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Background. Hepatocellular carcinoma often shows a resistance to transcatheter arterial embolization in the course of therapy repetition. Methods. Forty-four of 103 consecutive patients with hepatocellular carcinoma showing a resistance to repeated embolization therapy were treated with intra-arterial injection of the high-molecular weight antitumor agent styrene-maleic acid neocarzinostatin (Zinostatin) mixed with Lipiodol (group A). The remaining 59 patients received repeated embolization with epirubicin given in the same way (group B). Results. In group A, computerized tomography scans 3 months after the therapy showed "complete" accumulation of Lipiodol in 2 patients (4.5%), and "good" accumulation (50%–99%) in 11 (25.0%); 10%–49% accumulation was shown in 12 patients (41.9%), and less than 10% in 19 patients (32.6%). In group B, 1 patient (1.7%) showed complete accumulation, 4 (6.8%) showed "good" accumulation, 10 (16.9%) showed 10%–49% accumulation, and 44 (74.6%) showed less than 10%. Multivariate logistic regression analysis showed that factors affecting Lipiodol accumulation after therapy included tumor multiplicity (P < 0.0001), use of Zinostatin (P = 0.010), and decompensation of cirrhosis (P = 0.049). In the 44 patients with Zinostatin injection, tumor size was the only factor affecting Lipiodol accumulation. Survival rates in groups A and B were 70.4% and 45.8%, respectively, at the end of the first year, 36.8% and 17.3% at the end of the second year, and 24.5% and 13.0% at the end of the third year (P = 0.0087). Conclusion. Intra-arterial Zinostatin injection therapy increased the Lipiodol accumulation rate and the survival rate in patients with embolization-resistant HCC, and the best candidates for the treatment were patients with smaller liver cancer of 50 mm.

To elucidate the relationship between angiographic features and histological findings, an immunohistological study of alpha-smooth muscle actin was performed in 106 patients with small hepatocellular carcinoma. Arterial dominance or portal blood paucity were found in 73 patients (68.9%) on digital subtraction angiography, 88 (83.0%) on computerized tomographic arterial portography and 87 (82.1%) on carbon dioxide-enhanced ultrasonography. Among 73 patients with hypervascularity on angiography, 57 (78.1%) had thick-walled, nuclei-rich and slender-shaped vessels (type II), eight (11.0%) had thin-walled, nuclei-poor and oval-shaped vessels (type I) and the remaining eight had a mixed type of II and I. Conversely, among 33 patients without hypervascularity, five (15.2%) had a type II, 21 (63.6%) had a type I, five had a mixed type and two had no positive vessel. Tumour size, histological classification and amount of non-triadal vessels were also associated with the angiographic appearance of the tumours. Among varied aspects of the cancer including tumour size, tumour multiplicity, microscopic portal invasion, histological classification, amount of alpha-smooth muscle actin-positive vessels and shape of alpha-smooth muscle actin-positive vessels, multivariate logistic regression analysis demonstrated that the shape of alpha-smooth muscle actin-positive vessels was solely associated with angiographic hypervascularity independently (P

To assess the biochemical and histological characteristics of hepatitis G virus (HGV) infection we examined four patients who were infected with HGV only (HGV group), and compared them with 16 patients infected with both HGV and hepatitis C virus (HCV; HGV+HCV group) and 18 patients intecred with HCV only (HCV group). Biochemical examination showed a significantly low level of serum alanine aminotransferase (ALT) in the HGV group, and that the gamma-glutamyl transpeptidase (γ-GTP)/ALT ratio in the same group was significantly higher than in the other two groups.Although all three patient groups had a similar degree of liver fibrosis, both the degree of periportai inflammation and total histological activity index were significantly lower in the HGV group than in the other two groups. Fibrous enlargement of the portal tract without lymphoid infiltration and thin fibrous septa was characteristically observed in the HGV group. No significant difference was found between the HGV+HCV group and HCV group. Our results suggest that biochemical and histological changes in HGV infection are very mild and quite different from those of HCV infection.

Interferon (IFN) can reduce hepatitis C virus load and even eliminate the virus in 30-40% of patients. Several predictive factors for eradication of the virus have been reported and a higher dose of IFN tends to result in elimination of the virus. However, a small dose of IFN sometimes is as effective as a large dose in eradicating the virus. The predictive factors for such a response are not well established. We retrospectively analysed 50 patients with chronic hepatitis C who were treated with relatively small amounts of IFN (equal or less than 252 million units). Eleven patients were responders (elimination of hepatitis C virus (HCV) and normalization of alanine amino transferase (ALT) for at least 6 months), but the remaining 39 were non-responders. Multivariate analysis showed that the pretreatment viral load and total dose of IFN per kilogram of bodyweight were significant predictive factors of response to therapy. We also assessed the amino acid substitutions in the IFN sensitivity determining region (ISDR), NS5A codon 2209-2248, of HCV in serum samples obtained from 31 patients with HCV genotype 1b. The presence of more than one amino acid substitution in the ISDR tended to correlate with HCV genotype 1b elimination. As IFN is expensive and has a number of serious side effects, our study suggests that the optimal dose of IFN may vary from one patient to another and that more stringent criteria should be used to select the optimal dose for therapy.

