Loss of the orphan nuclear receptor SHP is more pronounced in fibrolamellar carcinoma than in typical hepatocellular carcinoma

Hepatocellular carcinoma (HCC) remains a major problem in oncology. The molecular mechanisms which underlie its pathogenesis are poorly understood. Recently the Small Heterodimer Partner (SHP), an orphan nuclear receptor, was suggested to be involved as a tumor suppressor in hepatocellular carcinoma development. To date, there are no such studies regarding fibrolamellar carcinoma, a less common variant of HCC, which usually affects young people and displays distinct morphological features. The aim of our project was to evaluate the SHP levels in typical and fibrolamellar hepatocellular carcinoma with respect to the levels of one of the cell cycle regulators, cyclin D1. We assessed the immunoreactivity levels of SHP and cyclin D1 in 48 typical hepatocellular carcinomas, 9 tumors representing the fibrolamellar variant, 29 non malignant liver tissues and 7 macroregenerative nodules. We detected significantly lower SHP immunoreactivity in hepatocellular carcinoma when compared to non malignant liver tissue. Moreover, we found that SHP immunoreactivity is reduced in fibrolamellar carcinoma when compared to typical hepatocellular carcinoma. We also found that SHP is more commonly lost in HCC which arises in the liver with steatosis. The comparison between the cyclin D1 and SHP expression revealed the negative correlation between these proteins in the high grade HCC. Our results indicate that the impact of loss of SHP protein may be even more pronounced in fibrolamellar carcinoma than in a typical form of HCC. Further investigation of mechanisms through which the loss of SHP function may influence HCC formation may provide important information in order to design more effective HCC therapy.     

肝癌患者经系统治疗后甲胎蛋白变化对预后的影响

目的:探讨肝癌患者经系统治疗后甲胎蛋白(AFP)变化对患者预后的影响。方法:对53例肝癌患者于入院后和系统治疗后分别于不同时间点测定血清AFP值后,将入院时AFP值作为基线,以变化50%作为标准分组,并进行无疾病进展生存时间(PFS)和总生存时间(OS)分析。结果:9例患者血清AFP值下降超过50%(A组),28例患者AFP值升高超过50%(B组),而AFP值变化小于50%(C组)的为16例。和C组相比,A组患者PFS明显延长(P<0.05),B组PFS明显缩短(P<0.05)。B组OS短于C组(P<0.01),而A组和C组间OS无明显差别(P>0.05)。结论:肝癌系统治疗前后的AFP值变化可作为临床上预后判断的标志之一。     

Alpha-fetoprotein concentrations measured by radioimmunoassay in diagnosing and excluding hepatocellular carcinoma.

Serum alpha-fetoprotein (AFP) concentrations were estimated by sensitive radioimmunoassay in 30 patients with cirrhosis complicated by hepatocellular carcinoma and in 100 patients with cirrhosis in whom malignancy was excluded. Twenty-nine of the 30 patients with hepatocellular carcinoma had concentrations above 10 IU/ml (10.5 ng/ml) (median 3500 IU/ml (3675 ng/ml)), whereas only one of the 100 patients with cirrhosis and no tumour development had a raised concentration. Eleven out of 20 patients in whom hepatocellular carcinoma had developed in an apparently normal liver had raised AFP concentrations. In this group the differential diagnosis is usually secondary carcinoma, and three of 50 such patients had AFP concentrations above 10 IU/ml. Noting raised AFP concentrations is thus of considerable value both in detecting and in excluding hepatocellular carcinoma in cirrhosis, for in this case such concentrations gave only 1% false-positive and 3% false-negative results. They are less useful, however, in distinguishing between primary tumours arising in patients without cirrhosis and secondary hepatic deposits, giving 6% false-positive and 45% false-negative results.     

Correlation between serum enzymes and serum unsaturated vitamin B12 binding proteins in primary liver carcinoma

The serum unsaturated vitamin B12-binding capacity (UBBC), unsaturated transcobalamin (UTC) I, UTC II, UTC III levels, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase activities and bilirubin concentration were estimated in 61 patients with liver diseases (31 with hepatoma, 30 with viral hepatitis). The levels of serum cobalamin, UTC I, UTC III, UBBC, alanine and aspartate aminotransferases, and bilirubin were raised in both hepatoma and viral hepatitis patients. Serum UTC II was reduced in both conditions. Alkaline phosphatase activity was significantly increased in hepatoma. Four significant correlations were observed among these parameters in the hepatoma patients while only one significant correlation was observed in viral hepatitis.     

