Analyses of proliferating cell nuclear antigen-positive cells in hepatocellular carcinoma: Comparisons with clinical findings

Abstract Proliferating tumour cells in 92 patients with hepatocellular carcinoma (HCC) were identified by an immunohistochemical method using a monoclonal antibody against proliferating cell nuclear antigen (PCNA). The rate of PCNA-positive cells in HCC tissues was positively correlated with histological grade and the tumour size and T factor of the tumour.
In order to analyse the relationship between prognostic factors and cumulative survival rate after obtaining tumour specimens, 49 patients whose clinical courses could be followed after needle biopsy were selected for evaluation. These patients were treated by medical therapy alone. Analyses of prognostic factors by Cox's proportional hazard model revealed that the patient's prognosis was significantly correlated with PCNA-positive rates as well as the tumour size and mode of therapy. Moreover, the cumulative survival rates were significantly ( P < 0.001) higher in patients with rates of PCNA-positive cells <15% than in those with ≧ 15%, even when tumour sizes were under 50 mm or tumours demonstrated the same degree of histological differentiation.
These findings indicate that the PCNA-positive rate in biopsied tissues provides useful prognostic information in patients with HCC treated only by medical therapy.     

慢性乙型肝炎患者肝组织内CD4+CD25+Foxp3+调节性细胞的作用

目的 观察慢性乙型肝炎患者肝内调节性T细胞(Treg)与病情活动、肝细胞损伤的关系.方法 对日本长崎大学医学院附属医齿学病院第一内科26例住院慢性乙型肝炎患者进行肝组织活检,采用免疫组织化学方法以单抗Foxp3、单抗CD3检测肝组织内Foxp3+细胞和CD3+细胞频率,计算Foxp3+/CD3+细胞比率,结合肝组织学炎症活动指数(HAI,采用Knodell评分系统)、ALT、AST、HBV DNA水平进行分析.数据用SPSS 13.0软件进行统计分析.结果 Foxp3+Treg主要分布在门脉区;Foxp3+/CD3+细胞(%)在肝实质重度炎症组明显高于轻度炎症组(P=0.007 6);Foxp3+/CD3+细胞(%)的增高与血清ALT、AST水平呈正相关,相关系数分别0.438、0.436(P值分别为0.025、0.026),与血清HBV DNA水平有相关趋势,但差异无统计学意义.结论 CD4+CD25+Foxp3+Treg在慢性乙型肝炎患者的发病机制中参与肝细胞的免疫损伤.     

Interleukin-6 functions as an autocrine growth factor in a cholangiocarcinoma cell line

Abstract The tumour cells of a human cholangiocarcinoma cell line, HuCC-T1, were found to express mRNA of interleukin-6 (IL-6) and to secrete a large amount of biologically active IL-6 in the culture medium at the concentration of 22.6 ng/mL. Interleukin-6 was demonstrated in the cytoplasm of the cells by immunohistochemical staining. Furthermore, these cells showed the presence of receptors for IL-6 on the surface, and DNA synthesis of the cells was stimulated by the exogenous addition of recombinant human IL-6 into the culture medium. The cell growth was significantly inhibited in the presence of anti-human IL-6 antibody in the culture medium. These findings indicate that IL-6 is one of the autocrine growth factors of this cell line in vitro.     

Echo-Doppler measurements of portal vein and superior mesenteric artery blood flow in humans: Inter- and intra-observer short-term reproducibility