Detection of hepatitis C virus (HCV) RNA by a second generation (ver 2) HCV bDNA-probe method (bDNA-probe) was compared with detection by the first generation (ver 1) assay. The two assays were performed simultaneously with the same serum samples of HCV genotypes 1b, 2a, 2b, 3a, and 3b. The positive rates with ver 1 were 82% for HCV genotype 1b (type 1b), 57.6% for HCV genotype 2a (type 2a), 75.0% for HCV genotype 2b (type 2b), 55.6% for HCV genotype 3a (type 3a), and 93.8% for HCV genotype 3b (type 3b). The positive rates with ver 2 were 95.0% for type 1b, 93.9% for type 2a, 83.3% for type 2b, 100% for type 3a, and 93.8% for type 3b. With Fisher's exact test, the detection rate for type 2a was significantly higher (P = 0.001) with ver 2 than with ver 1. We obtained regression lines using the HCV counts measured by bDNA-probe on the y axis and the HCV counts obtained by an HCV reverse transcriptase (RT)-competitive polymerase chain reaction method (competitive PCR) on the x axis. The gradients for types 1b, 2a, and 3b were greater with ver 2 compared to ver 1. The gradients for types 2a and 3b were the highest: for type 2a, y = 0.135x + 0.6 with ver 1 and y = 0.248x + 0.1 with ver 2; for type 3b, y = 0.366x + 0.1 with ver 1 and y = 0.727x + 0.3 for ver 2. In addition, HCV-RNA counts for all the genotypes tested in this study were significantly higher with ver 2 than with ver 1. Hence, we conclude that ver 2 of the bDNA-probe measures HCV-RNA counts closer to those obtained with competitive PCR than the ver 1 assay.

BACKGROUND To elucidate the influence of long term interferon administration on the rate of occurrence of hepatocellular carcinoma (HCC) in patients with hepatitis B virus (HBV)-related cirrhosis, the authors analyzed 313 consecutive patients with cirrhosis. METHODS Of the 313 patients, 94 underwent long term intermittent administration of interferon for ≥6 months, and the remaining 219 patients received no interferon or other antiviral drug. RESULTS Cumulative occurrence rates of HCC in the group treated with interferon and the untreated group were 4.5% and 13.3%, respectively, at the end of 3 years; 7.0% and 19.6%, respectively at the end of 5 years; and 17.0% and 30.8%, respectively, at the end of 10 years. The rate of HCC development in the treated group was significantly lower than that of the untreated group (P = 0.0124). The Cox proportional hazard model revealed that interferon treatment was an independent contributing factor in lowering the rate of carcinogenesis (odds ratio = 0.39; P = 0.031) even after correction by significant covariates in multivariate analysis. The virologic study showed that the role of interferon therapy from the viewpoint of cancer prevention was much more significant in patients with a HBV DNA concentration of ≥10 milliequivalents. CONCLUSIONS Interferon therapy for patients with HBV-related cirrhosis significantly decreased the HCC rate, especially in patients with a larger amount of serum HBV DNA. If interferon is administered properly for a selected group of patients, an effective strategy of cancer prevention can be achieved, even in patients with cirrhosis. Cancer 1998;82:827-35. © 1998 American Cancer Society.

In order to elucidate the influence of hepatitis B virus on the hepatocellular carcinogenesis in cirrhotic liver, quantitative analysis of precore region of hepatitis B virus was sequentially carried out. Among 191 patients who were diagnosed with cirrhosis in our hospital between 1974 and 1992 and had received no anti-viral therapy, 52 patients were selected from the following three patient groups: group A, 17 patients with liver cancer development within 5 years after the diagnosis of cirrhosis; group B, 12 patients with cancer development between 5 and 10 years after the diagnosis; and group C, 23 patients without cancer development during the observation period of 10 years or more. In group A, 16 out of 17 patients steadily showed a large amount of precore mutant virus with a high concentration of 10 4 copy/ml or more throughout the observation period. In group B, precore mutant virus was persistently high in eight of 12 patients. The other four patients showed an increase in mutant virus until the appearance of HCC, from a low concentration of less than 10 4 copy/ml to a high concentration. Of 23 patients in group C, no mutant virus was detected throughout the 10 year observation period in 11 patients and a complete disappearance of the mutant virus was found during the observation period in seven patients. Mutant virus was detected at least once in the clinical courses in the five remaining of group C. The amount of precore mutant virus at the endpoint of observation was significantly associated with HCC development (28/29 vs. 5/23, P

The glomerular changes of 188 consecutive autopsy cases with hepatitis C virus (HCV) infection were studied. The glomerular changes were classified as follows: Category I: membranoproliferative glomerulonephritis (MPGN; 21 cases, 11.2%), 2) Category II: membranous nephropathy (MN; 5 cases, 2.7%), 3) Category III: mesangial proliferative glomerulonephritis (MesGN; 33 cases, 17.6%), 4) Category IV: mesangial thickening type without proliferative mesangial cell (MT; 44 cases, 23.4%), and 5) Category V: almost normal glomeruli (85 cases, 45.2%). Glomerulonephritis was defined as glomeruli with an increase in mesangial matrix or a thickening of the capillary walls in the glomeruli; categories I-IV corresponded to glomerulonephritis in this study. Multivariate analysis, using a multiple logistic model, indicated that glomerulonephritis with HCV infection was the most strongly correlated to the existence of esophagogastric varices. Abnormal urinalysis, that is transient or continuous microhematuria or proteinuria, was observed in only 23 (12.2%) cases. These results showed that in HCV-RNA positive patients with esophagogastric varices the possibility of glomerulonephritis should be considered.

