Serum interleukin-8 levels in patients with hepatocellular carcinoma

Serum levels of interleukin-8 (IL-8) in 73 patients with hepatocellular carcinoma (HCC), 24 with liver cirrhosis (LC) and 18 with chronic hepatitis (CH), and in 20 healthy controls were measured by an enzyme-linked immunosorbent assay. The mean (± SD) level of serum IL-8 in patients with HCC was 48.4 ± 60.8 pg/ml, which were significantly (P < 0.01) higher than those in patients with LC (8.6 ± 8.6) and CH (2.7 ± 6.1), or that in controls (1.8 ± 5.8). It was revealed that serum IL-8 level was normalized after resection of HCC in patients whose serum IL-8 level had been high at the time of diagnosis. On the other hand, serum IL-8 levels were progressively increased with time in patients with HCC who had not received effective treatment. These findings suggest that IL-8 is produced by HCC cells in vivo, and serum IL-8 levels could be a marker for HCC. Moreover, significant correlation between serum IL-8 levels and the tumor sizes was found in patients with moderately differentiated HCC, which were mostly hypervascular on angiography. Serum IL-8 levels were significantly lower in patients with well-differentiated HCC than those with moderately differentiated HCC. Recent study demonstrated that IL-8 is an angiogenetic factor. Our present data thus suggest that IL-8 production from HCC may be involved in angiogenesis of this tumor.     

Plasma free amino acid pattern in chronic hepatitis as a sensitive and prognostic index

To evaluate the diagnostic and prognostic significance of plasma amino acid imbalance in chronic hepatitis (CH), plasma-free neutral amino acid levels were examined in 47 patients with CH, consisting of 8 chronic persistent hepatitis (CPH), 26 chronic aggressive hepatitis (CAH) 2A and 13 CAH 2B, compared with those of 58 patients with liver cirrhosis (LC) and of 12 healthy controls. Fischer’s ratio (a molar ratio of branched chain amino acids to aromatic amino acids) was found to be reduced in the order of normal subjects (3.5±0.4), CPH (3.0±0.2), CAH2A (2.7±0.3), CAH2B (2.1±0.3), compensated LC (LC-C, 1.5±0.4) and decompensated LC (LC-D, 1.1 ±0.2). Patients with CPH showed a significant decrease of the ratio compared with normal subjects (P<0.05). The ratio was significantly higher in patients with CAH 2B in comparison with LC-C (P<0.001). Especially, the ratio could be used to discriminate the three distinct stages of CH. Discriminant analysis, carried out using six amino acids, Fischer’s ratio and conventional liver function tests indicated that Fischer’s ratio was the most reliable parameter for differentiation of the three stages of CH. Furthermore, serial examinations of Fischer’s ratio in patients with CH remained unchanged in CPH, whereas was significantly reduced in CAH during 2-3 years follow-up. These results strongly suggest that Fischer’s ratio is a useful indicator for differential diagnosis and for prediction of the subsequent clinical course of CH as well as being a sensitive index for functional hepatic reserve in chronic active liver diseases.     

Evaluation of plasma tissue plasminogen activator (I-PA) levels in patients with liver diseases

Previous studies have demonstrated that plasma tissue plasminogen activator (t-PA) level was elevated in patients with liver disease. In this study, t-PA antigen levels were investigated in patients with acute hepatitis (AH; N = 12), chronic hepatitis (CH; N = 8), compensated liver cirrhosis (CLC; N = 40), decompensated liver cirrhosis (DLC; N = 23) and hepatocellular carcinoma (HCC; N = 35). The increased t-PA levels (higher than 14 ng/ml) were found in 33% (4/12) of AH on the early hospital days, 25% (2/8) of CH, 45% (18/40) of CLC and 91% (21/23) of DLC, and 60% (21/35) of Hcc cases. In patient with LC, the correlations between t-PA levels and serum total bilirubin (T.Bill) and hepatic synthetic functions were investigated. The results were that the t-PA levels correlated positively with T. Bil and negatively with liver synthetic functions such as albumin, protein C and choline-esterase, indicating that t-PA increased almost in proportion to the deterioration of hepatic function. Serial determination of t-PA in patients with HCC treated by transcatheter arterial embolization (TAE) revealed that TAE failed to normalize the t-PA levels. In one case of HCC complicated with disseminated intravascular coagulation (DIC), t-PA showed a marked increase at acute phase of DIC and subsequent decrease after the successful treatment for DIC by gabexate mesilate (FOY) infusion. These results suggest that increased t-PA in liver disease is due mainly to deterioration of hepatic function, and that secondary fibrinolytic state, such as DIC, is also a contributing factor.     

