1.Study on the method of quantitative analysis of serum ferritin and soluble transferrin receptor with protein microarray technology.
Ji Yong YIN ; Jing SUN ; Jian HUANG ; Wen Xian LI ; Jun Sheng HUO
Biomedical and Environmental Sciences 2012;25(4):430-439
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
2.Concentrations of serum iron and transferrin in children with nephrotic syndrome.
Hong-Zhu LU ; Yue-Sha YUAN ; Wan-Ming ZHANG ; Dan LIU ; Hong-Yan KUANG
Chinese Journal of Contemporary Pediatrics 2006;8(6):467-469
OBJECTIVENephrotic syndrome (NS) is characterized by marked urinary excretion of albumin and other intermediated-size plasma proteins such as transferrin. The aim of this study was to determine the changes of serum iron and transferrin and the relationship between the serum and urinary transferrin.
METHODSThe indexes related to iron metabolism, including serum iron, ferritin, transferrin, total iron-binding capacity, transferrin saturation and hematological parameters (Hb, MCV, MCH), and urinary transferrin were measured in 37 children with NS before treatment and at the remission stage. Thirty-five age-matched healthy children served as controls.
RESULTSSerum iron levels (18.8 +/- 3.8 micromol/L) in NS patients before treatment were significantly lower than in the healthy controls (22.2 +/-3.8 micromol/L) and those measured at the remission stage (21.0 +/- 3.5 micromol/L) (P < 0.01). Serum transferrin levels in NS patients before therapy (1.9 +/- 0.3 g/L) also decreased compared with those in the healthy controls (3.1 +/- 0.5 g/L) and those measured at the remission stage (2.9 +/- 0.6 g/L) (P < 0.01). In contrast, serum total iron-binding capacity and transferrin saturation were noticeably higher in NS patients before treatment than those in the healthy controls (total iron-binding capacity 56.4 +/- 9.2 micromol/L vs 50.7 +/- 6.8 micromol, P < 0.01; transferrin saturation 55.7 +/- 9.2 % vs 46.4 +/- 8.2%, P < 0.01) and were also higher than those measured at the remission stage (51.9 +/-7.7 micromol/L and 47.4 +/- 13.3%) (P < 0.01). Serum transferrin positively correlated to serum albumin (r = 0.609, P < 0.01) and negatively correlated to urinary transferrin (r = -0.550, P < 0.01) in NS patients before treatment.
CONCLUSIONSSerum iron and transferrin levels markedly decreased in NS patients, which may be partially related to the urinary loss of transferrin.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Iron ; blood ; Male ; Nephrotic Syndrome ; blood ; Transferrin ; analysis ; urine
3.The effect of transferrin detecting in digestive tract hemorrhage detection rate.
Chinese Journal of Medical Instrumentation 2011;35(6):462-464
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
4.Altered systemic iron metabolism in welders exposed to manganese.
Ling LU ; Long-lian ZHANG ; Guo-jun LI ; Wen-rui GUO ; Wan-nian LIANG ; Zheng WEI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(1):31-34
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
5.Changes of iron metabolism indices in children with various genotypes of thalassema.
Yu-Jun HUANG ; Shao-Guo WU ; Xiao-Bing OU ; Li ZHANG
Chinese Journal of Contemporary Pediatrics 2010;12(2):85-88
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
6.Experimental study of transferrin receptor molecular imaging in human hepatocellular carcinoma transplanted in nude mice.
Chinese Journal of Oncology 2006;28(2):96-98
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
8.Iron Supplementation Reverses the Reduction of Hydroxymethylcytosine in Hepatic DNA Associated With Chronic Alcohol Consumption in Rats.
Stephanie A TAMMEN ; Jung Eun PARK ; Phil Kyung SHIN ; Simonetta FRISO ; Jayong CHUNG ; Sang Woon CHOI
Journal of Cancer Prevention 2016;21(4):264-270
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*
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Alcohols
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Animals
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Blotting, Western
;
Cytosine
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Diet
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DNA Methylation
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DNA*
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Epigenomics
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Ferritins
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Humans
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Iron*
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Liver
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Male
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Methods
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Rats*
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Rats, Sprague-Dawley
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Receptors, Transferrin
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Spectrum Analysis
9.Clinical utility of reticulocyte hemoglobin content for the diagnosis of iron deficiency anemia in children.
Li-Sha DENG ; Hui-Min TENG ; Yan-Shan LI
Chinese Journal of Contemporary Pediatrics 2011;13(3):212-215
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
10.Low prealbumin levels are independently associated with higher mortality in patients on peritoneal dialysis.
Kyung Hee LEE ; Jang Hee CHO ; Owen KWON ; Sang Un KIM ; Ryang Hi KIM ; Young Wook CHO ; Hee Yeon JUNG ; Ji Young CHOI ; Chan Duck KIM ; Yong Lim KIM ; Sun Hee PARK
Kidney Research and Clinical Practice 2016;35(3):169-175
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
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Cardiovascular Diseases
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Comorbidity
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Creatinine
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Dialysis
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Follow-Up Studies
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Humans
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Inflammation
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Mortality*
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Multivariate Analysis
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Peritoneal Dialysis*
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Prealbumin*
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Renal Dialysis
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Risk Factors
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ROC Curve
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Transferrin