An Image Fusion System for Estimating the Therapeutic Effects of Radiofrequency Ablation on Hepatocellular Carcinoma.

BACKGROUND: During ultrasound-guided radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC), high echoic areas due to RFA-induced microbubbles can help calculate the extent of ablation. However, these areas also decrease visualization of target tumors, making it difficult to assess whether they completely cover the tumors. To estimate the effects of RFA more precisely, we used an image fusion system (IFS). PATIENTS AND METHODS: We enrolled patients with a single HCC who received RFA with or without the IFS. In the IFS group, we drew a spherical marker along the contour of a target tumor on reference images immediately after administering RFA so that the synchronized spherical marker represented the contour of the target tumor on real-time ultrasound images. When the high echoic area completely covered the marker, we considered the ablation to be complete. We compared outcomes between the IFS and control groups. RESULTS: We enrolled 25 patients and 20 controls, and the baseline characteristics were similar between the two groups. The complete ablation rates during the first RFA session were significantly higher in the IFS group compared with those in the control group (88.0% vs. 60.0%, P = 0.041). The number of RFA sessions was significantly smaller in the IFS group compared with that in the control group (1.1 ± 0.3 vs. 1.5 ± 0.7, P = 0.016). CONCLUSIONS: The study suggested that the IFS enables a more precise estimation of the effects of RFA on HCC, contributing to enhanced treatment efficacy and minimized patient burden.

[Pathogenesis of alcoholic chronic hepatitis].

In 1983, Nei et al. reported that alcoholic chronic hepatitis (ACH)(chronic hepatitis induced tal area. Recently, the number of alcoholics patients diagnosed with ACH has been increased In this review, we discussed the characteristics of liver histopathology and blood chemistry of ACH patients. In ACH, pericellular fibrosis, ballooned hepatocytes and/or bridging fibrosis, and infiltration of mononuclear lymphocytes is decreased after 6 to 8 weeks of abstinence from results suggest that ACH could be one type of alcoholic liver disease. The precise mechanism by alcohol) as one type of alcoholic liver disease. Since then, it has been discussed whether alcohol abuse, suggesting that alcohol may play a role in the infiltration of mononuclear lym ACH is one type of alcoholic liver disease, because there could be infection of unknown hepatitis virus in alcoholics and it is not clear why mononuclear lymphocytes infiltrate into the porphocytes in portal region. After abstinence of alcohol, serum levels of AST, ALT, and γ-GTP in patients with ACH returned to normal as in other types of alcoholic liver disease such as alcoholic fatty liver, alcoholic fibrosis, alcoholic hepatitis and alcoholic liver cirrhosis. These results suggest that ACH could be one type of alcoholic liver disease. The precise mechanism of the infiltration of mononuclear lymphocytes into portal areas of ACH patients is not known. We propose that the reason for the infiltration of natural killer (T) cells into portal areas could be due to the influx of endotoxin into portal vein resulting from the increased permeability of gut induced by alcohol.

Advanced ultrasonography technologies to assess the effects of radiofrequency ablation on hepatocellular carcinoma.

BACKGROUND: Radiofrequency ablation (RFA) is a curative therapy for hepatocellular carcinoma (HCC). In RFA, ultrasonography (US) is most commonly used to guide tumor puncture, while its effects are assessed using dynamic computed tomography or magnetic resonance. The differences in modalities used for RFA and assessment of its effects complicate RFA. We developed a method for assessing the effects of RFA on HCC by combining contrast-enhanced (CE) US and real-time virtual sonography with three-dimensional US data. PATIENTS AND METHODS: Before RFA, we performed a sweep scan of the target HCC nodule and the surrounding hepatic parenchyma to generate three-dimensional US data. After RFA, we synchronized multi-planar reconstruction images derived from stored three-dimensional US data with real-time US images on the same US monitor and performed CEUS and real-time virtual sonography. Using a marking function, we drew a sphere marker along the target HCC nodule contour on pre-treatment US- multi-planar reconstruction images so that the automatically synchronized sphere marker represented the original HCC nodule contour on post-treatment real-time CEUS images. Ablation was considered sufficient when an avascular area with a margin of several millimeters in all directions surrounded the sphere marker on CEUS. RESULTS: This method was feasible and useful for assessing therapeutic effects in 13 consecutive patients with HCC who underwent RFA. In 2 patients who underwent multiple sessions of RFA, HCC-nodule portions requiring additional RFA were easily identified on US images. CONCLUSIONS: This method using advanced US technologies will facilitate assessment of the effects of RFA on HCC.

