Serum paraoxonase and arylesterase activities for the evaluation of patients with chronic hepatitis

The sensitivity of standard biochemical tests for liver function is low and insufficient for a reliable determination of the presence or absence of liver disease. The aim of the present study was to investigate serum paraoxonase and arylesterase activities and lipid hydroperoxide (LOOH) levels, and to find out that whether the measurement of serum paraoxonase and arylesterase activities would be useful as an index of liver function status in chronic hepatitis (CH). Fourty-four patients with CH (24 CHB and 20 CHC) and 38 controls were enrolled. Serum paraoxonase and arylesterase activities were detected spectrophotometrically. LOOH levels were measured by the FOX-2 assay. Serum paraoxonase and arylesterase activities were significantly lower in patients with CH than controls (p < 0.001 for both), while LOOH levels were significantly higher (p < 0.001). Paraoxonase and arylesterase activities were inversely correlated with LOOH levels (r = -0.394, p < 0.05; r =-0.362, p < 0.05, respectively). Fibrosis scores of CH patients were significantly correlated with paraoxonase and arylesterase activities and LOOH levels (r =-0.276, p < 0.05; r = -0.583, p < 0.001 and r = 0.562, p < 0.001, respectively). Our results indicated that decrease in the activities paraoxonase and arylesterase may play a role in the pathogenesis of CH. In addition, serum paraoxonase and arylesterase activities measurement may add a significant contribution to the liver function tests.     

Serum paraoxonase and arylesterase activities in various forms of hepatitis B virus infection

The aim of this study was to determine and evaluate the activity of paraoxonase and arylesterase enzymes in various clinical forms of hepatitis B infection and to investigate the correlation between these parameters and chronic disease course/fibrosis. Overall, 40 patients diagnosed as hepatitis B carriers (CIHBV), 40 chronic active hepatitis B (CAHBV) patients, and 40 healthy adults (control group) between 18 and 65 years of age were enrolled the study. Serum paraoxonase and arylesterase activities were measured spectrophotometrically. Their activities were significantly lower in patients with CAHBV compared with CIHBV patients or with control group patients (P<0.001). There was a negative correlation between alanine aminotransferase levels and the activity of paraoxonase and arylesterase (r = -0.38, P = 0.001 and r = -0.28, P = 0.002, respectively). A statistically significant negative correlation was found between arylesterase activity in the sera of CAHBV patients and HBV DNA levels (ρ = -0.33, P = 0.03). On the contrary, no correlation was found between paraoxonase levels and HBV DNA levels (P>0.05). The histology activity index of CAHBV patients did not correlate with paraoxonase and arylesterase activities (P>0.05). In light of these findings, it may be assumed that during the progression of an inactive hepatitis B carrier to being actively infected, reduced paraoxonase and arylesterase activities may be observed.     

Maternal chronic hepatitis B virus is implicated with low neonatal paraoxonase/arylesterase activities

BACKGROUND: Paraoxonase/arylesterase activities are closely implicated with liver function and antiatherogenetic process. AIM: To evaluate whether maternal chronic hepatitis B virus, disease (HBV) affect serum neonatal paraoxonase/arylesterase activities. PATIENTS AND METHODS: 28 pregnant women with HBV and 28 healthy pregnant women (controls) in the delivery room and their newborns (cord blood) underwent laboratory examinations. Serological virus tests and liver function tests and paraoxonase (PON 1) activities were measured with the Siemens Advia 1650 Clinical Chemistry System, while total antioxidant capacity (TAC) levels and paraoxonase-arylesterase (PON-aryl) activities were measured spectrophotometrically. RESULTS: Serological HBV tests and HBV DNA showed chronic HBV (precore mutant G1896A) in the diseased mothers whereas anti-HBc and anti-HBe were detected in their neonates. Liver function parameters were found normal in controls and both groups of newborns. Moderately increased transaminase levels were measured in HBV mothers, whereas TAC levels were decreased in hepatic mothers and their newborns. Interestingly albumin levels did not differ among the studied groups. PON 1 and PON-aryl activities in the diseased mothers (148+/-14 U/mL/min, 130+/-16 KU/mL/min) and their infants (32+/-6 U/mL/min, 24+/-5 KU/mL/min) were significantly lower as compared to those of control mothers (217+/-16 U/mL/min, 196+/-14 KU/mL/min p<0.001) and their newborns (57+/-6 U/mL/min, 48+/-8 UK mL/min p<0.001). Inverse significant correlations were found between the studied enzyme activities and liver enzymes in all the groups of study except in infants born from HBV mothers and positive with TAC in all the studied groups. CONCLUSIONS: Decreased PON 1 and PON-aryl activities were measured in infants born from hepatic mothers probably as a consequence of their low TAC. Infants born from HBV mothers are at risk for developing LDL oxidation perinatally.     

