Histomorphometric analysis of patients with femoral neck fracture and 25-hydroxyvitamin D deficiency: a cross-sectional study.

INTRODUCTION: Vitamin D deficiency causes osteoporosis, bone mineralization disorders, and osteomalacia. Osteomalacia is diagnosed using blood biochemical tests, clinical symptoms, and imaging; however, accurate detection of mineralization disorders requires tissue observation. We investigated the prevalence of bone mineralization disorders and their relationship with serum 25-hydroxyvitamin D (25OHD) levels in patients with untreated osteoporosis with femoral neck fractures. MATERIALS AND METHODS: A non-demineralized specimen was prepared from the femoral head removed during surgery in 65 patients. Bone histomorphometry of cancerous bone in the femoral head center was conducted. Osteoid volume per bone volume (OV/BV) and osteoid thickness (O.Th) were measured as indicators of mineralization disorder. RESULTS: The mean serum 25OHD level (11.9 ± 5.7 ng/mL) was in the deficiency range (< 12 ng/mL). There were no clinically diagnosed cases of osteomalacia (OV/BV > 10% and O.Th > 12.5 µm); however, one case of mineralization disorder, considered histologically pre-osteomalacia (OV/BV > 5% and O.Th < 12.5 µm), was observed (OB/BV, 17.6%; O.Th, 12.3 µm). Excluding this case, those with severe (25OHD < 12 ng/mL, at risk of osteomalacia; n = 39) and non-severe deficiency (25OHD ≥ 12 ng/mL; n = 25) did not significantly differ in OV/BV (%; 0.77 ± 0.54 vs. 0.69 ± 0.38, p = 0.484) or O.Th (µm; 5.32 ± 1.04 vs. 5.13 ± 0.78, p = 0.410). Further, 25OHD and OV/BV were not significantly correlated (R = - 0.124, p = 0.327). CONCLUSION: This is the first study in the twenty-first century to examine serum 25OHD concentrations and bone mineralization disorders in Japanese patients with osteoporosis. The results indicate that vitamin D deficiency does not necessarily cause bone mineralization disorders and rarely leads to osteomalacia.

4D Flow MRI Reflects Physiological Hemodynamics for the Diagnosis and Management of Portosystemic Shunts.

A woman in her sixties with portosystemic shunt and hepatic encephalopathy underwent open mesenteric vein ligation, resulting in improved portal flow and blood ammonia. In this case, 4D flow MRI was a valuable diagnostic and follow-up tool, visualizing and quantifying physiological portal hemodynamics with features distinct from those of contrast-enhanced CT and digital subtraction angiography. Our case study highlights the value of 4D flow MRI for managing portosystemic shunts.

The Matsudai Knee Osteoarthritis Survey showed the longitudinal changes of knee phenotypes in alignment and structure during 23-28 years.

PURPOSE: The longitudinal changes in alignment and structure, including the joint line and cortical bone thickness (CBT) of the femur and tibia, and knee phenotype in patients with knee osteoarthritis (OA) remain unknown. The aim of this retrospective study was to clarify the longitudinal changes in matched healthy subjects. METHODS: The follow-up Matsudai Knee Osteoarthritis Survey was administered between 23 and 28 years. This study included 285 healthy knees from 235 females with an average age of 53 ± 6 years at baseline. The non-OA individuals, with an average age of 79 ± 4 years, were divided into three groups at baseline according to their follow-up radiographic results [the non-OA (n = 52), early OA (n = 131), and advanced OA groups (n = 102)]. Changes in alignment, joint line, CBT, and knee phenotype were assessed at baseline and at follow-up using standing anteroposterior radiographs. RESULTS: This study showed significant varus changes in the alignment (p < 0.001) and tibial and femoral joint line parameters (p < 0.05) in the OA group. Decreased CBT and increased mediolateral CBT ratios were observed in all groups (p < 0.001). The knee phenotypes in the OA groups were changed to varus angles, especially in the alignment and tibial joint line. CONCLUSIONS: The longitudinal changes of knee phenotypes in alignment and structure (CBT and joint line) from baseline to follow-up were shown in the OA groups. In addition, alignment and tibial structural factors at baseline are useful in predicting the incidence of knee OA in daily practice. LEVELS OF EVIDENCE: III.