To study the frequency of hepatitis G virus (HGV) infection after blood transfusion, we retrospectively examined serum samples obtained from 53 patients who received blood transfusion before, during and after resection of hepatocellular carcinoma. Whole blood or separated blood products (fresh frozen plasma, platelet and/or red blood cells) were administered to these patients. The total amount of fresh frozen plasma ranged from 10–248 U (median 38). A total of 15–80 U (median 40) of platelets were administered to 42 patients. Similarly, 1–15 U (median 4) of red blood cells were transfused into 22 patients. HGV RNA was extracted and detected by reverse transcription-nested polymerase chain reaction using 5′ noncoding region primers. At 1 month after transfusion, seven patients (13.2%) became positive for HGV RNA. Patients who acquired HGV were not different from those who did not with regard to clinical condition, the amount of blood transfusion, age, sex, co-infection with hepatitis C virus (HCV) or hepatitis B virus (HBV). Only one of the seven patients who was negative for HCV RNA and HBs antigen showed a flare up of alanine aminotransferase (ALT). In five of the patients, the virus persisted in the serum up to 2 years after transfusion. Our findings suggest that the incidence of HGV transmission by blood products is relatively high (13.2%) and that the disease, once acquired, is likely to become chronic.

BACKGROUND Hepatocellular carcinoma (HCC) occurs in patients with hepatitis C virus-RNA positive chronic liver disease. It is important to prevent HCC with drug administration. METHODS A retrospective study was undertaken to evaluate the long term preventive effect of Stronger Neo-Minophagen C (SNMC) on HCC development. SNMC is a Japanese medicine that is commonly administered to patients with chronic hepatitis C to improve the serum alanine aminotransferase (ALT) level. Of 453 patients diagnosed with chronic hepatitis C retrospectively in the study hospital between January 1979 and April 1984, 84 patients (Group A) had been treated with SNMC; SNMC was given at a dose of 100 mL daily for 8 weeks, then 2-7 times a week for 2-16 years (median, 10.1 years). Another group of 109 patients (Group B) could not be treated with SNMC or interferon for a long period of time (median, 9.2 years) and were given other herbal medicine (such as vitamin K). The patients were retrospectively monitored, and the cumulative incidence of HCC and risk factors for HCC were examined. RESULTS The 10th-year rates of cumulative HCC incidence for Groups A and B were 7% and 12%, respectively, and the 15th-year rates were 12% and 25%. By Cox regression analysis, the relative risk of HCC incidence in patients not treated with SNMC (Group B) was 2.49 compared with that of patients treated with SNMC (Group A). CONCLUSIONS In this study, long term administration of SNMC in the treatment of chronic hepatitis C was effective in preventing liver carcinogenesis. Cancer 1997; 79:1494-500. © 1997 American Cancer Society.

To explain the appearance rates of hepatocellular carcinoma (HCC) in patients treated with interferon (IFN) and to assess the risk factors for hepatocellular carcinogenesis, we studied 442 patients with chronic type C hepatitis for one to 7.6 years (median 2.8 years). Of these 442 patients, 135 (30.5%) showed a complete response with persistent HCV-RNA negativity for more than six months after completing IFN therapy. During the observation period, HCC developed in 16 patients. We divided the patients into three groups based on laparoscopic findings of the liver surface : smooth, irregular, and nodular. The cumulative appearance rates of HCC in these three liver types were zero, 6.6% and 14%, respectively, at the end of the fifth year. As to the efficacy of IFIM, the HCC appearance rate in patients who responded was lower than that in patients who did not (p

C型慢性活動性肝炎に対し天然型インターフェロン (以下IFN) α1回量6MUを2ないし8週間連日+週2～3回間歇で1年を越えて長期的に投与した26例につきIFN投与終了後の治療効果を検討した. IFN総投与量は762 (IU) ～1, 972 (IU) (中央値1, 236IU) であった. IFN終了後GPT正常かつHCV-RNA陰性が継続した著効は13例 (50%) であった. HCV-genotype 1bでみるとHCV-RNA量が106copy/ml以上では著効は23.1% (3/13) と低率であったが, HCV-RNA量が106copy/ml未満では著効は77.8% (7/9) と高率であった. HCV-genotypeが2a, 2bでは4例中3例 (75%) が著効であった. 従来難治といわれているHCV-genotype 1bでもHCV-RNAが106copy/ml未満であればIFN長期投与にて著効率が増加する可能性があると考えられた.

肝障害出現前または病初期から経過観察され, 肝障害発症時の状態が観察された原発性胆汁性肝硬変 (PBC) 3例について検討した. 症例1は慢性関節リウマチ治療開始11年後にd-ペニシラミンを開始した時よりALP上昇が始まり, 4年後にPBCの確定診断となり, 18年にわたる臨床経過が観察された. 症例2は, ネフローゼ症候群に対するステロイドの減量を契機に肝障害が出現した. ステロイドの増減により肝酵素の変動が認められたが, ウルソデオキシコール酸 (UDCA) 併用後は経過は安定している. 症例3は高血圧治療中に抗生剤投与をきっかけに発症したがウルソデオキシコール酸の投与により肝機能は安定した. いずれの例でも薬剤服用もしくはこれを必要とするような病態が肝障害顕在化の契機となっていた.

