated by using a densitometer, and its fatty acid composition was analyzed by using gas chromatography. Prostaglandin E2 concentration in the culture medium was measured by using radioimmunoassay. The expression of cyclooxygenase (COX)-1 and COX-2 was examined by using reverse transcription–polymerase chain reaction. In addition, after incubation with [1-14C] linoleic acid, radioactivities of both linoleic acid and arachidonic acid components of the PC fraction were counted. The effects of H. pylori broth culture filtrates on PC content, its fatty acid composition and prostaglandin (PG)E2 synthesis were also assessed. Results: Linoleic acid addition caused an increase in the composition of arachidonic acid, as well as linoleic acid, and also in PGE2 concentration. Cyclo-oxygenase-2 expression was induced in RGM-1 cells by the addition of linoleic acid. In addition, [1-14C] linoleic acid added to the culture medium was converted to [1-14C] arachidonic acid in RGM-1 cells. Helicobacter pylori broth culture filtrates decreased linoleic acid composition and increased arachidonic acid composition. Moreover, after incubation with H. pylori broth culture filtrates, PGE2 concentrations were higher than that of the controls. Conclusions: These findings suggest the presence of fatty acid elongase and Δ5- and Δ6-desaturases synthesize arachidonic acid from linoleic acid in RGM-1 cells. Thus, H. pylori infection may enhance PGE2 synthesis and accelerate n-6 fatty acid metabolism in gastric mucosal cells, which could make the gastric mucosal barrier more fragile.

Background and Aims: Whether Helicobacter pylori eradication alters gastric mucosal phospholipid contents and their fatty acid composition remains unclear. The aim of the present study was to clarify the effect of H. pylori eradication on gastric mucosal phosphatidylcholine (PC) content and its fatty acid composition. Methods: Endoscopic biopsy specimens were taken from the antrum and body of each of 19 asymtomatic male volunteers for detection of H. pylori, histopathological assessment of gastritis, phospholipid determination and fatty acid analysis. All the subjects with H. pylori infection were treated with eradication therapy. Endoscopy and tissue sampling were repeated again 1 and 6 months after all treatment. Results: In eight subjects, H. pylori infection was evident and was successfully eradicated. Pretreatment degrees of lymphocytes and plasma cells (inflammation) and polymorphonuclear neutrophils (activity) were greater in H. pylori-positive subjects compared with H. pylori-negative subjects (P < 0.001), whereas the degree of inflammation decreased (P < 0.001), and neutrophils had completely disappeared at 6 months after eradication. Moreover, the gastric mucosal PC contents at the antrum and body were unchanged within 1 month after cessation of treatment, but increased at 6 months after eradication (P < 0.05). At 6 months after cessation of treatment, H. pylori-eradicated subjects had an increase (+ 30% at antrum, + 18% at body) in linoleic acid composition and a decrease (− 37%, − 43%) in arachidonic acid composition of PC at the antrum and body, respectively. Conclusions: These findings suggest that H. pylori eradication reduces the production of various eicosanoids, resulting in the normalization of gastric mucosal PC content and its fatty acid composition, which may consequently cause the gastric mucosal hydrophobicity to be normalized.

Background: Although branched-chain aminoacids (BCAA) are reported to be effective in prolongation of the mean survival time of patients with liver cirrhosis, it is not clear whether BCAA could influence the immune function in those patients. Methods: Branched-chain amino acids were given as a supplement to carbon tetrachloride-induced cirrhotic rats, and an aminogram of the liver and kinetics of liver-associated lymphocytes (LAL) were then analysed. Results: Liver cirrhosis was established at the 12th week, and glutathione S-transferase placental form (GST-P)-positive lesions, which are known to be pre-neoplastic lesions, occupied 1.72 ± 0.84% of the liver at the 16th week in the controls. At this time the LAL showed an increase in the number of CD5-, CD8- and CD18-positive cells and augmentation of lectin-dependent cellular cytotoxicity (LDCC) activity. Furthermore, supplementation of BCAA increased the number of LAL, especially CD8-positive cells and natural killer cells, and augmented LDCC activity of LAL at the 16th week. The number of LAL was positively correlated with the valine concentration in the plasma and liver, and the area of GST-P-positive lesions tended to be decreased in the BCAA group. Conclusion: These findings suggest that BCAA administration has stimulatory effects on the local immune systems of the liver, which may have a potential to inhibit hepatocarcinogenesis. Moreover, among all amino acids valine might be an important amino acid for enhancing the immune function of LAL.

