OBJECTIVETo explore the role of interferon (IFN)-alpha/beta receptor beta subunit (IFNAR2) in the patients' response to IFN-alpha therapy as influenced by the grade of chronic hepatic inflammation, and understand the relation of IFNAR2 expression in the peripheral blood mononuclear cells (PBMCs) with HBV infection.

METHODSLiver tissue specimens were obtained from 21 patients with chronic hepatitis B for examination of the hepatic inflammation, and PBMCs were isolated from another 16 patients with chronic hepatitis B and 15 health control subjects. Both the hepatic tissues and PBMCs were examined for IFNAR2 expression using immunohistochemistry.

RESULTSThe 21 patients with chronic hepatitis B were divided into 3 groups according to the severity of hepatic inflammation, namely G(1) (n=3), G(2) (n=7) and G(3) (n=11) groups. The patients in G(3) group showed had significantly higher IFNAR2 expressions in liver (25.1307-/+7.0700) than those of the G(1) (5.6913-/+1.8422) and G(2) (7.4706-/+5.3572) groups (P=0.000). The IFNAR2 levels in the PBMCs, however, did not show significant difference between patients with chronic hepatitis B and the healthy control subjects.

CONCLUSIONIn patients with chronic hepatitis B, IFNAR2 expression level is positively correlated to the severity of hepatic inflammation, and increased IFNAR2 expression in severe hepatic inflammation is therefore likely to result in increased response rate to INF-alpha therapy. The expression of IFNAR2 in the PBMCs is not associated with HBV infection.

Female ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Leukocytes, Mononuclear ; metabolism ; Liver ; metabolism ; pathology ; Male ; Receptor, Interferon alpha-beta ; blood ; metabolism

OBJECTIVETo explore the expression of Toll-like receptor 4 (TLR4) and tumor necrosis factor-α (TNF-α) on peripheral-blood mononuclear cells (PBMCs) and their correlation with myocardial perfusion in patients with diabetic cardiomyopathy (DCM).

METHODSThe expression of TLR4 and TNF-α mRNA on PBMCs were examined by SYBR Green I real-time quantitative reverse transcription polymerase chain reaction (RT-PCR), the levels of TLR4 and TNF-α were examined by flow cytometric analysis and enzyme-linked immuno sorbent assay (ELISA) on DCM group (n = 20), Type 2 diabetic group (n = 22) and control group (n = 20). Myocardial perfusion was visualized by single-photon emission computed tomography (SPECT).

RESULTSThe expressions of TLR4 and TNF-α mRNA/protein on PBMCs in DCM group were significantly higher than in Type 2 diabetic group, and higher in Type 2 diabetic group than in control groups (P < 0.05); summed stress score (SSS) and summed rest score (SRS) of myocardial perfusion in DCM group were significantly higher than in Type 2 diabetic group, and higher in Type 2 diabetic group than in control groups (P < 0.01). The expression of TLR4, TNF-α was positively correlated with SSS (r = 0.75, P < 0.05; r = 0.931, P < 0.005) and SRS (r = 0.78, P < 0.005; r = 0.789, P < 0.005). SSS and SRS in DCM group were also positively correlated with soluble vascular cell adhesion molecule-1 (sVCAM-1) (r = 0.728, P < 0.005; r = 0.738, P < 0.005) but there was no correlation between SSS and SRS and brain natriuretic peptide, LVEF, E/A, HbA1c, FBG, FIN and LDL-C (P > 0.05).

CONCLUSIONThe increased expression of TLR4 and TNF-α mRNA/protein on PBMCs and increased serum sVCAM-1 is linked with reduced myocardial perfusion in DCM group. TLR4 and TNF-α may thus play a critical role in the myocardial perfusion inflammation injury in these patients.