Fine needle aspiration cytology of fibrolamellar hepatocellular carcinoma

The major cytologic features seen in fine needle aspirates from two cases of fibrolamellar hepatocellular carcinoma were: liver-like tumor cells, characterized by plump, polygonal forms with eosinophilic, granular cytoplasm; large oval nuclei with extremely prominent solitary nucleoli; and parallel bands of fibrous tissue and fibrocytes seen within the tumor fragments. Other helpful features included intracytoplasmic hyaline globules and well-delineated pale bodies. Clinically, the tumors occurred in young patients with noncirrhotic livers and ran a more favorable course than do other types of hepatocellular carcinoma.     

Evidence for hepatitis C viral infection in patients with primary hepatocellular carcinoma.

In testing for antibodies to the hepatitis C virus (anti-HCV) in 112 patients with primary hepatocellular carcinoma, 10 of 33 white patients (30%) and 15 of 79 Asian patients (19%) had a positive response to the antibody. The antibody profile to individual hepatitis C viral antigens and the presence of circulating hepatitis C viral RNA were determined in the 25 patients. The anti-HCV antibodies most frequently detected were toward the antigens from the core (C22) and NS3 regions. Serum hepatitis C viral RNA was present in 17 of the 25 patients (68%), and these patients tended to have serum levels of alanine and aspartate aminotransferases higher than those patients without viremia (136 +/- 22 U per liter versus 64 +/- 11 U per liter and 161 +/- 26 U per liter versus 79 +/- 14 U per liter, respectively, both P < .05). Of the 15 Asian patients with hepatocellular carcinoma and anti-HCV, 4 (27%) had coexisting hepatitis B surface antigen (HBsAg) and 13 (87%) had antibodies to either hepatitis B core or surface antigen. Of the 10 white patients with anti-HCV, however, only 1 (10%) had hepatitis B virus antibodies (P < .01). Among 4 Asian patients with coexisting anti-HCV and HBsAg, 1 was found to have serum hepatitis B viral DNA and the other 3 had hepatitis C viral RNA. A history of blood transfusion was obtained from 12 of the 25 patients with anti-HCV (48%); 20 (80%) had coexisting cirrhosis. Our findings support the hypothesis that hepatitis C virus is an important etiologic agent in the development of primary hepatocellular carcinoma in both white and Asian patients in the United States.     

Release of unsaturated vitamin B12 binding capacity from human granulocytes by the calcium ionophore A23187

The ionophore A23187, in the presence of calcium ions, was capable of eliciting the release of granule-associated unsaturated vitamin B₁₂ binding capacity from human granulocytes. The ionophore-induced extrusion of unsaturated vitamin B₁₂ binding capacity was concentration, time and temperature-dependent. The release was blocked by 2-deoxyglucose and was unaccompanied by cytotoxicity (trypan-blue uptake and lactate dehydrogenase release). The unsaturated vitamin B₁₂ binding capacity release was accompanied by the release of lysozyme, a specific granule marker enzyme.     

Physician's management of suspected vitamin B12 deficiency.

A retrospective study was undertaken to audit physician''s management of patients with a low serum level of vitamin B12 who were admitted to a university-affiliated teaching hospital during 1 year. Among the 34 patients 13 were proved to have pernicious anemia or vitamin B12 malabsorption, but for 12 of them there were unnecessary delays (several days or weeks) before initiation of investigation and therapy. An additional six patients, who had low serum levels of vitamin B12 and macrocytosis, most likely had true vitamin B12 deficiency, but proper investigation was not done and they did not receive any vitamin B12 or folic acid therapy. In another nine cases unexplained low serum levels of vitamin B12 were not properly investigated, and the patients either did not receive any vitamin B12 therapy or received it without proper documentation of a deficiency. Suggestions for facilitating early detection, investigation and treatment of megaloblastic anemia or vitamin B12 deficiency are given.     

