Ductular proliferation in liver tissues with severe chronic hepatitis B： An immunohistochemical study

AIM: To clarify the pathogenesis of ductular proliferation and its possible association with oval cell activation and hepatocyte regeneration. METHODS: Immunohistochemical staining and image analysis of the ductular structures in the liver tissues from 11 patients with severe chronic hepatitis B and 2 healthy individuals were performed. The liver specimens were sectioned serially, and then cytokeratin 8 (CK8), CK19, OV6, proliferating cell nuclear antigens (PCNA), glutathione-S-transferase (GST),α-fetal protein (AFP) and albumin were stained immunohistochemically. RESULTS: Typical and atypical types of ductular proliferation were observed in the portal tracts of the liver tissues in all 11 patients. The proliferating ductular cells were positive for CK8, CK19, OV6 and PCNA staining. Some atypical ductular cells displayed the morphological and immunohistochemical characteristics of hepatic oval cells. Some small hepatocyte-like cells were between hepatic oval cells and mature hepatocytes morphometri-cally and immunohistochemically. CONCLUSION: The proliferating ductules in the liver of patients with severe chronic liver disease may have different origins. Some atypical ductular cells are actually activated hepatic oval cells. Atypical ductular proliferation is related to hepatocyte regeneration and small hepatocyte-like cells may be intermediate transient cells between hepatic oval cells and mature hepatocytes.     

5-Fluorouracil concentration in blood, liver and tumor tissues and apoptosis of tumor cells after preoperative oral 5＇-deoxy-5-fluorouridine in patients with hepatocellular carcinoma

AIM: To study the levels of 5-fluorouracail (5-FU) in plasma, liver and tumor in patients with hepatoceilular carcinoma after oral administration of 5'-deoxy-5-fluorouridine (5'-DFUR). METHODS: Thirty-nine patients with hepatoceilular carcinoma were treated with oral 5'-DFUR for more than 4 d before operation. The contents of 5-FU in plasma, liver and tumor were measured by high performance liquid chromatography (HPLC) and apoptosis of tumor cells was evaluated by in-situ TUNEL after resection of tumor. RESULTS: The concentrations of 5-FU were 1.1 μg/mL, 5.6, 5.9, and 10.5 μg/g in plasma, the liver tissue, the center of tumor and the periphery of tumor, respectively. 5-FU concentration was significantly higher in the periphery of tumor than that in the liver tissue and the center of tumor (10.5±1.6 μg/g vs 5.6±0.8 μg/g, t = 21.38, P<0.05; 10.5±1.6 μg/g vs 5.9±0.9 μg/g, t = 20.07, P<0.05). 5-FU level was significantly lower in plasma than that in the liver and the tumor (1.1±0.3 μg/mL vs 5.6±0.8 μg/g, t = 19.63, P<0.05; 1.1±0.3 μg/mL vs 10.5±1.6 μg/g, t= 41.01, P<0.05). Apoptosis of tumor cells was significantly increased after oral 5'-DFUR compared to the control group without 5-DFUR treatment. CONCLUSION: There is a higher concentration of 5-FU distributed in the tumor compared with liver tissue and apoptosis of tumor cells is increased following oral 5'-DFUR compared with the control group. The results indicate that 5'-DFUR is hopeful as neo-adjuvant chemotherapy to prevent recurrence after resection of hepatoceilular carcinoma.     

Hepatic intra-arterial infusion of yttrium-90 microspheres in the treatment of recurrent hepatocellular carcinoma after liver transplantation： A case report

INTRODUCTION Strict criteria developed to select patients with hepatocellular carcinoma (HCC) for liver transplantation have increased the survival and decreased the recurrence of tumor after transplantation[1]. However, tumor recurrence and subsequent mo…     

Effect of bax,bcl—2 and bcl—xL on regulating apoptosis in tissues of normal liver and hepatocellular carcinoma