Case-Control Studies ; Diabetic Cardiomyopathies ; blood ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Myocardium ; metabolism ; pathology ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVEKawasaki disease (KD) is a febrile illness of childhood. The etiology of KD remains unknown. Multiple theories exist, including an infectious etiology and an immunological abnormality. Cardiac involvement ranges from myocarditis and pericarditis in the acute stage to the development of coronary artery aneurysms later in the course. The present study aimed to explore the effect of monocyte chemotactic protein-1 (MCP-1) in Kawasaki disease and its relationship with damage to the coronary arteries during the development of KD.

METHODSPlasma MCP-1 concentrations were measured by enzyme-linked immunosorbent assay (ELISA), and MCP-1 mRNA expression in peripheral blood mononuclear cells (PBMC) was measured by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) in comparison of three groups: 56 patients with KD, 60 age-matched patients with non-infectious diseases, and 66 age-matched febrile patients with various diseases.

RESULTSPlasma MCP-1 concentration and MCP-1 mRNA expression in PBMC of patients with active KD [(409.55 +/- 97.42) pg/ml] and (1.97 +/- 0.77) were higher than those of control group. Plasma MCP-1 levels and MCP-1 mRNA expression of inactive KD group [(301.64 +/- 71.55) pg/ml] and (1.31 +/- 0.39) were significantly higher than those of non-infectious diseases patients. There was a marked increase in patients with inactive KD than those of non-infective patients, but there were no significant differences between inactive KD and febrile patients. Plasma MCP-1 levels and MCP-1 mRNA expression were markedly increased in KD patients with coronary artery lesions than those in patients without coronary artery lesions.

CONCLUSIONPlasma MCP-1 concentration and MCP-1 mRNA expression in PBMC were significantly increased in patients with KD, and they were higher in KD with coronary artery lesions. It indicates that MCP-1 may be a useful parameter for monitoring disease activity in patients with KD.

Chemokine CCL2 ; blood ; genetics ; metabolism ; Child ; Child, Preschool ; Coronary Vessels ; pathology ; Female ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; Male ; Mucocutaneous Lymph Node Syndrome ; blood ; genetics ; pathology ; RNA, Messenger ; genetics

AIMTo explore the expression of heme oxygenase-1 (HO-1) in the peripheral blood mononuclear cell (PBMC) and relationship to ventilatory function in asthmatic patients.

METHODSEighteen asthmatic patients and eighteen healthy subjects were selected. HO-1 protein levels in PBMC were measured by immunohistochemical staining and PBMC HO-1 mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR), blood carbon monoxide Hb (COHb) percent value, serum total IgE concentration and pulmonary ventilatory function were observed in asthmatic patients and healthy subjects.

RESULTSThe percentage of cells in immunohistochemical staining positive staining of HO-1 were significantly higher in asthmatic patients (41.7 +/- 7.44%) compared with that of healthy subjects (10.5 +/- 4.36%, P < 0.01), the optical densities of PBMC HO-1 mRNA were higher in asthmatic patients (26.05 +/- 4.14) compared with that of healthy subjects (10.82 +/- 4.26, P < 0.01). The relation analysis showed PBMC HO-1 protein levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.56, -0.51, P < 0.01, respectively) and positive relation with COHb percent value, serum total I gE concentration (r = 0.80, 0.48, P < 0.05, respectively), and PBMC HO-1 mRNA levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.65, -0.67, P < 0.05, respectively) and positive relation with COHb percent value, serum total IgE concentration (r = 0.85, 0.62, P < 0.01, respectively).

CONCLUSIONThe expression of PBMC HO-1 protein and mRNA are increased significantly in asthmatic patients, HO-1 may play a significant role in the pathogenesis of asthma. The expression of HO-1 has relation with severity of asthma.

Adult ; Asthma ; blood ; pathology ; Case-Control Studies ; Female ; Heme Oxygenase-1 ; blood ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged

OBJECTIVETo investigate the expression and clinical significance of miRNA-155 and miRNA-146a in peripheral blood mononuclear cells (PBMC) and plasma of oral lichen planus (OLP) patients.