With a nitrosamine induced hamster pancreatic cancer cell line (HaP-T1), survival time and metastatic rates were compared between orthotopic cell implantation (OCI; n = 5) and orthotopic tissue implantation (OTI; n = 5) models. All the tumors were palpable (100% tumor take) after 1 week in both groups. Hamsters in the OCI group survived 71 +/- 2.17 days (range, 69-75 days), and in the OTI group, 73.8 +/- 4.03 days (range, 58-80 days). After necropsy, spontaneous metastases were noted in 100% of the animals. Direct invasion to adjacent organs was observed in four animals, and liver metastases, in three in the OTI group, which were significantly higher compared with the OCI group. On the other hand, peritoneal dissemination was observed only in the OCI group. Other metastatic sites showed no significant difference between the groups. All the histologically noted metastases had K-ras point mutation confirmed by polymerase chain reaction-restriction fragment length polymorphism (PCR/RFLP) analysis. We conclude that the homologous OTI model may be more useful than the OCI model. The OTI model may contribute to the development of therapeutic strategies in the field of pancreatic cancer research because of the capacity for invasion to adjacent organs, higher liver metastatic rate, and similarity to the clinical picture of the disease.

Background: Homologous orthotopic implantation models of pancreatic ductal cancer in Syrian golden hamsters can mimic the clinical features of the disease in humans. However, there have been no studies attempting a surgical approach to this animal model. Our aim was to clarify the possibility of curative resection in these tumor-bearing animals. Methods: A nitrosamine-induced pancreatic ductal adenocarcinoma cell line, HaP-T1, was used for the experiments. Hamsters underwent two surgeries: first, the cells or the tissue previously grown subcutaneously were implanted into the pancreas, and second, pancreatectomy and splenectomy were performed within the following 4 wk after implantation. Animals were examined until death, when necropsies were performed. Surgically resected and necropsied specimens were studied histopathologically and at the molecular level by detection of K-ras point mutation at codon 12. Results: At the second surgery, the success of implantation was 100%, and the curative resection rate was 75%. After observation for 400 d, 5 of 12 hamsters, which underwent second surgery within 15 d, were still alive. They were sacrificed and were found to be tumor-free. Conclusion: Orthotopic homologous implantation model of hamster pancreatic cancer combined with surgical resection may be useful for further research of adjuvant or neoadjuvant therapy for pancreatic carcinoma.

Background/Aims: Plasminogen activators and plasminogen activator inhibitors are important regulators of the balance between the proteolytic and antiproteolytic activities that determine extracellular matrix turnover. We examined the expression of plasminogen activator-plasmin system components in experimental liver fibrosis of rats. Methods: Liver fibrosis was produced in rats by injecting carbon tetrachloride for 6 to 12 weeks. Gene expression for plasminogen activator inhibitor-1 (PAI-1), urokinase and tissue plasminogen activators (uPA and tPA), urokinase plasminogen activator receptor (uPAR), and transforming growth factor-β 1 (TGF-β 1 ) was examined by Northern analysis. Western analysis was performed to detect protein expression of PAI-1, uPA and uPAR. An immunohistochemical study was performed to detect the localization of PAI-1. Additionally, primary cultured liver cells were examined by Northern and Western analyses for this protein with or without prior incubation with TGF-β 1 . Results: At 6 weeks, when fibrosis had occurred, uPA and uPAR mRNAs had increased 2.8-fold and 1.8-fold, respectively; PAI-1 and tPA mRNA levels were unchanged. At the cirrhotic stage (9 to 12 weeks), mRNA levels for PAI-1, uPA, uPAR and tPA were all increased. Western analysis also showed increased uPA and uPAR expressions in fibrotic liver, and increased PAI-1, uPA and uPAR expressions in cirrhotic liver. PAI-1 protein was also demonstrated immunohistochemically along sinusoids, vessels, and bile duct cells of normal and fibrotic liver. In liver cell cultures, Kupffer cells, hepatocytes, and especially stellate cells, expressed PAI-1. Expression was enhanced in stellate cells cultured from fibrotic or cirrhotic liver or stimulated in vitro with TGF-β 1 . Conclusion: Though increased uPA and uPAR may act on matrix degradation in fibrotic liver, increased PAI-1 together with uPA, uPAR and tPA are associated with overall inhibition of matrix degradation in cirrhotic liver. Hepatic stellate cells are an important source of PAI-1 during liver fibrosis.

