OBJECTIVETo establish and evaluate a protein microarray method for combined measurement of serum ferritin (SF) and soluble transferrin receptor (sTfR).

METHODSMicroarrayer was used to print both anti-SF antibodies I and anti-sTfR antibodies I on each protein microarray. Anti-SF antibodies II and anti-sTfR antibodies II were used as detection antibodies and goat antibodies coupled to Cy3 were used as antibodies III. The detection conditions of the quantitative analysis method for simultaneous measurement of SF and sTfR with protein microarray were optimized and evaluated. The protein microarray was compared with commercially available traditional tests with 26 serum samples.

RESULTSBy comparison experiment, mouse monoclonal antibodies were chosen as the probes and contact printing was chosen as the printing method. The concentrations of SF and sTfR probes were 0.5 mg/mL and 0.5 mg/mL respectively, while those of SF and sTfR detection antibodies were 5 μg/mL and 0.36 μg/mL respectively. Intra- and inter-assay variability was between 3.26% and 18.38% for all tests. The regression coefficients comparing protein microarray with traditional test assays were better than 0.81 for SF and sTfR.

CONCLUSIONThe present study has established a protein microarray method for combined measurement of SF and sTfR.

Animals ; Antibodies, Monoclonal ; analysis ; Ferritins ; blood ; Mice ; Protein Array Analysis ; methods ; Rabbits ; Receptors, Transferrin ; blood

OBJECTIVETo investigate the systemic changes of iron metabolism following manganese exposure.

METHODSNinety-seven welders and 91 workers with no history of exposure to manganese were recruited from the same factory in Beijing serving as the exposure group and the control group respectively. The welding rods used were type J422. The concentration of the manganese in the air of the work place was determined respectively with the national standard method. The serum iron and manganese, ferritin, transferrin and transferrin receptors were measured with the graphite furnace atomic absorption spectrophotometry and ELISA in both groups.

RESULTSThe permissible concentration-STEL of ambient Mn in welders' breathing zone ranged from 0.53 mg/m(3) to 2.19 mg/m(3), while the permissible concentration-TWA of ambient Mn was between 0.29 mg/m(3) and 0.92 mg/m(3) in the breathing zone of the workplace. Serum Mn and Fe concentrations in welders were about 1.40 times (P < 0.0l) and 1.2 times (P < 0.01), respectively, higher than those of control subjects. At the same time, the transferrin concentrations in serum were significantly higher (about 1.2 times, P < 0.05) in welders than in controls. In contrast, transferrin receptors were significantly lower (about 1.2 times) in exposed subjects than controls (P = 0.001). There was no difference in serum ferritin between the two groups (P = 0.112). Although there was no significant trend, the serum ferritin level was increased by 18% in comparison with that of the control. The abnormal percentage of serum Fe and Serum Mn in welders were 55.67% and 67.01% respectively, higher than those of control subjects. In addition, the correlations between all indicators and the duration of employment were not observed.

CONCLUSIONThe long term exposure to the manganese can induce the disorder of the iron metabolism, which is found in the expression of increase of the serum iron and transferrin as well as the decrease of transferrin receptors.

Female ; Ferritins ; blood ; Humans ; Iron ; metabolism ; Iron Metabolism Disorders ; chemically induced ; Male ; Manganese ; adverse effects ; Occupational Exposure ; adverse effects ; Receptors, Transferrin ; blood ; Transferrin ; analysis ; Welding

Extracellular Signal-Regulated MAP Kinases ; metabolism ; Glomerular Mesangium ; immunology ; Glycosylation ; Humans ; Immunoglobulin A ; metabolism ; Phosphorylation ; Receptors, Fc ; analysis ; genetics ; Receptors, Transferrin ; analysis ; genetics

OBJECTIVETo investigate the feasibility of (99m)Tc-transferrin ((99m)Tc-Tf) as an transferrin receptor imaging agent in nude mice bearing human hepatoma SMMC-7721 tumor.

METHODSBiodistribution of (99m)Tc-Tf in Balb/c mice was assayed, and transferrin receptor imaging in Balb/c nu/nu nude mice bearing human hepatocellular carcinoma SMMC-7721 xenografts was carried out, with or without inhibition of transferrin receptor with unlabeled transferrin. The receptor radioligand binding assay with (125)I-transferrin as ligand was established, and the number of receptors per cell and the K(d) were worked out.