To elucidate the epidemiology of infection with hepatitis C virus (HCV) subtype 3b (a rare subtype thought to have originated in Southeast Asia) in Japan, we examined the genotypic subtype in 1397 patients with HCV-related chronic liver diseases. Of 1330 patients with identified HCV RNA genotypes, 960 had subtype 1b, 243 had subtype 2a, 97 had subtype 2b, 14 (1.1%) had subtype 3b, and 16 had other types of HCV or mixed subtypes. The age, gender, and severity of liver disease in patients with HCV subtype 3b did not differ from these features in patients with other subtypes. Eleven of the 14 patients with the 3b subtype had once worked at Company A in Tokyo, Japan. Multivariate logistic analysis showed that working history at that company was independently associated with the incidence of the subtype; the risk ratio was 207.2 (P

高速MRIにて慢性肝疾患に合併した肝内小結節性病変の診断を試みた.対象は慢性肝疾患を合併した,腫瘤径20mm以下の肝細胞癌(HCC) 78結節,腺腫様過形成(AH) 16結節,粗大再生結節(LRN) 26結節,肝血管腫(CH) 29結節である.すべて息止め下で,T1強調画像,T2強説西像をファーストスピンエコー法で撮像し,引き続いてGadolinium-DTPA急速静注下でダイナミックMRI (Fast multiplanar spoiled GRASS法)を撮像した.それぞれ20秒前後の息止めにて施行し,検査時間は約15分であった.T1強調画像,T2強調画像のいずれかで高信号もしくは低信号病変として検出されたのはHCC 92%, AH 56%, LRN 37%, CH 100%,ダイナミックMRIで早期濃染はHCC 74%, AH 0%, LRN 0%, CH 3%, 5分後の晩期濃染はHCC 0%, AH 0%, LRN 0%, CH 97%であった.高速MRIは短時間施行でき,侵襲も少なく,肝内小結節性病変の診断には感度,特異度共に高く有用な検査法と考えられた.

C型慢性活動性肝炎に対し天然型α-インターフェロン(以下IFN)1回量6MUを8週間連日+16週間週2～3回間歇投与を施行しIFN終了後2年以上経過した252例につき血清トランスアミナーゼ(以下GPT), HCV-RNA,肝組織像の変化および肝細胞癌発生率につき検討した.252例中IFN投与を予定スケジュールにて終了しえたのは236例であり,この236例中IFN終了後GPT正常かつIFN終了6カ月後のHCV-RNA陰性の著効は105例(44.5%)であった.さらにIFN終了後の肝生検像の変化については著効例ではgradeの改善はIFN終了後1～3年で22/23 (95.7%)にみられるのに比し,stageの改善は1～3年では4/23 (17.4%),3～7年では5/8 (62.5%)とstageの改善にはより長期間を要すると考えられた.一方不完全著効及び無効例でにはgrade及びstageの改善率は著効例に比し低率であった.またIFN終了後の肝細胞癌の発生は236例中2例(0.8%)で肝細胞癌がみられた.

Background Concanavalin A (Con A) affinity electrophoresis of serum alpha-fetoprotein (AFP) can distinguish hepatocellular carcinoma (HCC) from other malignancies when the serum AFP concentration is elevated. However, Con A has not been able to distinguish HCC from benign chronic liver disease such as cirrhosis or chronic hepatitis. Methods The Con A affinity electrophoresis of serum AFP was analyzed in patients with a serum AFP concentration greater than 50 ng/mL by antibody affinity electrophoresis and Western blotting in an attempt to distinguish hepatocellular carcinoma from benign chronic liver disease. Before the assay, the serum AFP concentrations were adjusted between 100 ng/ml and 300 ng/ml by concentrating or diluting the samples. Results Of 180 patients with HCC, 44 (24%) had a single band and 91 (51%), 35 (19%), and 10 (6%) had 2, 3, and 4 bands, respectively. All 35 patients with chronic hepatitis had a single band. All but 1 of 72 patients with cirrhosis had a single band. Multiple AFP bands on Con A affinity electrophoresis appear to be diagnostic of HCC. This method has a sensitivity of 76%, a specificity of 99%, a positive predictive value of 99%, and a negative predictive value of 71% for detecting HCC. The number of AFP bands correlated with serum AFP concentration and tumor size in patients with HCC. Conclusions This assay is useful for distinguishing HCC from benign chronic liver diseases.

We analyzed the pre-S and S genes of hepatitis B virus (HBV) obtained from a pair of siblings, who tested positive for antibody to hepatitis B surface antigen, and identified novel HBV variants. While Patient 1 tested positive for hepatitis B surface antigen (HBsAg) by radioimmunoassay, Patient 2 tested negative for the same antigen. Two different mutants were obtained from Patient 1. The first strain showed three amino acid mutations in the S region at positions 130 (G → N), 138 (C → S) and 145 (G → R). Another strain had mutations at 133 (M → T) and 134 (F → I). Although one strain obtained from Patient 2 had a mutation at 145 (G → K), the second strain showed no mutation in the S region. The different genetic and serological states found in these patients suggest that further diagnostic and vaccine strategies should be considered in eliminating HBV infection.

A prospective randomized trial was conducted to evaluate the efficacy of long-term oral administration of low-dose tegafur combined with uracil as an adjuvant chemotherapy, following transcatheter arterial embolization (TAE) in 40 patients with hepatocellular carcinoma (HCC). Forty eligible patients were randomized into two groups: 20 with TAE plus UFT (a compound of tegafur 200 mg and uracil 448 mg per day) and 20 with TAE alone. A good necrosis rate or decrease in size of more than 70% of the original tumor mass was attained in 10 by TAE plus UFT arm and in 12 by TAE arm alone. As for the "responded" patients, there was no significant difference in the time from tumor response to tumor regrowth between the two groups. The appearance rate of ascites and/or encephalopathy in patients with chemotherapy was slightly higher than that in control patients. The median survival time was 22.7 months for TAE plus UFT arm and 28.2 months for TAE arm alone. There was no significant difference in the cumulative survival curves. In conclusion, these results indicated no substantial benefit for this chemotherapy regimen, as an adjuvant therapy for patients with HCC during repeated TAE.