Pancreatic ductal adenocarcinoma with acinar-to-ductal metaplasia-like cancer cells shows increased cellular proliferation.

BACKGROUND/OBJECTIVES: Acinar-to-ductal metaplasia (ADM) has been shown to contribute to the development of pancreatic ductal adenocarcinoma (PDAC) in genetically engineered mouse models, but little is known about whether acinar cell plasticity contributes to carcinogenesis in human PDAC. We aimed to assess whether cancer cells that stain positive for amylase and CK19 (ADM-like cancer cells) are present in human resected PDAC and to investigate their role in tumor progression. METHODS: We immunohistochemically investigated the presence of ADM-like cancer cells, and compared the clinical and histological parameters of PDAC patients with and without ADM-like cancer cells. RESULTS: ADM-like cancer cells were detected in 16 of 60 (26.7%) PDAC specimens. Positive staining for anterior gradient protein 2 (AGR2) was observed in 14 of 16 (87.5%) PDAC specimens with ADM-like cancer cells. On the other hand, the intensity of AGR2 expression (negative, low/moderate or high) was lower in PDAC with ADM-like cancer cells (9/7) than in PDAC without these cells (11/33) (P = 0.032). The presence of ADM-like cancer cells was significantly correlated with increased cell proliferation (P = 0.012) and tended to be associated with MUC1 expression (P = 0.067). CONCLUSIONS: These results indicated that acinar cells may act as the origin of human PDAC, and that their presence may be useful for the stratification of human PDAC to predict prognosis.

Normal coronal kinematics of dynamic alignment and bony positions relative to the ground in three-dimensional motion analysis during gait: A preliminary study.

BACKGROUND: During gait, healthy knee coronal kinematics of each bony axis and lower extremity alignment are important because they could be useful as reference data for several surgeries and provide clarification of the etiology of diseases around the knee in healthy participants; however, it remains unknown. OBJECTIVE: The objective of this study was to clarify the kinematics of lower extremity alignment and the bony axes relative to the ground during gait, focused on the coronal plane, in healthy individuals by applying our unique three-dimensional (3D) motion analysis. METHODS: The study included 21 healthy individuals, including 9 healthy females and 12 healthy males with an average age of 36 ± 17 years. Knee kinematics were calculated in a gait analysis by combining the data from a motion-capture system and a 3D lower-extremity alignment assessment system on biplanar long-leg radiographs by using a 3D-2D registration technique. The main kinematic parameters were the dynamic position change relative to the ground, applying the femoral anatomical axis (FAA), tibial anatomical axis (TAA), and dynamic alignment in the coronal plane during the stance phase of gait. RESULTS: The average changes in FAA, TAA, and dynamic varus alignment were 3.7° ± 1.2°, 3.5° ± 0.8°, and 3.0° ± 1.2°, respectively. The TAA tilted laterally during the loading response and a plateau area appeared afterwards; the FAA gradually inclined laterally until the terminal stance phase, and the dynamic alignment showed varus angular change during the loading response. CONCLUSIONS: The tibia and femur were found to change approximately 2-5° of the position of the bony axes relative to the ground. In terms of clinical relevance, our findings can be used to clarify the etiology of diseases around the knee joint and as reference data for surgeries.

A case of successful treatment with antiretroviral therapy for HIV in a patient with marked liver dysfunction.

Background: Human immunodeficiency virus (HIV) infection is often complicated with hepatitis virus infection. Antiretroviral therapy (ART) should be initiated with caution for patients with severe virus- or drug-induced acute hepatitis while considering factors that might interfere with the initiation of therapy. Case report: Herein, we present a case of a 67-year-old woman who presented with symptoms of severe liver injury of unknown cause. Laboratory examinations revealed HIV infection. The HIV viral load was high, and treatment with ART was considered. However, a liver biopsy could not be performed because of hyperbilirubinemia and the risk of severe hepatic damage. After assessing the risk of further liver damage, ART was safely administered despite hyperbilirubinemia. Treatment with ART could successfully reduce the viral load and bilirubin levels. Conclusion: ART treatment could be safely used for patients with HIV to reduce the viral load and bilirubin levels while avoiding the risk of liver failure.

