Purpose: We present results of patients with hepatocellular carcinoma (HCC) treated with proton beam therapy. Experimental Design: We reviewed 162 patients having 192 HCCs treated from November 1985 to July 1998 by proton beam therapy with or without transarterial embolization and percutaneous ethanol injection.The patients in the present series were considered unsuitable for surgery for various reasons, including hepatic dysfunction, multiple tumors, recurrence after surgical resection, and concomitant illnesses. The median total dose of proton irradiation was 72 Gy in 16 fractions over 29 days. Results: The overall survival rate for all of the162 patients was 23.5% at 5 years.The local control rate at 5 years was 86.9% for all 192 tumors among the 162 patients.The degree of impairment of hepatic functions attributable to coexisting liver cirrhosis and the number of tumors in the liver significantly affected patient survival. For 50 patients having least impaired hepatic functions and a solitary tumor, the survival rate at 5 years was 53.5%.The patients had very few acute reactions to treatments and a few late sequelae during and after the treatments. Conclusions: Proton beam therapy for patients with HCC is effective, safe, well tolerable, and repeatable. It is the useful treatment mode for either cure or palliation for patients with HCC irrespective of tumor size, tumor location in the liver, insufficient feeding of the tumor with arteries, presence of vascular invasion, impaired hepatic functions, and coexisting intercurrent diseases.

Inchin-ko-to (ICKT), an herbal medicine, and its ingredients exert potent choleretic effects by a "bile acid-independent" mechanism. The current study was designed to determine whether ICKT or its ingredients potentiate multidrug resistance-associated protein 2 (Mrp2; Abcc2)-mediated choleresis in vivo. Biliary secretion of Mrp2 substrates and the protein mass, subcellular localization, and messenger RNA (mRNA) level of Mrp2 were assessed in rat liver after infusion of genipin, an intestinal bacterial metabolite of geniposide, a major ingredient of ICKT. The function of Mrp2 was also assessed by the adenosine triphosphate (ATP)-dependent uptake of Mrp2-specific substrates using canalicular membrane vesicles (CMVs) from the liver. Infusion of genipin increased bile flow by 230%. It also increased biliary secretion of bilirubin conjugates and reduced glutathione (GSH) by 513% and 336%, respectively, but did not increase bile acid secretion. The ATP-dependent uptake of estradiol 17-␤-D-glucuronide (E 2 17␤G; by 265%), leukotriene C4 (LTC 4 ; by 161%), taurolithocholate-3-sulfate (TLC-3S; by 266%), and methotrexate (MTX; by 234%) was significantly stimulated in the CMVs from the liver. These effects were not observed in Mrp2-deficient rats. Under these conditions, genipin treatment increased the protein mass of Mrp2 in the CMVs but not the mRNA level. In immunoelectron microscopic studies, a marked increase in Mrp2 density in the canalicular membrane (CM) and microvilli was observed in the genipin-treated liver tissue sections when compared with the vehicle-treated liver tissue sections. In conclusion, genipin may enhance the bile acid-independent secretory capacity of hepatocytes, mainly by stimulation of exocytosis and insertion of Mrp2 in the bile canaliculi. ICKT may be a potent therapeutic agent for a number of cholestatic liver diseases. (HEPATOLOGY 2004;39:167-178.)

A new member of the UDP-N-acetylglucosamine: bgalactose b1,3-N-acetylglucosaminyltransferase (b3Gn-T) family having the b3-glycosyltransferase motifs was identified using an in silico method. This novel b3Gn-T was cloned from a human colon cancer cell line and named b3Gn-T8 based on its position in a phylogenetic tree and enzymatic activity. b3Gn-T8 transfers GlcNAc to the non-reducing terminus of the Galb1-4GlcNAc of tetraantennary N-glycan in vitro. HCT15 cells transfected with b3Gn-T8 cDNA showed an increase in reactivity to both LEA and PHA-L4 in a flow cytometric analysis. These results indicated that b3Gn-T8 is involved in the biosynthesis of poly-Nacetyllactosamine chains on tetraantennary (b1,6-branched) N-glycan. In most of the colorectal cancer tissues examined, the level of b3Gn-T8 transcript was significantly higher than in normal tissue. b3Gn-T8 could be an enzyme involved in the synthesis of poly-N-acetyllactosamine on b1-6 branched N-glycans in colon cancer.