RESULTS(99m)Tc-Tf was mainly distributed in the liver, blood and kidneys. At 18 h post injection, the ratios of tumor/blood, tumor/liver and tumor/muscle were 24.2, 1.7 and 35.5, respectively. The ratio of tumor uptake at 18 h without unlabeled transferrin pretreatment to that with pretreatment was 16.3. The number of receptors per cell was 2.4 x 10(5) and the K(d) was found to be 4.46 nmol/L.

CONCLUSIONAll those results indicate that (99m)Tc-Tf is specifically localized in tumor lesions of nude mice. (99m)Tc-Tf may be useful in detection of hepatocellular carcinoma.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Organotechnetium Compounds ; pharmacokinetics ; Random Allocation ; Receptors, Transferrin ; analysis ; Tissue Distribution ; Transferrin ; pharmacokinetics

BACKGROUND: Alcohol is known to affect two epigenetic phenomena, DNA methylation and DNA hydroxymethylation, and iron is a cofactor of ten-eleven translocation (TET) enzymes that catalyze the conversion from methylcytosine to hydroxymethylcytosine. In the present study we aimed to determine the effects of alcohol on DNA hydroxymethylation and further effects of iron on alcohol associated epigenetic changes. METHODS: Twenty-four male Sprague-Dawley rats were fed either Lieber-DeCarli alcohol diet (36% calories from ethanol) or Lieber-DeCarli control diet along with or without iron supplementation (0.6% carbonyl iron) for 8 weeks. Hepatic non-heme iron concentrations were measured by colorimetric assays. Protein levels of hepatic ferritin and transferrin receptor were determined by Western blotting. Methylcytosine, hydroxymethylcytosine and unmodified cytosine in DNA were simultaneously measured by liquid chromatography/mass spectrometry method. RESULTS: Iron supplementation significantly increased hepatic non-heme iron contents (P < 0.05) but alcohol alone did not. However, both alcohol and iron significantly increased hepatic ferritin levels and decreased hepatic transferrin receptor levels (P < 0.05). Alcohol reduced hepatic DNA hydroxymethylation (0.21% ± 0.04% vs. 0.33% ± 0.04%, P = 0.01) compared to control, while iron supplementation to alcohol diet did not change DNA hydroxymethylation. There was no significant difference in methylcytosine levels, while unmodified cytosine levels were significantly increased in alcohol-fed groups compared to control (95.61% ± 0.08% vs. 95.26% ± 0.12%, P = 0.03), suggesting that alcohol further increases the conversion from hydroxymethylcytosine to unmodified cytosine. CONCLUSIONS: Chronic alcohol consumption alters global DNA hydroxymethylation in the liver but iron supplementation reverses the epigenetic effect of alcohol.
Alcohol Drinking* ; Alcohols ; Animals ; Blotting, Western ; Cytosine ; Diet ; DNA Methylation ; DNA* ; Epigenomics ; Ferritins ; Humans ; Iron* ; Liver ; Male ; Methods ; Rats* ; Rats, Sprague-Dawley ; Receptors, Transferrin ; Spectrum Analysis

OBJECTIVETo study trend of dynamic change in level of serum transferrin receptor (sTfR) in the process of iron supplementation to provide evidence for sTfR in evaluating the efficacy of iron supplementation.

METHODSTotally, 942 child-bearing-age women aged 18 to 45 years were selected from Longfang City, Hebei Province and Shunyi County, Beijing. Biochemical indicators of iron metabolism were measured for all of them, including serum levels of ferritin (SF) and zinc protoporphyrin (ZPP), and hemoglobin (Hb). According to the current criteria for assessing iron status, women were screened for iron deficiency erythropoiesis (IDE) or iron deficiency anemia (IDA). Seventy-two women agreed to participate in the study, and 59 of them finished whole dynamic observations with signed informed consent. Four capsules of ferrous L-threonate (containing 7 mg of iron element per capsule) were administered for women with IDE every other day and for women with IDA every day, respectively, for 12 weeks. Serum biochemical indicators and level of sTfR were measured in 0 wk, 3 wk, 6 wk, 9 wk and 12 wk, respectively, during the process of iron supplementation, and their dynamic changes were observed.