Long-term phlebotomy successfully alleviated hepatic iron accumulation in a ferroportin disease patient with a mutation in SLC40A1: a case report.

BACKGROUND: Hereditary hemochromatosis is a heterogenous group of inherited iron-overload conditions that is characterized by increased intestinal absorption and deposition in vital organs. Hepcidin is a soluble regulator that acts to attenuate both intestinal iron absorption and iron release from reticuloendothelial macrophages through internalization of ferroportin-1, an iron exporter. Ferroportin disease is hereditary hemochromatosis which is affected by SLC40A1, a gene coding ferroportin-1, and phenotypically classified into two forms (classical and nonclassical). In nonclassical form, ferroportin mutations are responsible for a gain of function with full iron export capability but insensitivity to downregulation by hepcidin. Here, we report a case of nonclassical ferroportin disease. CASE PRESENTATION: A 46-year-old Japanese man showed elevated serum iron (284 µg/dl), ferritin (1722 ng/ml), transferrin saturation ratio (91.3%), and hepcidin-25 level (139.6 ng/ml). Magnetic resonance imaging (MRI) demonstrated a marked reduction in the signal intensity of the liver in T1- and T2-weighted images. The liver histology exhibited a large amount of iron that had accumulated predominantly in hepatocytes. We identified a heterozygous 1520A > G (p.H507R) mutation in the SLC40A1 gene. Phlebotomy (400 ml at a time) was monthly performed for 3 years in this patient. Importantly, the serum hepcidin level (1.0 ng/ml) was normal when the serum ferritin level was normal and hepatic iron accumulation was remarkably reduced after 3 years of phlebotomy. CONCLUSIONS: The present case demonstrated for the first time that there was a correlation between hepatic iron levels as measured by MRI and serum hepcidin levels through long-term phlebotomy in a patient with ferroportin disease with the p.H507R mutation of in SLC40A1.

The majority of patellar avulsion fractures in first-time acute patellar dislocations included the inferomedial patellar border that was different from the medial patellofemoral ligament attachment.

PURPOSE: Accurate assessment of the locations of patellar avulsion fractures in acute patellar dislocations is clinically relevant for decision making for treatment. The study aim was to classify the locations of patellar avulsion fractures with a focus on the ligament attachments of medial stabilizing structures. METHODS: Out of 131 first-time acute traumatic patellar dislocations, 61 patients had patellar fractures. Subsequently, 10 patients with isolated osteochondral fractures of the articular surface in the patella were excluded. Finally, 51 patients (34 females and 17 males, average age: 18.5 years, 95% CI 16.1-20.9) were included in the study cohort. Based on the locations of the patellar attachment, the patients were divided into three groups: the superior group [medial patellofemoral ligament (MPFL) attachment], inferior group [medial patellotibial ligament (MPTL)/medial patellomeniscal ligament (MPML) attachment], and mixed group. RESULTS: In the patellar avulsion group (51 patients), the superior group, mixed group, and inferior group contained 8/51 (16%), 12/51 (24%), and 31/51 (61%) patients, respectively. CONCLUSIONS: This study showed that 84% of the patellar avulsion fractures were located in the inferomedial patellar border, which consisted of MPTL/MPML attachments that were clearly different from the true "MPFL" attachment at the superomedial patellar border. In terms of the clinical relevance, the acute surgical repair of MPTL/MPML attachments in the inferomedial patellar border may not sufficiently control the patella if optimal management of the MPFL is not performed. LEVEL OF EVIDENCE: IV.

Dipeptidyl Peptidase 4 Inhibitors Reduce Hepatocellular Carcinoma by Activating Lymphocyte Chemotaxis in Mice.