METHODSTwenty-five female and seven male OLP patients (OLP group) aged 25 to 54 years were selected from January 2012 to May 2013. The diagnosis was confirmed by pathology and the lesions were divided into two non-erosive OLP group (18 cases) and erosive OLP group (14 cases). Twenty healthy sex and age matched volunteers served as control. miRNA-155 and miRNA-146a expressions in PBMC and plasma were examined by real-time PCR. The difference between OLP group and control group was statistically analyzed.

RESULTSThe expressions of PBMC and plasma miRNA-155 were higher in OLP patients than those in the healthy control (median, 0.07 vs 0.03, P < 0.05; 5.84 vs 1.32, P < 0.01). The median expression level of miRNA-146a in PBMC and plasma of OLP patients and healthy controls were (1.26 vs 0.58, P < 0.05) and (412.60 vs 238.42, P < 0.01). The plasma miRNA-155 and miRNA-146a expressions were significantly higher in erosive OLP group than those in non-erosive OLP group. There were no significant differences in the expression of PBMC miRNA-155 and miRNA-146a between the two groups.

CONCLUSIONSThe expressions of PBMC and plasma miRNA-155 and miRNA-146a are higher in OLP patients. The expressions of plasma miRNA-155 and miRNA-146a are associated with OLP severity. The over expression of miRNA-155 and miRNA-146a in OLP may play a role in the pathogenesis of OLP.

Adult ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Lichen Planus, Oral ; metabolism ; pathology ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Real-Time Polymerase Chain Reaction

This study was aimed to analyze the different proteomes between human acute leukemia (AL) cells and normal white blood cells by proteomic technology in order to lay the basis for diagnosing AL and understanding the mechanism of leukemogenesis. The proteins from AL cells of 40 AL patients identified by FAB classification and proteins from normal lymphocytes and granulocytes of 20 normal volunteers were separated by two-dimensional electrophoresis (2-DE), and the differentially expressed proteins between the two groups were identified by both matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and electronspray ionization (ESI)-MS/MS. The results showed that among the differentially expressed proteins between AL cells and normal lymphocytes and granulocytes, some proteins involved in the process of malignant transformation (such as Op18, NM23-H1), cell proliferation (such as PCNA) and apoptosis inhibition (such as tumor necrosis factor inhibitor protein) were found to be up regulated in AL cells. However, some proteins involved in differentiation and physiological functions of normal cells were down regulated in AL cells. It is concluded that there are many events involved in the process of leukemogenesis, expression of some proteins relating to the malignant transformation, cell proliferation and apoptosis inhibition are up-regulated in AL cells. The proteome analysis may provide a new approach to explaining the molecular mechanism underlying the pathogenesis of AL.
Adolescent ; Adult ; Bone Marrow Cells ; chemistry ; pathology ; Female ; Humans ; Leukocytes ; chemistry ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Proteome ; analysis

OBJECTIVETo investigate peripheral blood monocyte (PBMC) gene expression profile in patients with fulminant hepatic failure (FHF) by cDNA microarray.

METHODSMicroarrays consisting of 8,192 human cDNAs and labelled cDNAs prepared from PBMC in both 10 FHF patients and 10 asymtomatic surface antigen carriers (ASC) were applied to analyze gene expression. Relative ratios of gene expression in individuals were obtained by comparing the hybridization results, by GenePix 4000B scanning and by ImaGene3.0 software analysis, of Cy5-labelled cDNA from FHF patients with those of Cy3-labelled cDNA from ASC.

RESULTS249 genes out of 8,192 were identified differently, at least two times. Most of the genes (79%) involved in cell signaling transduction, cell cycles, metabolism, inflammatory response and apoptosis, whose mRNAs were differently regulated.

CONCLUSIONSThese results suggest that HBV infection alters a broad range of cellular genes expression during developing into FHF and provide a framework for future functional study on the genes expressed differently.

