Serum miR-379 expression is related to the development and progression of hypercholesterolemia in non-alcoholic fatty liver disease.

INTRODUCTION: Non-alcoholic fatty liver disease (NAFLD) has a wide spectrum, eventually leading to cirrhosis and hepatic carcinogenesis. We previously reported that a series of microRNAs (miRNAs) mapped in the 14q32.2 maternally imprinted gene region (Dlk1-Dio3 mat) are related to NAFLD development and progression in a mouse model. We examined the suitability of miR-379, a circulating Dlk1-Dio3 mat miRNA, as a human NAFLD biomarker. METHODS: Eighty NAFLD patients were recruited for this study. miR-379 was selected from the putative Dlk1-Dio3 mat miRNA cluster because it exhibited the greatest expression difference between NAFLD and non-alcoholic steatohepatitis in our preliminary study. Real-time PCR was used to examine the expression levels of miR-379 and miR-16 as an internal control. One patient was excluded due to low RT-PCR signal. RESULTS: Compared to normal controls, serum miR-379 expression was significantly up-regulated in NAFLD patients. Receiver operating characteristic curve analysis suggested that miR-379 is a suitable marker for discriminating NAFLD patients from controls, with an area under the curve value of 0.72. Serum miR-379 exhibited positive correlations with alkaline phosphatase, total cholesterol, low-density-lipoprotein cholesterol and non-high-density-lipoprotein cholesterol levels in patients with early stage NAFLD (Brunt fibrosis stage 0 to 1). The correlation between serum miR-379 and cholesterol levels was lost in early stage NAFLD patients treated with statins. Software-based predictions indicated that various energy metabolism-related genes, including insulin-like growth factor-1 (IGF-1) and IGF-1 receptor, are potential targets of miR-379. CONCLUSIONS: Serum miR-379 exhibits high potential as a biomarker for NAFLD. miR-379 appears to increase cholesterol lipotoxicity, leading to the development and progression of NAFLD, via interference with the expression of target genes, including those related to the IGF-1 signaling pathway. Our results could facilitate future research into the pathogenesis, diagnosis, and treatment of NAFLD.

Antifibrotic effects of ambrisentan, an endothelin-A receptor antagonist, in a non-alcoholic steatohepatitis mouse model.

AIM: To examine the effects of the endothelin type A receptor antagonist ambrisentan on hepatic steatosis and fibrosis in a steatohepatitis mouse model. METHODS: Fatty liver shionogi (FLS) FLS-ob/ob mice (male, 12 wk old) received ambrisentan (2.5 mg/kg orally per day; n = 8) or water as a control (n = 5) for 4 wk. Factors were compared between the two groups, including steatosis, fibrosis, inflammation, and endothelin-related gene expression in the liver. RESULTS: In the ambrisentan group, hepatic hydroxyproline content was significantly lower than in the control group (18.0 µg/g ± 6.1 µg/g vs 33.9 µg/g ± 13.5 µg/g liver, respectively, P = 0.014). Hepatic fibrosis estimated by Sirius red staining and areas positive for α-smooth muscle actin, indicative of activated hepatic stellate cells, were also significantly lower in the ambrisentan group (0.46% ± 0.18% vs 1.11% ± 0.28%, respectively, P = 0.0003; and 0.12% ± 0.08% vs 0.25% ± 0.11%, respectively, P = 0.047). Moreover, hepatic RNA expression levels of procollagen-1 and tissue inhibitor of metalloproteinase-1 (TIMP-1) were significantly lower by 60% and 45%, respectively, in the ambrisentan group. Inflammation, steatosis, and endothelin-related mRNA expression in the liver were not significantly different between the groups. CONCLUSION: Ambrisentan attenuated the progression of hepatic fibrosis by inhibiting hepatic stellate cell activation and reducing procollagen-1 and TIMP-1 gene expression. Ambrisentan did not affect inflammation or steatosis.

Nimesulide, a cyclooxygenase-2 selective inhibitor, suppresses obesity-related non-alcoholic fatty liver disease and hepatic insulin resistance through the regulation of peroxisome proliferator-activated receptor γ.