Background & Aims: CD26, a multifunctional transmembrane glycoprotein, is expressed in various cancers and functions as dipeptidyl peptidase 4 (DPP4). We investigated whether CD26 expression is associated with hepatocellular carcinoma (HCC) progression and whether DPP4 inhibitors exert antitumor effects against HCC. Methods: CD26 expression was examined in 41 surgically resected HCC specimens. The effects of DPP4 inhibitors on HCC were examined by using HCC cell lines (Huh-7 and Li-7), xenograft tumors in nude mice, and a nonalcoholic steatohepatitis-related HCC mouse model. Results: CD26 expression in HCC specimens was associated with increased serum DPP4 activity, as well as a more advanced stage, less tumor immunity, and poorer prognosis in HCC patients. The HCC cell lines and xenograft tumors exhibited CD26 expression and DPP4 activity. The DPP4 inhibitors did not exhibit antitumor effects in vitro, but natural killer (NK) and/or T-cell tumor accumulation suppressed growth of xenograft tumor and HCC in vivo. The antitumor effects of DPP4 inhibitors were abolished by the depletion of NK cells or the neutralization of CXCR3, a chemokine receptor on NK cells. EZ-TAXIScan, an optical horizontal chemotaxis apparatus, identified enhanced NK and T-cell chemotaxis by DPP4 inhibitors ex vivo in the presence of Huh-7 cells and the chemokine CXCL10, which binds to CXCR3. The DPP4 inhibitors prevented the biologically active form of CXCL10 from being truncated by Huh-7 cell DPP4 activity. DPP4 inhibitors also suppressed tumor angiogenesis. Conclusions: These results provide a rationale for verifying whether DPP4 inhibitors clinically inhibit the progression of HCC or augment the antitumor effects of molecular-targeting drugs or immunotherapies against HCC.

Wisteria floribunda agglutinin-positive Mac-2 binding protein predicts the development of hepatocellular carcinoma in patients with non-alcoholic fatty liver disease.

AIM: As it is not practical to perform regular screening for hepatocellular carcinoma (HCC) in all patients with non-alcoholic fatty liver disease (NAFLD), there is a need to identify NAFLD patients who are at high risk for HCC. Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA+ -M2BP) has been shown to be a surrogate marker for predicting HCC as well as a liver fibrosis marker in patients with chronic hepatitis B and C. The aim of this study was to investigate whether WFA+ -M2BP predicts HCC development in NAFLD patients. METHODS: Serum WFA+ -M2BP was retrospectively measured in 331 patients with histologically proven NAFLD, 51 of whom developed HCC. The association of WFA+ -M2BP and HCC development in NAFLD patients was investigated. RESULTS: The WFA+ -M2BP values were significantly greater in NAFLD patients with HCC than in those without HCC among patients with liver fibrosis ≥stage 3. Multivariate analysis identified WFA+ -M2BP as one of the predictive factors for HCC development (odds ratio, 1.57; 95% confidence interval, 1.083-2.265; P = 0.017). The optimal cut-off index of WFA+ -M2BP for predicting HCC was 1.255 with specificity of 78.4% and sensitivity of 70.4%. The area under the receiver operating characteristic curve value for the prediction of HCC development was 0.806. The cumulative incidence rate of HCC was significantly greater in patients with WFA+ -M2BP ≥ 1.255 (n = 61) than in those with WFA+ -M2BP < 1.255 (n = 137) among patients who were followed up for more than 2 years after the diagnosis of NAFLD. CONCLUSIONS: Wisteria floribunda agglutinin-positive Mac-2 binding protein predicts HCC development and is a useful surrogate marker for identifying NAFLD patients who are at a high risk for HCC.

Assessment of hepatic fibrosis with superb microvascular imaging in hepatitis C virus-associated chronic liver diseases.