To evaluate portal blood flow in small hepatocellular carcinomas (HCCs) by means of computed tomography during arterial portography (CTAP) and CTAP with hepatic arterial obstruction (CTP-HAO) achieved by means of balloon occlusion.Thirteen patients with small HCC (20 mm in diameter) underwent CTAP, CTP-HAO, carbon dioxide-enhanced ultrasound (CEUS), and digital subtraction angiography (DSA). The imaging findings were correlated with histologic features.The first group of patients (n = 3) had tumors with portal blood flow at both CTAP and CTP-HAO and no hypervascularity at CEUS. The second group (n = 3) had tumors with portal blood flow at CTP-HAO but not at CTAP and hypervascularity at CEUS only. The third group (n = 7) had tumors without portal blood flow at CTAP or CTP-HAO and hypervascularity at DSA and CEUS. The first and second groups had well-differentiated HCCs; six of seven patients in the third group had moderately differentiated HCCs.Lack of portal blood flow was reversible in well-differentiated HCCs but irreversible in the other tumors.

To elucidate the detectability of small hepatocellular carcinoma by various imaging modalities, we performed digital subtraction angiography, computed tomographic arterioportography and carbon dioxide-enhanced ultrasonography. Of 76 patients with a small hepatocellular carcinoma of 2 cm or less in maximum diameter, 61 underwent digital subtraction angiography, computed tomographic arterioportography and enhanced ultrasonography at the same time. Concerning the 61 patients undergoing all the procedures, the characteristics of hepatocellular carcinoma were found in 57.4% (35 of 61) by digital subtraction angiography, 75.4% (46 of 61) by computed tomographic arterioportography and 72.1% (44 of 61) by enhanced ultrasonography. Among them, four hepatocellular carcinomas were detected only by enhanced ultrasonography, three were diagnosed only by computed tomographic arterioportography and two were diagnosed by both of them. Except for six hemangioma nodules that were easily diagnosed only with angiography, four of 55 benign hepatic nodules (7.3%) showed false-positive findings suggestive of hepatocellular carcinoma with either computed tomographic arterioportography or enhanced ultrasonography. In conclusion, computed tomographic arterioportography and enhanced ultrasonography could complementarily detect a small hepatocellular carcinoma more sensitively than digital subtraction angiography.

Interferon has been shown to be an effective treatment for some patients with chronic hepatitis C. In this study, the value of retreatment of nonresponders to interferon was investigated. Thirty-eight patients with hepatitis C virus (HCV)-RNA-positive chronic hepatitis C who had been treated with beta-interferon but still showed an alanine aminotransferase (ALT) level50 KU upon completion of therapy were retreated with alpha-interferon. Eight patients (21.1%) had normalization of ALT levels for at least 6 months after the completion of retreatment. The factors related to normalization of ALT levels after interferon retreatment were studied. Of 16 patients with transient HCV-RNA negativity 1 month after the initial interferon therapy, 7 (43.8%) had a complete response, with normalization of ALT levels and undetectable HCV-RNA, more than 6 months after interferon retreatment. On the other hand, of the 22 patients with HCV-RNA activity 1 month after the initial interferon therapy, only 1 (4.5%) had a complete response. Multivariate analysis, using a multiple logistic model, indicated that a complete response to readministration of interferon was most strongly correlated to transient negative conversion for HCV-RNA after the initial course of treatment.

To determine the patient-dependent and virus-related factors that may predict sustained response to interferon-α therapy, we prospectively evaluated 60 consecutive patients with chronic hepatitis C who received a standardized treatment schedule of interferon-α. Twenty-eight patients achieved a long-term sustained remission, 14 patients had temporary responses during treatment but relapsed after completing it and 14 patients did not respond. Four patients dropped out because of severe side effects and were excluded from evaluation of efficacy. Twenty-one variables were chosen as possible predictors of sustained response and were analyzed by means of multi-variate analysis. Variables related to the hepatitis C virus included genotype and concentration in serum before treatment. The former was determined by means of the polymerase chain reaction with genotype-specific primers for genotypes PT, K1, K2a and K2b, which were deduced from nonstructural region 5 of the hepatitis C virus genome. The latter was measured with a competitive polymerase chain reaction technique. Three variables were statistically significant (p

To study the antiviral effect of lymphoblastoid alpha interferon (IFN) on hepatitis C virus (HCV) we conducted a randomized, controlled trial on 80 patients with chronic hepatitis C using three different doses. Patients were randomly assigned to treatment with 1, 3 or 6 million units of lymphoblastoid IFN-alpha daily for 2 weeks. To assess the antiviral effect of IFN, the amount of HCV present in the serum was estimated by competitive nested polymerase chain reaction (PCR) before and after 2 weeks of treatment. The multiple logistic analysis was used to evaluate factors associated with virus clearance, adjusting the imbalance in predictive factors among patients. Hepatitis C virus became negative as assessed by nested PCR after therapy in 26, 50 and 63% of patients receiving 1, 3 and 6 mega units, respectively. Hepatitis C virus was cleared more often in patients having initially low (< 10(5)/mL) amounts of virus. No significant decrease in the amount of virus was observed in the untreated, control group. Patients without bridging fibrosis in liver histology and with HCV genotypes other than K1 (type II) tended to respond well. These results indicate that lymphoblastoid IFN-alpha suppresses HCV in a dose dependent manner. Higher initial virus amounts, bridging fibrosis and genotype K1 were factors associated with poor response.

reverse transcription (RT) -competitive nested PCRによるC型肝炎ウイルスRNAの測定法について,各種standardを用いて検討した.各種DNA standard間の比較では,38bp, 110bpのinsersionを導入したstandardでは,34bpのdeletionを導入したstandardに比べ100.5copy/ml, 103.5copy/ml多い値を示した.制限酵素切断点を導入したstandardでは,34bpのdeletionを導入したものより100.5copy/ml,多い値を示した.DNA stantardとRNA standardを比較すると,DNA standardはRNA standardより102copy/ml少ない値を示した.しかし,DNA standardを使用した場合,試薬,所要時間はRNA standardに比べ約1/5であった.34bpのdeletionを導入したDNA standardを用いて再現性の検討を行ったところ,日差再現性,術者間再現性は良好であり,添加回収試験の成績も良好であった.以上より,34bpのdeletionを導入したDNA standardを用いた測定法は簡易さ,再現性とも優れていた.