Oxidant and antioxidant parameters in patients with Brucella canis

ObjectivesThe aim of this study was to evaluate the oxidative status in patient with Brucella canis (B. canis) and healthy controls by measurement of total antioxidant capacity (TAC) and total oxidant capacity (TOS), and calculation of oxidative stress index (OSI).Design and methodsSix patients with B. canis and 6 healthy control subjects were included in this study. Diagnosis of B. canis was made by rapid slide agglutination test (RSAT) and Modified Plate Agglutination Test (MPAT). The serum samples of B. canis positive patients and controls were used for measurement of TAC and TOS levels.ResultsThe TOS and OSI index were significantly higher in brucellosis positive patients than control group (p < 0.001 and p < 0.0001, respectively), while TAC level was found similar in patient and control groups.ConclusionFindings of this study have shown that B. canis generate low TOS and OSI index in man compared with smooth Brucella species infection. This difference may help to provide understanding about the pathogenesis of B. canis infection in man.     

Ghrelin,paraoxonase and arylesterase levels in depressive patients before and after citalopram treatment

ObjectivesThe purpose of this study was to examine alterations in lipid profiles and in the serum concentrations of acylated and desacylated ghrelin, paraoxonase and arylesterase in psychiatric patients before and after treatment with 40 mg citalopram daily for 3 months.Design and methodsSamples were collected from 22 healthy controls and 24 psychiatric patients before and after citalopram treatment. Blood levels of acylated and desacylated ghrelin were measured by radioimmunoassay. Paraoxonase and arylesterase activities were determined spectrophotometrically. Lipid parameters were measured on the OLYMPUS-AU400.ResultsIt was found that the levels of acylated, desacylated ghrelin, paraoxonase arylesterase, total cholesterol and triglyceride were lower in depressive patients before citalopram treatment than in the control group. Those parameters were not restored after citalopram treatment except for the arylesterase level.ConclusionDecreased PON1 and ghrelin levels as well as fluctuations in lipid profiles may be involved in the etiology of depressive disorders.     

福建省慢性丙型肝炎患者丙型肝炎病毒基因分型的分析

目的观察福建省慢性丙型肝炎患者基因型分布特点以及与患者性别、年龄、丙型肝炎病毒（HCV） RNA 之间的关系。方法应用反转录聚合酶链反应（RT-PCR）和 SANGER 测序法对155例慢性丙型肝炎患者的HCV 进行基因分型。结果在155例标本中,HCV 基因1型占55．48％,基因2型占18．71％,基因3型13．51％,基因6型12．26％,未见基因4、5型;男女慢性 HCV 感染均以1型为主要基因型,在其他基因型中,男性3型、6型多于2型,女性则以2型为主,差异有统计学意义（P ＜0．05）;基因2型平均年龄为（49岁）,3型平均年龄为（35岁）,差异有统计学意义（P＜0．05）。结论福建地区 HCV 基因型以1b 型为主,其次是2a 型,未见基因4、5型;基因2型的感染者平均年龄较基因3型大。     