Circulating vascular endothelial growth factor (VEGF) is a possible tumor marker for metastasis in human hepatocellular carcinoma

Vascular endothelial growth factor (VEGF) is closely related to angiogenesis in various human cancers. However, little is known of its circulating levels in hepatocellular carcinoma (HCC). We examined circulating VEGF levels in chronic liver disease to assess their clinical significance. Plasma VEGF concentrations were determined, by enzyme immunoassay, in patients with chronic hepatitis (CH; n = 36), liver cirrhosis (LC; n = 77), and HCC (n = 86) for a cross-sectional study. Plasma VEGF levels in healthy controls (n = 20) and CH, LC, and HCC patients were 17.7 ± 5.4 (mean ± SD), 30.6 ± 22.8, 34.4 ± 27.0, and 51.1 ± 71.9 pg/ml, respectively. The levels were significantly elevated in the HCC group, compared with the control, CH, and LC groups. Plasma VEGF levels in stage I, II, III, IVA, and IVB HCC patients were 27.6 ± 16.1, 26.5 ± 13.7, 35.8 ± 15.3, 45.4 ± 39.4, and 103.1 ± 123.2 pg/ml, respectively. The stage IVB patients with remote metastasis showed significantly marked elevation compared with the patients at the other stages. Platelet numbers were weakly correlated with plasma VEGF levels in the HCC group. Plasma VEGF level was highly elevated in patients with HCC, particularly those with metastatic disease. We consider that plasma VEGF is a possible tumor marker for metastasis of HCC. Circulating VEGF may be derived mainly from the large burden of tumor cells, and partly from platelets activated by the vascular invasion of HCC cells. (Received June 30, 1997; accepted Oct. 30, 1997)     

High serum levels of vascular endothelial growth factor in patients with human hepatocellular carcinoma

Vascular endothelial growth factor (VEGF) is intimately involved in neovascularization. In addition, it is know that in human hepatocellular carcinoma (HCC), angiogenesis is indispensable for tumor growth. In this study, we measured the serum VEGF levels of patients with HCC and studies the relationship between the serum VEGF level and maximum tumor diameter as well as that between the serum VEGF level and the serum α-fetoprotein (AFP) level. Mean serum VEGF level were 5.33 ± 0.77, 3.97 ± 0.68, 2.64 ± 0.78, and 2.57 ± 0.97 ng/ml for patients with HCC, chronic hepatitis (CH), or liver cirrhosis (LC) and normal controls (NC), respectively, with that of the HCC patients being significantly (P < 0.05) higher than that of the LC patient or NC. In addition, the serum VEGF level was significantly (r = 0.53, P < 0.05) correlated with the maximum tumor diameter in the HCC patients, and the sera of the patients with hypervascular HCC showed a significantly (P < 0.01) higher VEGF titer than the sera of the patients with isovascular or hypovascular HCC. However, there was no significant correlation between serum VEGF level and serum AFP level. These findings suggest that VEGF may play an important role, apart from that in AFP production, in the development of HCC.     

