Hiroaki Haga

Potential therapeutic application of intravenous autologous bone marrow infusion in patients with alcoholic liver cirrhosis.

The present study was conducted to evaluate the application and efficacy of autologous bone marrow infusion (ABMi) for improvement of liver function in patients with alcoholic liver cirrhosis (ALC). Five subjects and 5 control patients with ALC who had abstained from alcohol intake for 24 weeks before the study were enrolled. Autologous bone marrow cells were washed and injected intravenously, and the changes in serum liver function parameters, and the level of the type IV collagen 7S domain as a marker of fibrosis, were monitored for 24 weeks. The distribution of activated bone marrow was assessed by indium-111-chloride bone marrow scintigraphy. The number of cells infused was 8.0±7.3×10(9) (mean±standard error). The serum levels of albumin and total protein and the prothrombin time were significantly higher during the follow-up period after ABMi than during the observation period in treated patients, whereas no such changes were observed in the controls. In the patients who received ABMi, the Child-Pugh score decreased in all 3 who were classified as class B; the serum levels of type IV collagen 7S domain improved in 4 of the 5 patients; and bone marrow scintigraphy demonstrated an increase of indium-111-chloride uptake in 3 of the 4 patients tested. ABMi for patients with ALC helps improve liver function parameters in comparison with observation during abstinence and ameliorates the degree of fibrosis in terms of serum markers and bone marrow activation in most cases.

Impact of metabolic syndrome on elevated serum alanine aminotransferase levels in the Japanese population.

Measurement of the serum alanine aminotransferase (ALT) level is used as an initial test for detection of liver diseases, and recent studies have also highlighted its potential value as a measure of overall health and survival as a marker of an increased risk of metabolic disorder. This study was designed to clarify the prevalence of elevated ALT levels in the Japanese population and to assess factors associated with ALT elevation. The subjects were 2165 individuals aged 40 to 85 years who participated in a Japanese community-based study referred to as the Takahata Study. Serum ALT levels and factors associated with ALT elevation were investigated. Among 2087 subjects who were negative for hepatitis B and C, the rates of elevated ALT greater than 30 U/L in men and greater than 25 U/L in women were 217 (22.7%) of 957 and 239 (21.2%) of 1130, respectively. These ALT cutoff levels had a specificity of more than 80% for exclusion of subjects with none or 1 of 3 metabolic risk factors: hypertension, lipid metabolism abnormality, and hyperglycemia. Multivariate analysis revealed 5 factors with a significant association with ALT elevation in men (n = 957): high gamma-glutamyltranspeptidase, low adiponectin, high low-density lipoprotein cholesterol, high body mass index, and high homeostasis model assessment insulin resistance index. Similarly, 4 factors were significantly associated with ALT elevation in women (n = 1130): high gamma-glutamyltranspeptidase, low adiponectin, high body mass index, and high homeostasis model assessment insulin resistance index. These results suggest that elevated ALT levels in the Japanese population older than 40 years have a strong association with metabolic syndrome-related features including obesity and insulin resistance.

Characteristics of rat bone marrow cells differentiated into a liver cell lineage and dynamics of the transplanted cells in the injured liver

BackgroundBone marrow cells (BMCs) have been shown to differentiate into a liver cell lineage, but little is known about their dynamics following transplantation. BMCs were cultured to investigate the expression of liver-specific genes in vitro and transplanted into in vivo liver-injury models to elucidate their dynamics in the liver.MethodsThe mRNA expression of various liver-specific genes in BMCs cocultured with hepatocytes was analyzed using reverse transcription-polymerase chain reaction. BMCs from transgenic rats expressing green fiuorescent protein were transplanted into the spleen of rat liver-injury models induced with 2-acetylaminofiuorene (2-AAF) or carbon tetrachloride (CCl4). BMCs were also transplanted directly into livers treated with CCl4 to determine which route is better for transplantation.ResultsBMCs differentiated into a liver cell lineage in vitro and expressed mRNAs consistent with mature hepatocytes, including albumin. The transplanted BMCs were found in the liver in the CCl4-induced injury model, but not in the 2-AAF-induced model. The hepatocyte growth factor and fibroblast growth factor mRNA levels in the liver were significantly higher in the CCl4-induced model than in the 2-AAF-induced model. Migration of BMCs to the liver was more effective following injection into the liver, rather than into the spleen.ConclusionsCultured BMCs differentiated into a liver cell lineage are a potential source for cell transplantation. Transplantation is successful in the severely injured liver with a high level of expression of mRNAs for growth factors. Injection of BMCs directly into the liver is the preferred route of administration.