Intrahepatic lymphoid follicle (ILF) formation is one of the most characteristic and commonly observed histological features in patients with chronic hepatitis C. However, little is known regarding whether follicles in the liver belong to functional lymphoid tissues, where B cells are activated, differentiated, and proliferated, or if the lymphocytes are merely infiltrated after recruitment from the secondary lymphoid organs. To ascertain this possibility, we examined the expression of markers for B-cell activation, differentiation, and proliferation in ILFs in patients with chronic hepatitis C using surgically resected specimens, and compared them with specimens of perihepatic lymph nodes by an immunohistochemical technique. Germinal center (GC) formation in the ILFs was frequently found in HCV-positive cases. The distribution of immunoglobulin M (IgM)-, IgD-, and IgG-positive cells and the expression patterns of Ki-67, CD23, or bcl-2 and bcl-6 gene products in the follicles with GC formation in the liver of patients with chronic hepatitis C were similar to those of lymph nodes, indicating that B cells are activated, proliferated, and differentiated in the ILFs with GC formation in patients with chronic hepatitis C. Oligoclonal expansion of B cells in the livers with ILFs was confirmed by an analysis of immunoglobulin heavy chain (IgH) gene rearrangement determined by polymerase chain reaction (PCR). These data strongly suggest that ILFs with GC formation, which are frequently found in patients with chronic hepatitis C, may functionally be the same as those found in lymph nodes with respect to B-cell expansion and maturation.

We report a 36–year-old female patient with small cell carcinoma of the gallbladder with liver metastasis, that was associated with pancreaticobiliary maljunction (PBM) without bile duct dilatation. On admission, a gallbladder mass and multiple liver tumors were detected by US, CT and ERCP. Moreover, ERCP clearly demonstrated an abnormally long common channel between the pancreatic and bile ducts, indicating PBMwithout choledochal dilatation. US-guided biopsy of the tumor histopathologically confirmed small cell carcinoma. The serum neuron-specific enolase (NSE) level was very high. Transarterial chemo-embolization followed by systemic chemotherapy resulted in marked tumor reduction, but this was followed by rapid exacerbation. The patient died of the tumor 145 days after admission. The proposed mechanism of the car-cinogenesis of this rare gallbladder malignancy under the preexisting condition of PBM was discussed. (Dig Endosc 1999, 11: 246–249)

Hepatic stellate cells (HSCs) play an important role in fibrogenesis of the liver through their activity of biosynthesizing a widely variety of extracellular matrix proteins. However, in vitro experiments using primary cultured HSCs are limited, because HSCs in primary culture tend to dedifferentiate quickly. We recently established several HSC lines by transfection with SV 40 large T antigen (SV40 TAg) without any culture crisis. The cell lines have both morphological and functional characteristics of HSCs. Immunohistochemically, these cell lines showed positive staining of SV 40 TAg in nuclei and desmin and type IV collagen in cytoplasms. Cell lines also had messenger RNA expressions of type I collagen α1 chain, α-smooth muscle actin, hepatocyte growth factor, transforming growth factor-β1. These cell lines would be useful for in vitro studies in regard to the mechanism of differentiation or transformation of rat HSCs.

Plasma levels of docosahexaenoic acid (DHA, C22:6 omega 3) were found to be decreased in 11 patients with alcoholic and non-alcoholic liver cirrhosis depending on the severity of liver damage. In this reduction, we found impaired metabolism of omega-3 long-chain polyunsaturated fatty acids in the cirrhotic liver and poor dietary intake of DHA to involved in the reduction of DHA plasma levels. The deficiency of this fatty acid, which is concentrated in the nervous tissues, may be related to the impaired neural function observed in hepatic encephalopathy of these patients. Oral DHA supplementation was supplied in the form of a polyunsaturated fatty acid-enriched soft oil capsule (omega 3/omega 6 ratio = 0.91, and P/S ratio = 1.87). Twelve capsules per day (containing 408 mg DHA, which corresponds to one-fourth of the DHA content in a normal daily diet) improved the DHA contents in the plasma phospholipid fractions of 5 alcoholic patients with low DHA levels.