The reproducibility of echo-Doppler measurements of portal vein and superior mesenteric artery blood flow has not been extensively studied. In the present study, two groups of subjects were examined to test inter- and intra-observer reproducibility. Each study population consisted of 15 non-portal hypertensive and 15 portal hypertensive subjects. With a standardized technique, the cross-sectional area and velocity of blood flow in the portal vein and superior mesenteric artery were recorded in triplicate by skilled operators. The flow volume of each vessel was calculated by multiplying the cross-sectional area by the velocity of blood flow. The measurements were performed in a blind fashion over a 60 min period. The reproducibility of measurements was assessed by calculation of intraclass correlation coefficients and coefficients of variation. The intra-observer intraclass correlation coefficient was 0.77 for portal vein blood flow and 0.84 for superior mesenteric artery blood flow, suggesting good reproducibility. The intra-observer coefficient of variation was 11 and 9%, respectively. In contrast, the interobserver intraclass correlation coefficient was calculated to be 0.49 for portal blood vein blood flow and 0.57 for superior mesenteric artery blood flow, indicating fair reproducibility. In addition, the interobserver coefficients of variation were calculted to be 20 and 18%, respectively. These data suggest that intra-observer reproducibility in echo-Doppler measurements of portal vein and superior mesenteric artery blood flow is acceptable but inter-observer reproducibility is not. Examination by a single operator, rather than multiple operators, is therefore advisable. Even when measurements are performed by a single investigator an approximate variance of 10% in the measurement in a single subject should be expected.     

Changes of serum aminotransferase activities in relation to serum hepatitis Be antigen levels in chronic type B hepatitis

The purpose of this study was to determine the level of serum hepatitis Be antigen (HBeAg) during hepatitis B virus carriage and its clinical significance. The mean level of serum HBeAg, quantitated by solid-phase enzyme immunoassay, was 5924 units in asymptomatic carriers (s.d. = 5534, n= 100), 3044 units in patients with chronic persistent hepatitis (s.d. = 4826, n= 34), 3599 units in chronic active hepatitis (s.d. = 4953, n= 45) 1348 units in liver cirrhosis (s.d. = 2379, n= 25) and 766 units in hepatocellular carcinoma (s.d. = 1257, n= 18). In the 10 HBeAg-positive patients with chronic active hepatitis, the fluctuation in HBeAg level was followed by changes in alanine aminotransferase (ALT) activity. Among the 36 peaks of HBeAg and ALT, HBeAg peaks preceded ALT peaks in 22 and simultaneous peaks were present in 14. The changes of HBeAg level were closely related to increase in disease activity as estimated by ALT activities; therefore, HBeAg quantitation can be a useful predictor of disease activity in chronic hepatitis B.     

Protection by Glycyrrhizin against Warm Ischemia‐Reperfusion–Induced Cellular Injury and Derangement of the Microcirculatory Blood Flow in the Rat Liver

Background/Aim: The mechanism by which ischemia‐reperfusion (I/R)‐induced derangement of the hepatic microcirculation leads to tissue injury is not fully understood. We postulated that alterations to the hepatic microcirculation, including hemodynamic derangement and increased leukocyte‐endothelium interaction, play a role, and that glycyrrhizin exerts its hepatoprotective effects, in part, by reducing these microcirculatory changes. Materials and Methods: Wistar rats were subjected to 30–60 minutes segmental hepatic ischemia, followed by 120 minutes of reperfusion. Glycyrrhizin was administered prior to ischemia. Using intravital fluorescence microscopy, the administration of fluorescein isothiocyanate–conjugated erythrocytes allowed the measurement of erythrocyte‐velocity (RBC_vel), lobular, and sinusoidal perfusion. Bleb formation was observed by electron microscopy. Blood and tissue were taken for the assessment of liver injury. Results: Glycyrrhizin reduced I/R‐induced liver injury (histology, liver enzymes) and reduced hepatocyte apoptosis (TUNEL, caspase‐3 activity). Glycyrrhizin inhibited hepatocyte bleb formation and reversed the I/R‐induced reductions in lobular perfusion and RBC_vel. Leukocyte rolling and adherence in postsinusoidal venules and neutrophil infiltration were reduced by glycyrrhizin. I/R‐induced elevation in HMGB1 was prevented by glycyrrhizin. Conclusions: Early bleb formation with deranged microcirculatory flow and leukocyte‐endothelium interaction would appear to contribute to I/R‐induced hepatocellular injury. Glycyrrhizin exerts its hepatoprotective effect by preventing these changes, in addition to a direct cellular effect.