RESULTSLevel of sTfR in women with IDE and IDA was (26.62 +/- 10.57) nmol/L and (41.25 +/- 21.96) nmol/L, respectively, significantly higher than normal level. During the process of iron supplementation, level of sTfR changed as the following characteristics. In women with IDE, level of sTfR kept stable within the first 3 weeks of iron supplementation, then dropped gradually and progressively, reached to normal, with (17.86 +/- 5.57) nmol/L, in the 12 wk after iron supplementation. In women with IDA, level of sTfR dropped quickly within the first 3 wk of iron supplementation, then dropped slowly until the 9th wk and kept stable, and reached to normal level in the 12 wk, with (19.54 +/- 5.94) nmol/L and a ratio of sTfR/SF of 12.23 +/- 4.34. Ratio of sTfR/SF changed as level of sTfR during the process of iron supplementation. Level of sTfR correlated reversely with levels of Hb and SF and positively with level of ZPP.

CONCLUSIONSerum level of sTfR in child-bearing age women gradually decreased to normal with the restoration of their normal iron status during the process of iron supplementation and could be used as a specific indicator for assessing efficacy of iron supplementation.

Adolescent ; Adult ; Anemia, Iron-Deficiency ; blood ; drug therapy ; Biomarkers ; blood ; Dietary Supplements ; Female ; Hemoglobins ; analysis ; Humans ; Iron ; administration & dosage ; therapeutic use ; Middle Aged ; Protoporphyrins ; blood ; Receptors, Transferrin ; blood

OBJECTIVETo study the clinical utility of measuring reticulocyte hemoglobin content (CHr) in the diagnosis of iron deficiency anemia (IDA) in children.

METHODSOne hundred children with IDA at ages of 1 to 6 years and 50 healthy children were enrolled. Red blood cell parameters, CHr, hemoglobin (Hb), red blood count (RBC) and mean corpusular volume (MCV), were determined using the Blood Cell Analyzer. Serum ferritin (SF) levels were determined using radioimmunoassay double antibody techique. Soluble serum transferrin (sTfR) levels were determined using ELISA.

RESULTSThe values of Hb (100 ± 6 g/L vs 126 ± 8 g/L) and CHr (18 ± 5 pg vs 31 ± 3 pg) in the IDA group were significantly lower than normal controls (P<0.01). SF levels (11 ± 4 μg/L) in the IDA group were also lower than normal controls (59 ± 36 μg/L) (P<0.01). In contrast, the values of sTfR in the IDA group were significantly higher than normal controls (4.8 ± 2.1 mg/L vs 1.4 ± 0.6 mg/L; P<0.01). In both groups, there was a positive correlation between the values of CHr and Hb [r=0.540 (control group), r=0.734 (IDA group); P<0.01]. In the IDA group, CHr was positively correlated with SF(r=0.464; P<0.01) and negatively correlated with sTfR(r=-0.450; P<0.01). When the cut-off value of CHr was 27.8 pg, the sensitivity and specificity for the diagnosis of IDA were 88.0% and 90.0%, respectively and the area under the ROC curve was 0.948.

CONCLUSIONSCHr can be used as an index for the diagnosis of IDA in children.

Anemia, Iron-Deficiency ; blood ; diagnosis ; Child ; Child, Preschool ; Female ; Hemoglobins ; analysis ; Humans ; Infant ; Male ; ROC Curve ; Receptors, Transferrin ; blood ; Reticulocytes ; chemistry

The phenotypes of the bone marrow cells in various subtypes of myelodysplastic syndromes (MDS) and its clinical implication were explored. The antigen expression of a panel of antigens expressed in marrow cells from 30 patients with subtypes of MDS was assayed by alkaline phosphatase anti-alkaline phosphatase method. The results showed that the expression of myeloid antigens appeared abnormality, CD13 and CD33, found on granulocyte and macrophage precursors, increased, and CD15 decreased. There were no significant changes for monocytic antigen CD14 and lymphoid antigens CD7 and CD10. CD34 was increased in RAEB/RAEB-t and was not increased in RA/RAS patients. CD71, expressed by erythroblast and proliferative cells, was higher in all subtypes of MDS than that in control group. It is suggested that the bone marrow cells from MDS patients showed abnormality of more than two series of immunophenotypes, detection of immunophenotype in MDS cells might be contributed to the diagnosis and predicting prognosis.
Adult ; Aged ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Antigens, CD7 ; analysis ; Antigens, Differentiation, B-Lymphocyte ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Bone Marrow Cells ; immunology ; CD13 Antigens ; analysis ; Female ; Humans ; Immunophenotyping ; Lewis X Antigen ; analysis ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology ; Neprilysin ; analysis ; Receptors, Transferrin ; Sialic Acid Binding Ig-like Lectin 3

