Significance of Serum Adiponectin and Insulin Resistance Levels in Diagnosis of Egyptian Patients with Chronic Liver Disease and HCC

One possible hypothesis for pathogenesis of hepatocellular carcinoma is deregulated expressed adipokines(adipose tissue cytokines). Chronic inflammation in the cirrhotic liver adipose tissue is associated with a modificationin adipokine secretion. Changes in serum levels of adiponectin are known to be associated with the development ofinsulin resistance. Increased insulin resistance is a pathophysiological feature of nonalcoholic fatty liver disease(NAFLD), one of the most common causes of chronic liver disease. In addition, it was suggested that liver cancerdevelopment is probably connected with insulin resistance. The aim of this study is to evaluate the significance ofserum Adiponectin level and insulin resistance in patients with chronic liver disease and hepatocellular carcinoma.Patient and Methods: 100 patients were enrolled in this cross sectional study and divided as following: Group I: 52HCV patients with chronic liver disease (CLD).Group II: 48 patients with hepatocellular carcinoma (HCC). For allsubjects, Serum Adiponectin and Insulin Resistance parameters (Fasting serum Insulin, Fasting serum Glucose, HOMAIR) were measured. Results: Serum Adiponectin was significantly lower in patients with hepatocellular carcinoma(p=0.000 ) and it is inversely correlated to tumor size and the number (p= 0.0001).Meanwhile, Insulin Resistanceparameters (Fasting s. Insulin, Fasting s. Glucose, HOMA IR) were significantly higher in HCC patients than CLDpatients (p= 0.0001). Conclusion: Insulin Resistance is significantly associated with the development of HCC. Serumlevel of Adiponectin may guard against HCC development among patients with chronic liver disease.     

Differential expression of haptoglobin isoforms in chronic active hepatitis, cirrhosis and HCC related to HBV infection

The three main complications of hepatitis B virus (HBV) infection are chronic active hepatitis (CAH), liver cirrhosis, and hepatocellular carcinoma (HCC). The aim of this study was to identify differentially expressed serum proteins among the three liver complications in patients with HBV infection. Differentially expressed proteins have been shown to be potential biomarkers for disease diagnosis, prognosis and therapy guidance. Two-dimensional polyacrylamid gel electrophoresis (2DE) combined with liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed on sera from CAH, cirrhosis and HCC patients with HBV infection, as well as those obtained from healthy individuals. Of 54 differentially expressed (≥1.5-fold and p<0.05) protein spots, 35 spots were identified by LC-MS/MS. The identified spots correlated to 13 proteins. The proteins included haptoglobolin α-2 and β?isoforms, haptoglobin cleaved β isoforms, retinol-binding protein, transthyretin, ficolin, leucine-rich-α-2-glycoprotein, α-1-antitrypsin and clusterin. Of particular interest is the significant increase of haptoglobin α-2 isoforms in HCC patients compared to cirrhosis ones. In contrast, a significant decrease of the isoforms was noted among cirrhosis patients.     

Application of SELDI-TOF-MS to identify serum biomarkers for renal cell carcinoma

Purpose

Early diagnosis is critical for improving the outcome of patients with renal cell carcinoma (RCC). In this study, we applied a proteomic approach to identify serum biomarkers associated with different stages of renal tumor development.

Materials and methods

The protein expression profiles in patient serum samples were analyzed using surface enhanced laser desorption/ionization time-of-flight mass spectroscopy (SELDI-TOF-MS). The subjects included 65 patients with renal cell carcinomas, 34 with benign renal tumors, and 69 normal controls. A diagnostic decision tree was developed and validated based on the differentially expressed proteins between the serum of patients with small (?3 cm) RCC tumors and normal controls.

Results

The numbers of proteins differentially expressed in the serum samples were 29 between the patients with RCC and normal controls, 18 between patients with RCC and benign renal tumors, and 35 between patients with small RCC tumors and normal controls, respectively. The diagnostic decision tree generated from the differentially expressed proteins between patients with small RCC tumors and normal controls proved efficient in the early diagnosis of renal cell carcinoma, with a sensitivity of 81.8% and specificity of 100%. The eukaryotic initiation factor 2B delta subunit (eIF2B-delta) was identified as the most highly up-regulated protein in the serum of patients with RCC.