Cyclooxygenase (COX)-2 selective inhibitors suppress non-alcoholic fatty liver disease (NAFLD); however, the precise mechanism of action remains unknown. The aim of this study was to examine how the COX-2 selective inhibitor nimesulide suppresses NAFLD in a murine model of high-fat diet (HFD)­induced obesity. Mice were fed either a normal chow diet (NC), an HFD, or HFD plus nimesulide (HFD-nime) for 12 weeks. Body weight, hepatic COX-2 mRNA expression and triglyceride accumulation were significantly increased in the HFD group. Triglyceride accumulation was suppressed in the HFD-nime group. The mRNA expression of hepatic peroxisome proliferator-activated receptor Î³ (PPARγ) and the natural PPARγ agonist 15-deoxy-Δ12,14-prostaglandin J2 (15d­PGJ2) were significantly increased in the HFD group and significantly suppressed in the HFD-nime group. Glucose metabolism was impaired in the HFD group compared with the NC group, and it was significantly improved in the HFD-nime group. In addition, the plasma insulin levels in the HFD group were increased compared with those in the NC group, and were decreased in the HFD-nime group. These results indicate that HFD-induced NAFLD is mediated by the increased hepatic expression of COX-2. We suggest that the production of 15d-PGJ2, which is mediated by COX-2, induces NAFLD and hepatic insulin resistance by activating PPARγ. Furthermore, the mRNA expression of tissue inhibitor of metalloproteinases-1 (TIMP­1), procollagen-1 and monocyte chemoattractant protein-1 (MCP-1), as well as the number of F4/80-positive hepatic (Kupffer) cells, were significantly increased in the HFD group compared with the NC group, and they were reduced by nimesulide. In conclusion, COX-2 may emerge as a molecular target for preventing the development of NAFLD and insulin resistance in diet-related obesity.

A Series of microRNA in the Chromosome 14q32.2 Maternally Imprinted Region Related to Progression of Non-Alcoholic Fatty Liver Disease in a Mouse Model.

BACKGROUND & AIMS: Simple steatosis (SS) and non-alcoholic steatohepatitis (NASH) are subtypes of non-alcoholic fatty liver disease (NAFLD), and the pathogenic differences between SS and NASH remain unclear. MicroRNAs (miRNAs) are endogenous, non-coding, short RNAs that regulate gene expression. The aim of this study was to use animal models and human samples to examine the relationship between miRNA expression profiles and each type of NAFLD (SS and NASH). METHODS: DD Shionogi, Fatty Liver Shionogi (FLS) and FLS ob/ob mice were used as models for normal control, SS and NASH, respectively. Microarray analysis and real-time PCR were used to identify candidate NAFLD-related miRNAs. Human serum samples were used to examine the expression profiles of these candidate miRNAs in control subjects and patients with SS or NASH. RESULTS: Fourteen miRNAs showed clear expression differences among liver tissues from SS, NASH, and control mice with good reproducibility. Among these NAFLD candidate miRNAs, seven showed similar expression patterns and were upregulated in both SS and NASH tissues; these seven candidate miRNAs mapped to an miRNA cluster in the 14q32.2 maternally imprinted region delineated by delta-like homolog 1 and type III iodothyronine deiodinase (Dlk1-Dio3 mat). Software-based predictions indicated that the transforming growth factor-ß pathway, insulin like growth factor-1 and 5' adenosine monophosphate activated protein kinase were potential targets of theses Dlk1-Dio3 mat NAFLD candidate miRNAs. In addition, serum samples from patients with SS or NASH differed markedly with regard to expression of the putative Dlk1-Dio3 mat miRNAs, and these differences accurately corresponded with NAFLD diagnosis. CONCLUSION: The expression profiles of seven miRNAs in 14q32.2 mat have high potential as biomarkers for NAFLD and for improving future research on the pathogenesis and treatment of NASH.

Therapeutic effects of the dipeptidyl peptidase-IV inhibitor, sitagliptin, on non-alcoholic steatohepatitis in FLS-ob/ob male mice.

