OBJECTIVE: This study was designed to evaluate hematological disorders and the orchestrating roles of hepcidin and IL-6 in rat models of thioacetamide (TAA) and carbon tetrachloride (CCl4) hepatotoxicity. METHODS: Rats were intraperitoneally injected with TAA (10 mg/100 g rat weight dissolved in isosaline) or CCl4 (100 μL/100 g rat weight diluted as 1:4 in corn oil) twice weekly for eight consecutive weeks to induce subchronic liver fibrosis. Blood and tissue samples were collected and analyzed. RESULTS: CCl4 but not TAA significantly decreased the RBCs, Hb, PCV, and MCV values with minimal alterations in other erythrocytic indices. Both hepatotoxins showed leukocytosis, granulocytosis, and thrombocytopenia. By the end of the experiment, the erythropoietin level increased in the CCl4 model. The serum iron, UIBC, TIBC, transferrin saturation%, and serum transferrin concentration values significantly decreased, whereas that of ferritin increased in the CCl4 model. TAA increased the iron parameters toward iron overload. RT-PCR analysis revealed increased expression of hepatic hepcidin and IL-6 mRNAs in the CCl4 model and suppressed hepcidin expression without significant effect on IL-6 in the TAA model. CONCLUSION These data suggest differences driven by hepcidin and IL-6 expression between CCl4 and TAA liver fibrosis models and are of clinical importance for diagnosis and therapeutics of liver diseases.
Animals ; Blood Chemical Analysis ; Carbon Tetrachloride ; toxicity ; Hepcidins ; pharmacology ; Injections, Intraperitoneal ; Interleukin-6 ; pharmacology ; Iron ; blood ; metabolism ; Leukocytosis ; chemically induced ; therapy ; Liver Cirrhosis ; chemically induced ; therapy ; Male ; Rats ; Thioacetamide ; toxicity ; Thrombocytopenia ; chemically induced ; therapy ; Transferrin ; metabolism

BACKGROUND: Prealbumin, a sensitive marker for protein–energy status, is also known as an independent risk factor for mortality in hemodialysis patients. We investigated the impact of prealbumin on survival in incident peritoneal dialysis (PD) patients. METHODS: In total, 136 incident PD patients (mean age, 53.0 ± 15.8 years) between 2002 and 2007 were enrolled in the study. Laboratory data, dialysis adequacy, and nutritional parameters were assessed 3 months after PD initiation. Patients were classified into 2 groups according to prealbumin level: high prealbumin (≥ 40 mg/dL) and low prealbumin (< 40 mg/dL). RESULTS: The patients in the low-prealbumin group were older and had more comorbidities such as diabetes and cardiovascular diseases compared with the patients in the high-prealbumin group. Mean subjective global assessment scores were lower, and the high-sensitivity C-reactive protein levels were higher in the low-prealbumin group. Serum creatinine, albumin, and transferrin levels; percent lean body mass; and normalized protein catabolic rate were positively associated, whereas subjective global assessment scores and high-sensitivity C-reactive protein levels were negatively associated with prealbumin concentration. During the median follow-up of 49 months, patients in the lower prealbumin group had a higher mortality rate. Multivariate analysis revealed that prealbumin < 40 mg/dL (hazard ratio, 2.30; 95% confidence interval, 1.14–4.64) was an independent risk factor for mortality. In receiver operating characteristic curves, the area under the curve of prealbumin for mortality was the largest among the parameters. CONCLUSION: Prealbumin levels were an independent and sensitive predictor for mortality in incident PD patients, showing a good correlation with nutritional and inflammatory markers.
C-Reactive Protein ; Cardiovascular Diseases ; Comorbidity ; Creatinine ; Dialysis ; Follow-Up Studies ; Humans ; Inflammation ; Mortality* ; Multivariate Analysis ; Peritoneal Dialysis* ; Prealbumin* ; Renal Dialysis ; Risk Factors ; ROC Curve ; Transferrin

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

A 39-year-old woman with end-stage renal disease, which was maintained on haemodialysis, developed secondary haemochromatosis after receiving blood transfusions and intravenous iron supplementation without sufficient serum ferritin concentration monitoring. The patient received intravenous deferoxamine three times a week, combined with high-dose recombinant human erythropoietin therapy and haemodialysis. After three months, improvements in biochemical indicators and iron overload were noted.
Adult ; Chelating Agents ; chemistry ; Erythropoietin ; therapeutic use ; Female ; Ferritins ; blood ; Hemochromatosis ; complications ; Hemoglobins ; analysis ; Humans ; Kidney Failure, Chronic ; complications ; therapy ; Recombinant Proteins ; therapeutic use ; Renal Dialysis ; adverse effects ; Sequence Analysis, DNA ; Tomography, X-Ray Computed ; Transferrin ; chemistry ; Transfusion Reaction ; Treatment Outcome

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 compare the sensitivity, the specificity and the anti-jamming of several excrement occult blood experimental techniques. To evaluate the effect of transferrin (Tf) in the excrement in the digestive tract hemorrhage detection rate.

METHODSFor 600 patients of clinical suspicious digestive tract hemorrhage, take their excrement specimen, using the chemical process (pyramidon semi-quantitative examination law) to detect hemoglobin (Hb), and using monoclonal antibody colloidal gold method to detect Hb and Tf.