AIM: Superb microvascular imaging (SMI) is an ultrasound Doppler technique using a unique algorithm that allows visualization of minute vessels with slow velocity and minimal motion artifacts. The aim of this preliminary study was to investigate whether SMI could predict liver fibrosis by visualizing the vessels present in the vicinity of the liver surface because the morphology of the peripheral hepatic vasculature is affected by the progression of liver fibrosis. METHODS: We recruited 29 patients with biopsy-proven chronic hepatitis C or liver cirrhosis C, and 36 patients without liver disease as controls. Using an Aplio 500 ultrasound system with a 7-MHz or 12-MHz linear probe, we assessed the vascular shapes and the bifurcation angles of five randomly selected vessels in the vicinity of the liver surface. The vascular shape was scored based on the number of winding and/or irregular vessels. RESULTS: The mean vascular score and the mean bifurcation angle were significantly greater in patients with advanced liver fibrosis (3.5 ± 1.1 and 90.5 ± 14.3) than in those with mild-to-moderate liver fibrosis (1.3 ± 1.4 and 68.0 ± 16.1) and controls (0.6 ± 0.7 and 62.2 ± 10.5). The area under the receiver-operating curve of the vascular score and the bifurcation angle were 0.88 with 76.5% sensitivity and 83.3% specificity, and 0.87 with 94.1% sensitivity and 75.0% specificity, respectively. CONCLUSION: The present results indicate that SMI potentially predicts the extent of liver fibrosis by detecting small vessels present in the vicinity of the liver surface.

Fibroblast activation protein-α-expressing fibroblasts promote the progression of pancreatic ductal adenocarcinoma.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive desmoplastic stromal response. Fibroblast activation protein-α (FAP) is best known for its presence in stromal cancer-associated fibroblasts (CAFs). Our aim was to assess whether FAP expression was associated with the prognosis of patients with PDAC and to investigate how FAP expressing CAFs contribute to the progression of PDAC. METHODS: FAP expression was immunohistochemically assessed in 48 PDAC specimens. We also generated a fibroblastic cell line stably expressing FAP, and examined the effect of FAP-expressing fibroblasts on invasiveness and the cell cycle in MiaPaCa-2 cells (a pancreatic cancer cell line). RESULTS: Stromal FAP expression was detected in 98% (47/48) of the specimens of PDAC, with the intensity being weak in 16, moderate in 19, and strong in 12 specimens, but was not detected in the 3 control noncancerous pancreatic specimens. Patients with moderate or strong FAP expression had significantly lower cumulative survival rates than those with negative or weak FAP expression (mean survival time; 352 vs. 497 days, P = 0.006). Multivariate analysis identified moderate to strong expression of FAP as one of the factors associated with the prognosis in patients with PDAC. The intensity of stromal FAP expression was also positively correlated to the histological differentiation of PDAC (P < 0.05). FAP-expressing fibroblasts promoted the invasiveness of MiaPaCa-2 cells more intensively than fibroblasts not expressing FAP. Coculture with FAP-expressing fibroblasts significantly activated cell cycle shift in MiaPaCa-2 cells compared to coculture with fibroblasts not expressing FAP. Furthermore, coculture with FAP expressing fibroblasts inactivated retinoblastoma (Rb) protein, an inhibitor of cell cycle progression, in MiaPaCa-2 cells by promoting phosphorylation of Rb. CONCLUSIONS: The present in vitro results and the association of FAP expression with clinical outcomes provide us with a better understanding of the effect of FAP-expressing CAFs on the progression of PDAC.

Multicenter comparative study of laparoscopic and open distal pancreatectomy using propensity score-matching.

BACKGROUND: Laparoscopic distal pancreatectomy has been shown to be associated with favorable postoperative outcomes using meta-analysis. However, there have been no randomized controlled studies yet. This study aimed to compare laparoscopic and open distal pancreatectomy using propensity score-matching. METHODS: We retrospectively collected perioperative data of 2,266 patients who underwent distal pancreatectomy in 69 institutes from 2006-2013 in Japan. Among them, 2,010 patients were enrolled in this study and divided into two groups, laparoscopic distal pancreatectomy and open distal pancreatectomy. Perioperative outcomes were compared between the groups using unmatched and propensity matched analysis. RESULTS: After propensity score-matching, laparoscopic distal pancreatectomy was associated with favorable perioperative outcomes compared with open distal pancreatectomy, including higher rate of preservation of spleen and splenic vessels (P < 0.001); lower rates of intraoperative transfusion (P = 0.020), clinical grade of pancreatic fistula (International Study Group on Pancreatic Fistula grade B and C; P < 0.001), and morbidity (P < 0.001); and shorter hospital stay (P = 0.001), but a longer operative time (P < 0.001). CONCLUSIONS: Laparoscopic distal pancreatectomy was associated with more favorable perioperative outcomes than open distal pancreatectomy.