Non-alcoholic steatohepatitis is characterized by hepatic fat accumulation, inflammation and varying degrees of fibrosis. The dipeptidyl peptidase­IV enzyme is important in glucose metabolism, as well as lipid accumulation, extracellular matrix metabolism and immune stimulation. Furthermore, the enzyme activity of dipeptidyl peptidase­IV is known to be increased in non­alcoholic steatohepatitis. Therefore, dipeptidyl peptidase­IV inhibitors are potential therapeutic agents for non­alcoholic steatohepatitis. The present study assessed the therapeutic effects of sitagliptin, a dipeptidyl peptidase­IV inhibitor, on non­alcoholic steatohepatitis using fatty liver Shionogi­ob/ob male mice. Sitagliptin (2 mg/kg/day; n=10) or placebo (control; n=10) was orally administered to fatty liver Shionogi­ob/ob mice for 12 weeks, and hepatic steatosis, fibrosis, inflammation and oxidative stress were assessed in comparison with the controls. Sitagliptin administration reduced body weight and blood glucose levels, and improved hepatic fibrosis. It also inhibited the gene expression levels of fatty acid synthase, transforming growth factor­ß1, tissue inhibitor of metalloproteinases­1, procollagen­type 1, tumor necrosis factor­α, monocyte chemoattractant protein­1 and enhanced peroxisome proliferator activated receptor­α. Furthermore, a marked attenuation of hepatic stellate cell activation and Kupffer cells was observed in the sitagliptin group. A decrease in oxidative stress and apoptosis was also observed. Sitagliptin attenuated the progression of hepatic fibrosis by improving lipid metabolism, inflammation and oxidative stress in non-alcoholic steatohepatitis.

Clinical usefulness of the ablative margin assessed by magnetic resonance imaging with Gd-EOB-DTPA for radiofrequency ablation of hepatocellular carcinoma.

BACKGROUND & AIMS: The aim of this study was to investigate the feasibility of ablative margin (AM) grading by magnetic resonance imaging (MRI) with Gd-EOB-DTPA administered prior to radiofrequency ablation (RFA), and to identify factors for achieving a sufficient AM and predictors for local tumor progression. METHODS: A total of 124 hepatocellular carcinomas (HCCs) were treated by RFA after Gd-EOB-DTPA administration. MRI and enhanced CT were performed within seven hours and one month after RFA. The AM assessment was categorized using three grades: AM (+), low-intensity area with continuous high-intensity rim; AM zero, low-intensity area with discontinuous high-intensity rim; and AM (-), low-intensity area extends beyond the high-intensity rim. Patients were followed and local tumor progression was observed. RESULTS: AM (+), AM zero, AM (-), and indeterminate were found in 34, 33, 26, and 31 nodules, respectively. The overall agreement rate between MRI and enhanced CT for the diagnosis of AM was 56.8%. The κ coefficient was 0.326 (p<0.001), indicating moderate agreement. Multivariate logistic regression analysis showed that a significant factor for the achievement of AM (+) on MRI was no contiguous vessels. The cumulative local tumor progression rates (0% at 1, 2, and 3 years) in 33 AM (+) nodules were significantly lower than those (3.6%, 11.5%, and 18.3% at 1, 2, and 3 years respectively) in 32 AM zero nodules. A multivariate Cox proportional hazards model identified tumor size as an independent predictor for local tumor progression. CONCLUSION: Gd-EOB-DTPA-MRI enabled an early assessment of RFA effectiveness in the majority ofHCC nodules. Local tumor progression was not detected in AM (+) nodules during the follow-up.

Rapid and early α-fetoprotein and des-γ-carboxy prothrombin responses to initial arterial infusion chemotherapy predict treatment outcomes of advanced hepatocellular carcinoma.

The aim of the present study was to predict the effects of transarterial infusion (TAI) chemotherapy based on early changes in α-fetoprotein (AFP) and des-γ-carboxy prothrombin (DCP) in patients with advanced hepatocellular carcinoma (HCC). Seventy-four patients who underwent TAI with cisplatin, 5-fluorouracil, mitomycin C and epirubicin for advanced HCC were enrolled. Antitumor responses were evaluated 6 months after TAI. Rapid and early responses were defined as the ratio of AFP or DCP after 1 week and 1 month compared to baseline. A total of 5, 10, 17 and 42 patients had complete response (CR), partial response (PR), stable disease (SD) or progressive disease (PD), respectively. Early AFP response was significantly lower in the CR+PR compared to the SD+PD groups (P<0.01). The early DCP response was significantly lower in the CR+PR compared to the SD+PD. The sensitivity and specificity of rapid and early AFP responses in the CR+PR were 0.78 and 0.72, and 0.80 and 0.73, respectively, and those of rapid and early DCP responses were 0.67 and 0.65, and 0.77 and 0.71, respectively. The combination of AFP and DCP responses had higher specificity compared to AFP or DCP alone responses. Patients were divided into responder and non-responder groups to evaluate the prediction of survival outcome. Early responders of AFP, DCP and AFP+DCP, who were divided based on the cut-off values of CR+PR survived significantly longer than the non-responders (P<0.05). In conclusion, rapid or early responses of AFP and/or DCP levels 1 and 4 weeks after TAI chemotherapy helped to predict the treatment effects.

