Masaharu Tada

Fluorine-18 Fluorodeoxyglucose Positron Emission Tomography Predicts Tumor Differentiation, P-glycoprotein Expression, and Outcome after Resection in Hepatocellular Carcinoma

Purpose: To investigate the diagnostic value of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) for prediction of tumor differentiation, P-glycoprotein (P-gp) expression, and outcome in hepatocellular carcinoma (HCC) patients. Experimental Design: Seventy HCC patients who underwent curative resection were prospectively enrolled in the study. FDG-PET was done 2 weeks preoperatively, and the standardized uptake value (SUV) and the tumor to nontumor SUV ratio (TNR) were calculated from FDG uptake. Tumor differentiation and P-gp expression were examined with H&E and immunohistochemical staining, respectively. Results: SUV and TNR were significantly higher in poorly differentiated HCCs than in well-differentiated (P = 0.001 and 0.002) and moderately differentiated HCCs (P < 0.0001 and P < 0.0001). The percentage P-gp–positive area was significantly higher in well-differentiated HCCs than in poorly differentiated (P < 0.0001) and moderately differentiated HCCs (P = 0.0001). Inverse correlations were found between SUV and P-gp expression (r = −0.44; P < 0.0001) and between TNR and P-gp expression (r = −0.47; P = 0.01). Forty-three (61.4%) patients had postoperative recurrence. The overall and disease-free survival rates in the high TNR (≥2.0) group were significantly lower than in the low TNR (<2.0) group (P = 0.0001 and 0.0002). In multivariate analysis, a high α-fetoprotein level (risk ratio, 5.46; P = 0.003; risk ratio, 8.78; P = 0.006) and high TNR (risk ratio, 1.3; P = 0.03; risk ratio, 1.6; P = 0.02) were independent predictors of postoperative recurrence and overall survival. Conclusions: The results suggest that preoperative FDG-PET reflects tumor differentiation and P-gp expression and may be a good predictor of outcome in HCC.

CHOP deficiency attenuates cholestasis-induced liver fibrosis by reduction of hepatocyte injury

CCAAT/enhancer-binding protein (C/EBP) homologous protein (CHOP) is a key component in endoplasmic reticulum (ER) stress-mediated apoptosis. The goal of the study was to investigate the role of CHOP in cholestatic liver injury. Acute liver injury and liver fibrosis were assessed in wild-type (WT) and CHOP-deficient mice following bile duct ligation (BDL). In WT livers, BDL induced overexpression of CHOP and Bax, a downstream target in the CHOP-mediated ER stress pathway. Liver fibrosis was attenuated in CHOP-knockout mice. Expression levels of alpha-smooth muscle actin and transforming growth factor-beta1 were reduced, and apoptotic and necrotic hepatocyte death were both attenuated in CHOP-deficient mice. Hepatocytes were isolated from WT and CHOP-deficient mice and treated with 400 microM glycochenodeoxycholic acid (GCDCA) for 8 h to examine bile acid-induced apoptosis and necrosis. GCDCA induced overexpression of CHOP and Bax in isolated WT hepatocytes, whereas CHOP-deficient hepatocytes had reduced cleaved caspase-3 expression and a lower propidium iodide index after GCDCA treatment. In conclusion, cholestasis induces CHOP-mediated ER stress and triggers hepatocyte cell death, and CHOP deficiency attenuates this cell death and subsequent liver fibrosis. The results demonstrate an essential role of CHOP in development of liver fibrosis due to cholestatic liver damage.

Inhibition of c-Jun NH2-terminal kinase switches Smad3 signaling from oncogenesis to tumor- suppression in rat hepatocellular carcinoma†