Human plasma R-type vitamin B12-binding proteins. II. The role of transcobalamin I, transcobalamin III, and the normal granulocyte vitamin B12-binding protein in the plasma transport of vitamin B12.

The normal human granulocyte vitamin B12-binding protein, transcobalamin I, and transcobalamin III, have been labeled with 125I-labeled N-succinimidyl 3-(4-hydroxyphenyl)propionate and utilized for plasma clearance studies performed with rabbits. Both moieties of 125I-labeled granulocyte vitamin B12-binding protein-[57Co]vitamin B12 were cleared rapidly from the plasma (is less than 90% by 5 min) by the liver. After 30 min, the bulk of the 125I reappeared in the plasma in small molecular weight (less than 1000) form and was rapidly excreted in the urine. After 60 min the bulk of the [57Co]vitamin B12 reappeared in the plasma bound to rabbit transcobalamin II and was subsequently taken up by a variety of tissues. Approximately 15% of the 125I-labeled granulocyte vitamin B12-binding protein-[57Co-a1vitamin B12 was excreted intact into the bile during the period from 10 to 80 min after injection. The hepatic uptake of the protein-vitamin B12 complex was blocked by the prior injection of desialyzed fetuin but not by native fetuin. Similar results were obtained with 125I-labeled transcobalamin III-[57Co]vitamin B12. Approximately 90% of both moieties of 125I-labeled transcobalamin I-[57Co]vitamin B12 had prolonged plasma survivals similar to that of 125I-labeled bovine serum albumin. After treatment with neuraminadase, both moieties of the 125I-labeled transcobalamin I-[57Co]vitamin B12 complex were cleared rapidly from the plasma by the liver in a manner that was indistinguishable from that observed in the case of untreated granulocyte vitamin B12-binding protein and transcobalamin III. These observations indicate that desialyzed transcobalamin I and the native forms of the granulocyte vitamin B12-binding protein and transcobalamin III are cleared from plasma by the mechanism elucidated by Ashwell and Morell (Ashwell, G., and Morell A. G. (1974) Adv. Enzymol. 41, 99-128) that is capable of clearing a wide variety of asialoglycoproteins. These observations have implications concerning the function of the human R-type vitamin B12-binding proteins, the nature of the enterohepatic circulation of vitamin B12, the biological significance of the mechanism described by Ashwell and Morell, and the etiology of the increased plasma concentration of human R-type protein that occurs frequently in chronic myelogenous leukemia and occasionally in hepatocellular carcinoma and other solid tumors.     

Sex steroid binding patterns in primary biliary cirrhosis complicated by hepatocellular carcinoma.

Owing to recent findings of certain unusual sex steroid binding in liver disease--particularly an allosteric biphasic pattern (pattern A) unique to the serum of patients with hepatocellular carcinoma--the serum binding characteristics for 5 alpha-dihydrotestosterone were examined in serum samples from six patients with primary biliary cirrhosis who had developed hepatocellular carcinoma. In all serum samples taken after the development of tumour pattern A binding only was obtained, and in four cases in which earlier samples were also examined there was a transformation from the normal, non-specific binding pattern, or an allosteric plateau pattern seen in non-malignant liver disease (designated D and C respectively), to pattern A coincident with the rise in serum alpha fetoprotein. In one patient chemotherapy leading to a fall in alpha fetoprotein abolished pattern A binding, showing further its close association with tumour growth. The value of pattern A binding as a tumour marker in hepatocellular carcinoma warrants further study.     

Fucosylation of serum alpha-fetoprotein in patients with primary hepatocellular carcinoma

alpha-Fetoprotein specimens were prepared from the sera of four patients with hepatocellular carcinoma. The lentil lectin-reactive and lectin-nonreactive variants of this glycoprotein were also prepared from the serum of one of the four patients by affinity chromatography with immobilized lectin. The correlation between the carbohydrate structure of these compounds and their reactivity in crossed immuno-affinoelectrophoresis with lentil lectin was studied by chemical analysis and affinity chromatography of the glycopeptides with lectin columns. It was found that the lentil lectin-reactive variant contained a carbohydrate chain of the fucosylated biantennary complex type. These data together with previous findings indicate that most of the patients with hepatocellular carcinoma have an elevated serum concentration of fucosylated alpha-fetoprotein.     