Plasma Endotoxin and Serum Cytokine Levels in Patients With Alcoholic Hepatitis: Relation to Severity of Liver Disturbance

Background: Endotoxin plays an important role in the initiation and aggravation of alcoholic liver disease. In this study, we evaluated plasma endotoxin levels and serum concentrations of cytokines and lipopolysaccharide binding protein (LBP) during the acute and recovery phase of patients with alcoholic hepatitis; we also explored the prognostic factors associated with a fatal outcome. Methods: Fourteen patients, consisting of eight patients with alcoholic hepatitis (AH), five cirrhotics with superimposed AH (LC+AH), and one patient with severe alcoholic hepatitis (SAH), were studied. Among these, two with LC+AH died of hepatic failure. Results: Plasma endotoxin levels in the acute phase were higher in patients with AH (184.4 ± 159.4 pg/ml) and LC+AH (206.9 ± 174.9 pg/ml) than in healthy subjects (10.4 ± 5.5 pg/ml, p < 0.001). In particular, in one patient with SAH and one of two nonsurvivors, plasma endotoxin levels were markedly high relative to the other cases. In most survivors, plasma endotoxin levels decreased in the recovery phase, whereas they further increased at the terminal stage in one of two nonsurvivors. Serum interleukin (IL)-6 and IL-8 levels in the acute phase were significantly higher in patients with AH and LC+AH as compared with healthy subjects. These levels were especially high in nonsurvivors and in one patient with SAH. IL-10 increased in two nonsurvivors, one patient with SAH, and one with LC+AH. In the recovery phase, these cytokine levels in survivors tended to decrease, but in nonsurvivors, IL-6 remained high, and IL-8 and IL-10 further increased. Tumor necrosis factor-α levels were below the detection limit throughout the course in all patients. Serum lipopolysaccharide binding protein (LBP) generally was elevated in the acute phase and decreased in the recovery phase in all survivors, but in one of the nonsurvivors, LBP was elevated markedly at the terminal stage. In the acute phase, plasma endotoxin levels were correlated positively with white blood cell counts, neutrophil counts, and serum IL-8. IL-8 was correlated positively with neutrophil counts and negatively with serum Cholinesterase, hepaplastin test, and serum albumin levels. IL-6 was correlated positively with white blood cell and neutrophil counts, C-reactive protein, and serum total bilirubin and negatively with hepaplastin test and serum total protein levels. Serum LBP was correlated positively with white blood cell and neutrophil counts. Conclusions: Endotoxemia and related elevation of IL-8 may play an important role in the activation and migration of neutrophils in patients with alcoholic hepatitis. Marked elevation of inflammatory cytokines, IL-6 and IL-8, are related to severity and poor prognosis of alcoholic hepatitis. Serum LBP may serve as an index of inflammatory reaction in alcoholics.     

Contribution of Hepatitis C Virus to the Progression of Alcoholic Liver Disease

Background: We determined hepatitis C virus (HCV) antibody, HCV RNA, and genotype in patients with alcoholic liver disease and studied the involvement of HCV in alcoholic liver disease. Additionally, we used the histological activity index (HAI) to study the influence of HCV on the severity of inflammation. Methods: The subjects were 143 patients with alcoholic liver disease: 7 with fatty liver (FL), 18 with hepatic fibrosis (HF), 24 with alcoholic hepatitis (ALH), 39 with chronic hepatitis (CH), 42 with liver cirrhosis (LC), and 13 with hepatocellular carcinoma (HCC). The HCV RNA positivity rate in each type of disease was 0/7 (0%), 1/18 (6%), 2/24 (8%), 27/39 (69%), 24/42 (57%), and 7/13 (54%), respectively. It was high in the advanced hepatic lesions. Results: Clinically, the serum hepatic function tests after abstinence from drinking improved significantly in the HCV RNA negative patients compared with the positive patients. The proportion of genotype II in each type of disease was 0/0, 0/1 (0%), 1/2 (50%), 18/27 (67%), 18/24 (75%), and 7/7 (100%), respectively. It became high with the advance of pathophysiology. The HCV RNA amount stood at 7.5 ± 0.4 [log (copies/ml)] in CH, 7.9 ± 0.4 in LC, and 8.4 ± 0.8 in HCC, with a statistically significant difference between CH and HCC. However, we found no changes in the HCV RNA amount due to abstinence from drinking. The HAI score was high in the HCV RNA positive patients, but several cases in the HCV RNA negative group showed severe inflammatory changes. Therefore, judging the presence or absence of HCV RNA with the HAI score alone was considered difficult. Conclusions:: These results suggest that HCV, particularly genotype II, plays an important role in the advance of disease to LC and HCC in heavy drinkers.     