Association between functional promoter polymorphisms of macrophage migration inhibitory factor (MIF) gene and ulcerative colitis in Japan.

Macrophage migration inhibitory factor (MIF) is a key proinflammatory mediator. Two functional polymorphisms were identified in the promoter region of MIF gene. We attempted to clarify the associations between these polymorphisms and ulcerative colitis (UC). The study was performed in 111 patients with UC and 209 subjects without UC. We employed the PCR-SSCP method to detect gene polymorphisms. Overall, 5/5-CATT genotype was a decreased risk for the development of UC (OR, 0.51; 95% CI, 0.26-0.99). In addition, 7/7-CATT genotype was significantly associated with chronic continuous phenotype and distal colitis phenotype (OR, 5.49; 95% CI, 1.19-25.3, and OR, 6.10; 95% CI, 1.32-28.2, respectively), whereas 5/5-CATT genotype had an inhibitory effect on the development of UC after 20years of age (OR, 0.33; 95% CI, 0.14-0.82). On the other hand, G-173C polymorphism did not affect the susceptibility to and the phenotypes of UC. Our results suggested that tetranucleotide CATT repeat of MIF gene promoter may be associated with the development of UC and the severity of inflammation in patients with UC.

Inhibition of γ-glutamyltransferase ameliorates ischaemia-reoxygenation tissue damage in rats with hepatic steatosis.

BACKGROUND AND PURPOSE: Hepatic steatosis may be associated with an increased γ-glutamyltransferase (γ-GT) levels. Ischaemia-reoxygenation (IR) injury causes several deleterious effects. We evaluated the protective effects of a selective inhibitor of γ-GT in experimentally induced IR injury in rats with obesity and steatosis. EXPERIMENTAL APPROACH: Otsuka Long-Evans Tokushima Fatty (OLETF) rats with hepatic steatosis were used in the current study. The portal vein and hepatic artery of left lateral and median lobes were clamped to induce ischaemia. Before clamping, 1 ml of saline (IR group) or 1-ml saline containing 1 mg·kg-1 body weight of GGsTop (γ-GT inhibitor; IR-GGsTop group) was injected into the liver via the inferior vena cava. Blood flow was restored after at 30 min of the start of ischaemia. Blood was collected before, at 30 min after ischaemia and at 2 h and 6 h after reoxygenation. All the animals were killed at 6 h and the livers were collected. KEY RESULTS: Treatment with GGsTop resulted in significant reduction of serum ALT, AST and γ-GT levels and hepatic γ-GT, malondialdehyde, 4-hydroxy-2-nonenal and HMGB1 at 6 h after reoxygenation. Inhibition of γ-GT retained normal hepatic glutathione levels. There was prominent hepatic necrosis in IR group, which is significantly reduced in IR-GGsTop group. CONCLUSION AND IMPLICATIONS: Treatment with GGsTop significantly increased hepatic glutathione content, reduced hepatic MDA, 4-HNE and HMGB1 levels and, remarkably, ameliorated hepatic necrosis after ischaemia-reoxygenation. The results indicated that GGsTop could be an appropriate therapeutic agent to reduce IR-induced liver injury in obesity and steatosis.

[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.

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.

Serum endocan as a survival predictor for patients with liver cirrhosis.

BACKGROUND: The relationship between endocan expression and outcome in patients with chronic liver disease is not fully understood. OBJECTIVE: To examine whether serum endocan level is predictive of outcome in patients with liver cirrhosis. METHODS: A total of 68 patients with liver cirrhosis were enrolled. Outcome predictors were analyzed using the Cox proportional hazards model. The overall survival rates were calculated using the Kaplan-Meier method, and differences were evaluated using the log-rank test. RESULTS: During the median follow-up period (7.1 years), nine patients had hepatocellular carcinoma (HCC) and 10 patients died. Of the deceased patients, nine died due to hepatic decompensation or associated conditions. No significant factors were found to be predictive of the occurrence of HCC. In contrast, an elevated serum endocan level (≥2.0 ng/mL; HR 2.34 [95% CI 1.05 to 7.03]; P=0.037) and high Child-Pugh grade B/C (HR 2.65 [95% CI 1.30 to 6.89; P=0.006) were predictive of poor survival. Kaplan-Meier analysis revealed that the respective cumulative survival rates at five and 10 years were 97.1% and 87.4% in patients with serum endocan levels <2.0 ng/mL and 85.8% and 64.4% in patients with levels ≥2.0 ng/mL (P=0.009), respectively. Moreover, the cumulative survival rates were significantly different among the patient groups divided according to serum endocan level and Child-Pugh grade (P=0.002). CONCLUSION: These findings suggest that serum endocan level may be a survival predictor for patients with liver cirrhosis.