Transforming growth factor beta (TGF‐β) signaling involves both tumor‐suppression and oncogenesis. TGF‐β activates the TGF‐β type I receptor (TβRI) and c‐Jun N‐terminal kinase (JNK), which differentially phosphorylate the mediator Smad3 to become COOH‐terminally phosphorylated Smad3 (pSmad3C) and linker‐phosphorylated Smad3 (pSmad3L). TβRI‐dependent pSmad3C transmits a tumor‐suppressive TGF‐β signal, while JNK‐dependent pSmad3L promotes carcinogenesis in human chronic liver disorders. The aim of this study is to elucidate how SP600125, a JNK inhibitor, affected rat hepatocellular carcinoma (HCC) development, while focusing on the domain‐specific phosphorylation of Smad3. The rats received subcutaneous injections of either SP600125 or vehicle 11 times weekly together with 100 ppm N‐diethylnitrosamine (DEN) administration for 56 days and were sacrificed in order to evaluate HCC development 28 days after the last DEN administration. The number of tumor nodules greater than 3 mm in diameter and the liver weight/body weight ratio were significantly lower in the SP600125‐treated rats than those in the vehicle‐treated rats (7.9 ± 0.8 versus 17.7 ± 0.9: P < 0.001; 6.3 ± 1.2 versus 7.1 ± 0.2%: P < 0.05). SP600125 significantly prolonged the median survival time in rats with DEN‐induced HCC (113 versus 97 days: log‐rank P = 0.0018). JNK/pSmad3L/c‐Myc was enhanced in the rat hepatocytes exposed to DEN. However, TβRI/pSmad3C/p21WAF1 was impaired as DEN‐induced HCC developed and progressed. The specific inhibition of JNK activity by SP600125 suppressed pSmad3L/c‐Myc in the damaged hepatocytes and enhanced pSmad3C/p21WAF1, acting as a tumor suppressor in normal hepatocytes. Conclusion: Administration of SP600125 to DEN‐treated rats shifted hepatocytic Smad3‐mediated signal from oncogenesis to tumor suppression, thus suggesting that JNK could be a therapeutic target of human HCC development and progression. (HEPATOLOGY 2009.)

Fluorine-18 fluorodeoxyglucose positron emission tomography predicts lymph node metastasis, P-glycoprotein expression, and recurrence after resection in mass-forming intrahepatic cholangiocarcinoma.

BACKGROUND Patients with intrahepatic cholangiocarcinoma (ICC) have a poor prognosis, and lymph node metastasis is an important prognostic factor. In this study, we investigated the usefulness of fluorodeoxyglucose positron emission tomography (FDG-PET) as a marker for lymph node metastasis, P-glycoprotein (P-gp) expression, and recurrence in ICC. METHODS The subjects were 35 patients who underwent FDG-PET. Detectability of lymph node metastasis using FDG-PET was compared with that using computed tomography (CT) or magnetic resonance imaging (MRI). In patients who underwent resection, expression of P-gp was examined immunohistochemically, and the relationship between P-gp expression and the standardized uptake value (SUV) in FDG-PET was investigated. Survival rates were analyzed using clinical and pathologic factors. RESULTS Of the 35 patients, 5 did not undergo surgery based on FDG-PET findings (2 with extrahepatic metastasis, and 3 with para-aortic lymph node metastasis) and 3 underwent laparotomy only (2 with peritoneal dissemination and 1 with para-aortic lymph node metastasis). The diagnostic accuracies of FDG-PET, CT, and MRI for detection of lymph node metastasis were 86%, 68%, and 57%, the sensitivities were 43%, 43% and 43%, and the specificities were 100%, 76%, and 64%, respectively. A negative correlation was found between SUV and P-gp expression (P = .002; r = -0.62). The disease-free survival rates in the high SUV group (>or=8.5) were significantly lower than in the low SUV group (<8.5; P = .04), and a high SUV was an independent predictor of postoperative recurrence in multivariate analysis (risk ratio, 1.3; P = .03). CONCLUSIONS FDG-PET is useful for prediction of lymph node metastasis, P-gp expression and recurrence in ICC.

High volume hydrodynamic injection of plasmid DNA via the hepatic artery results in a high level of gene expression in rat hepatocellular carcinoma induced by diethylnitrosamine.

Hydrodynamic injection of naked plasmid DNA (pDNA) via the tail vein is a safe and effective method of gene transfer to the liver. However, successful gene transfer has yet to be shown for hepatocellular carcinoma (HCC); therefore, we investigated the feasibility and efficacy of hydrodynamic injection via the tail vein and hepatic artery in a diethylnitrosamine (DEN)‐induced HCC model in rats.

Loss of Sept4 exacerbates liver fibrosis through the dysregulation of hepatic stellate cells

BACKGROUND/AIMS Septins are ubiquitous and multifunctional scaffold proteins involved in cytoskeletal organization, exocytosis and other cellular processes. We disclose the quiescent hepatic stellate cells (HSCs)-specific expression of a septin subunit Sept4 in the liver, and explore the significance of the septin system in liver fibrosis. METHODS We analyzed the expression of alpha-smooth muscle actin (alpha-SMA), collagens and other markers in primary cultured HSCs derived from wild-type and Sept4(-/-) mice. We compared susceptibility of these mice to liver fibrosis induced by either carbon tetrachloride treatment, bile duct ligation or methionine/choline-deficient diet. Collagen deposition, the principal parameter of liver fibrosis, was quantified both histochemically (Massons trichrome stain) and biochemically (hydroxyproline content). RESULTS In vitro, Sept4 mRNA/protein was remarkably downregulated in HSCs through myofibroblastic transformation. Sept4(-/-) HSCs showed normal morphology and proliferation, while myofibroblastic transformation as monitored by the upregulation of alpha-SMA and collagen was accelerated compared to wild-type HSCs. In vivo, liver fibrosis was consistently more severe in Sept4(-/-) mice than in wild-type littermates in all of the three paradigms of hepatitis/liver fibrosis. CONCLUSIONS These data concordantly indicate that the HSC-specific septin subunit Sept4 and perhaps the septin system are involved in the suppressive modulation of myofibroblastic transformation and fibrogenesis associated with liver diseases.