Branched-chain amino acids reduce hepatic iron accumulation and oxidative stress in hepatitis C virus polyprotein-expressing mice.

BACKGROUND & AIMS: Branched-chain amino acids (BCAA) reduce the incidence of hepatocellular carcinoma (HCC) in patients with cirrhosis. However, the mechanisms that underlie these effects remain unknown. Previously, we reported that oxidative stress in male transgenic mice that expressed hepatitis C virus polyprotein (HCVTgM) caused hepatic iron accumulation by reducing hepcidin transcription, thereby leading to HCC development. This study investigated whether long-term treatment with BCAA reduced hepatic iron accumulation and oxidative stress in iron-overloaded HCVTgM and in patients with HCV-related advanced fibrosis. METHODS: Male HCVTgM were fed an excess-iron diet that comprised either casein or 3.0% BCAA, or a control diet, for 6 months. RESULTS: For HCVTgM, BCAA supplementation increased the serum hepcidin-25 levels and antioxidant status [ratio of biological antioxidant potential (BAP) relative to derivatives of reactive oxygen metabolites (dROM)], decreased the hepatic iron contents, attenuated reactive oxygen species generation, and restored mitochondrial superoxide dismutase expression and mitochondrial complex I activity in the liver compared with mice fed the control diet. After 48 weeks of BCAA supplementation in patients with HCV-related advanced fibrosis, BAP/dROM and serum hepcidin-25 increased and serum ferritin decreased compared with the pretreatment levels. CONCLUSIONS: BCAA supplementation reduced oxidative stress by restoring mitochondrial function and improved iron metabolism by increasing hepcidin-25 in both iron-overloaded HCVTgM and patients with HCV-related advanced fibrosis. These activities of BCAA may partially account for their inhibitory effects on HCC development in cirrhosis patients.

Hepatitis C virus core protein suppresses mitophagy by interacting with parkin in the context of mitochondrial depolarization.

Hepatitis C virus (HCV) causes mitochondrial injury and oxidative stress, and impaired mitochondria are selectively eliminated through autophagy-dependent degradation (mitophagy). We investigated whether HCV affects mitophagy in terms of mitochondrial quality control. The effect of HCV on mitophagy was examined using HCV-Japanese fulminant hepatitis-1-infected cells and the uncoupling reagent carbonyl cyanide m-chlorophenylhydrazone as a mitophagy inducer. In addition, liver cells from transgenic mice expressing the HCV polyprotein and human hepatocyte chimeric mice were examined for mitophagy. Translocation of the E3 ubiquitin ligase Parkin to the mitochondria was impaired without a reduction of pentaerythritol tetranitrate-induced kinase 1 activity in the presence of HCV infection both in vitro and in vivo. Coimmunoprecipitation assays revealed that Parkin associated with the HCV core protein. Furthermore, a Yeast Two-Hybrid assay identified a specific interaction between the HCV core protein and an N-terminal Parkin fragment. Silencing Parkin suppressed HCV core protein expression, suggesting a functional role for the interaction between the HCV core protein and Parkin in HCV propagation. The suppressed Parkin translocation to the mitochondria inhibited mitochondrial ubiquitination, decreased the number of mitochondria sequestered in isolation membranes, and reduced autophagic degradation activity. Through a direct interaction with Parkin, the HCV core protein suppressed mitophagy by inhibiting Parkin translocation to the mitochondria. This inhibition may amplify and sustain HCV-induced mitochondrial injury.

Hepatic oxidative stress in ovariectomized transgenic mice expressing the hepatitis C virus polyprotein is augmented through suppression of adenosine monophosphate-activated protein kinase/proliferator-activated receptor gamma co-activator 1 alpha signaling.