About 90% of human pancreatic cancers carry K-ras point mutation, which may play an important role in tumorigenesis. We investigated the inhibitory effects of anti-sense oligonucleotides targeting K-ras point mutation on the growth of cultured human pancreatic cancer cells. Eight human pancreatic cancer cell lines were screened for K-ras codon 12 point mutations by PCR-RFLP analysis and direct sequencing. Then, 3 cell lines with the major types of K-ras point mutation, i.e.,HuP-T1, HuP-T3 and PANC-1, and 1 without mutation, BxPC-3, were used for the experiments. Seventeen mer anti-sense oligonucleotides were designed, targeting the point mutation of K-ras codon 12, and transfected into the cells by the liposome-mediated method. Cell-growth activities were estimated by MTT assay. Levels of K-ras mRNA expression were determined using quantitative RT-PCR, and K-ras p21 protein synthesis was evaluated with Western blotting. Mutation-matched anti-sense oligonucleotides effectively inhibited the growth of these pancreatic cancer cell lines, except for BxPC-3, by suppressing K-ras mRNA expression and K-ras p21 protein synthesis. Moreover, mutation-matched anti-sense oligonucleotides showed stronger anti-proliferative effects than did mutation-mismatched ones. Our results suggest that anti-sense therapy specific to point mutations of K-ras mRNA is a practical approach to selective suppression of tumor growth, with little effect on normal cells. Int. J. Cancer 80:553–558, 1999. © 1999 Wiley-Liss, Inc.

Hepatic perisinusoidal cell population consists of hepatic stellate cells, Kupffer cells, endothelial cells, and Pit cells. These cells are isolated by enzymic digestion and purified by density gradient centrifugation. With isolation of stellate cells, conventional method is unable to eliminate the contamination of Kupffer cells because the densities of these two cells are similar. We report here an improved method for isolation of highly purified hepatic stellate cells, using dichloromethylene diphosphate (CL2MDP), which has selective cytotoxicity of Kupffer cells. Three days after the single intravenous administration of liposome-encapsulated CL2MDP, the Kupffer cells disappeared almost completely from the liver. Following Percoll density gradient centrifugation, the purity of the hepatic stellate cells exceeded 98% without any contamination of the Kupffer cells. Kupffer cells are reported to affect the physiological functions of stellate cells. The availability of highly purified stellate cells will facilitate the investigation of their functions in primary culture.

We report the cases of three patients with anorexia nervosa (AN) who each recovered rapidly after experiencing a life-threatening episode with severe thrombocytopenia. All three cases were the typical restricting-type of AN, occurring in adolescence. They refused to be admitted to a hospital until their general condition had been severely deteriorated. Their lowest platelet counts were 2.9, 4.6, and 2.3 x 10(4)/mm3, respectively. Apparent hemorrhagic tendencies, such as purpura, gingival and nasal bleeding, and gastrointestinal bleeding were observed. The bone marrow examination showed apparent hypoplasia in two patients. No evidence of disseminated intravascular coagulation or autoantibody to platelets was detected. The platelet counts recovered rapidly by water and nutritional supplementation. The recovery from the AN itself was excellent in all three patients without specific psychotherapy.

Hepatic encephalopathy (HE) accompanied by an impairment of consciousness from orientation disorder (grade II) to coma (grade IV) is considered to be overt HE and is treated as an emergency. However, subclinical hepatic encephalopathy (SHE) can be detected by sensitive and quantitative neuropsychological examinations in cirrhotic patients without overt HE. The introduction of the SHE concept is clinically important for preventing the deterioration of SHE (grades 0 and I) to overt HE (grade II and more severe), prolonging the compensated state of cirrhosis without its deterioration to hepatic failure, and the continuation of patient treatment at home. We developed a new diagnostic method for SHE using a quantitative neuropsychological test, with the computerization of all operations. Evaluations of cerebral function and morphology are useful for the determination of the pathophysiology of HE, and assist the diagnosis of SHE. The latencies of the P3 wave in the visually evoked potential and the P300 wave in the event-related potential are prolonged in cirrhotic patients with SHE and are well expressed in three-dimensional coloured topograms (brain mapping). Automated polysomnographic analysis is useful for continuous-monitoring electroencephalograms (EEG) and for the detection of the sleep disturbance observed in cirrhotic patients with SHE. Brain atrophy in computed tomography (CT), magnetic resonance imaging (MRI) and high signals in the basal ganglia in the MR-T1-weighted images have frequently been observed in patients with SHE. The reduction of regional cerebral blood flow (rCBF) by 99m technetium-1, 1-ethylcysteinate dimer ( 99m Tc-ECD)-single photon emission computed tomography (SPECT) and the choline/N -acetylaspartic acid ratio by 1 proton-magnetic resonance spectroscopy ( 1 H-MRS) were observed in the hippocampus in patients with SHE. These approaches (cerebral function tests and imaging diagnoses of the brain) can also be used to evaluate the effectiveness of treatments for HE; for example, branched-chain amino acid (BCAA) was shown by automated continuous polysomnographic analysis to be a psychotropic drug which acts directly on the central nervous system and the clinical significance of choline administration to HE patients is now being evaluated by 'H-MRS and neuropsychological tests.