OBJECTIVES: Immunologic studies have characterized the numbers and types of inflammatory cells in diseased inflammatory bowel disease (IBD) mucosa but have yielded conflicting results regarding intestinal lymphocytes activation in IBD. We investigated the levels of lymphocytes subsets, interleukin-2 receptor, transferrin receptor, and T cell receptors in mainly isolated lamina propria lymphocytes. Including intraepithelial lymphocytes of normal colonic mucosa or IBD (ulcerative colitis and Crohn's disease) mucosa to understand the pathogenesis of IBD. We have results from this study. RESULTS: 1) In comparing ulcerative colitis with control, IL-2R (p < 0.05), TR (p < 0.01), and CD3/HLA-DR (<0.05) showed a significant increase. 2) In comparing Crohn's disease with control, CD3 (P < 0.05), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. 3) In comparing Crohn's disease with ulcerative colitis, CD19 (p < 0.01), TR (p < 0.01), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. CONCLUSION: From these results, there are increased T cell markers, IL-2R, TR, and CD3/HLA-DR in UC, but differently, decreased CD3, TCR alpha/beta and TCR gamma/delta in CD compared with control. In addition, definitive differences in lymphocytes markers, CD19, TR, TCR alpha/beta and TCR gamma/delta, which are higher in UC than in CD, may elucidate the different immunopathogenesis between UC and CD.
Adult ; Antigens, CD3/analysis ; Colitis, Ulcerative/pathology ; Colitis, Ulcerative/immunology ; Colitis, Ulcerative/diagnosis* ; Comparative Study ; Crohn Disease/pathology ; Crohn Disease/immunology ; Crohn Disease/diagnosis* ; Diagnosis, Differential ; Female ; HLA-DR Antigens/analysis ; Human ; Immunophenotyping* ; Intestinal Mucosa/pathology ; Intestinal Mucosa/immunology* ; Male ; Middle Age ; Receptors, Antigen, T-Cell/analysis ; Receptors, Interleukin-2/analysis ; Receptors, Transferrin/analysis ; Sensitivity and Specificity ; Tissue Culture

Transferrin receptor (TR) performs the major function of binding and internalizing its specific iron-loaded ligand, transferrin, and its expression is closely linked to the proliferation status of the cell. This study was undertaken to elucidate TR expression in the hyperplastic lesion of hepatocyte in chemically induced hepatic carcinogenesis. The resistant hepatocyte model was chosen for a rat model of carcinogenesis and Sprague-Dawley rats were divided into the following groups: the control groups of normal diet and iron-rich diet with or without hydroxyquinoline and the groups of carcinogen alone and carcinogen plus iron-rich diet with or without administration of hydroxyquinoline. Microscopic changes in the liver, expression of transferrin receptor and glucose-6-phosphatase were studied. The hepatocyte of the control group showed both cytoplasmic and membranous expression of TR. The liver of rats fed on high iron diet accumulated iron and the expression of TR was down regulated by intrahepatic iron accumulation. In the carcinogen administered group the resistant hepatocyte of hyperplastic lesion revealed strong membranous expression of TR and failed to accumulate iron in spite of high iron diet but in contrast the surrounding non-resistant hepatocyte expressed TR in both the membrane and cytoplasm and stored iron when fed on high iron diet. The strong membranous expression of TR is one of the characteristics of the resistant hepatocyte of hyperplastic lesion and it seems to be related to the inability to accumulate iron in spite of a high iron diet.
Animal ; Glucose-6-Phosphatase/metabolism ; Immunohistochemistry ; Iron/analysis/pharmacology ; Liver/chemistry/enzymology/pathology ; Liver Neoplasms, Experimental/enzymology/*ultrastructure ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Transferrin/*biosynthesis ; Support, Non-U.S. Gov't