AIM: Oxidative stress plays an important role in hepatocarcinogenesis of hepatitis C virus (HCV)-related chronic liver diseases. Despite the evidence of an increased proportion of females among elderly patients with HCV-related hepatocellular carcinoma (HCC), it remains unknown whether HCV augments hepatic oxidative stress in postmenopausal women. The aim of this study was to determine whether oxidative stress was augmented in ovariectomized (OVX) transgenic mice expressing the HCV polyprotein and to investigate its underlying mechanisms. METHODS: OVX and sham-operated female transgenic mice expressing the HCV polyprotein and non-transgenic littermates were assessed for the production of reactive oxygen species (ROS), expression of inflammatory cytokines and antioxidant potential in the liver. RESULTS: Compared with OVX non-transgenic mice, OVX transgenic mice showed marked hepatic steatosis and ROS production without increased induction of inflammatory cytokines, but there was no increase in ROS-detoxifying enzymes such as superoxide dismutase 2 and glutathione peroxidase 1. In accordance with these results, OVX transgenic mice showed less activation of peroxisome proliferator-activated receptor-γ co-activator-1α (PGC-1α), which is required for the induction of ROS-detoxifying enzymes, and no activation of adenosine monophosphate-activated protein kinase-α (AMPKα), which regulates the activity of PGC-1α. CONCLUSION: Our study demonstrated that hepatic oxidative stress was augmented in OVX transgenic mice expressing the HCV polyprotein by attenuation of antioxidant potential through inhibition of AMPK/PGC-1α signaling. These results may account in part for the mechanisms by which HCV-infected women are at high risk for HCC development when some period has passed after menopause.

Risk factors for survival and the development of hepatocellular carcinoma in patients with primary biliary cirrhosis.

OBJECTIVE: Early diagnosis of hepatocellular carcinoma (HCC) is critical in the management of patients with primary biliary cirrhosis (PBC), since the prognosis of PBC has improved. The aim of this study was to investigate whether HCC development affects the prognosis of PBC and to identify the risk factors for HCC in Japanese patients with PBC. METHODS: We compared the survival rates between patients with HCC and those without and analyzed the risk factors for HCC development in 210 patients with PBC who were followed up for a median period of 8.5 years. RESULTS: HCC developed during follow-up in 11 patients (5.2%) and was diagnosed simultaneously at the time of diagnosis of PBC in five patients (2.4%) who were excluded from the analysis. A Kaplan-Meier analysis showed a significant difference in overall survival between the patients who did and did not develop HCC (p<0.001). A multivariate analysis revealed age (OR: 1.08, 95% confidence interval [CI]: 1.03-1.13, p=0.001), the albumin level (OR: 0.24, 95% CI: 0.10-0.56, p=0.001), the total bilirubin level (OR: 1.60, 95% CI: 1.09-2.36, p=0.017) and HCC development (OR: 2.97, 95% CI: 1.24-7.15, p=0.015) to be significant prognostic factors and identified only an advanced histological stage (Scheuer's classification III or IV, OR: 6.27, 95% CI: 1.80-21.83, p=0.004) to be a risk factor associated with HCC. CONCLUSION: HCC development significantly affects the survival of patients with PBC, and an advanced histological stage is the only risk factor associated with HCC development. These results highlight the important role of liver fibrosis in hepatocarcinogenesis in patients with PBC.

Clinical usefulness of non-protein respiratory quotient measurement in non-alcoholic fatty liver disease.

AIM: Little is known about the effects of non-alcoholic fatty liver disease (NAFLD) on energy metabolism, although this disease is associated with metabolic syndrome. We measured non-protein respiratory quotient (npRQ) using indirect calorimetry, which reflects glucose oxidation, and compared this value with histological disease severity in NAFLD patients. METHODS: Subjects were 32 patients who were diagnosed with NAFLD histopathologically. Subjects underwent body composition analysis and indirect calorimetry, and npRQ was calculated. An oral glucose tolerance test was performed, and plasma glucose area under the curve (AUC glucose) was calculated. RESULTS: There were no differences in body mass index, body fat percentage or visceral fat area among fibrosis stage groups. As fibrosis progressed, npRQ significantly decreased (stage 0, 0.895 ± 0.068; stage 1, 0.869 ± 0.067; stage 2, 0.808 ± 0.046; stage 3, 0.798 ± 0.026; P < 0.005). Glucose intolerance worsened and insulin resistance increased with fibrosis stage. npRQ was negatively correlated with AUC glucose (R = -0.6308, P < 0.001), Homeostasis Model of Assessment - Insulin Resistance (R = -0.5045, P < 0.005), fasting glucose (R = -0.4585, P < 0.01) and insulin levels (R = -0.4431, P < 0.05), suggesting that decreased npRQ may reflect impaired glucose tolerance due to insulin resistance, which was associated with fibrosis progression. Estimation of fibrosis stage using npRQ was as accurate as several previously established scoring systems using receiver-operator curve analysis. CONCLUSION: npRQ was significantly decreased in patients with advanced NAFLD. Our data suggest that measurement of npRQ is useful for the estimation of disease severity in NAFLD patients.