Conclusions

SELDI-TOF-MS is a simple, sensitive and highly reproducible technique that can be used to identify serum biomarkers. The serum biomarkers identified by this study may facilitate early diagnosis of RCC and offer new targets for mechanistic understanding and clinical therapy of this disease.     

Clinical significance of type III procollagen peptide in sera of patients with liver cancer

We assayed type III procollagen peptide in the sera of 213 patients with various liver diseases and 23 normal controls by radioimmunoassay. The non-cancerous limit of the serum level of type III procollagen peptide was defined as the mean +/- 2 SD of the patients with chronic hepatitis, liver cirrhosis and alcoholic liver disease; it was 50 ng/ml. The percentage of type III procollagen peptide in sera exceeding this limit was 22.2% in patients with hepatocellular carcinoma and 17.4% in metastatic liver cancer. Only patients with liver cirrhosis accompanied by alcoholic hepatitis exceeded this limit. In patients with hepatocellular carcinoma with peptide concentrations above 50 ng/ml, the serum level of GOT, GPT, LDH, T. Bil., LAP, gamma-GTP and T. Chol. was significantly higher than in patients with hepatocellular carcinoma whose serum peptide level was below 20 ng/ml.     

Comparative analysis of the liver and plasma proteomes as a novel and powerful strategy for hepatocellular carcinoma biomarker discovery

The extraordinary developments made in the past decade in proteomic technologies, in particular in mass spectrometry, have enabled investigators to consider designing studies to search for diagnostic and therapeutic biomarkers by scanning complex proteome samples. We developed a method based on extensive fractionation of intact proteins, to comprehensively and quantitatively profile the liver and plasma proteomes in health and disease. We have applied this method to samples collected from patients with early hepatocellular carcinoma (HCC) and from patients with liver cirrhosis as well as to samples collected from three mouse models of HCC. This method allowed for the identification of proteins that differ in expression levels in liver tissue or in plasma with disease progression from liver fibrosis, cirrhosis or steatohepatitis to HCC. The comparative analysis of the liver and plasma proteomes generated from human and mouse specimens, constitutes a novel and powerful strategy for HCC biomarker discovery.     

CD44和CD133在慢性乙肝和肝细胞癌患者组织中的表达及其临床意义

目的:研究CD44和CD133在HBV诱导的慢性肝病和肝细胞癌（HCC）患者肝组织中的表达,并评价其与炎症活动度、纤维化分期（慢性肝炎合并肝硬化或无肝硬化）及肝细胞癌分级的相关性。方法:选取2015年1月至2017年12月期间,我院收治的慢性乙肝未发生肝硬化的患者24例,慢性乙肝伴有肝硬化的患者24例和肝细胞癌患者24例,分为组Ⅰ、组Ⅱ、组Ⅲ,另收集病理肝脏组织正常,血清HBV抗体和HBsAg均为阴性的患者10例,根据METAVIR评分系统对非肿瘤性肝活检中的坏死性炎症活动度和纤维化分期进行评分,采用免疫组化SABC法检测CD44和CD133在各组中的表达水平。结果:各组中的CD44和CD133表达量均随疾病进展而显著增加,差异有统计学意义（P＜0.05）。与A1F1评分相比,患者CD44和CD133的表达在炎症活动评分和纤维化达到A3F4评分时,有显著升高（P＜0.05）;且随肝细胞癌分级的升高,CD44和CD133的表达显著增加（P＜0.05）。结论:在慢性肝炎、肝硬化和肝细胞癌中均有CD44和CD133的表达。CD44和CD133的表达与炎性活动、纤维化分期和肿瘤分级增加有显著相关性。因此肿瘤干细胞标记物CD44和CD133可以用于开发新的肝细胞癌靶向药物和预防性治疗药物。     

Isoelectric focusing of alphafetoprotein in patients with hepatocellular carcinoma--frequency of specific banding patterns at non-diagnostic serum levels.