Seventy-five cirrhotic patients with hyperammonemia in the past or at the time of the study were randomly divided into two groups (treated with lactulose or nontreated) in 14 hospitals in Japan. Thirty-six cirrhotic patients were diagnosed as having subclinical hepatic encephalopathy (SHE), and 39 were diagnosed as non-SHE. SHE was diagnosed when the results of all three of the quantitative psychometric tests used (number connection test, and symbol digit and block design tests of the Wechsler adult intelligence scale [revised]) were abnormal as compared with age-matched normal values. The mean number of abnormal psychometric test results and the prevalence of SHE were used for a quantitative evaluation of the efficacy of the lactulose treatment. Twenty-two of the SHE patients were treated with lactulose (45 mL/d) for 8 weeks, and the other 14 SHE patients did not receive lactulose. In the SHE patients administered lactulose, the results of the quantitative psychometric evaluation were significantly improved at 4 and 8 weeks after the beginning of the lactulose administration. The SHE had disappeared in 10 (50%) of the 20 treated patients at week 8, but it persisted in 11 (85%) of the untreated 13 patients. We concluded that lactulose treatment in cirrhotic patients with SHE is effective with respect to psychometric tests.

We examined the relationship between the values of plasma protein induced by vitamin K absence or antagonist II (PIVKA-II) and serum α-fetoprotein (AFP) and the presence of AFP mRNA in the mononuclear cells of the peripheral blood in 50 patients with hepatocellular carcinoma (HCC) . AFP mRNA in the blood was detected in 26 of 50 patients with HCC (52.0%). The frequency of positive cases in each TNM stage was: none of 5 in stage I, 3 of 14 (21.4%) in stage II, 6 of 10 (60.0%) in stage III and 17 of 21 (81.0%) in stage IV. AFP mRNA was detected more frequently in patients with high serum AFP level (400 ng/ml and more than 400 ng/ml) (12/16, 75.0%) than in those with low AFP level (less than 400 ng/ml) (14/34, 41.2%) (P

Background/Aims: To examine the T-cell repertoire which is involved in the immunopathogenesis of chronic hepatitis, we analyzed the T-cell receptor Vβ gene usage in liver-infiltrating lymphocytes by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical technique. Methods: Complementary DNA was synthesized from RNA which was extracted from 26 liver biopsy specimens and from peripheral blood lymphocytes from eight subjects, and amplified by RT-PCR. Radioactivity of each amplified product using 32 P-labeled primers was measured and the percentage of each Vβ expression was calculated. Results: The mean frequency of Vβ5.1 (11.1%) in liver-infiltrating lymphocytes of chronic hepatitis C was highest among those of all Vβ regions, and was significantly higher than that in both peripheral blood lymphocytes of chronic hepatitis C and liver-infiltrating lymphocytes of chronic hepatitis B. In the immunohistochemical analysis, Vβ5.1-positive cells were mostly observed in portal areas where inflammatory reactions occurred. The sequence of the complementarity determining region (CDR)3 on T-cell receptor expressing Vβ5.1 were examined in six patients with chronic hepatitis C. The sequences were similar to each other and all had one common amino acid (valine) irrespective of different HLA haplotype. Conclusions: These data suggest that Vβ5.1-positive cells are preferentially accumulated in the liver of chronic hepatitis C and are involved in the immunopathogenesis of the disease. Sequence analysis showed that Vβ5.1-positive cells recognize a common conventional antigen and valine recognized at the same position of the CDR3 may be a key residue in determining an antigen/major histocompatibility complex contact point.