DNA, Complementary ; genetics ; Female ; Gene Expression ; Gene Expression Profiling ; Hepatitis B ; genetics ; pathology ; Hepatitis B, Chronic ; genetics ; pathology ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo study the expression and localization of co-stimulators in the mucosa of patients with ulcerative colitis (UC), and to explore its role in the pathogenesis of UC.

METHODSExpression of co-stimulators CD86 and inducible co-stimulator (ICOS) was studied by immunohistochemistry on paraffin-embedded mucosal tissue from patients with active UC (64 cases), inactive UC (51 cases) and normal controls (20 cases). Immunostaining for CD28 was also carried out on frozen fresh mucosal tissue sampled from patients with active UC (7 cases), inactive UC (2 cases) and normal controls (5 cases). In addition, expression of CD4, CD8 and CD20 were also examined.

RESULTSIn active UC, increased expression of CD86 was not only observed in lamina propria mononuclear cells but also in the intestinal epithelial cells, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in lamina propria mononuclear cells was detected in active UC, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in intestinal epithelial cells was also seen in active UC, as compared with that of inactive UC (P < 0.01). The expression of CD86 was higher in inactive UC than in the normal controls (P < 0.05 or P < 0.01). However, the expression of ICOS showed no statistically significant difference between inactive UC and normal controls. Increased expression of CD28 in active UC, compared with that in inactive UC and normal controls, was also noticed (P < 0.05 or P < 0.01). The number of CD4 or CD8-positive intraepithelial lymphocytes and lymphocytes infiltrating in the lamina propria and small vessel walls was much higher in active UC than in inactive UC and normal controls (P < 0.01). Moreover, the ratio of CD4/CD8 was highest in active UC (P < 0.01). The number of CD20-positive B lymphocytes in lamina propria was also higher in active UC than in inactive UC and normal controls (P < 0.01).

CONCLUSIONSIn active UC, CD86 and ICOS were over-expressed in the intestinal epithelial cells and lamina propria mononuclear cells. The phenomenon suggests that abnormal expression of co-stimulators may contribute to the deregulation of acquired immune responses in UC.

Adult ; Aged ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; B7-2 Antigen ; metabolism ; CD28 Antigens ; metabolism ; CD4-CD8 Ratio ; Case-Control Studies ; Colitis, Ulcerative ; metabolism ; pathology ; Epithelial Cells ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Inducible T-Cell Co-Stimulator Protein ; Intestinal Mucosa ; metabolism ; pathology ; Leukocytes, Mononuclear ; metabolism ; pathology ; Male ; Middle Aged ; Mucous Membrane ; metabolism ; pathology ; Young Adult

OBJECTIVEViral hepatitis remains a major public health problem and the most common type of liver disease worldwide. There are an increasing number of patients with chronic hepatitis B who develop acute hepatitis on chronic condition (AOC) and die of acute hepatic failure both as a result of lack of understanding of the pathogenesis of the disease and lack of effective treatment. The hallmark of AOC is the extreme rapidity of the necromicroinflammatory process resulting in widespread or total hepatocellular necrosis in weeks or even days. Our previous studies have shown in an experimental animal model of fulminant viral hepatitis caused by murine hepatitis virus strain 3, the importance of macrophage activation, and expression of a unique gene mfgl 2 which encodes a serine protease capable of directly cleaving prothrombin to thrombin, resulting in widespread fibrin deposition within the liver and hepatocyte necrosis. The undergoing study in this report is designed to identify the role of hfgl 2 (human fibrinogen like protein 2) /fibroleukin in patients with viral hepatitis.