Serum levels of alphafetoprotein are raised in 60-80% of patients with hepatocellular carcinoma. Although widely used as a serum marker, frequent false-positive results in patients with benign liver disease, result in poor specificity. This occurs particularly when levels of alphafetoprotein fall between 50-500 ng ml-1, the so-called ''grey area''. Recent reports suggest that isoelectric focusing of alphafetoprotein demonstrates certain bands that are more specific for hepatocellular carcinoma. Our aim was to determine whether the apparent specificity of this new approach is gained at the expense of decreased sensitivity. Sera from 110 patients with a ''non-diagnostic'' serum alphafetoprotein level (50-500 ng ml-1) were examined by isoelectric focusing and quantified by densitometric scanning. Ten patients with chronic liver disease and a raised serum alphafetoprotein level (50-500 ng ml-1), but with no evidence of hepatocellular carcinoma, were also studied. Isoelectric focusing revealed characteristic hepatocellular carcinoma bands (bands +II and +III) in 96% patients overall, and 100% of those with levels of total alphafetoprotein greater than 100 ng ml-1. No such bands were seen among ten subjects with cirrhosis but without hepatocellular carcinoma. Bands that are characteristic of hepatocellular carcinoma (bands +II or +III) are seen in the great majority of hepatocellular carcinoma patients; their absence makes a diagnosis of hepatocellular carcinoma extremely unlikely.     

蛋白芯片技术对甲胎蛋白阴性肝癌诊断的意义

 目的　探讨表面增强激光解吸离子化飞行时间质谱（简称SELDI）技术在甲胎蛋白（AFP）阴性肝癌患者中的诊断价值。方法　应用美国Ciphergen公司SELDI仪和CM10芯片检测57例血清AFP阴性（<20 μg／L）肝癌患者和55例健康对照外周血清。采用Biomarker Wizard软件分析,筛选出特征性的蛋白峰,结合临床病理资料分析该蛋白峰的诊断价值。结果　筛选出的特异蛋白峰质荷比为4.2×103、4.1×103、6.7×103、5.7×103、6.5×103、6.9×103、5.8×103,利用差异蛋白峰对17例AFP阴性的肝癌患者和13例健康对照进行盲筛,其灵敏度和特异度分别为88.23 ％和92.31 ％。筛选出的特异蛋白峰与患者年龄、性别、肿瘤大小以及是否合并肝硬化等临床资料无关。结论　应用SELDI技术筛选肝癌患者血清特异性肿瘤标志物的方法快速、有效,对血清AFP阴性患者的正确诊断有较大的辅助作用。     

Evaluation of CA 125 as a serum marker of hepatocellular carcinoma

Serum CA 125 concentrations were raised in 90.4% of 115 southern African black patients with hepatocellular carcinoma. Seventy-four percent of 62 patients with amebic hepatic abscess, 60% of 40 patients with chronic hepatic parenchymal disease (chronic hepatitis or cirrhosis), and 60.9% of 41 patients with acute viral hepatitis also had raised values. The median serum CA 125 concentration for patients with hepatocellular carcinoma differed significantly from the benign hepatic disease groups analysed (p less than 0.0002). Serum alpha-fetoprotein levels were raised in 90.4% of the 115 hepatocellular carcinoma patients. CA 125 is thus a highly sensitive marker for hepatocellular carcinoma, but lacks specificity.     

应用SELDI-TOF-MS技术分析南通地区食管癌血清差异表达蛋白

目的:应用SELDI-TOF-MS技术寻找食管鳞癌血清中相关差异表达蛋白。方法:应用表面增强激光解析离子化飞行时间质谱(SELDI-TOF-MS)技术,用WCX2芯片分析了24对食管鳞癌和正常对照的血清蛋白表达谱。结果:与正常对照相比,发现其中6个血清蛋白在食管鳞癌和正常对照组的差异有统计学意义(P<0.05),其中在食管鳞组低表达的有M/Z为4623,5012,5809和9422,而高表达的有5910,11681。结论:SELDI-TOF-MS技术是一种快速、简便易行且高通量的分析方法,不仅能直接筛选出食管癌患者血清中差异表达的潜在标记物,而且可能具有较好的临床应用前景。     

Hepatocellular carcinoma in the era of immunotherapy

Hepatocellular carcinoma is a common malignancy which usually emerges on a background of chronic liver disease. Unfortunately, with contemporary management, patients with advanced hepatocellular carcinoma have few treatment options, and prognosis is poor. The emergence of immunotherapy has afforded new therapeutic opportunities. This article reviews the clinical evidence for immunotherapy in advanced hepatocellular carcinoma and presents ideas for future drug development.     