AIM: To investigate the expression of bax, bcl-2 and bcl-xL mRNA in the tissues of normal liver and hepatocellular carcinoma (HCC), and analyze the relationship between the expression of bax, bcl-2 and bcl-xL mRNA and clinical parameters of HCC patients. METHODS: The expression of bax, bcl-2 and bcl-xL mRNA of normal liver and HCC was measured by Northern blot. Statistical analyses were made by t test and correlation analysis. RESULTS: A very low mRNA level was indicated at bax, bcl-2 and bcl-xL in the HCC tissues in contrast to the tissues of normal liver by Northern blot analysis. The analyses of mRNA level revealed that HCC tissues exhibited a mean 7.6-fold decrease in bax, 4.2-fold in bcl-2 and 3.5-fold in bcl-xL in comparison with normal control tissues, respectively. Positive correlation was found between bax and bcl-xL (r=0.7061, P<0.01). There was no significance between the mRNA expression of these three genes and age, gender, tumor differentiation and tumor stage of HCC patients. CONCLUSION: The results are consistent with the fact that apoptosis rarely occurs in normal livers but increases in HCC, indicating that bcl-2 and bcl-xL may play a very important role in regulating the apoptosis of normal liver and HCC.     

Usefulness of serum des-γ-carboxy prothrombin in detection of hepatocellular carcinoma

AIM: Des-gamma-carboxy prothrombin (DCP) has been reported to be more sensitive and specific in diagnosing hepatocellular carcinoma (HCC) when compared with alpha-fetoprotein (AFP). However, its ability to identify small HCC still remains unclear. Thus, we conducted a cross-sectional case control study to evaluate whether DCP is better than AFP for differentiating HCC from nonmalignant liver disease and further evaluate the usefulness of DCP in early diagnosis of small HCC. METHODS: Serum DCP and AFP levels were determined in 127 patients. Among these patients, 32 were with non-cirrhotic chronic hepatitis, 34 were with compensated cirrhosis, and 61 were with HCC. The cut-off value for the DCP and AFP were set as 40 mAU/mL and 20 ng/mL, respectively. To compare the diagnostic value of DCP and AFP in distinguishing HCC from nonmalignant chronic liver disease, receiver operating characteristic (ROC) curves were constructed for each assay. RESULTS: The accuracy, sensitivity and specificity of DCP were higher than AFP in detecting HCC (81.9%, 77% and 86.4% vs 68.5%, 59% and 77.3%, respectively). The area under the ROC (AUROC) curves revealed that DCP had a better accuracy than AFP in diagnosis of HCC (0.85 [95%CI, 0.78-0.91] vs 0.73 [95%CI, 0.65-0.81], P = 0.013). In 39 patients with solitary HCC, the positive rates of DCP were 100% in patients with tumor size larger than 3 cm, 66.7% in patients with tumor size 2-3 cm and 50% in patients with tumor size less than 2 cm. The positive rates of AFP in patients with tumor size larger than 3 cm, 2-3 cm and less than 2 cm were 55.6%, 50%, and 33.3%, respectively. The median level of DCP in HCC patients with tumor size larger than 3 cm was significantly higher than those with tumor size 2-3 cm and those with the size of less than 2 cm. CONCLUSION: Our study indicates that DCP has a better diagnostic value than AFP in differentiating HCC from nonmalignant chronic liver disease. DCP has not only a stronger correlation with HCC than AFP in tumor size but also more effectiveness than AFP in detecting small size of HCC.     

肝癌组织和血清中钙囊素的表达及其意义

目的 研究人肝癌组织及血清中钙囊素(S100A11)的表达及其临床意义.方法 免疫组织化学法检测46例肝癌及其癌旁组织中S100A11表达,比较肝癌与癌旁组织中S100A11的阳性率,分析肝癌组织中S100A11表达水平与各临床参数之间的关系;同时检测肝癌(62例)、肝硬化(32例)、慢性肝炎(30例)患者及健康体检者(28名)血清中S100A11浓度,比较S100A11浓度与甲胎蛋白(AFP)、γ-谷氨酰转肽酶同工酶Ⅱ(GGT-Ⅱ)诊断肝癌的灵敏性及特异性.结果 S100A11在肝癌组织中的表达阳性率(78.3%)显著高于癌旁组织(19.6%,P<0.01),其表达水平与分化程度相关,分化程度越低表达水平越高.根据ROC曲线,当确定诊断界值为7.3μg/L时,S100A11在肝癌患者血清中的阳性率为30.6%,明显高于肝硬化、慢性肝炎及健康人(P值均<0.05).肝癌患者血清中S100A11与AFP、GGT-Ⅱ均无相关性,联合检测三项指标对肝癌诊断有互补性,可使诊断敏感性提高至84.5%.结论 S100A11可能与肝癌的发生发展有关.联合检测S100A11与AFP和GGT-Ⅱ可提高对肝癌的诊断敏感度.