Clinical evaluation of tissue plasminogen activator (t-PA) levels in patients with liver diseases.

Tissue plasminogen activator (t-PA) levels in plasma or serum were studied in 416 patients with liver diseases: acute hepatitis (AH, n = 30); fulminant hepatitis (FH, n = 36); chronic inactive hepatitis (CIH, n = 57); chronic active hepatitis (CAH, n = 39); compensated liver cirrhosis (cLC, n = 78); decompensated liver cirrhosis (dLC, n = 84); hepatocellular carcinoma (HCC, n = 64); advanced hepatocellular carcinoma (aHCC, n = 28); and compared with that of a control group (n = 106) of healthy subjects. The t-PA levels showed significant increase in patients with AH, FH, CAH, cLC, dLC and HCC, compared with normal controls. The abnormal rates in t-PA levels (higher than 8.3 ng/ml) for each type of liver diseases were 86.1% in FH, 46.2% in CAH, 50% in cLC, 85.7% in dLC, 67.2% in HCC, and 89.3% in aHCC. t-PA levels tended to be higher in more advanced liver diseases. t-PA levels significantly correlated positively with plasminogen activator inhibitor (PAI-1) in AH, cLC, dLC, HCC and aHCC, and negatively with plasmin alpha 1-plasmin inhibitor complex (PIC), plasminogen (Plg), FDP, AT III and alpha 2-plasmin inhibitor (alpha 2-PI) in dLC, prothrombin time (PT) and fibrinogen (Fbg) in HCC. t-PA levels in patients with FH, CAH and dLC were significantly higher than those in patients with AH, CIH and cLC, respectively. Moreover, the changes of t-PA levels in the clinical courses of various liver diseases revealed that t-PA levels increased sensitively with progression of liver diseases or in advanced liver diseases.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum hepatocyte growth factor levels in liver diseases: Clinical implications

Although recent studies have shown that hepatocyte growth factor (HGF) is a potent mitogen in vivo, the significance of serum HGF in liver diseases remains unclear. To clarify clinical significance of serum HGF in liver diseases, serum HGF was measured in 127 patients with liver diseases and in 200 healthy individuals, using a highly sensitive immunoradiometric assay (IRMA). This assay is specific for HGF and is sensitive enough to detect 0.1 ng/mL of HGF. Mean values for serum HGF in acute hepatitis (AH), chronic hepatitis (CH), liver cirrhosis (LC), hepatocellular carcinoma (HCC), primary biliary cirrhosis (PBC), fulminant hepatic failure (FHF), and normal controls were 0.45, 0.40, 1.05,1.06, 0.44, 16.40, and 0.27 ng/mL, respectively. Serum HGF levels in these diseases were significantly increased compared with those in the controls (P < .001), and exhibited a positive correlation with total bilirubin, indocyanine green (ICG) test (R15), asparate aminotransferase (AST), and a negative correlation with albumin and prothrombin time (P < .001). Cirrhotic patients with modified Child class C had higher levels of serum HGF than those graded as modified Child class A or B (P < .001). In CH, serum HGF levels were significantly related to the histological activity index (HAI) score (P < .002). Seven patients with HCC who underwent transcatheter arterial embolization (TAE) exhibited a gradual increase in serum HGF levels up to day 4 after treatment; these higher levels were maintained until day 7, although AST reached a peak on day 2 and then decreased gradually. During clinical courses of patients with AH and CH, serum HGF was increased immediately after elevations of aminotransferases, and decreased as clinical symptoms improved. Serum HGF levels in survivors with FHF or AH were decreased during the illness (P = .0156), whereas serum HGF levels in nonsurvivors with FHF were increased. These findings suggest that serum HGF reflects the degree of liver dysfunction in chronic hepatic failure, and that serial measurement of serum HGF levels in acute hepatic injury serves as a prognostic factor.     