Oxidant, antioxidant status and metabolic data in patients with beta-thalassemia

BACKGROUND: In beta-thalassemia major impaired biosynthesis of beta globin leads to accumulation of unpaired alpha globin chain. An iron overload, usually observed, generates oxygen-free radicals and peroxidative tissue injury. AIM: To investigate hematological parameters, oxidative stress and the antioxidant capacity in beta-thalassemia patients compared to control subjects in order to determine their impact in several organs functions. METHODS: This study was conducted on 56 beta-thalassemia major patients compared to 51 healthy subjects. We determined metabolic parameters (glycaemia, lipid parameters, electrolytes, iron indices, hepatic, renal and heart functions tests), plasmatic thiobarbituric acid reactive substances (TBARS), plasmatic peroxyl radical trapping potential (TRAP), plasmatic superoxide dismutase (SOD), erythrocyte gluthathione peroxidase (GPX), plasmatic vitamin E, vitamin A and trace elements. RESULTS: Except triglycerides, lipid fractions were significantly decreased in beta-thalassemia compared to controls. Serum ferritin, iron, TBARS concentrations, SOD and GPX activities were significantly increased. But TRAP, vitamin E and zinc concentrations were significantly decreased. CONCLUSION: Our findings confirm the peroxidative status generated by iron overload in beta-thalassemia major patients and highlight the rapid formation of marked amounts of TBARS and the increase of SOD and GPX activity. Our study suggested that in beta-thalassemia the first organ impaired is the liver.     

Detection of hepatitis C virus RNA in patients with chronic hepatitis C virus infections during and after therapy with alpha interferon.

In 24 patients with hepatitic C virus (HCV) infection who participated in a randomized trial with alpha 2B interferon, HCV RNA analysis by the polymerase chain reaction with two separate primer sets was performed at weeks 0, 4, and 24 and during a follow-up period of 6 to 9 months. Prior to therapy all patients were HCV RNA positive. During therapy, HCV RNA decreased to an undetectable level (< 1 chimpanzee infectious dose per ml) in nine patients at week 4. After week 4, no additional cases of HCV RNA disappearance (< 1 chimpanzee infectious dose per ml) were observed. During follow-up, HCV RNA could not be detected in four of the six patients with a sustained alanine aminotransferase response. These results suggest the probable predictive value of HCV RNA levels for detecting the failure of therapy in an early stage of HCV infection.     

Phylogenetic analysis of hepatitis GB virus type C/hepatitis G virus in Spanish patients with chronic hepatitis B or C virus infection.

The nucleotide sequence of hepatitis GB virus type C (HGBV-C)/hepatitis G virus (HGV) NS3/helicase and 5'-untranslated regions from 23 Spanish patients were analyzed to assign the HGV isolates one of the proposed HGBV-C/HGV genotypes. The analysis of the evolutionary distance frequency showed that the distances among all sequences in NS3/helicase region were distributed around a single peak of 0.20, suggesting that all included sequences belonged to the same HGBV-C/HGV genotype. By contrast, in the 5'-untranslated region, all the distances corresponding to our sequences and those of the HGBV-C/HGV types 2 and 3 were distributed around a major peak of 0.03. The remaining distances corresponding to the HGBV-C/HGV type 1 sequences were distributed around a minor peak of 0.11. The phylogenetic tree and pairwise comparison of evolutionary distances among the 5'-untranslated region of the infected patients and each HGBV-C/HGV genotype demonstrated that our HGBV-C/HGV isolates belonged to subtype 2a (17/23; 78%) and 2b (5/23; 22%). No relation was found between HGBV-C/HGV subtype and hepatitis B or C virus infection.     

丙型肝炎病毒感染者17种自身抗体的检测及其临床意义

目的：观察丙型肝炎病毒（HCV）感染者的自身抗体情况,了解 HCV 感染后疾病严重程度与抗核抗体（ANA）谱阳性率的关系。方法收集123例 HCV感染患者的血清标本作为 HCV组,根据病毒感染的严重程度将其分为3组：A组,急性丙型肝炎17例;B组,慢性丙型肝炎64例;C组,肝硬化42例。将排除病毒感染和自身免疫性疾病的健康者52例作为对照组。采用间接酶联免疫吸附测定法检测抗 HCV 抗体,线性免疫印迹法检测123例 HCV 感染者血清中抗 dsDNA、核小体、组蛋白、SmD1、增殖细胞核抗原（PCNA）、P0、SSA/Ro60kD、SSA/Ro52kD、SSB/La、CENP-B、Scl70、U1-snRNP、AMA-M2、Jo-1、PM-Scl、Mi-2和Ku 抗体。结果47．2％（58/123）HCV 感染者至少1种自身抗体阳性,明显高于对照组的9．6％,差异有统计学意义（P＜0．01）。ANA谱17种自身抗体阳性率由高到低依次为：抗SSA/Ro52kD抗体（23．6％）、抗SSB/La抗体（6．5％）、抗 Mi-2抗体（5．7％）、抗PCNA抗体（4．9％）、抗核小体抗体（3．3％）、抗 Ku 抗体（3．3％）,抗 dsDNA、SmD1、AMA-M2、PM-Scl 抗体均为1．6％,抗组蛋白、P0、SSA/Ro60kD、CENP-B、U1-snRNP、Jo-1抗体均为0．8％,未发现抗Scl-70抗体。ANA谱阳性率与 HCV感染者的性别和年龄无关（P＞0．05）。急性丙型肝炎患者、慢性丙型肝炎患者与肝硬化患者的 ANA 谱阳性率分别为47．1％、40．6％和57．1％,差异无统计学意义（P＝0．25）。结论 HCV感染者可检测到多种自身免疫性疾病的特异度抗体,其ANA谱阳性率与性别、年龄及 HCV感染疾病的严重程度均无关。     