Usefulness of contrast-enhanced ultrasound with Sonazoid for evaluating liver abscess in comparison with conventional B-mode ultrasound.

AIM: The purpose of this study was to evaluate the usefulness of contrast-enhanced ultrasound (CEUS) with Sonazoid (perfluorobutane) in patients with liver abscess. Sonazoid is a contrast agent with a low mechanical index and is phagocytosed by Kupffer cells. METHODS: Twenty-two patients with liver abscess were evaluated with conventional US, real-time CEUS with Sonazoid, and enhanced computed tomography (CT). After 0.5 mL of Sonazoid was administrated i.v., CEUS images in the vascular and post-vascular phases were observed. RESULTS: Conventional US showed hypoechoic lesions in 13 (59.1%), isoechoic in four (18.2%), hyperechoic in two (9.1%), mixed echoic in two (9.1%) and undetected in one (4.8%) patient. CEUS showed perilesional enhancement in 19 (86.4%) lesions in the vascular phase and well-defined unenhanced areas in 22 (100%) lesions in the post-vascular phase. CEUS revealed that 18 abscesses were cystic type and three were honeycomb type. Twenty-one abscesses (95.5%) had clearer appearances on CEUS than on conventional US in regard to the extent of necrotic or liquefied lesions seen. We could confirm reduction of the lesions after therapy in 13 (92.9%) of 14 patients followed up by CEUS. CONCLUSION: Most of the liver abscesses showed perilesional enhancement in the vascular phase and unenhanced areas in the post-vascular phase. The appearance of liver abscesses was clearer on CEUS than on conventional US. CEUS with Sonazoid can be a more effective diagnostic and therapeutic tool for liver abscess.

Therapeutic effects of the direct renin inhibitor, aliskiren, on non-alcoholic steatohepatitis in fatty liver Shionogi ob/ob male mice.

AIM: Non-alcoholic steatohepatitis (NASH) is a manifestation of metabolic syndrome in the liver that is characterized by hepatic fat accumulation, inflammation and varying degrees of fibrosis. The renin-angiotensin system (RAS) appears to play important roles in NASH. Direct renin inhibitors (DRI) reduce plasma renin activity (PRA) through interaction with the active site of the enzyme and reduce the formation of angiotensin-II (AT-II). Therefore, the DRI aliskiren may further suppress the RAS. This study examined the effects of aliskiren on NASH in fatty liver Shionogi (FLS)-ob/ob male mice that are the closest animal model of metabolic syndrome-related NASH in humans. METHODS: Aliskiren (100 mg/kg per day, aliskiren group) or a placebo (control group) was p.o. administrated to eight FLS-ob/ob mice each for 16 weeks and factors including steatosis, fibrosis, inflammation and oxidative stress were compared between the two groups. RESULTS: Amounts of hepatic fibrosis were significantly lower in the aliskiren group than in the control group. Areas of α-smooth muscle actin positivity, the numbers of F4/80 positive, 8-hydroxy-2-deoxyguanosine positive cells and immunohistochemical staining of 4-hydroxynonenal were also significantly decreased in the aliskiren group. Levels of RNA expression for transforming growth factor-ß1, connective tissue growth factor and monocyte chemoattractant protein-1 were significantly lower in the aliskiren group. CONCLUSION: Aliskiren attenuated the progression of hepatic fibrosis by inhibiting the activation of hepatic stellate and Kupffer cells and by reducing oxidative stress.

Assessment of track microbubble flow signals on contrast-enhanced ultrasound with perflubutane following percutaneous liver biopsy.

OBJECTIVE: It is important to detect post-liver biopsy hemorrhage early and confirm hemostasis in the clinical setting. Contrast-enhanced ultrasound (CEUS) is a sensitive and highly specific tool for detecting active bleeding. The aim of this study was to investigate the rate of detection of track microbubble flow signals on CEUS with perflubutane following liver biopsy and to assess the disappearance of these signals. METHODS: Microbubble flow signals along the needle track on CEUS were examined in 100 patients who underwent percutaneous US-guided liver biopsies. The microbubble flow signals were examined repeatedly until their disappearance. The patients were followed up with clinical and laboratory data to detect clinically significant hemorrhaging. RESULTS: Microbubble flow signals on CEUS following percutaneous liver biopsy were seen in 33% of the patients. There were no significant differences in the platelet count, prothrombin time or length of the biopsy specimen between the patients with and those without microbubble flow signals on CEUS. The microbubble flow signals disappeared over time in all patients. There were no cases of clinically significant hemorrhaging in the present study. CONCLUSION: Track microbubble flow signals on CEUS are frequently observed after biopsies. The disappearance of a microbubble flow signal is a useful index for confirming hemostasis of postbiopsy hemorrhaging.