Cholestasis is the main determinant of abnormal CA 19-9 levels in patients with liver cirrhosis

BACKGROUND/AIMS: Altered CA19-9 levels are commonly found in patients with liver cirrhosis though a clear explanation for this finding has not yet been given. The aim of this study was to investigate whether CA19-9 levels might be related to alterations in biochemical parameters and/or to functional impairment in cirrhotic patients with and without hepatocellular carcinoma. METHODS: We studied 126 patients with liver cirrhosis, 60 of whom also had hepatocellular carcinoma. CA19-9 values were related to clinical, biochemical and functional parameters. In half of the patients CA19-9 levels were related to the monoethylglycinexylidide test, which is a dynamic liver function test. RESULTS: In more than half the cases CA19-9 values were above the upper limit. Liver function worsening as assessed by Child-Pugh's score and monoethylglycinexylidide test did not seem to influence the alteration of the marker. By contrast, in univariate analysis CA19-9 correlated with aminotransferases, gamma-glutamyltransferase and alkaline phosphatase. Multivariate analysis showed that besides alkaline phosphatase also the presence of hepatocellular carcinoma might influence the alteration of CA19-9, although the marker was of no use for the diagnosis of liver cancer in patients with altered though not diagnostic alpha-fetoprotein levels. CONCLUSIONS: In our study we confirmed the correlation of CA19-9 levels with cholestasis and cytolysis parameters. Moreover, we found no association between CA19-9 levels and impaired liver function as assessed by means of the Child-Pugh's score and the monoethylglycinexylidide test, which is cholestasis-independent and explores liver metabolic and clearance activities. The cholestatic picture that characterizes liver cirrhosis might enhance the expression and passage of the marker from the bile to the blood. The addition of CA19-9 assessment is not useful for the diagnosis of hepatocellular carcinoma in patients with non-diagnostic levels of alpha-fetoprotein. Caution should therefore be used when evaluating CA19-9 in cirrhotic patients with cholestasis, since false positive results may occur.     

Thalassaemia,iron, and pregnancy.

Haematological values of 35 pregnant women with beta-thalassaemia trait were followed during pregnancy. The discriminant function, calculated from haematological indices, was of no value in diagnosing beta-thalassaemia trait during pregnancy. Initially patients were given iron supplements only if the serum iron and total iron binding capacity levels indicated iron deficiency, but bone marrow biopsies performed in the first 22 patients at 32 weeks indicated deficient iron stores. These patients were therefore given iron irrespective of their serum iron level. All subsequent patients with beta-thalassaemia were also put on iron routinely at booking. Retrospectively the patients were divided into two groups. Patients in group 1 (18 patients) had received iron for less than 12 weeks, and their haemoglobin levels fell significantly during pregnancy (P less than 0-001). Haemoglobin levels in 16 patients who had received iron for more than 12 weeks (group 2), however, did not fall significantly during pregnancy (P less than 0-6). It is suggested (contrary to common practice) that patients with beta-thalassaemia trait should be given iron supplements during pregnancy. Serum folate and vitamin B12 levels did not change significantly in these patients and there was no increase in the incidence of maternal or fetal complications.     

Malabsorption of protein bound vitamin B12.

Patients with subnormal serum vitamin B12 concentrations were tested for absorption of protein bound vitamin B12 and compared with controls. Absorption of the protein bound vitamin appeared to decrease with increasing age in healthy subjects. Differences between the result of this test and the result of the Schilling test in patients who had undergone gastric surgery were confirmed; such differences were also seen in some patients who had iron deficiency anaemia, an excessive alcohol intake, or folate deficiency. Defective absorption was also found in six patients with an adequate dietary intake of vitamin B12, normal Schilling test results, low serum vitamin concentrations, and tissue changes responding to treatment with vitamin B12. Malabsorption of the vitamin from protein bound sources, which is not detected by the Schilling test, may produce vitamin B12 deficiency of clinical importance.     