METHODSLiver tissues were obtained from 23 patients with AOC hepatitis B, and from 13 patients with inactive chronic hepatitis B (CHB) and 14 patients with chronic hepatitis B with cirrhosis during the year of 1995 to the end of 2001. Liver biopsies were performed within 30 min after the patients were diagnosed with death as a result of acute hepatic failure. Liver samples were also obtained from 4 liver donors as normal controls. In addition, peripheral blood mononuclear cells (PBMC) were isolated from 30 patients (unpaired) with AOC hepatitis B and 10 patients with CHB during the May of 2001 to March of 2002 and 10 healthy volunteer as negative control. PBMCs were freshly isolated and smeared on slides and kept at -80 degree C for further use. Histological sections were stained with hemotoxylin and eosin. A 169 bp of hfgl 2 cDNA probe and a polyclonal or monoclonal antibody against hfgl 2 were used to detect the expression of hfgl 2 mRNA and protein in liver samples as well as PBMC by immune histochemistry separately.

RESULTSLiver tissues from the patients with acute on chronic hepatitis had classical pathological features of acute necroinflammation. Hfgl 2 was detected by immune histochemistry in 21 of 23 patients (91.3%) in liver sections from patients with acute on CHB, while only 1 of 13 patients (7.7%) with CHB and cirrhosis and no evidence of active disease had hfgl 2 mRNA or protein expression. 28 of 30 patients (93.3%) with acute on CHB and 1 of 10 with CHB were detected with hfgl 2 expression in PBMC. There was no hfgl 2 expression in either the liver tissue or the PBMC from the normal donors. There was positive correlation of hfgl 2 expression and the severity of the disease displayed by the value of bilirubin and PT.

CONCLUSIONThe molecular and cellular results reported here in patients with acute on chronic hepatitis and who died of acute hepatic failure correlates with previous report in 8 patients with fulminant hepatic failure (FHF) and mimic closely the changes observed in the murine model of fulminant viral hepatitis in which the pathogenesis of the disease has been studied in a stepwise fashion. This study further suggests that virus induced hfgl 2 prothrombinase/fibroleukin expression and the potent function of the protein it encodes plays a pivotal role in initiating acute severe hepatitis on the baseline of chronic hepatitis. The measurement of hfgl 2/fibroleukin expression in PBMC may serve as a useful marker to monitor the severity of disease in patients with the AOC hepatitis B and a target for therapeutic intervention.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Fibrinogen ; biosynthesis ; genetics ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; Liver ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Severity of Illness Index ; T-Lymphocytes ; metabolism

OBJECTIVETo study the expression level and intracellular localization of APOBEC3G in peripheral blood mononuclear cells (PBMCs) and liver tissues of chronic HBV patients.

METHODSThe expression level and intracellular localization of APOBEC3G in PBMCs and liver tissues were detected using the western blot and confocal laser scanning microscope (CLSM).

RESULTSWestern-blot showed that the expression level of APOBEC3G in PBMCs of healthy controls was very low. The relative expression levels of APOBEC3G in PBMC of patients with chronic hepatitis B, chronic severe hepatitis, liver cirrhosis, or liver cancer were 4.12+/-0.21, 4.07+/-0.28, 4.16+/-0.36 or 4.21+/-0.39 respectively, which were higher than that in the healthy controls. However, there was no significant difference in APOBEC3G expression among different chronic HBV patients (q = 0.931, 0.744, 1.675, 1.675, 2.606 or 0.931, respectively, all P values more than 0.05). In addition, there was no significant difference on APOBEC3G in liver tissues between chronic hepatitis B patients and hepatocellular carcinoma patients (4.40+/-0.34 vs 4.34+/-0.43, q = 0.588, P more than 0.05). CLSM indicated that the localization of APOBEC3G protein was in cytoplasm of PBMCs and hepatocytes.

CONCLUSIONAPOBEC3G is upregulated in the PBMCs of chronic hepatitis B patients.

APOBEC-3G Deaminase ; Blotting, Western ; Case-Control Studies ; Cytidine Deaminase ; genetics ; metabolism ; Cytoplasm ; metabolism ; Hepatitis B, Chronic ; metabolism ; pathology ; virology ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; metabolism ; pathology ; virology ; Liver Neoplasms ; metabolism ; pathology ; virology ; Microscopy, Confocal ; methods ; RNA, Messenger ; genetics ; metabolism