Ablative margin states by magnetic resonance imaging with ferucarbotran in radiofrequency ablation for hepatocellular carcinoma can predict local tumor progression.

BACKGROUND: Our aim was to determine how well ablative margin (AM) grading assessed by magnetic resonance imaging (MRI) with ferucarbotran administered prior to radiofrequency ablation (RFA) predicts local tumor progression in comparison with enhanced computed tomography (CT). METHODS: 101 hepatocellular carcinomas were treated by RFA after ferucarbotran administration. We performed T2*-weighted MRI after 1 week and enhanced CT after 1 month. The assessment was categorized in three grades: AM(+): high-intensity area with continuous low-intensity rim; AM zero: high-intensity area with discontinuous low-intensity rim; and AM(-): high-intensity area extending beyond the low-intensity rim. RESULTS: AM(+), AM zero, AM(-) and indeterminable were found in 47, 36, 8 and 10 nodules, respectively. The overall agreement rate between MRI and enhanced CT for the diagnosis of AM was 71.3%. The κ coefficient was 0.523 (p < 0.001), indicating moderate agreement. Multivariate logistic regression showed that a significant factor for the achievement of AM(+) on MRI was only segment location (odds ratio 5.9, non-segment 4 + 8 vs. segment 4 + 8). The cumulative local tumor progression rates (4.4, 7.6, and 7.6% in 1, 2, and 3 years) in 47 AM(+) nodules were significantly lower than those (13.9, 33.4, and 41.8% in 1, 2, and 3 years) in 36 AM zero nodules. A multivariate Cox proportional hazards model identified contiguous vessels (odds ratio 12.0) and AM(+) on MRI (odds ratio 0.19) as independent factors for local tumor progression. CONCLUSION: AM assessment by MRI using ferucarbotran can predict local tumor progression after RFA and enable early and less invasive diagnosis.

Fatty liver Shionogi-ob/ob mouse: A new candidate for a non-alcoholic steatohepatitis model.

AIM: The fatty liver Shionogi (FLS) mouse develops hereditary fatty liver without obesity. The FLS-ob/ob mouse made by transferring the leptin(ob) gene demonstrates several metabolic disorders and marked fat deposition in the liver. The aim was to evaluate which mouse model, the FLS or FLS-ob/ob, is more useful for non-alcoholic steatohepatitis research. METHODS: FLS (n = 40) and FLS-ob/ob (n = 40) mice were fed a standard diet for 12, 24, 36 and 48 weeks, and then killed. The degree of fat deposition, oxidative stress, fibrosis and apoptosis were analyzed in the liver. Hepatic mRNA expression of fibrogenic and pro-inflammatory cytokines was measured. RESULTS: FLS mice developed hepatic steatosis and slight fibrosis without obesity between 12 and 48 weeks of age. Conversely, FLS-ob/ob mice developed severe steatosis at 12 weeks of age, and steatohepatitis with increased oxidative stress and advanced fibrosis between 24 and 36 weeks of age. At 48 weeks of age, some FLS-ob/ob but not FLS mice, progressed to cirrhosis. Transforming growth factor-ß1, connective tissue growth factor and tumor necrosis factor-α mRNA expression levels were greater in FLS-ob/ob than FLS mice between 24 and 48 weeks of age. The number of apoptotic cells in the liver was greater at 12 weeks of age in FLS mice and at 48 weeks of age in FLS-ob/ob mice. CONCLUSION: FLS-ob/ob mice developed more severe steatohepatitis with fibrosis than FLS mice, and had increased oxidative stress and apoptosis. These findings indicate that the FLS-ob/ob mouse is a more useful model for steatohepatitis research.