Serum Iron, Vitamin B12 and Folic Acid Levels in Parkinson’s Disease

We aimed to investigate possible associations between systemic iron metabolism deficiency and Parkinson's disease, and also to research any possible correlations between stage of the disease and vitamin B12 and folic acid levels. 33 male and 27 female patients diagnosed with idiopathic Parkinson's disease and 22 male and 20 female age- and sex-matched controls were enrolled in the study. Having the diagnosis of secondary Parkinsonism or Parkinson plus syndromes, and for the females, not being in the menopausal stage were considered as exclusion criteria. Recordings of blood samples of both groups collected after 8 h fasts were assessed in terms of serum iron, ferritin levels and iron-binding capacity, vitamin B12 and folic acid levels. The Hoehn and Yahr scale was used to determine the stage of the disease. No statistically significant difference was found with respect to mean serum iron, median serum ferritin levels and median serum iron-binding capacity between the groups. A statistically significant but inverse correlation was found between symptoms' duration and serum iron and ferritin levels. There was no statistically significant difference between the groups with respect to vitamin B12 and folic acid levels. However, a statistically significant but inverse correlation was determined between the patients' vitamin B12 levels and the Hoehn and Yahr scores. As Parkinson's disease progresses, serum iron, ferritin and vitamin B12 levels may decrease. The lower levels of these parameters may be the cause of the progression or may be the result of it.     

Clinical evaluation of lentil lectin-reactive alpha-fetoprotein-L3 in histology-proven hepatocellular carcinoma

INTRODUCTION: Serum alpha-fetoprotein (AFP) is a useful marker of hepatocellular carcinoma (HCC), although the serum AFP concentration is also increased in patients with chronic liver diseases (CLD). The analysis of AFP glycoforms has been known to be of diagnostic value. We applied the lectin-affinity electrophoresis and antibody-affinity blotting techniques to HCC patients in Vietnam in order to better understand the role of lentil lectin-affinity AFP-L3 in the diagnosis and differential diagnosis of HCC, and its relationship with the biological characteristics of HCC. METHODS: Lens culinaris agglutinin-reactive AFP (AFP-L3) was measured in 65 patients with histologically proven HCC and 25 patients with CLD. All patients had serum AFP levels above 54 ng/mL. AFP-L3 levels were determined by lectin affinity electrophoresis coupled with antibody-affinity blotting. The diagnosis of HCC was confirmed histologically by ultrasound-guided biopsy. RESULTS: The mean value of AFP-L3 in the HCC patients was 49.6 +/- 21.6%, which was significantly higher (p<0.001) than that in the 25 CLD patients (10.7 +/- 4.3%). When the cutoff level for AFP-L3 was set at 15% (mean +/- SD), the sensitivity was 96.9%, the specificity 92.0% and the accuracy 95.5% in the 65 HCC patients. There was no clear correlation between serum AFP level and AFP-L3 percentage (r=0.16). There was no correlation between AFP-L3 and the maximum diameter of HCC nodules (r=0.05). However, the mean AFP-L3 value was higher in moderately or poorly differentiated HCC than in well differentiated tumors (p<0.001). CONCLUSIONS: AFP-L3 is potentially a clinically useful marker for the differentiation of increased AFP levels in hepatocellular carcinoma and chronic liver diseases. The AFP-L3 percentage is closely related to HCC differentiation. We consider the analysis of AFP-L3 a useful adjunct in the diagnosis of HCC.     

全反式维甲酸对急性早幼粒细胞白血病患者血清学的影响

目的：探讨全反式维甲酸（All trans retinoic acid,ATRA）作为辅助剂在急性早幼粒细胞白血病（Acute promyelocytic leukemia, APL）治疗中,对患者血清促红细胞生成素（Erythropoietin,EPO）、血清铁蛋白、叶酸和维生素B12水平的影响。方法：回顾性分析 我院2011 年6 月-2015 年6 月接诊的50 例急性早幼粒细胞白血病患者的临床资料。根据患者的治疗方法不同将患者分为两组, A 组（亚砷酸钠治疗组）和B 组（亚砷酸钠联合ATRA）。对比两组患者治疗后血清铁蛋白、EPO、叶酸和维生素B12 恢复情况。结 果：两组患者入院时所有血清EPO、铁蛋白等比较,差异无统计学意义（P>0.05）;治疗后两组患者血清各指标均有所改善（P<0. 05）;治疗后B 组患者中EPO、血清B12、血清铁蛋白和叶酸恢复正常水平者明显多于A 组,差异具有统计学意义（P<0.05）。结论： 全反式维甲酸辅助治疗急性早幼粒细胞白血病患者能够更好的改善患者血清EPO、铁蛋白、叶酸和维生素B12 的异常,对于疾病 的治疗有一定的效果。     