Expression of hepatocyte growth factor (HGF) and HGF receptor (c-met) proteins in liver diseases: an immunohistochemical study

BACKGROUND: Hepatocyte growth factor (HGF) is a potent mitogen for hepatocytes in vivo as well as in vitro. Serum levels of HGF vary in liver diseases, reflecting liver damage and dysfunction. However there are no studies reporting expression of HGF and HGF receptor (c-met protein) simultaneously in various liver diseases. METHODS: To clarify the clinical significance of HGF/c-met protein expression in liver diseases, liver tissues from 62 patients consisting of 7 with acute hepatitis (AH), 20 with chronic hepatitis (CH), 9 with liver cirrhosis (LC) and 26 with hepatocellular carcinoma (HCC) were immunohistochemically examined. RESULTS: Intense staining of HGF was observed in patients from AH, CH and LC, although no immunoreactivity was seen in HCC. The expression of c-met protein was higher in patients with HCC and AH than in those with CH (p < 0.05). A correlation of immunoreactivity between HGF and c-met protein was not observed expect in patients with LC (p < 0.01). The extent of c-met expression had no correlation with differentiation of HCC, tumour size, presence of portal invasion, or serum AFP levels. CONCLUSION: The results of the present study suggest that HGF plays an important role in human liver diseases, mostly in a manner independent of c-met protein expression.     

Clinical evaluation of serum tissue inhibitor of metalloproteinases-1 levels in patients with liver diseases

Serum levels of the tissue inhibitor of metalloproteinases-1 (TIMP-1) were measured in 268 patients with liver diseases by means of a one-step sandwich enzyme immunoassay. In the cases of acute hepatitis, chronic active hepatitis (CAH), liver cirrhosis (LC) and hepatocellular carcinoma (HCC), the levels of TIMP-1 were higher than those of the control group. Tissue inhibitor of metalloproteinases-1 levels correlated with type III procollagen peptide and with type IV collagen, indicating TIMP-1 as a useful marker for hepatic fibrosis. Levels of TIMP-1 also correlated with aspartate aminotransserase and alanine aminotransferase levels and showed the highest levels in acute hepatitis. Thus, TIMP-1 might also reflect hepatic inflammation. Serum levels of α-fetoprotein and TIMP-1 had a significant positive correlation in patients with HCC. A cut-off level of TIMP-1 between LC and HCC was set at 440 ng/mL, having a low sensitivity and a high specificity. These results suggest the usefulness of TIMP-1 as a tumour marker in cases of HCC where α-fetoprotein levels are not elevated.     

Soluble CD163 in patients with liver diseases: very high levels of soluble CD163 in patients with fulminant hepatic failure

Background The levels of several cytokines and chemokines are elevated in various liver diseases, especially in fulminant hepatic failure (FHF). Activated macrophages may have a role in the production of these immune modulators. CD163 is a member of a scavenger receptor family and is expressed mainly on activated macrophages, and a soluble form of CD163 (sCD163) is released from activated macrophages. The aim of this study was to assess sCD163 levels in patients with FHF and to evaluate their clinical significance.Methods The levels of sCD163 in the sera were measured in 21 patients with FHF, 17 patients with acute hepatitis (AH), 22 patients with chronic hepatitis (CH), and 14 normal healthy controls (NC), by an enzyme-linked immunosorbent assay. The levels of sCD163 were observed serially in patients with FHF and AH.Results The levels of sCD163 in the sera from patients with FHF were significantly higher than those in patients with AH and CH and the NC group (P < 0.0001). There was a good correlation between serum levels of sCD163 and prothrombin time (r = –0.677; P < 0.0001). A kinetic study revealed that the levels of sCD163 decreased in patients with AH and in survivors of FHF, whereas the levels of sCD163 progressively increased in nonsurvivors of FHF.Conclusions This study shows that the products of activated macrophages may be involved in the pathogenesis of FHF. This study also inspires optimism that sCD163 may possess prognostic importance in FHF.     