Regulation of CYP2E1 by ethanol and palmitic acid and CYP4A11 by clofibrate in primary cultures of human hepatocytes.

CYP2E1 and CYP4A11 are cytochrome P450 enzymes that are regulated by physiological conditions including diabetes and fasting. In addition, the xenochemical clofibrate has been reported to induce both rodent CYP2E1 and CYP4A. These findings suggest similar modes of regulation. Also in common to both enzymes is the ability to metabolize fatty acids such as laurate and arachidonic acid. Here, we used primary cultures of human hepatocytes to determine if certain xenochemicals could regulate CYP2E1 and CYP4A11. Ethanol significantly (p < 0.05) increased expression of CYP2E1 mRNA by 216 +/- 32% of control, but did not alter CYP4A11 mRNA accumulation (145 +/- 22% of control). In contrast, hepatocytes exposed to ethanol exhibited only a slight elevation in CYP2E1 protein (122 +/- 13% of control) and a negligible effect on CYP4A11 protein. Clofibrate significantly (p < 0.05) enhanced both CYP4A11 mRNA and protein by 239 +/- 30% and 154 +/- 10% of control, respectively, but did not increase CYP2E1. Because rodent CYP4A is reportedly regulated by fatty acids through peroxisome proliferator activated receptor alpha (PPARalpha) and CYP2E1 is induced by high fat diets, we examined the effects of a medium chain fatty acid, palmitate on CYP2E1 mRNA content. Palmitic acid significantly (p < 0.05) increased CYP2E1 mRNA to 326 +/- 57% of control. Collectively, results presented here identify agents that enhance CYP2E1 and CYP4A11 at the transcription level and suggest that fatty acids may represent a similar mode of regulation for these P450 enzymes. The lack of induction of CYP2E1 protein by ethanol in human hepatocytes indicates that for certain P450 enzymes, isolated hepatocytes may not be an adequate tool for predicting in vivo responses.

Serum endocan as a novel prognostic biomarker in patients with hepatocellular carcinoma.

Endocan is a vascular endothelium-derived factor regulated by angiogenic factors. The aim of this study was to determine whether serum endocan levels are prognostic for survival in patients with hepatocellular carcinoma (HCC). Serum endocan levels were measured in 64 HCC patients who were naïve to treatment, eight apparently healthy subjects, and 68 patients with liver cirrhosis; the latter two groups served as controls. Prognostic factors for the survival of HCC patients were examined using a Cox proportional hazards model. The median serum endocan levels were 1.145 ng/mL (range, 0.93-1.68 ng/mL) in healthy subjects, 1.93 ng/mL (range, 0.45-8.47 ng/mL) in liver cirrhosis patients, and 3.73 ng/mL (range, 0.74-10.95 ng/mL) in HCC patients (P = 0.0001). In HCC patients, elevated serum endocan levels were significantly associated with poor hepatic function (P = 0.015), a greater number of tumors (P = 0.034), and vascular invasion (P = 0.043). The median follow-up period was 23.0 months, and 33 HCC patients died during follow up. Multivariate analysis showed that serum endocan levels ≥ 2.20 ng/mL (hazard ratio 2.36, 95% confidence interval 1.22-5.36, P = 0.008) as well as elevated serum α-fetoprotein and des-γ-carboxy prothrombin levels were independent prognostic biomarkers for poor survival. The combination of serum endocan and these two additional markers was significantly predictive of worse survival (P < 0.0001). Thus, serum endocan may be a prognostic biomarker for survival in HCC patients, and the combination of serum endocan, α-fetoprotein, and des-γ-carboxy prothrombin levels can result in better prognostic stratification of these patients.

-449 C>G polymorphism of NFKB1 gene, coding nuclear factor-kappa-B, is associated with the susceptibility to ulcerative colitis.