The role of hepatitis C virus RNA detection in various substrates of patients with chronic hepatitis C

Puncture biopsy of the liver and blood count were made in 72 patients with chronic hepatitis C (CHC). Morphological alterations in the liver were assessed by Knodell index. The blood serum, lymphocytes and hepatic tissue were examined for a genome form of hepatitis C virus (HCV) RNA, blood lymphocytes and hepatic tissue--for a relevant replication form. HCV RNA was detected using "nested" RT-PCR. Only 26% patients had symptoms of asthenovegetative and dyspeptic syndromes. Normal alaninaminotransferase (ALT) level was observed in 24% patients, the rest had it high. HCV RNA was encounted more frequently in hepatic tissue than lymphocytes or serum (83, 68 and 46%, respectively). A replication form of HCV RNA was present in hepatic tissue of 31% patients and was absent in the lymphocytes. The incidence of the RNA detection was not related either to the disease symptoms or morphological alterations in hepatic tissue. The occurrence of the genome and replication forms in hepatic tissue does not correlate to ALT level. HCV RNA occurs more often in the serum, blood lymphocytes and in three substrates simultaneously in patients with hyperalatemia.     

Removal of hepatitis C virus by G-1 beads in sera from patients with chronic hepatitis C

Recently, a new method of extracorporeal granulocyte depletion apheresis has been developed to treat inflammatory systemic diseases using an Adacolumn (Japan Immunoresearch Laboratories, Takasaki, Japan) that is filled with acetate cellulose beads (G-1 beads) to adsorb the granulocytes. We examined whether hepatitis C virus (HCV) is adsorbed after incubation of the Adacolumn with the sera from patients with HCV-RNA-positive chronic hepatitis C. PATIENTS AND METHODS: A total of 10 patients with chronic hepatitis C, whose levels of HCV RNA were greater than 800 kIU/ml were examined. The serum was incubated with 500 G-1 beads in a syringe at 37 degrees C for 1 h. After removal of the serum, the beads were washed with RNase-free water. The G-1 beads were removed from the syringe after centrifugation. RNA was extracted from 200 microl of the wash waste and from 10, 50, 100 and 200 beads, respectively, using TRIZol regent. Detection of HCV RNA was performed using the nested PCR method. RESULTS: HCV RNA was detected from as few as 10 G-1 beads. HCV RNA was not detected from waste fluid collected after the last wash from any of the patients. Further, HCV RNA was detected in the initial waste fluid after the 37 degrees C incubation with serum in all of the patients. Since HCV RNA was detected on the G-1 beads, but not from the last washing solution in the current examination, these results suggest that the G-1 beads adsorbed HCV RNA. CONCLUSIONS: Our in vitro study confirmed that G-1 beads adsorbed HCV; therefore, apheresis using a column filled with G-1 beads may reduce the HCV RNA load in the blood of patients with chronic hepatitis C.     

戊型肝炎病毒IgM抗体阳性患者系列血清3种生物标志物结果分析

目的分析戊型肝炎病毒（HEV）IgM抗体阳性患者的入院时、住院中和出院时的系列血清中3种生物标志物的变化情况及其诊断意义。方法收集临床常规检测的HEVIgM抗体阳性患者的入院时、住院中和出院时的血清,用酶联免疫（EIA）试剂检测HEVIgG抗体和IgM抗体,用实时荧光PCR方法检测核酸,分析其变化情况。结果在25例HEVIgM抗体阳性患者系列血清中,HEVIgM抗体为4例高值无明显变化、15例高值明显下降变化、1例明显升高变化、5例低值无明显变化,HEVIgG抗体为3例高值无明显变化、15例高值明显升高变化、4例高值下降后维持高值、1例低值转为阴性、2例始终阴性,HEVRNA为入院第1天22例（88％）阳性、入院后1周15例（62．5％）阳性、入院后2周1例（5．6％）阳性。结论戊型肝炎患者发病过程中,系列血清HEV IgG抗体和IgM抗体及HEV RNA联合检测有助于戊型肝炎的明确诊断。     