To investigate pathophysiological diversities in the repairing process of gastric ulcer, distribution density of basic fibroblast growth factor (bFGF)-positive fibroblasts and myofibroblasts and vascular endothelial cells, mucosal haemoglobin content, PAS-positive mucus amount and glandular index were compared in the peripheral region of an open ulcer (the unhealed group ; n = 17), the central region of a red scar (the red scar group ; n = 32) and the central region of the white scar (white scar group ; n = 32). Density of bFGF-positive fibroblasts and myofibroblasts and vascular endothelial cells was highest in the unhealed group, followed by the red scar group, while the white scar group showed a low value close to control. Mucosal haemoglobin content was high in the red scar and unhealed groups. PAS-positive mucus amount in the unhealed and red scar groups showed lower values compared with that in the white scar group. Glandular index in the unhealed group was the lowest, followed by the red scar group, while the white scar group neared control values. Statistically significant correlations were observed between the density of bFGF-positive 'fibroblasts and myofibroblasts' and density of bFGF-positive vascular endothelial cells, between the density of bFGF-positive vascular endothelial cells and mucosal haemoglobin content and between the PAS-positive mucus amount and glandular index. Discriminant analysis demonstrated that the unhealed group could be distinguished from the red and white scar groups, based on glandular index, density of bFGF-positive 'fibroblasts and myofibroblasts', mucosal haemoglobin content and PAS-positive mucus amount, while the red scar group could be discriminated from the white scar group based on the density of bFGF-positive 'fibroblasts and myofibroblasts', density of bFGF-positive vascular endothelial cells, glandular index, haemoglobin content and PAS-positive mucus amount. The white scar group was difficult to discriminate from control. Our present results show that the recovery of glandular formation and mucus production continues throughout the repairing process, whereas the acceleration of angiogenesis and granulation formation is observed only in unhealed ulcers and at the red scar stage.

Glycyrrhizin, a major component of a herb (licorice), has been intravenously used for the treatment of chronic hepatitis B in Japan and improves liver function with occasional complete recovery from hepatitis. This substance modifies the intracellular transport and suppresses sialylation of hepatitis B virus (HBV) surface antigen (HBsAg) in vitro. This study was designed to clarify the pharmacological basis for its effectiveness. The structure-bioactivity relationship of glycyrrhizin, glycyrrhetic acid 3-O-monoglucuronide and glycyrrhetic acid was determined, and glycyrrhetic acid was found to be the most active of them. The amounts of three substances bound to the liver were evaluated in guinea pigs after intravenous administration of glycyrrhizin. Glycyrrhizin and glycyrrhetic acid 3-O-monoglucuronide were detected at concentrations of 31.8-1.3 micrograms/g of liver, but glycyrrhetic acid was not detected. When glycyrrhizin attained these concentrations in the cellular fraction of the PLC/PRF/5 cell culture, it suppressed the secretion of HBsAg as reported previously. These results indicated that glycyrrhizin administered intravenously might bind to hepatocytes at the concentration at which glycyrrhizin could modify the expression of HBV-related antigens on the hepatocytes and suppress sialylation of HBsAg.

We designed and produced a new instrument that measures the color of the gastrointestinal mucosa during conventional endoscopic examination. The instrument consists fundamentally of an optical fiber sensor and a light source. One end of the optical fiber sensor is connected to the light source and a commercially available spectrophotometric colorimeter, while the other end is passed through the biopsy channel of a commercially available endoscope and placed in contact with the gastrointestinal mucosa. The color of the mucosa is then measured. To evaluate the accuracy of color measurement, the instrument was used to measure the color of 108 color chips belonging to the color range of the gastrointestinal mucosa or surrounding regions, selected from the Munsell Book of Color. The instrument correctly determined the relative hue, chroma and value of all 108 color chips tested. It also had the advantages of consistently measuring the color chips under standardized conditions and of promptly displaying the color readings during endoscopic examination. This instrument was also used to measure the color of the mucosa at different sites of the upper gastrointestinal tract in 20 patients. There were no noteworthy problems in using the instrument clinically. Mucosal color (hue, value and chroma) of the upper gastrointestinal tract was demonstrated to have specific characteristics according to the region measured. The instrument provided highly accurate, objective information that facilitated the distinction of slight differences in mucosal color, an important factor in the endoscopic diagnosis of gastrointestinal diseases.