Off-On Characteristics of Magnetically Controlled Superconducting Switch

This paper describes structure of an experimental switch, the experimental results and the theoretical approach for the off-on characteristics of the switch. The experimental magnetically controlled superconducting switch, which is composed of superconducting gate wire (NbZr/CuNi) and superconducting field coil (NbTi/CuNi), has been made. Some experiments on the switching characteristics of the switch have been carried out. The experimental results show special characteristics of switching: l)at failure of switching-on, the gate current at zero magnetic induction is independent of the initial current and is constant and 2)the gate current at the off-on transient state, that is, from the normal state to the superconducting state of the superconducting gate wire, does not depend on the voltage across the gate wire, but depends uniquely on the magnetic field around the gate wire. The gate current can be explained to be a minimum normal-zone propagation current given by the equal-area theorem on cryogenic stabilization.

Difficulty Achieving a Preoperative Diagnosis of IgG4-Related Sclerosing Cholangitis

A 75-year-old male was admitted to our hospital because of bile duct stenosis. He had no medical history of autoimmune disease. The level of tumor markers, serum IgG, and IgG4 were within normal ranges. Computed tomography showed perihilar and distal bile duct stenosis and wall thickening without swelling or abnormal enhancement of the pancreas. Endoscopic retrograde cholangiopancreatography showed perihilar and distal bile duct stenosis. A biopsy and cytology from the distal bile duct stenosis suggested adenocarcinoma, and cytology from the perihilar bile duct also suggested adenocarcinoma. A preoperative diagnosis of perihilar and distal bile duct cancer was made, and the patient underwent left hepatectomy and pancreaticoduodenectomy. Resected specimens showed wall thickening in the perihilar and distal bile duct; however, tumors were unclear. A histopathological examination revealed lymphoplasmacytic infiltration, storiform fibrosis, and obliterative phlebitis in the perihilar and distal bile ducts. Immunohistochemistry revealed diffuse infiltration of IgG4-positive plasma cells in the perihilar and distal bile ducts. Lymphoplasmacytic infiltration, inflammatory change, storiform fibrosis, and obliterative phlebitis were shown in the pancreas. A final diagnosis of IgG4-related sclerosing cholangitis (IgG4-SC) with autoimmune pancreatitis was made. We herein report a case in which a preoperative diagnosis of IgG4-SC was difficult due to normal serum IgG4 levels and no obvious pancreatic lesion.

AB061. P033. A comparison of delayed gastric emptying and nutritional status after pylorus-preserving versus stomach-preserving pancreaticoduodenectomy

Background: This study was performed to compare the incidence of delayed gastric emptying (DGE), postoperative outcome and long-term nutritional status between pyloruspreserving pancreaticoduodenectomy (PPPD) and subtotal stomach-preserving pancreaticoduodenectomy (SSPPD). Methods: We retrospectively analyzed 133 patients who undergoing PPPD (n=89) or SSPPD (n=44) between March 2011 and December 2015. All cases of duodenojejunostomy in PPPD and gastrojejunostomy in SSPPD were performed antecolically. The postoperative nutritional status was explored by changes ratio in the body weight, serum total protein and serum albumin for 1 year after surgery. Results: The overall incidence of the DGE was 12%. The incidence of DGE was 13.5% (grade A: 5.6%, grade B: 4.5%, grade C: 3.4%) in PPPD group and 9.1% (grade A: 4.5%, grade B: 4.5%, grade C: 0%) in SSPPD group, and was no significant differences in both groups. The mean postoperative hospital stay was 42.8 days in the PPPD group and 37.2 days in the SSPPD group, and was no significant differences in both groups. The body weight ratio was decreased at 6 months after surgery in the SSPPD group, whereas it continued to decrease at 9 months after surgery in the PPPD group. It was gradually increased 9 months later after surgery in SSPPD group, and it was increased 12 months later after surgery in PPPD group. The serum total protein ratio and serum albumin ratio were decreased at 3 months after surgery and were gradually increased 6 months later after surgery in both groups. There were no significant differences with regard to postoperative body weight ratio, serum total protein ratio and serum albumin ratio in both groups for 1 year after surgery. Conclusions: SSPPD is equivalent outcomes in incidence of DGE and in postoperative long-term nutritional status comparing PPPD.