RESULTSFinally the hemoglobin chemical process (hereafter refers to as chemical process) to detect upper gastrointestinal hemorrhage with the positive rate 57.3%, and the detection of hemorrhage of lower digestive tract's positive rate is 44.8%; Hemoglobin monoclonal antibody colloidal gold method (hereafter refers to as colloid gold law) to examine upper gastrointestinal hemorrhage with a positive rate 60.4%, under examination hemorrhage with positive rate 77.6%; transferrin monoclonal antibody colloidal gold method (hereafter refer to as transferrin law) to examine upper gastrointestinal hemorrhage with a positive rate 82.3%, examination hemorrhage of lower digestive tract with a positive rate 66.4%; The union examination law (hemoglobin and transferrin to be detected twice, once positive that is positive) examines upper gastrointestinal hemorrhage the positive rate is 90.8%, hemorrhage of lower digestive tract's positive rate is 97.6%.

CONCLUSIONExcrement transferrin has the high detection rate in the upper gastrointestinal hemorrhage; Hb and the Tf combined examination may obviously raise the digestive tract hemorrhagic disease's positive detection rate.

Feces ; chemistry ; Gastrointestinal Hemorrhage ; diagnosis ; Gold Colloid ; Humans ; Occult Blood ; Transferrin ; analysis

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

BACKGROUND: Carbohydrate-deficient transferrin (CDT) levels have rarely been determined in an Asian population. We evaluated the analytical performance of a test for measuring CDT levels by using capillary electrophoresis (EP). METHODS: We determined the precision of CDT measurement by using capillary EP and nephelometry and compared the CDT values obtained using both the methods. We included healthy control subjects, abstinent patients with liver disease, and individuals consuming varying amounts of alcohol. RESULTS: The CDT measurement by using capillary EP were correlated well with those CDT measurement by using nephelometry, N Latex CDT assay, Y=0.5706X+1.581, R=0.930. The results obtained from both methods showed good qualitative agreement with each other (kappa coefficient=0.61). Genetic variants of transferrin isoforms were detected in 4.1% of the tested population. Both the CDT and gamma-glutamyl transpeptidase (GGT) levels in the abstinent patients with liver disease were significantly higher than those in healthy abstinent individuals (0.9% vs. 0.5%, 109.5 mg/dL vs. 28.5 mg/dL, respectively), but the difference in CDT values in the 2 groups was less pronounced for the CDT values. Individuals who had a mean daily alcohol intake of more than 60 g/day showed significantly higher CDT levels than those who had a mean daily alcohol intake of less than 60 g/day (1.9% vs. 0.7%, P=0.03). CONCLUSIONS: The CDT test using capillary EP showed good performance, and this method has several advantages such as automation and detection of variant forms. Thus, CDT can be a more useful marker than GGT for monitoring alcohol abstinence, especially in patients with liver disease.
Adolescent ; Adult ; Aged ; Automation ; Electrophoresis, Capillary/*methods ; Female ; Gene Frequency ; Humans ; Liver Diseases, Alcoholic/diagnosis ; Male ; Middle Aged ; Nephelometry and Turbidimetry/methods ; Protein Isoforms/analysis ; ROC Curve ; Republic of Korea ; Transferrin/*analogs & derivatives/analysis ; gamma-Glutamyltransferase/analysis

OBJECTIVETo study the value of iron metabolism indices, serum iron (SI), total iron blinding capacity (TIBC) and transferring (Tf), in thalassema.

METHODSThe serum samples from 9 children with silent alpha thalassema, 56 with standard alpha thalassema, 26 with HbH disease, 40 with beta+ thalassema, 56 with beta0 thalassema, 45 with iron deficiency anemia (IDA) and 70 healthy children were detected for SI, TIBC and Tf levels.

RESULTSThe SI level increased (p<0.01), while the TIBC level decreased significantly in the beta0 thalassema group compared with those in the other groups (p<0.05 or 0.01), but the Tf level was not different. The Tf level of both the silent alpha thalassema and the standard alpha thalassema groups was statistically lower than that of the healthy group (p<0.01), but the levels of SI and TIBC were similar to the healthy group. Though the SI level of the HbH disease group was similar to the healthy group, the TIBC and Tf levels were statistically lower (p<0.01).

CONCLUSIONSCompared with Tf, SI and TIBC are better indices for monitoring iron loading in children with thalassema. The increased SI level and decreased TIBC level are two indices for the diagnosis of beta(0) thalassema in children with cellule anaemia.

Adolescent ; Anemia, Iron-Deficiency ; metabolism ; Child ; Child, Preschool ; Female ; Genotype ; Humans ; Infant ; Iron ; metabolism ; Male ; Thalassemia ; genetics ; metabolism ; Transferrin ; analysis

The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide (NO) secretion of mouse macrophages stimulated by lipopolysaccharide (LPS) for 24 h was assayed by Griess reagent system. We found that RCE had potent anti-inflammatory effects on mice. RCE had little cell toxic effect on mouse lymphocytes and macrophages. RCE strongly inhibited the excessive production of inflammatory mediators (NO, CD69, CD25, CD71), in a dose-dependent manner, like cyclosporine A injection. RCE could inhibit proliferation of CD3+ T lymphocytes. These data suggested that RCE might exhibit anti-inflammatory effect by inhibiting the activation and proliferation of mouse lymphocytes and the NO secretion of mouse macrophages.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; CD3 Complex ; analysis ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Lectins, C-Type ; Lymphocyte Activation ; drug effects ; Macrophages ; cytology ; secretion ; Male ; Mice ; Mice, Inbred BALB C ; Nitric Oxide ; secretion ; Plants, Medicinal ; chemistry ; Receptors, Transferrin ; metabolism ; T-Lymphocytes ; cytology ; drug effects ; metabolism ; Trifolium ; chemistry