Focal nodular hyperplasia-like nodule with reduced expression of organic anion transporter 1B3 in alcoholic liver cirrhosis.

We report a patient with alcoholic liver cirrhosis who had a 15 mm focal nodular hyperplasia (FNH)-like nodule in the liver. This FNH-like nodule was diagnosed as hepatocellular carcinoma (HCC) mainly based on hypervascularity during the hepatic arterial phase, washout pattern during the equilibrium phase and low signal intensity during the hepatobiliary phase in gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced MRI; it was surgically resected. Its histology exhibited hepatocyte hyperplasia, fibrous septa containing unpaired small arteries accompanied by reactive bile ductules, remarkable iron deposits and sinusoidal capillarization, and was compatible with the diagnosis of an FNH-like nodule. When we analyzed the images of the present nodule retrospectively, low signal intensity on in-phase and isosignal intensity on opposed-phase T1-weighted MRI may have reflected iron deposits in the FNH-like nodule. In addition, a low signal intensity on T2-weighted MRI and no detection in diffusion-weighted MRI may help in distinguishing FNH-like nodules from HCC, since these image findings are inconsistent with typical HCC. Immunohistochemical analysis revealed a markedly reduced expression of organic anion transporter (OATP) 1B3 in this nodule, which implied decreased Gd-EOB-DTPA uptake by hepatocytes and accounted for the low signal intensity during the hepatobiliary phase on Gd-EOB-DTPA-enhanced MRI. To the best of our knowledge this is the first report in which an FNH-like nodule was assessed for OATP1B3 expression.

Type 1 interferon receptor in peripheral blood mononuclear cells may predict response to intra-arterial 5-fluorouracil + interferon therapy for advanced hepatocellular carcinoma.

BACKGROUND: Type 1 interferon alpha receptor 2 (IFNAR2) in the liver has been reported to be a predictive factor for the response to intra-arterial 5-fluorouracil (5-FU) + systemic interferon (IFN)-alpha combination therapy in patients with advanced hepatocellular carcinoma. We tested whether IFNAR2 expression in peripheral blood mononuclear cells could predict the response to 5-FU + IFN. METHODS: Predictive factors for survival and response to therapy were determined in 30 patients with advanced hepatocellular carcinoma who underwent treatment with 5-FU + IFN. IFNAR2 expression in peripheral blood mononuclear cells was measured in 11 of the 30 patients. RESULTS: With a mean number of 4.2 courses of combination therapy, one patient (3%) showed a complete response, eight (27%) showed partial responses, 13 (43%) had stable disease, and eight (27%) showed progressive disease. The median survival time of responders (complete response/partial response) was 12.7 months and that of nonresponders (stable disease/progressive disease) was 7.5 months. The one-year and two-year cumulative survival rates of responders and nonresponders were 87/69% and 40/11%, respectively (P = 0.019). Multivariate analysis identified response to therapy (P = 0.037) as the sole independent determinant of survival. The expression level of IFNAR2 in peripheral blood mononuclear cells was significantly (P = 0.012) higher in responders (6.5 ± 2.4) than in nonresponders (2.4 ± 0.6), even though no clinical factors were identified as being associated with the response to the combination therapy. CONCLUSION: IFNAR2 expression in peripheral blood mononuclear cells may predict the response to 5-FU + IFN therapy in patients with advanced hepatocellular carcinoma, although these data are preliminary.