A cell kinetic study of liver cells in various liver diseases by the detection of DNA polymerase alpha

DNA polymerase alpha (DNA-P alpha) in the nuclei of hepatocytes was visualized by the immunoperoxidase method to study the number of liver cells which were at the stage of G1, S, and G2 stage in the cell cycle. Seven liver specimens from patients with acute hepatitis (AH), 17 with chronic persistent hepatitis (CPH), 32 with chronic active hepatitis (CAH), 6 with liver cirrhosis (LC), 4 with hepatocellular carcinoma (HCC) and 4 with hospital controls were studied. The number of DNA-P alpha-positive hepatocytes in 1000 hepatocytes were as follows: 19.1 +/- 18.0 in AH, 8.8 +/- 6.1 in CPH, 27.3 +/- 23.8 in CAH, 21.8 +/- 14.3 in LC, 545.3 +/- 184.0 in HCC and 1.1 +/- 1.1 in hospital controls. The number of DNA-P alpha-positive hepatocytes in HCC were significantly increased compared with other liver diseases. Likewise, those in CAH and LC were higher than those in CPH and hospital controls. The liver cell necrosis was thought to be one of the secondary stimulators for cell division of hepatocytes.     

肝病患者一氧化氮合酶表达异常及其临床意义

目的 探讨肝病患者抗氧化能力及一氧化氮合酶（NOS） 表达的临床价值。方法 收集26例急性肝炎（AH）、36例慢性肝炎（CH）、13例肝炎后肝硬化（LC）和23例原发性肝癌（HCC）患者的血清,分别检测其总抗氧化能力（TAO）、NOS浓度及一氧化氮（NO）水平,分析它们在肝脏疾病中的改变机制。结果 患者血清TAO在AH、CH中异常率为80％,在LC和HCC为50％;血清中NOS活性在肝病患者中的异常率在70％ 左右;NO水平在AH、CH和LC中异常率为70％,HCC组为48％。AH组、CH组TAO平均水平明显高于对照组,但LC和HCC组的差异不明显;肝病患者NO和NOS平均浓度显著高于对照组,但HCC患者NOS和NO平均浓度均低于AH、CH和LC组患者。结论 肝病患者血清NOS水平与NO浓度呈高度正相关,NO增加可能 对肝细胞起保护作用。     

乙型肝炎病毒X基因的原核表达及血清抗HBx抗体的检测

目的 应用表达蛋白检测乙型肝炎患者血清抗-HBx抗体水平并探讨其临床意义。方法 通过PCR扩增获得HBVX基因，与原核表达载体Pet32a＋连接构建PET32a-HBX原核表达载体，转化E．coli BL21表达获得重组融合蛋白。经切胶透析纯化后，应用重组蛋白HBx建立检测血清中抗-HBx抗体的间接ELISA方法，分别检测正常人组、急性肝炎组、慢性肝炎组、肝硬化组和肝细胞癌组患者血清中的抗-HBx抗体。结果 获得具有免疫原性的HBx融合蛋白；ELISA检测表明，慢性肝炎组、肝硬化组和肝细胞癌组的抗-HBx抗体的水平均高于急性肝炎组，差异具有显著性；在三组之问，慢性肝炎组高于肝硬化组和肝细胞癌组，差异具有显著性，肝硬化组和肝细胞癌组的抗-HBx抗体水平无显著性差异。结论 HBV患者血清中抗-HBX抗体是乙型肝炎病毒感染的一种特异性抗体，是HBV感染的血清学指标之一，可以反映乙型肝炎肝炎患者病情的变化。     

Incremento en las concentraciones de ligando CD40 soluble plaquetario en pacientes con síndrome antifosfolipídico primario