1liver-infiltrating lymphocytes 4 of patients with chronic hepa-A cytotoxic T lymphocyte (CTL) response to the hepatitis C, the role of CTL responses in HCV infection is untitis C virus (HCV) nucleoprotein residues 88-96 that are known. HCV infection frequently persists and is implicated the minimal and optimal epitope for human leukocyte in the development of chronic hepatitis, cirrhosis, and hepatoantigen (HLA) B44-restricted CTLs was assessed in 27 cellular carcinoma. 5 Posttransfusion HCV infection has de-HLA B44-positive patients with chronic HCV infection.creased substantially after introduction of an anti-HCV assay Serum HCV RNA concentration and the amino acid sefor blood screening, but community-acquired HCV infection quence of the residues 81-100 were also determined.still occurs. Interferon therapy is effective in less than 50% Three patients were infected with HCV with uncommon of patients with chronic hepatitis C. 6 Understanding the role amino acid substitutions within the epitope. One was of CTL responses in HCV infection may contribute to the infected with HCV with an amino acid substitution in development of strategies for the prevention of HCV infection the flanking residues of the epitope. To stimulate CTLs and the elimination of HCV from infected individuals. in the peripheral blood, 9-mer peptides that correRecently, we showed the presence of CTLs that recognize sponded to the residues 88-96 of the individual patients endogenously synthesized HCV antigen in the peripheral were synthesized and used. Seven of the 27 patients demblood of some patients with HCV infection by stimulation onstrated a CTL response to the residues 88-96 with of peripheral blood lymphocytes (PBLs) with HCV synthetic specific cytotoxic activities higher than 20%. The CTL peptides.3 activities were significantly higher in patients with a The CTLs recognized an epitope in the HCV nucleoprotein low titer of serum HCV RNA than in those with a high residues 81-100 in association with human leukocyte antigen titer of serum HCV RNA (P Å .0006). Some of the patients (HLA) B44. The minimal and optimal epitope was further dethat demonstrated a CTL response to the residues 88-fined to be the residues 88-96. 7 The 9-mer peptide of the resi-96 also demonstrated a CTL response to a newly identidues 88-96 was recognized by and stimulated the CTLs more fied HLA B44-restricted CTL epitope or a known HLA efficiently than the 20-mer peptide of the residues 81-100. A11-restricted CTL epitope or both. No apparent associa-In a characteristic antiviral CTL response in vivo, immunotion was observed between the CTL response and the dominant CTLs recognize only one or a few multiple immunostage of disease, or between the CTL response and the genic CTL epitopes in the antigen, 8,9 but as many as five grade of necroinflammatory activity. The results suggest different epitopes for HCV-specific CTLs were detected in a that the HLA B44-restricted CTLs together with other single individual. 10 CTL activities to HCV could not be dem-HCV-specific CTLs ...

The accumulation of various 25-hydroxylated C 27 -bile alcohols in blood and their excretion in urine are characteristic features of cerebrotendinous xanthomatosis (CTX) a recessively inherited inborn error of bile acid synthesis caused by mutations in the mitochondrial sterol 27-hydroxylase (CYP27) gene. These bile alcohols may be intermediates in the alternative cholic acid side chain cleavage pathway. The present study was undertaken to identify enzymes and reactions responsible for the formation of these bile alcohols and to explain why Cyp27 ؊/؊ mice do not show CTX-related abnormalities. Microsomal activities of 5␤-cholestane-3␣,7␣,12␣-triol 25-and 26-hydroxylases, 5␤-cholestane-3␣,7␣,12␣,25-tetrol 23R-, 24S-, and 27-hydroxylases and testosterone 6␤-hydroxylase, a marker enzyme for CYP3A, in Cyp27 ؊/؊ mice livers were markedly up-regulated (5.5-, 3.5-, 6.5-, 7.5-, 2.9-, and 5.4-fold, respectively). In contrast, these enzyme activities were not increased in CTX. The activities of 5␤-cholestane-3␣,7␣,12␣-triol 25-and 26-hydroxylases and 5␤-cholestane-3␣,7␣,12␣,25-tetrol 23R-, 24R-, 24S-, and 27-hydroxylases were strongly correlated with the activities of testosterone 6␤-hydroxylase in control human liver microsomes from eight unrelated donors. Troleandomycin, a specific inhibitor of CYP3A, markedly suppressed these microsomal side chain hydroxylations in both mouse and human livers in a dosedependent manner. In addition, experiments using recombinant overexpressed human CYP3A4 confirmed that these microsomal side chain hydroxylations were catalyzed by a single enzyme, CYP3A4. The results demonstrate that microsomal 25-and 26-hydroxylations of 5␤-cholestane-3␣,7␣,12␣-triol and microsomal 23R-, 24R-, 24S-, and 27-hydroxylations of 5␤-cholestane-3␣,7␣,12␣,25-tetrol are mainly catalyzed by CYP3A in both mice and humans. Unlike Cyp27 ؊/؊ mice, CYP3A activity was not up-regulated despite marked accumulation of 5␤-cholestane-3␣,7␣,12␣-triol in CTX.