To elucidate the appearance rates of hepatocellular carcinoma in cirrhosis and to assess the risk factors for hepatocellular carcinogenesis, we prospectively studied 795 consecutive patients with viral or alcoholic cirrhosis for 2 to 17 yr (median of 5.8 yr). During the observation period, hepatocellular carcinoma developed in 221 patients. Cumulative appearance rates of hepatocellular carcinoma were 19.4%, 44.3% and 58.2% at the end of the fifth, tenth and fifteenth years, respectively. When classified by the type of hepatitis virus infection, the appearance rates of hepatocellular carcinoma in 180 patients with only HBsAg and in 349 patients with only antibodies to hepatitis C virus were 14.2% and 21.5% at the fifth yr, 27.2% and 53.2% at the tenth yr and 27.2% and 75.2% at the fifteenth yr, respectively. Cox proportional hazard model identified that α-fetoprotein levels (p = 0.00001), age (p = 0.00067), positive hepatitis C virus antibodies (p = 0.00135), total alcohol intake (p = 0.00455) and indocyanine green retention rate (p = 0.04491) were independently associated with the appearance rates of hepatocellular carcinoma. Whereas age and indocyanine green retention rate were independent predictors for the appearance rate of liver tumor in the subgroup of HBsAg-positive patients, α-fetoprotein levels, age and past alcohol consumption were independent predictors in the group of hepatitis C virus antibody–positive patients. These epidemiological results suggest that some differences exist in the activity and modes of cancer promotion between hepatitis B virus infection and hepatitis C virus infection. (HEPATOLOGY 1993;18:47–53).

Although biochemical and virological responses to corticosteroid withdrawal therapy for chronic hepatitis B have been extensively studied, long term changes in liver histology have not been well documented.We retrospectively analyzed 45 paired liver biopsy specimens taken before and after treatment from 40 patients who persistently showed biochemical remission and an absence of HBe antigen (RIA) for up to 20 yr.The grading scores for necroinflammatory and fibrotic activity in the liver specimens decreased significantly after corticosteroid withdrawal therapy. Histological scores graded according to Knodell's components improved significantly in every category after corticosteroid withdrawal therapy. However, inflammatory cell infiltrates remained within the liver for long periods. The disappearance rate of necroinflammation in the periportal, lobular, and portal regions of the liver were 25%, 7.4%, and 7.4%, respectively, at yr 5 after therapy, and were 84.4%, 78.2%, and 58.7%, respectively, at yr 10 after therapy. The cumulative disappearance rate, calculated using the Kaplan-Meier method, was significantly lower for portal inflammation than for periportal necroinflammation.Our results show that: 1) despite clinical remission of chronic hepatitis B virus infection, long periods are needed for histological resolution of necroinflammation in the liver; and 2) by a Cox proportional hazard analysis of the factors contributing to histopathological disappearance of disease-related inflammation, the degree of fibrosis of liver biopsy specimens from pretherapy patients was the most statistically significant factor (p = 0.049).

Background/Aims/Methods: The aim of this study was to elucidate the rate of development to cirrhosis and the rate of development to cirrhosis and the rate of appearance of hepatocellular carcinoma in chronic viral hepatitis and to assess the risk factors for the development of disease in 2215 consecutive patients with viral hepatitis who were prospectively studied for a median observation period of 4.1 years. Results: The rates of development to cirrhosis were 7.6%, 21.7%, and 32.2%, at the 5th, 10th, and 15th year, respectively. The carcinogenesis rates were 3.4%, 10.5%, and 22.4% at the 5th, 10th, and 15th year, respectively. The appearance rates of cancer in 645 patients with only hepatitis B surface antigen and in 1500 patients with only anti-hepatitis C virus antibodies were 2.1% and 4.8% at the 5th year, 4.9% and 13.6% at the 10th year, and 18.8% and 26.0% at the 15th year, respectively. The proportional hazard model identified that the amount of alcohol intake ( p =0.0002) and the indocyanine green retention rate ( p =0.022) were independently associated with carcinogenesis in hepatitis type B; and stage of hepatitis ( p p =0.0046), history of blood transfusion ( p =0.0093), albumin ( p =0.012), and amount of alcohol intake ( p =0.031) were independently associated with the carcinogenesis rate in hepatitis type C. Although the severity of portal fibrosis was closely correlated with the future disease development and carcinogenesis in chronic hepatitis C, it was not a good predictor in chronic hepatitis B. Conclusion: These epidemiological results suggest that there are some differences in the activity and modes of disease progression and cancer promotion between hepatitis B virus infection and hepatitis C virus infection.

Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However, the emergence of lamivudine-resistant mutant virus, with amino acid substitution in the YMDD motif of DNA polymerase, has been reported. We report the emergence and takeover of YMDD mutant and re-takeover by wild type during and after long-term lamivudine therapy. YMDD mutants were detected in five patients who showed DNA breakthrough (HBV DNA becoming detectable after a period of DNA negativity), which occurred after 9 to 14 months of lamivudine therapy. Four of five mutants had amino acid sequence YIDD, and the remaining mutant had YVDD. Patients with high HBV-DNA titer and/or hepatitis B e antigen tended to develop breakthrough (P = .038). Using a sensitive and specific polymerase chain reaction (PCR)-based method developed in this study, the emergence of YMDD mutants was detected 1 to 4 months before DNA breakthrough, but not detected in any of the pretreatment sera. The mutants were predominant at breakthrough, but were replaced by wild-type virus 3 to 4 months after cessation of therapy in the two patients who discontinued therapy. One of these patients had a relapse of hepatitis. Mutant continued to replicate in the remaining three patients who continued to receive treatment, and relapse occurred in only one of these patients. Our results suggest that the replication of YMDD mutant viruses is less than wild type and is re-overtaken by wild type after cessation of therapy. Re-administration of lamivudine, possibly combined with other antiviral therapy, might be useful in some patients experiencing hepatitis with lamivudine-resistant variants.

To assess the efficacy of the zinostatin derivative, the anti-tumor agent, styrene-maleic acid neocarzinostatin, in treating multiple small liver cancers, 29 patients with multiple hepatocellular carcinoma of 3 cm or less in diameter were treated with intraarterial injections of this high molecular weight agent, mixed with Lipiodol. Computed tomography 3 months after the first therapy showed complete deposition of Lipiodol in the entire area of the original tumor in 8 patients (27.6%), 50%–99% deposition in 4 (13.8%), 10%–49% in 10 (34.5%), and less than 10% in 7 (24.1%). After repeated injections, Lipiodol deposition in the entire area of the original tumor was found in 11 patients (37.9%). The degree of Lipiodol deposition depended on the angiographic vascularity of the tumor and on the images of the computed tomogram during arterial protography. Although complete deposition of Lipiodol was found in all tumors in 10 (58.8%) of the 17 patients with well demarcated round hypervascularity, only 1 (8.3%) of 12 patients with ill demarcated tumors showed complete deposition of Lipiodol in the tumors. Taking into account that hypervascularity on angiograms was closely correlated with the degree of Lipiodol accumulation on computed tomograms taken later, it appears that well demarcated round-shaped liver cancer is the best candidate for styrene-maleic acid neocarzinostatin therapy.

Background/Aims: Although biochemical and virological responses to interferon-α therapy for chronic hepatitis C virus infection have been extensively studied, long-term changes in liver histology have not been well documented. Methods: We retrospectively analyzed 105 paired liver biopsy specimens taken before and after treatment from 93 patients who persistently showed biochemical remission and an absence of viremia for up to 68 months. Results: The grading scores for necroinflammatory and fibrotic activity in the liver biopsy specimens decreased significantly after interferon-α therapy. Histological scores graded according to Knodell's components improved significantly in every category after interferon-α therapy. However, inflammatory cell infiltrates remained within the portal tracts for long periods: necroinflammation in the periportal lobular regions were absent in most of the post-therapy specimens. The cumulative disappearance rate, calculated using the Kaplan-Meier method, was significantly lower for portal inflammation than for periportal or lobular necroinflammation but was equivalent to that for histological disease activity. On univariate analysis, age and fibrosis at the onset of treatment were significant factors influencing the response of histological disease activity to interferon-α therapy ( p =0.025 and 0.49, respectively). Using Cox's proportional hazard analysis, age was the only significant independent predictor of histological response to treatment ( p =0.035). Conclusions: Clinical remission of chronic hepatitis C virus infection is associated with histological resolution of necroinflammation in the periportal and lobular regions. Host-related factors are likely to influence whether early remission of inflammation after interferon-α therapy occurs.

A prospective randomized trial was conducted to evaluate the efficacy of long-term oral administration of low-dose 5'-deoxy-5-fluorouridine (5'-DFUR) as an adjuvant chemotherapy, following transcatheter arterial embolization (TAE) in 40 patients with hepatocellular carcinoma (HCC). Forty eligible patients were randomized into two groups: 20 with TAE plus 5'-DFUR (400 mg/day) and 20 with TAE alone. A good necrosis rate or decrease in size of more than 70% of the original tumor mass was attained in 14 by the TAE plus 5'-DFUR arm, and in 12 by the TAE arm at 3 months after the first TAE. Although all five patients with HCC and 70-99% necrosis rate after the first TAE in the TAE alone arm showed a less than 70% necrosis rate at 12 months, four of the seven patients with a 70-99% necrosis in the TAE plus 5'-DFUR arm retained a necrosis of more than 70% at 12 months after the first TAE. The appearance rate of ascites and/or encephalopathy in patients with chemotherapy was not different from that of patients with TAE alone. One-year survival rates in the TAE plus 5'-DFUR arm and the TAE alone arm were 75.0% and 85.0%, 2-year rates were 64.2% and 66.2%, and 3-year rates were 64.6% and 49.7%, respectively. There was no significant difference in the survival curves. In conclusion, adjuvant therapy with 5'-DFUR was well tolerated without significant side effects, and it might maintain a good necrosis state of HCC after TAE. In order to confirm a beneficial effect of the chemotherapy on the survival period, a study using more patients and longer observation periods will be required.