ObjectiveTo determine the concentrations of sCD40L in patients with PAPS, and establish its association with the number of thrombosis.Patients and methodsWe included patients with PAPS and healthy controls of the same age and sex. For analysis, patients with PAPS were divided into 2 groups: 1) patients with 1 thrombosis, and 2) patients with >1 thrombosis. Soluble CD40L concentrations were determined by ELISA method.ResultssCD40L concentrations were significantly higher in patients with PAPS compared with the controls (9.72 ng ± 11.23 ng/ml vs. 4.69 ± 4.04 ng/ml) (P=.04) There was no association between serum levels of sCD40L and the number of thrombosis (1 thrombosis: 9.81 ± 9.87 ng/ml vs 9.63 ± 12.75 ng/ml in ≥ 1 thrombosis (P=.13). In women with pregnancy and abortions, (13 patients) concentrations of sCD40L were higher than in those patients without a history of abortion (26 patients) but without statically significant difference (12.11 ± 16.46 ng/ml vs. 8.80 ± 8.61 ng/ml) (P=.33). There was no correlation between levels of sCD40L and the total number of thrombosis.ConclusionsPatients with PAPS have higher concentrations of sCD40L compared with healthy subjects, although this is not associated with a greater number of thrombosis. Among patients with PAPS, there is a tendency to higher concentrations of sCD40L in women with pregnancy and history of abortion. Since the platelet is the main cellular source of sCD40L, is possible that this pathway plays a pathogenic role in patients with PAPS.     

Simultaneous measurements of serum alpha-fetoprotein and protein induced by vitamin K absence for detecting hepatocellular carcinoma. South Tohoku District Study Group

OBJECTIVE: We evaluated the measurements of serum alpha-fetoprotein (AFP) and the protein induced by vitamin K absence (PIVKA-II) in 734 patients with chronic hepatitis (CH) and liver cirrhosis (LC) who had been followed-up for the development of hepatocellular carcinoma (HCC). METHODS: Serum AFP and PIVKA-II were measured every month and abdominal ultrasonography was performed every 3 months. Youden's index (sensitivity + specificity -1) was calculated. RESULTS: On an average follow-up period of 374.5 days, HCC was detected in three HBsAg-positive LC patients (10.0%/yr), four anti-HCV-positive CH patients (1.35%/yr), 21 anti-HCV-positive LC patients (7.8%/yr), and one patient with both HBsAg- and anti-HCV-positive LC (22.7%/yr). At the time of HCC detection, the size of HCC was 4.7+/-0.6 (mean +/- SD) cm in HBsAg-positive patients and 2.4+/-1.3 cm in anti-HCV-positive patents. Cut-off values of 20 ng/ml for AFP (Youden's index = 0.422) and 60 mAU/ml for PIVKA-II (Youden's index = 0.316) gave the highest index for each marker. When these two markers were combined, cut-off values of 40 ng/ml for AFP and 80 mAU/ml for PIVKA-II gave the highest index (Youden's index = 0.500, sensitivity = 65.5%, specificity = 85.5%, positive predictable value = 14.8%, negative predictable value = 98.3%). The levels of AFP or PIVKA-II increased within three months before the detection of HCC. CONCLUSIONS: Simultaneous measurements of serum AFP and PIVKA-II levels that are performed every 3 months are useful for detecting a developing HCC. The optimal cut-off values for AFP and PIVKA-II may be 40 ng/ml and 80 mAU/ml, respectively.     

氯吡格雷抑制急性冠状动脉综合征患者血小板释放可溶性CD40配体

目的探讨在非ST段抬高急性冠状动脉综合征（non—ST-segment elevation acute coronary syndromes，NSTEACS）患者短期应用氯吡格雷是否有抑制ADP诱导血小板释放可溶性CD40配体（sCD40L）的作用。方法NSTEACS42例，患者服用氯吡格雷6—8d，治疗前后采静脉血。提取富含血小板血浆（platelet rich plasma，PRP）并用二磷腺苷诱导血小板聚集和释放sCD40L，在不同时间点终止反应，用酶联免疫法测量sCD40L浓度，进行自身前后对照。结果治疗前后血浆sCD40L分别为（0．20±0．16）μg／L和（0．19±0．18）μg/L（P〉0．05）；治疗前后PRP受ADP诱导后20min释放的sCD40L浓度分别为（4．3±2．5）μg／L和（2.8±1．9）μg／L（P〈0．001），诱导后40min释放的sCD40L浓度分别为（5．3±3．1）μg／L和（2．9±1．6）μg／L（P〈0．001）。结论短期应用氯吡格雷可能对非sT段抬高急性冠状动脉综合征患者血小板炎症因子sCD40L释放具有抑制作用，提示氯吡格雷很可能具有抗炎效应．     