At least three hepatitis C virus strains implicated in Swedish and Danish patients with intravenous immunoglobulin-associated hepatitis C

BACKGROUND: Three reported Swedish cases of hepatitis C in patients receiving an intravenous immunoglobulin (Gammagard, Baxter Healthcare, Deerfield, IL) were among the first to bring to light a worldwide outbreak of hepatitis C associated with non-solvent/detergent (SD)- treated Gammagard. In February 1994, all implicated batches of Gammagard were recalled and exposed patients traced. STUDY DESIGN AND METHODS: Sera from all identified and hepatitis C-viremic Swedish and Danish patients (n = 14) exposed to the implicated batches underwent hepatitis C virus genotyping and sequencing of the core region and hypervariable region 1 of E2. Genomic amplification was also done on 15 non-SD-treated batches of Gammagard. RESULTS: Twelve patients were infected with subtype 1a and surprisingly, two with subtype 2b. Analysis of the core region showed identical sequences in four patients and the only consistently positive batch. Five patients shared another sequence, whereas three other subtype 1a patients each manifested unique sequences. The two subtype 2b isolates were identical. Genomic fingerprinting of the hypervariable region confirmed identity within each group with great stringency. Amplification with isolate-specific primers showed mixed infection in one patient whose exposure was confined to a single batch. CONCLUSION: The few batches implicated presumably were contaminated with several strains.     

Oxidizability of apolipoprotein B-containing lipoproteins and serum paraoxonase/arylesterase activities in preeclampsia

OBJECTIVES: Lipoprotein oxidation has been implicated in the pathogenesis of preeclampsia and paraoxonase, an antioxidant enzyme shown to protect lipoproteins from being oxidized. The aim of the present study was to evaluate oxidizability of apolipoprotein B-containing lipoproteins and serum paraoxonase/arylesterase activities in preeclampsia. DESIGN AND METHODS: Twenty-one women with mild preeclampsia, 21 women with severe preeclampsia, and 20 women with normal uncomplicated pregnancy were included in this study. Serum malondialdehyde (MDA) level, an indicator of lipid peroxidation, was measured by high-performance liquid chromatography (HPLC). The oxidizability of apolipoprotein B-containing lipoproteins was evaluated by copper-induced in vitro peroxidation of the isolated fraction of apolipoprotein B-containing lipoproteins coupled with the thiobarbituric acid-reactive substances assay and expressed as the difference between copper-treated MDA and basal MDA (DeltaMDA). The serum paraoxonase and arylesterase activities were determined spectrophotometrically. RESULTS: Serum MDA and DeltaMDA levels of apolipoprotein B-containing lipoproteins were significantly higher in both mild and severe preeclampsia groups than in the normal pregnant group. Serum paraoxonase and arylesterase activities were not significantly different among the study groups. CONCLUSIONS: Enhancement in oxidizability of apolipoprotein B-containing lipoproteins accompanying with dyslipidemia and increased serum MDA levels suggests that those lipoproteins play a role in the pathogenesis of preeclampsia. Further studies are needed to investigate serum paraoxonase activity in women with normal pregnancies and preeclampsia.     

血液透析患者丙型肝炎病毒型特异性PCR基因分型

目的 建立一种丙型肝炎病毒型特异性PCR基因分型方法。方法 对47份血液透析患者HCV RNA阳性的标本,用5’非编码区(5'-NCR)1、2、3、1b型特异性引物,进行逆转录巢式PCR扩增分型。结果 47份HCV RNA样本43例可以分型;优势株为1b亚型,1b及1b相关型占总样本数的87．23％(41／47);7例为混合感染,占可分型数的16．27％(7／43)。结论 本组血液透析患者感染的HCV基因型的分布可能与地区流行及医源性传播有关,其同源性有待核苷酸序列分析加以印证。