Clinical manifestations of liver injury in patients with anorexia nervosa

The number of Japanese patients with anorexia nervosa (AN) is increasing as society changes. Mild liver injury is a complication of AN in around 30% of cases. In some rare instances, patients present with severe liver injury similar to acute liver failure. However, there are numerous uncertainties over the clinical characteristics of this condition. The objective of the present study was to clarify the clinical characteristics of AN complicated by liver injury and to investigate the factors related to hepatic complications.

Serum levels of stem cell factor and thrombopoietin are markedly decreased in fulminant hepatic failure patients with a poor prognosis.

Background and Aim:  Hematopoietic growth factors including stem cell factor (SCF), thrombopoietin (TPO) and granulocyte colony stimulating factor (G‐CSF) have a potential role in inducing bone marrow hematopoietic stem cells to move into the circulation, and the association of these factors with liver regeneration has received a lot of attention recently. The aim of this study was to determine the serum levels of such factors in patients with acute liver injury.

Successful percutaneous radiofrequency ablation of adrenal metastasis from hepatocellular carcinoma.

To the Editor: Recently, radiofrequency ablation (RFA) has been widely used as therapy for hepatocellular carcinoma (HCC), because it is safe and only minimally invasive to the human body.1,2 However, the therapeutic efficacy of RFA for extrahepatic metastasis of HCC is still unclear. The adrenal gland is one of the organs often affected by hematogenous metastasis of HCC. As a recent report has indicated that percutaneous RFA is a safe and well-tolerated procedure for the treatment of unresectable adrenocortical carcinoma,3 it appears that RFA could also be applicable for the treatment of HCC metastasis to the adrenal gland. So far, surgical procedures have been employed for the removal of adrenal metastasis from HCC, but the optimal treatment regimen is still inconclusive. Here, we describe the use of ultrasonography (US)-guided percutaneous RFA for the treatment of a patient with adrenal metastasis of HCC. We found that this procedure allowed complete ablation of the metastasis without any severe adverse events. The patient was a 69-year-old woman suffering from Child’s class A, compensated cirrhosis due to hepatitis C. Computed tomography (CT) scan showed that an HCC, 20 mm in diameter, in the right lobe of the liver, had metastasized to the right adrenal gland and right portal vein, resulting in a 20-mm adrenal tumor mass and tumor emboli, respectively. The HCC was treated by arterial infusion of the chemotherapeutic agents, fluorouracil and cisplatin, according to the planned regimen, via a subcutaneously implanted injection port. Six months after the start of chemotherapy, CT imaging showed that both the primary HCC in the liver and the tumor emboli were reduced markedly, to an undetectable level, and the serum alpha-fetoprotein (AFP) level had a decreased from 29 740 ng/ml before therapy to 6280 ng/ml after. However, the metastatic lesion in the adrenal gland was found to have increased in diameter, to 40 mm (Fig. 1a). Histological examination of a biopsy sample of the adrenal tumor, taken using a 21-gauge needle, showed the features of well-differentiated HCC. Immunohistochemical analysis of the tumor cells revealed positive staining for AFP. After local anesthesia was carried out with intradermal and subcutaneous lidocaine (1%), percutaneous transhepatic thermoablation of the metastatic tumor was performed twice, under US guidance, using a cool-tip needle radiofrequency system (Radionics, Burlington, MA, USA), comprising a 17-gauge, 3-cm active-tip RF electrode and radiofrequency generator, with an allowable output power of 120 W for 12 min. The first RFA was performed on the upper side of the tumor, and the second RFA was performed on the lower side of the tumor. There were no procedure-related complications. Both the plasma cortisol and catecholamine levels were measured 1 week before and after RFA. The plasma cortisol levels before and after RFA were 13.6 and 10.8 mg/dl, respectively. The plasma catecholamine levels before and after RFA were 16.0 and 10.0 pg/ml (adrenaline), 188.0 and 94.0 pg/ml (noradrenaline), and 8.0 and 5.0pg/ml (dopamine), respectively. All levels of adrenal hormones were within the normal ranges. CT scan 6 months after the RFA showed that the adrenal metastatic HCC had been completely ablated and devascularized, with loss of contrast enhancement (Fig. 1b). The serum AFP level returned to the normal range, below 10 ng/ml, 4 months after the RFA. To our knowledge, this is the first report of the successful treatment of adrenal metastasis of HCC using percutaneous RFA. This procedure was safe and well-tolerated and allowed complete ablation of the localized adrenal metastasis in this patient. Recently, it has been reported that unintended injury to normal adrenal tissue during RFA of adrenal tumors can lead to hypertensive crisis, a potentially catastrophic complication.4 Therefore, we must pay sufficient attention to serious adverse

Bone marrow cell-based regenerative therapy for liver cirrhosis.