Plasma free amino acid pattern in chronic hepatitis as a sensitive and prognostic index

To evaluate the diagnostic and prognostic significance of plasma amino acid imbalance in chronic hepatitis (CH), plasma-free neutral amino acid levels were examined in 47 patients with CH, consisting of 8 chronic persistent hepatitis (CPH), 26 chronic aggressive hepatitis (CAH) 2A and 13 CAH 2B, compared with those of 58 patients with liver cirrhosis (LC) and of 12 healthy controls. Fischer's ratio (a molar ratio of branched chain amino acids to aromatic amino acids) was found to be reduced in the order of normal subjects (3.5 +/- 0.4), CPH (3.0 +/- 0.2), CAH2A (2.7 +/- 0.3), CAH2B (2.1 +/- 0.3), compensated LC (LC-C, 1.5 +/- 0.4) and decompensated LC (LC-D, 1.1 +/- 0.2). Patients with CPH showed a significant decrease of the ratio compared with normal subjects (P less than 0.05). The ratio was significantly higher in patients with CAH 2B in comparison with LC-C (P less than 0.001). Especially, the ratio could be used to discriminate the three distinct stages of CH. Discriminant analysis, carried out using six amino acids, Fischer's ratio and conventional liver function tests indicated that Fischer's ratio was the most reliable parameter for differentiation of the three stages of CH. Furthermore, serial examinations of Fischer's ratio in patients with CH remained unchanged in CPH, whereas was significantly reduced in CAH during 2-3 years follow-up. These results strongly suggest that Fischer's ratio is a useful indicator for differential diagnosis and for prediction of the subsequent clinical course of CH as well as being a sensitive index for functional hepatic reserve in chronic active liver diseases.     

Detection and clinical significance of acetone-insoluble liver cell membrane antigen in sera of patients with chronic active liver diseases

An enzyme-linked immunosorbent assay was developed to detect insoluble liver cell membrane antigen (LMAg) which gives rise to serum LMA (anti-LM) in HBsAg-negative patients. The optical density (OD) ratio of the average LMAg level of normal subjects was less than 1.2. In HBsAg-positive cases, high LMAg levels (OD ratio greater than 2.4) were noted in 8 of 8 patients with acute hepatitis (AH), 3 of 8 with chronic persistent hepatitis (CPH), 5 of 10 with moderate chronic aggressive hepatitis (CAH), 7 of 10 severe CAH and 4 of 8 with liver cirrhosis (LC). In HBsAg-negative cases, however, high LMAg levels were noted in only 6 of 8 patients with AH, 1 of 10 with CPH, 1 of 10 with moderate CAH, 1 of of 10 with severe CAH, 0 of 8 with LC, 0 of 8 with fatty liver and 5 of 10 with alcoholic hepatitis. In micro-immunodiffusion experiments, intensively absorbed rabbit anti-rat LM precipitated two organ-specific components of rat liver homogenate, one of which was identical to liver specific protein (LSP). In immunohistochemical demonstrations of LMAg and LSP, anti-LM, prepared from the serum of a HBsAg-negative CAH patient, bound to both human and rat acetone-fixed liver cell membranes, but not to those of human or rat kidneys. Absorbed rabbit anti-rat LM also bound to liver cell membranes, but absorbed anti-rat LSP lacked organ-specificity when assayed with the immunofluorescence technique using acetone-fixed liver sections. In conclusion, the appearance of serum LMAg was associated with high-SGPT patients and HBsAg-positive CAH patients.