Serum proteome profiling of primary breast cancer indicates a specific biomarker profile

Non-invasive biomarkers for early breast cancer detection are urgently needed, as the risk of recurrent morbidity and mortality is closely related to the stage of the disease at the time of primary surgery. Currently, there are no established clinical biomarkers for breast cancer. Evaluation of protein expression patterns in body fluids using proteomic technologies can be used to discover new biomarkers for the detection of breast cancer. The aim of this study was to identify a biomarker signature identifying primary non-metastatic breast cancer and healthy controls. We screened 91 serum samples including 45 breast cancer patients and 46 healthy women using a proteomic approach. We found 14 biomarkers whose combination detects breast cancer patients from non-cancer controls with a sensitivity of 89% and specificity of 67%. Five biomarkers were comparable with previously identified proteins from published data using similar approaches. This biomarker panel allows accurate discrimination between breast cancer and healthy individuals. In addition, it could distinguish subgroups of breast cancer based on patterns of several specific biomarkers. Further validation of biomarkers could potentially facilitate the early diagnosis of breast cancer as an aid to imaging diagnostics.     

Clinicopathological Criteria for Multicentricity of Hepatocellular Carcinoma and Risk Factors for Such Carcinogenesis

Multicentric occurrence is an important characteristic of hepatocellular carcinoma. We evaluated clinicopathological criteria for multicentric hepatocellular carcinoma and identified risk factors for such carcinogenesis. Subjects were 251 consecutive patients undergoing liver resection for hepatocellular carcinoma. One kind of multicentric hepatocellular carcinoma had at least one tumor consisting of well-differentiated hepatocellular carcinoma, together with moderately or poorly differentiated hepatocellular carcinoma located in a separate region. The other kind had an area of well-differentiated component around hepatocellular carcinoma with less differentiation in all occurrences. The outcome of patients with tumors classified in this way was studied. Univariate and multivariate analyses were done to identify risk factors for multicentric hepatocellular carcinoma. The cumulative survival rate was significantly higher in patients with multicentric hepatocellular carcinoma than in patients with hepatocellular carcinoma associated with intrahepatic metastasis. Analysis by Cox's proportional hazard model showed that multicentricity was not a factor in the outcome. The risk of multicentric occurrence increases with progression of chronic liver disease. Univariate analysis showed hepatitis C virus marker and hepatitis B core antibody to be risk factors. By multivariate analysis, the odds ratio for multicentric occurrence in patients infected with hepatitis C virus and with serum hepatitis B virus core antibody compared with patients without either hepatitis C virus or hepatitis B virus was 10.86. This ratio in patients with hepatitis C virus alone was 4.30. These criteria for multicentric hepatocellular carcinoma seem to be clinically useful. Hepatitis C virus infection with or without former infection by hepatitis B virus is a strong risk factor for multicentric hepatocarcinogenesis.     

飞行时间质谱技术在乳腺癌诊断中的应用

 目的 采用表面增强激光解吸电离飞行时间质谱（SELDI-TOF-MS）技术检测乳腺癌患者血清蛋白质指纹图谱，分析健康人与乳腺癌患者以及乳腺癌患者手术前后特异性标志蛋白变化，探讨血清蛋白质指纹图谱在乳腺癌疗效评价及复发监测中的临床意义。方法 用SELDI-TOF-MS技术检测30例乳腺癌患者（术前和术后第7天）及20名健康人的血清蛋白质指纹图谱，筛选乳腺癌特异性蛋白标志物，结合支持向量机软件建立诊断模型。比较乳腺癌手术前后特异性蛋白的变化。结果 与健康人血清蛋白质谱相比，术前乳腺癌血清中有3个差异蛋白，质荷比为2043、3938的标志分子低表达，质荷比为 5639的标志分子高表达。术后质荷比为2043、3938的标志分子稳定上调，质荷比为 5639的标志分子稳定下调，差异具有统计学意义。以筛选3个特异性蛋白质峰的数据构建的诊断模型经交叉验证，灵敏性和特异性均为100 %。结论 SELDI-TOF-MS检测血清蛋白质组学图谱在乳腺癌的早期诊断、术后病情转归及监测复发等方面具有一定的临床指导意义     