Is vitamin D important for preserving cognition? A positive correlation of serum 25-hydroxyvitamin D concentration with cognitive function

This study investigates the association of vitamin D status with cognitive function and discusses potential mechanisms for such an effect. The relationship of vitamin B12 with cognition was also assessed. A retrospective review of older adults presenting to a university-affiliated clinic providing consultative assessments for memory problems was performed. Charts of all patients (n=80) presenting for initial visits were reviewed to identify those who had serum 25-hydroxyvitamin D (25(OH)D), vitamin B12, and mini-mental state examination score (MMSE) all obtained on their first visit (n=32). Correlation analyses between MMSE and 25(OH)D and vitamin B12 levels were performed. Serum 25(OH)D concentration and MMSE showed a (p=0.006) positive correlation; no (p=0.875) correlation was observed between serum B12 concentration and MMSE. In conclusion, the positive, significant correlation between serum 25(OH)D concentration and MMSE in these patients suggests a potential role for vitamin D in cognitive function of older adults.     

Impact of maternal serum alpha-fetoprotein screening on antenatal diagnosis.

An analysis has been made of indications for amniocentesis in the Edinburgh area from 1979 to 1981. About 5% of all mothers underwent the procedure. Among 2137 amniocenteses, 37% were performed on mothers 35 years old or more, and 30% on patients with raised serum alpha-fetoprotein. The total number of amniocenteses and the categories have been stable for the past three years. As a result of amniocentesis 104 pregnancies were terminated, 66 of which (63%) followed a raised maternal serum alpha-fetoprotein indication, while only 10 (9.6%) were in mothers aged 35 or more. There were a further 12 terminations based on raised serum alpha-fetoprotein but where no amniocentesis had been thought necessary. Even when figures for anencephaly are excluded from the analysis, maternal serum alpha-fetoprotein screening was responsible for detecting 35 out of 63 (56%) abnormal fetuses. This constitutes a strong case for the continuation of alpha-fetoprotein screening programmes.     

血清Hcy，叶酸和维生素B12在胃癌及其癌前疾病中的作用

目的:探讨血清同型半胱氨酸(Hcy)、叶酸以及维生素B12在胃癌及癌前疾病中的水平及临床意义。方法:收集2014年1月至2016年8月我院收治的100例胃癌患者(胃癌组),及100例胃良性病变患者包括41例胃炎、34例胃溃疡、25例胃息肉(癌前病变组),并于同期随机选择200例健康体检者为对照组,采用循环酶法测定三组的血清Hcy,电化学发光免疫分析法测定叶酸及维生素B12水平,并分析各指标与胃癌临床病理特征的关系。结果:胃癌组、癌前病变组血清Hcy水平均高于对照组,叶酸及维生素B12水平均低于对照组,并且胃癌组血清Hcy水平高于癌前病变组,叶酸及维生素B12水平低于癌前病变组,差异有统计学意义(P0.05)。Ⅲ+Ⅳ期胃癌患者Hcy水平高于Ⅰ+Ⅱ期,进展期患者Hcy水平高于早期,有淋巴结转移患者Hcy水平高于无转移者,差异有统计学意义(P0.05);Hcy表达与性别、年龄、病变位置以及分化程度无关,差异无统计学意义(P0.05)。叶酸、维生素B12的表达在胃癌患者中与各临床病理特征(性别、年龄、TNM分期、肿瘤浸润深度、病变位置、有无淋巴结转移、分化程度)无明显关系,差异无统计学意义(P0.05)。结论:血清Hcy在胃癌患者中呈高水平表达,而叶酸及维生素B12呈低水平表达,联合检测三种指标有助于早期区分胃癌及癌前病变,同时血清Hcy还可能参与了胃癌的发生发展过程。Hcy、叶酸及维生素B12可作为早期鉴别诊断胃癌及其癌前病变的重要指标。