We performed serial hemodynamics in 15 patients with 21 well-differentiated hepatocellular carcinomas. The total length of the observation period ranged from 129 to 678 days (median, 368). We investigated both arterial and portal blood flow at intervals of at least 4 months. Arterial blood flow was measured with carbon dioxide-enhanced ultrasonography (US), and portal blood flow was measured with computed tomographic arterial portography (CTAP). None of the tumors were hypervascular on the initial study; however, by the final study nine nodules (42.9%) had become homogeneously hypervascular, two (9.5%) had become partially hypervascular, and the others did not change. Of 10 nodules without portal blood supply on any study, 8 became hypervascular by the final study. Of 11 nodules with portal blood supply on the initial study, a follow-up study showed no change in portal flow in 6 and the loss of portal flow in 5. Four of these five became hypervascular, with a documented loss of portal flow before the increase in arterial flow. The doubling time of tumors with a homogeneous increase in arterial blood flow ranged from 89 to 333 days (median, 172), whereas the doubling time of other tumors ranged from 227 to 607 days (median, 392). Thus, growth rate and vascularity in well-differentiated hepatocellular carcinoma are closely correlated. When the well-differentiated hepatocellular carcinoma has portal blood flow and is not hypervascular, it grows slowly. On the other hand, when it loses portal blood flow and becomes hypervascular, it grows rapidly.

Background/Aims: Although intraspousal transmission of hepatitis C virus (HCV) has been speculated, there is no direct evidence. Methods: To investigate whether transmission of HCV occurs by this route, 295 spouses of persons diagnosed with HCV were studied. Of these, 25 (8.8%) tested positive for anti-HCV. Next, the HCV genotype was determined by polymerase chain reaction using a mixed primer set, and cDNA was obtained in 17 of the 25 couples for comparison of the genotypes. Results: Of these 17, 14 (82.4%) spouses were shown to be infected with HCV of the same genotype. Analysis of the nucleotide sequence of putative E1 gene of eight couples having the same HCV genotype revealed that five couples had remarkably high nucleotide sequence homologies (>97%), whereas samples obtained from the remaining three couples showed relatively low homology (91–92%). Nucleotide sequence homologies were significantly higher between spouses than between non-spouse pairs of isolates. Phylogenetic analysis using the neighbor-joining method suggests that infection in these five couples probably occurred after marriage. Furthermore, none of the five couples had shared other possible transmission routes such as intravenous drug use, dental treatment or acupuncture. Conclusion: These data strongly suggest the occurrence of intraspousal transmission of HCV

We examined 613 Toranomon Hospital patients with HCV RNA-positive chronic liver disease to elucidate the viral genotype in the Tokyo metropolitan area. An epidemiological and clinical study was conducted in the 565 patients whose HCV genotype could be determined. The HCV genotypes found were type II, in 414 patients (67.5%); type III, in 103 (16.8%); type IV, in 37 (6.0%); type V, in 4 (0.7%); mixed genotype II and III, in 5 (0.8%); and mixed genotype II and IV, in 2 (0.3%). The HCV genotype could not be determined in 48 patients. Type II was most prevalent. The HCV genotype I was not found at all. There were no significant differences between genotypes in relation to sex, age, history of blood transfusion, or the progression of the disease. It was uncommon to find a history of blood transfusion in the patients with mixed genotypes; however, a high incidence of hepatic disorders was noted among the family members of these patients. Ninety-two percent of the patients with HCV genotype II tested positive for C100-3, while 70.9% of those with type III, and 43.2% of those with type IV tested positive for this antibody. HCV genotype II was most prevalent, and the positivity rate for anti C100-3 in patients with this HCV genotype was significantly higher (P

Interferon has been shown to clear hepatitis C virus (HCV) from serum and normalize ALT levels in patients with hepatitis C. To study various predictive factors, serum samples were analyzed from 26 patients treated with IFN-α and 26 control patients. Patients with genotype II initially had higher viremia and showed slower viral clearance compared with patients with genotype III or IV. Relapse was related to a slow titer decrease (P < 0.04). A high degree of nucleotide sequence variability in the E2/NS1 region was found in three patients with genotype II who responded poorly. HCV sequences from one of the persistent nonresponders were analyzed to study changes in the nucleotide sequences of the HVR. Additional amino acid substitutions were seen after the first treatment, and a single substrain eventually overtook others. We suggest that genotype II patients often have faster replication, which may explain their poor response to IFN treatment.

Four subtypes of hepatitis C virus (HCV), Pt(I), K1(II), K2a(III) and K2b(IV), have been suggested based on the nucleotide sequences of the non-structural (NS) 5 region. A fifth subtype from Japanese patients, Tr(V), which shows a less than 68% homology in nucleotide sequence when compared with other subtypes has been identified. A one-step method which enables a quick determination of subtype using polymerase chain reaction with a mixed primer set deduced from the sequence of each subtype has been developed. Using this technique, the subtypes of 418 out of 478 Japanese patients (87.4%) were determined. The incidence of each subtype in Japan was as follows: K1(II), 307 (73.4%); K2a(III), 74 (17.7%); K2b(IV), 28 (6.7%); and Tr(V), 3 (0.7%). This one-step subtyping technique should be useful for studying the epidemiology or biology of the HCV.

A novel technique for separating and concentrating metals with liquid surfactant membranes containing a mobile carrier has recently attracted practical interest through nuclear, hydrometallurgical and waste-water treating processes. In this work, to obtain the basic information for development of a novel separation technique, the extraction of copper was carried out using liquid surfactant membranes containing benzoylacetone as a mobile carrier. The effects of the processes of chelating complex formation and diffusion on extraction rate were examined experimentally and theoretically on the basis of a simple model for facilitated transport. The experimental results could be explained approximately by considering that the overall extraction rates of copper were controlled by the chelating complex formation and diffusion processes in the aqueous phase close to the interface of the liquid membrane.