Abstract：

Objective To explore the expression of calcium binding protein (S100A11) and its clinical significances in human hepatocellular carcinoma (HCC) tissue and blood plasma. Methods The expressions of S100A11 in 46 cases of HCC tissues and their paracancerous tissues were detected by immunohistochemistry. The relationship between S100A11 expression level in HCC tissues and clinical parameters was analyzed. The S100A11 expression levels in blood plasma of HCC patients (62 cases), liver cirrhosis patients (32 cases), chronic hepatitis patients (30 cases) and healthy subjects (30 cases) were detected. The sensitivity and specificity of S100A11, alpha fetoprotein (AFP) and γ-glutamyl transpeptidase Ⅱ (GGT-Ⅱ ) in HCC diagnosis were compared. Results The positive rate of S100A11 in HCC tissue (78. 3%) was significantly higher than that in paracancerous tissues (19. 6%) (P<0. 05). The expression level was correlated with the degree of differentiation, the lower differentiation degree with the higher expression level. According to ROC curve, if the cutoff points for diagnosis was set at 7. 3 μg/L, the positive rate of S100A11 in HCC patients blood plasma was 30. 6% , which was significantly higher than that in the blood plasma of patients with liver cirrhosis, patients with chronic hepatitis and healthy persons (P<0. 05). There was no correlation between S100A11 and AFP or GGT-Ⅱ in the blood plasma of HCC patients. These three indicators were complementary in HCC diagnosis, and the diagnostic sensitivity increased to 84.5% with combined detection. Conclusions S100All may be related to HCC genesis and development. The HCC diagnostic sensitivity may be increased with combined detection of S100All ,AFP and GOT- Ⅱ.     

Systemic chemotherapy for hepatocellular carcinoma in non-cirrhotic liver： A retrospective study

AIM： To investigate the efficacy and toxicity of systemic chemotherapy in a retrospective study of patients with hepatocellular carcinoma （HCC） occurring in normal or fibrotic liver without cirrhosis.
METHODS： Twenty-four patients with metastatic or locally advanced HCC in a normal or a fibrotic liver were given systemic chemotherapy （epirubicin, cisplatin and 5-fluorouracil or epirubicin, cisplatin and capecitabine regimens）. Tumor response, time to progression, survival, and toxicity were evaluated.
RESULTS： There were 7 women and 17 men, mean age 54 ± 10 years; 18 patients had a normal liver and 6 had a fibrotic liver （F1/F2 on biopsy）. Mean tumor size was 14 cm, 5 patients had portal vein thrombosis and 7 had metastasis. Patients received a median of 4 chemotherapy sessions. Overall tolerance was good. There were 5 partial responses （objective response rate = 22%）, and tumor control rate was 52%. Second line surgical resection was possible in two patients. Median survival was 11 mo, and 1- and 2-year overall survival rates were 50% ± 10% and 32 ± 11%, respectively.
CONCLUSION： In patients with HCC in a non-cirrhotic liver, chemotherapy was well tolerated and associated with an objective response rate of 22%, including two patients who underwent secondary surgical resection.     

Measurement of immunoreactive gamma-glutamyl transpeptidase in human sera and liver tissues of patients with various liver diseases

Immunoreactive gamma-glutamyl transpeptidase in human serum and liver tissue was measured by a solid phase enzyme-linked immunosorbent assay. The immunoreactive gamma-glutamyl transpeptidase was significantly elevated in the sera of patient with hepatocellular carcinoma. On the other hand, in sera of patients with non-neoplastic diseases, including chronic hepatitis, acute hepatitis, fatty liver and hemangioma, the immunoreactive gamma-glutamyl transpeptidase was not elevated. In hepatocellular carcinoma and metastatic liver tumor tissues, the immunoreactive gamma-glutamyl transpeptidase content was also elevated, showing good correlation with the enzyme protein content in sera. However, no correlation was found between the activity of gamma-glutamyl transpeptidase determined by an enzymatic assay and the content determined by an enzyme-linked immunosorbent assay. On immunohistochemical examination, the immunoreactive enzyme protein without enzymatic activity was detected only in the cytoplasm of cancer cells. This suggested that there is an increased level of the immunologically active but enzymatically inactive form of gamma-glutamyl transpeptidase in hepatoma tissues.     