Bone marrow cells are capable of differentiation into liver cells. Therefore, transplantation of bone marrow cells has considerable potential as a future therapy for regeneration of damaged liver tissue. Autologous bone marrow infusion therapy has been applied to patients with liver cirrhosis, and improvement of liver function parameters has been demonstrated. In this review, we summarize clinical trials of regenerative therapy using bone marrow cells for advanced liver diseases including cirrhosis, as well as topics pertaining to basic in vitro or in vivo approaches in order to outline the essentials of this novel treatment modality.

Impaired mitochondrial β-oxidation in patients with chronic hepatitis C: relation with viral load and insulin resistance

BackgroundHepatic steatosis is often seen in patients with chronic hepatitis C (CH-C). It is still unclear whether these patients have an impaired mitochondrial β-oxidation. In this study we assessed mitochondrial β-oxidation in CH-C patients by investigating ketogenesis during fasting.MethodsThis study consisted of thirty patients with CH-C. Serum levels of insulin and hepatitis C virus (HCV) core protein were measured by chemiluminescence enzyme immunoassay. The subjects were then fasted, and venous blood samples were drawn 12 h and 15 h after the start of fasting. The levels of blood ketone bodies were measured by an enzymatic cycling method. The rate of change in total ketone body concentration was compared with that in eight healthy volunteers.ResultsThe rate of change in total ketone body concentration between 12 h and 15 h after the start of fasting was significantly lower in CH-C patients than in healthy volunteers (129.9% (8.5-577.3%) vs. 321.6% (139.6-405.4%); P <0.01). The rate of change in total ketone body concentration in patients with a serum level of HCV core protein of 10000 fmol/L or higher was significantly lower than in patients with a level of less than 10000 fmol/L (54.8% (8.5-304.3%) vs. 153.6% (17.1-577.3%); P <0.05). The rate of change in total ketone body concentration in patients with a homeostasis model assessment of insulin resistance (HOMA-IR) of 2.5 or higher was significantly lower than in patients with a HOMA-IR of less than 2.5 (56.7% (8.5-186.7%) vs. 156.4% (33.3-577.3%); P <0.01).ConclusionsThese results suggest that mitochondrial β-oxidation is impaired, possibly due to HCV infection in patients with CH-C.

MiR-139-5p is associated with inflammatory regulation through c-FOS suppression, and contributes to the progression of primary biliary cholangitis

Primary biliary cholangitis (PBC) is a chronic cholestatic liver disease characterized pathologically by destruction of intrahepatic bile ducts. PBC is largely classified into three subtypes based on clinical course: (i) gradually progressive, (ii) portal hypertension, and (iii) hepatic failure. Previous studies have indicated that serum levels of the pro-inflammatory cytokine TNF-α, is elevated in PBC patients with fibrosis. Although the severity of cholangitis might also be related to the PBC subtype, its etiology has been unclear. Several studies have shown that microRNAs (miRNAs) demonstrate specific expression patterns in various diseases. In the present study, we evaluated miRNA expression patterns among the PBC subtypes using comprehensive deep sequencing. We also carried out histologic examination by laser capture microdissection and investigated how the identified miRNAs were involved in PBC clinical progression using the miRNA transfection method. On average, ~11 million 32-mer short RNA reads per sample were obtained, and we found that the expression levels of 97 miRNAs differed significantly among the four groups. Heat mapping demonstrated that the miRNA profiles from hepatic failure and portal hypertension type were clustered differently from those of the gradually progressive type and controls. Furthermore, we focused on miR-139-5p, which has an adequate number of total short reads. Quantitative reverse transcription PCR showed that miR-139-5p was significantly downregulated in clinically advanced PBC. Also, examination of liver tissues demonstrated that the expression of lymphocyte-derived miR-139-5p was significantly higher in hepatocytes. In vitro, the level of TNF-α was significantly elevated in supernatant of cells with upregulation of miR-139-5p. Furthermore, c-FOS gene transcription was repressed. Thus, we have demonstrated a novel inflammation-regulatory mechanism involving TNF-α and c-FOS transcription through miR-139-5p in the NF-κB signaling pathway. We conclude that the specific miRNA miR-139-5p might be involved in the pathogenesis of PBC, especially during clinical progression.

Expression of the RNA‐binding protein Musashi1 in adult liver stem‐like cells

Aim:  Musashi1 is an RNA‐binding protein that regulates the Notch signaling pathway in stem cells. Our previous study revealed that Musashi1 is expressed in early hepatocytes during liver development in the mouse. However, whether this unique protein is expressed with Notch signaling markers in adult liver stem‐like cells remains unknown.