Therapeutic effects of angiotensin II type 1 receptor blocker, irbesartan, on non-alcoholic steatohepatitis using FLS-ob/ob male mice.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of a metabolic syndrome characterized by accumulation of hepatic fat, inflammation and varying degrees of fibrosis. Angiotensin (AT)-II has been reported to play a role in the establishment of NASH. This study examined the effects of an AT-II receptor blocker, irbesartan, on NASH using fatty liver Shionogi (FLS)-ob/ob male mice as the closest animal model of human metabolic syndrome-related NASH. Irbesartan (30 mg/kg/day) was orally administered to FLS-ob/ob mice for 12 weeks (irbesartan group). The effects of irbesartan on steatohepatitis were examined using factors including steatosis, fibrosis, inflammation and oxidative stress. The areas of hepatic fibrosis and hepatic hydroxyproline content were significantly lower in the irbesartan group compared to controls. The areas of α-smooth muscle actin-positivity and F4/80-positive cells were significantly decreased in the irbesartan group. The percentage of 8-hydroxy-2-deoxyguanosine (8-OHdG)-positive cells and 8-OHdG DNA content were significantly decreased in the irbesartan group compared to controls. Levels of RNA expression for procollagen I, transforming growth factor ß1, tumor necrosis factor-α, sterol regulatory element-binding protein 1c and fatty acid synthase were significantly lower in the irbesartan group compared to controls. In contrast, the gene expression of peroxisome proliferator activated receptor-α was significantly higher in the irbesartan group compared to controls. Irbesartan administration improved hepatic steatosis and attenuated the progression of hepatic fibrosis by inhibiting the activation of hepatic stellate cells and Kupffer cells and reducing oxidative stress.

Assessment of ablative margin by unenhanced magnetic resonance imaging after radiofrequency ablation for hepatocellular carcinoma.

PURPOSE: The aim of this study was to evaluate the feasibility of magnetic resonance imaging (MRI) without a contrast agent to visualize the ablative margin after radiofrequency ablation (RFA) for hepatocellular carcinoma (HCC), compared with enhanced CT. METHODS: Twenty-five HCCs in 19 patients were treated by RFA. T1-weighted MRI was performed before and after RFA, and the signal intensities of the tumors and surrounding liver tissues were measured. Treatment efficacy was assessed based on three grades: margin (+), a continuous high-intensity rim around the index tumor; margin zero, a partially discontinuous high-intensity rim; margin (-), the tumor extends beyond the high-intensity rim. RESULTS: Twelve (86%) of fourteen low-intensity tumors on the pre-MRI were visualized as low-intensity tumors on post-MRI, and the ablative margins were visualized as high-intensity rims. Two (67%) of three high-intensity tumors on pre-MRI were visualized as higher-intensity tumors in the high-intensity ablative margin. Because the signal intensities of tumors and surrounding tissues in 14 tumors that were low- or high-intensity tumors on pre-MRI increased to the same extent, the tumors and ablative margin could be distinguished on post images. In 6 (75%) of the 8 iso-intensity tumors on pre-MRI, the ablative margin and tumor could also not be discriminated on post-MRI. The overall agreement between MRI and CT for the ablative margin was good (κ coefficient=0.716, p=0.00002). CONCLUSION: In 82% of low- or high-intensity tumors on pre-MRI, post-MRI without a contrast agent enabled visualization of the ablative margin as a high-intensity rim, and it was possible to evaluate the ablative margin earlier and easier than with enhanced CT.

[A case of AFP-producing gastric cancer responding to the combination of systemic chemotherapy, transcatheter arterial embolization and hepatic infusion chemotherapy].

We report a case of a 64-year-old male with a-fetoprotein(AFP)-producing gastric cancer accompanied by large liver metastases and multiple lymph node metastases. The patient's serum AFP level was 42,307 ng/mL and a biopsy specimen showed AFP-positive tumor cells immunohistochemically. Systemic chemotherapy by tegafur gimeracil oteracil potassium(S-1)and local therapy for the hepatic metastases consisting of transcatheter arterial embolization (TAE)and infusion of epirubicin(EPI)to the hepatic arteries decreased the serum AFP level and reduced the gastric cancer and metastases. Due to the increase of AFP and lymph node metastases, we had to successively change the regimen to paclitaxel(PTX), a combination of cisplatin(CDDP)/irinotecan(CPT-11)and S-1. Continuous systemic chemotherapy in combination with various drugs for gastric cancer treatment followed by TAE and hepatic infusion chemotherapy for hepatic metastases proved effective. The patient survived for 3 years and 2 months.