Utility of gamma-glutamyl transpeptidase and mean corpuscular volume in alcoholic liver disease

The present study was designed to establish the role of Gamma-glutamyl transpeptidase (gamma-GT) and mean corpuscular volume (MCV) in alcoholic liver disease (ALD). Serum gamma-GT, total and direct bilirubin, albumin, total protein, AST, ALT and ALP were assayed by standard methods in a clinical chemistry autoanalyser. MCV, Hb, PCV and RBC were measured by an automated cell counter. Activity of gamma-GT and MCV levels were significantly higher in the patients with ALD compared to controls. A gamma-GT level of > or = 25 U/l was found to be significantly associated with ALD. MCV level > or = 100 fl/l showed a significant association with ALD. An AST to ALT ratio > 1 was found in 92% of the patients. None of the patients showed an ALT level > or = 300 IU/l. The degree of AST elevation in the patients with ALD was higher (3.7 times) then ALT (3.2 times). A gamma-GT level > or = 25 IU/L and an MCV level > or = 100 fl/l stand as markers of heavy alcohol consumption in this study. An AST to ALT ratio > 1 was present in most of the patients with ALD. The degree of elevation of AST was higher than ALT in the patients with ALD.     

Mature hepatocytes actively divide and express gamma-glutamyl transpeptidase after D-galactosamine liver injury

Abstract: Aims/Background: We studied the fate of hepatocytes in the rat liver after D-galactosamine injury by genetic labeling using recombinant retroviruses carrying the Escherichia coli lacZ gene coupled to a nuclear localization signal. Methods: Hepatocytes were either labeled by direct injection of 2.5 ml high-titer retrovirus-containing medium in the regenerating liver parenchyma after administration of a single dose of D-galactosamine. Alternatively hepatocytes were pre-labeled, 24 h after a two-thirds hepatectomy, by injecting the same volume of retroviral solution in the portal vein and D-galactosamine was administered 15 days later. Gammaglutamyl transpeptidase and β-galactosidase activities were assessed on cryostat sections, along with localization of the hepatocyte-specific HES6 antigen. Results: Morphological observations, as well as β-galactosidase activity detection, showed that hepatocytes actively divide as early as 1 day after D-galactosamine injection. Gamma-glutamyl transpeptidase activity was detected in biliary cells, but also in mature hepatocytes, prelabeled with β-galactosidase before D-galactosamine administration. Conclusions: These experiments demonstrate that hepatocytes can divide to restore the liver mass after D-galactosamine liver injury. Furthermore, we also show that gamma-glutamyl transpeptidase, which has been reported to be expressed only by fetal or preneoplastic hepatocytes, can be re-expressed by mature hepatocytes during the recovery process.     

The diagnostic value of the ratio of serum gamma-glutamyl transpeptidase to alkaline phosphatase in alcoholic liver disease

A prospective study of the initial liver laboratory tests was carried out in the following patients: 55 patients with alcoholic liver disease, 53 with cholangitis, 41 with hepatocellular carcinoma, 65 with acute viral hepatitis, and 49 with hepatitis-B surface antigen-positive chronic active hepatitis. There was considerable overlap in the levels of the serum gamma-glutamyl transpeptidase (GT) and alkaline phosphatase (AP) among the five groups. However, the ratio of GT to AP was significantly higher in the group with alcoholic liver disease than in any of the other four groups. When the ratio was higher than 1.4, the diagnostic efficiency for distinguishing the alcohol group from the other four groups was 78% (the normal upper limit for GT and AP being 35 and 115 U/1, respectively). A possible explanation for this higher ratio in alcoholic liver disease is suggested. We conclude that when the GT and AP is greater than 1.4, it is of greater diagnostic value than either variable alone in differentiating alcoholic from other liver diseases.