Preclinical and post-treatment changes in the HCC-associated serum proteome

SELDI-based proteomic profiling of body fluids is currently in widespread use for cancer biomarker discovery. We have successfully used this technology for the diagnosis of hepatocellular carcinoma (HCC) in hepatitis C patients and now report its application to serial serum samples from 37 hepatitis C patients before development of HCC, with HCC and following radiofrequency ablation of the tumour. As with alpha-fetoprotein, an accepted biomarker for HCC, we hypothesised that HCC-associated proteomic features would 'return to normal' following successful treatment and the primary aim of our study was to test this hypothesis. Several SELDI peaks that changed significantly during HCC development were detected but they did not reverse following treatment. These data may be interpreted to suggest that the characteristic SELDI profile is not linearly related to tumour burden but may result from the progression of underlying liver disease or from the emergence of precancerous lesions. beta2-Microglobulin, a protein previously reported to be markedly elevated in patients with HCV related HCC, was also the most significantly HCC associated proteomic feature (m/z 11720) in this study.     

Analysis of intrahepatic lymphocyte subsets in a transgenic mouse model of immune-mediated hepatocarcinogenesis

Long-term, persistent liver cell injury increases the risk for hepatocellular carcinoma (HCC) development in chronic viral hepatitis. In support of this notion, we have developed a unique animal model of chronic immune-mediated liver disease that induces hepatocellular carcinogenesis using HBV transgenic mice; however, the intrahepatic inflammatory response was not precisely evaluated. The current study demonstrated that hepatitis B surface antigen (HBsAg)-specific cytotoxic T lymphocytes (CTLs) were detected at a frequency of 0.05% of CD8+ T lymphocytes in the liver, and that monocytes/macrophages were remarkably increased as the disease developed. These results suggest that a minimal number of intrahepatic virus-specific CTLs and the recruited monocytes/macrophages may contribute to the process of chronic liver inflammation.     

Clinical aspects of intrahepatic bile duct carcinoma including hilar carcinoma: a study of 57 autopsy-proven cases.

The clinical features of 57 autopsied cases of intrahepatic bile duct carcinoma including 28 cases of the peripheral type (cholangiocarcinoma in the narrow sense) and 29 cases of the hilar type are described in comparison with those of hepatocellular carcinoma, with a review of the literature on the clinicopathological aspects of intrahepatic bile duct carcinoma. As compared with hepatocellular carcinoma, the average age of the patients was older; the male predominance was not obvious, chronic parenchymal liver disease was infrequent in the past history, association of primary cirrhosis was seldom, cholestatic features were frequently the early signs and more pronounced during the course, the liver was enlarged to a lesser extent, ascites was less common, signs of portal hypertension were absent or minimal, and extrahepatic metastases were less frequent. In many respects, the hilar type resembled extrahepatic bile duct carcinoma, and the peripheral type was somewhat between it and hepatocellular carcinoma. Although the overall survival was not much different from that for hepatocellular carcinoma, early diagnosis is emphasized; this would make surgical management possible. Differential diagnosis from hepatocellular carcinoma may be possible in the majority with direct cholangiography, liver scan, celiac angiography, determination of alpha-fetoprotein and hepatitis B antigen, and blood chemistry such as SGOT, SLDH, serum bilirubin and alkaline phosphatase. Illustrative cases are given including one patient with a hilar carcinoma who survived for more than 2 years after transhepatic biliary drainage.     

肝星状细胞和肝细胞癌的相互作用

肝星状细胞(HSC)是肝纤维化及正常肝脏中细胞外基质的主要来源.慢性肝损伤时,HSC发生表犁变化转变成肌纤维母细胞,引起肝纤维化.肝细胞癌是多因素疾病,基质中有明显星状细胞浸润.探讨肝细胞癌与HSC相互作用关系可为肝细胞癌的治疗提供新思路.