AIM: To clarify the association between a polymorphism -449 C>G (rs72696119) in 5'-UTR of NFKB1 with ulcerative colitis (UC). METHODS: The studied population comprised 639 subjects, including patients with UC (UC cases, n = 174) and subjects without UC (controls, n = 465). We employed polymerase chain reaction-single strand conformation polymorphism to detect the gene polymorphism. RESULTS: The rs72696119 G allele frequencies in controls and UC cases were 33.4% and 38.5%, respectively (P = 0.10). Genotype frequency of the GG homozygote in UC cases was significantly higher than that in controls (P = 0.017), and the GG homozygote was significantly associated with susceptibility to UC [odds ratio (OR), 1.88; 95%CI, 1.13-3.14]. In male subjects, the GG homozygote was associated with an increased risk for UC (OR, 3.10; 95%CI, 1.47-6.54; P = 0.0053), whereas this association was not found in female subjects. In addition, the GG homozygote was significantly associated with the risk of non-continuous disease (OR, 2.06; 95%CI, 1.12-3.79; P = 0.029), not having total colitis (OR, 2.40; 95%CI, 1.09-3.80, P = 0.040), disease which developed before 20 years of age (OR, 2.80; 95%CI, 1.07-7.32, P = 0.041), no hospitalization (OR, 2.28; 95%CI, 1.29-4.05; P = 0.0090) and with a maximum of 8 or less on the UCDAI score (OR, 2.45; 95%CI, 1.23-4.93; P = 0.022). CONCLUSION: Our results provide evidence that NFKB1 polymorphism rs72696119 was significantly associated with the development of UC. This polymorphism influences the susceptibility to and pathophysiological features of UC.

Association between common genetic variant of HRH2 and gastric cancer risk.

Histamine plays important physiological roles in the upper gastrointestinal tract and acts via the H2 receptor. The -1018 G>A (rs2067474) in an enhancer element of the promoter and non-synonymous rs79385261 (Asn46Thr) were identified in HRH2. We attempted to clarify the associations of these polymorphisms with gastric carcinogenesis. The study was performed in 321 patients with gastric cancer and 599 subjects with no evidence of gastric malignancies on upper gastroduodenal endoscopy. The genotypes were determined using a one-tube multiplex PCR-SSCP method. The degree of gastritis was assessed in 496 subjects and serum pepsinogen (PG) I/II levels were measured in 124 subjects without gastric cancer. The minor allele of Asn46Thr could not be detected. The frequencies of the -1018 A allele in the non-GC and GC groups were 13.5% and 8.26%, respectively (p=0.00077). Overall, -1018 GG homozygotes had an increased risk for developing gastric cancer (OR 1.68; 95% CI 1.17-2.42; p=0.0052), especially intestinal type cancer (OR 1.94; 95% CI 1.23-3.08; p=0.0047). In subjects aged >60 years, the adjusted risk for gastric cancer among individuals who were -1018 GG homozygotes was 1.87 (range 1.19-2.93; p=0.0065) compared with A carriers. In the gastric cancer cases located in the antrum and at comparative advanced stage, -1018 GG homozygosity was a significantly increased risk factor. In subjects >60 years, the metaplasia score was significantly higher in -1018 GG homozygotes than A carriers. Both atrophy and metaplasia scores were significantly increased with age only in -1018 GG homozygotes. The PG I/II ratio was significantly decreased in H. pylori positive GG homozygotes than negative GG homozygotes and positive A carriers. Our results suggest that -1018 GG homozygosity of HRH2 may be associated with the severity of gastric mucosal atrophy. This genotype has an increased risk for the subsequent development of gastric cancer, especially intestinal type, at advanced age.

Functional promoter polymorphisms of macrophage migration inhibitory factor in peptic ulcer diseases.

Macrophage migration inhibitory factor (MIF) is a key proinflammatory mediator, which plays a pivotal role in inflammatory and immune diseases. We attempted to clarify the association of functional polymorphisms of MIF gene promoter with the development of gastro-duodenal ulcer. The study was performed in 471 stocked DNAs obtained from the subjects, including 93 healthy volunteers, with no evidence of gastric malignancy. We employed the PCR-SSCP method to detect gene polymorphisms. In all 471 DNAs, 92 and 43 were obtained from gastric and duodenal ulcer patients, respectively. By an unadjusted analysis, infection with Helicobacter pylori (H. pylori), male gender and non-steroidal anti-inflammatory drug (NSAID/aspirin) use were significantly associated with a risk for developing a gastric ulcer, whereas MIF promoter polymorphisms were not. On the other hand, infection with H. pylori, male gender and 7-CATT repeat at position -794 were significantly associated with the development of a duodenal ulcer, whereas NSAID/ aspirin use was not. By the analysis after adjustment for age, gender, NSAID/aspirin use and H. pylori infection status, 7/7-CATT homozygote had a significantly increased risk for the development of duodenal ulcers (OR, 6.31; 95% CI, 1.50-26.6; p=0.012). No factors were significantly associated with the development of peptic ulcers in NSAID/aspirin users. Our results suggested that tetranucleotide repeat polymorphism of MIF gene promoter might be associated with the development of duodenal ulcers.

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.

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.