High performance liquid chromatography (HPLC) is currently the mainstream technology for detecting hemoglobin. Glycated hemoglobin (HbA1c) is a gold indicator for diagnosing diabetes, however, the accuracy of HbA1c test is affected by thalassemia factor hemoglobin F (HbF)/hemoglobin A2 (HbA2) and variant hemoglobin during HPLC analysis. In this study, a new anti-interference hemoglobin analysis system of HPLC is proposed. In this system, the high-pressure three-gradient elution method was improved, and the particle size and sieve plate aperture in the high-pressure chromatography column and the structure of the double-plunger reciprocating series high-pressure pump were optimized. The system could diagnose both HbA1c and thalassemia factor HbF/HbA2 and variant hemoglobin, and the performance of the system was anti-interference and stable. It is expected to achieve industrialization. In this study, the HbA1c and thalassemia factor HbF/HbA2 detection performance was compared between this system and the world's first-line brand products such as Tosoh G8, Bio-Rad Ⅶ and D10 glycosylated hemoglobin analysis system. The results showed that the linear correlation between this system and the world-class system was good. The system is the first domestic hemoglobin analysis system by HPLC for screening of HbA1c and thalassemia factor HbF/HbA2 rapidly and accurately.
Chromatography, High Pressure Liquid ; Fetal Hemoglobin/analysis* ; Glycated Hemoglobin A/analysis* ; Hemoglobin A2/analysis* ; Hemoglobins

β-thalassaemias are inherited hemoglobin disorders caused by defects in the β-globin gene. In recent years, researches have re-mentioned the therapeutic significance of drug-induced fetal hemoglobin (HbF), which can reduce the imbalance of α and β chains and improve the severity of anemia by increasing the expression of γ chain. Drug trials, such as hydroxyurea, thalidomide and desitabine have shown elevated hemoglobin, decreased blood transfusion dependence, and reduced symptoms other than anemia after treatment. In addition, in vitro experiments suggested that HbF can also induce by other drugs, which providing important clues for safe and effective HbF inducers. Therefore, this article reviews the current research progress so as to expect beneficial to clinical treatment.
Blood Transfusion ; Fetal Hemoglobin ; Humans ; Hydroxyurea ; beta-Globins ; beta-Thalassemia

BACKGROUND: Fetal hemoglobin (HbF) levels in different hemoglobin variants in Osogbo, Nigeria, were estimated using two principal methods of estimation using existing information for HbF concentration and distribution of various hemoglobin variants in the area, as well as diagnosed cases of thalassemia. Two hundred and sixty samples collected from HbSS, HbSC, HbAA, HbAS, and HbAC subjects were analyzed. HbF level and hemoglobin type were determined in this study. METHODS: The hemoglobin type was determined using cellulose acetate electrophoresis at an alka-line pH, and HbF was determined by the acid elution and alkaline denaturation methods. RESULTS: The mean+/-SD of HbF in the respective hemoglobin variants was as follows: HbSS, 2.09+/-1.94%; HbSC, 0.85+/-0.54%; HbAA, 0.69+/-0.46%; HbAS, 0.52+/-0.31%; and HbAC, 0.57+/-0.26%. The mean HbF level across the hemoglobin variants was statistically significant (P<0.05). Investigating the sex distribution of the HbF level in the studied population revealed a slightly higher mean HbF level in females than in their male counterparts. CONCLUSION: Within the study population, the HbF level was found to be highest in HbSS and lowest in HbAS. The two methods of estimating HbF are equally reliable, since there was no significant difference between the results obtained from the two methods.
Cellulose ; Electrophoresis, Cellulose Acetate ; Female ; Fetal Hemoglobin ; Hemoglobin A ; Hemoglobins ; Humans ; Hydrogen-Ion Concentration ; Male ; Nigeria ; Sex Distribution ; Thalassemia

BACKGROUND: The Apt test is used for differentiating neonatal blood from the maternal blood in the meconium. The test requires a sufficient amount of blood to be detectable to the naked eyes. But many of the specimens in the hospital laboratory contain only a small amount of blood and is not detectable to the naked eyes and usually the results are reported 'impossible to determine' due to the small amount of blood. We developed a latex agglutination test to solve this problem and to differentiate the neonatal blood from the maternal blood in a small amount that was not detectable with the naked eyes. METHODS: Latex reagents for hemoglobin A (Hb A) and hemoglobin F (Hb F) were made. Ten milligrams of meconium were dissolved in 1mL of deionized water (DW). Ten milligrams of meconium that had shown negative results for both of the above reagents were mixed, each with 10microL of whole blood (WB) from 10 pregnant women and 10 neonates who had various hemoglobin concentrations (10-17 g/dL). Each of the 20 mixtures was dissolved in 1 mL of DW (WB 10microL/mL). Then, serial dilutions were made at a ratio of 1:10 until the final concentration of 10 pL/mL. Each of the six dilutes were tested with the two latex reagents. RESULTS: The dilutes of WB 10microL/mL looked red, WB 1microL/mL looked pink and all other dilutes showed no colors to the naked eyes. The reagent for Hb A showed agglutination in dilutes from WB 1microL to 1 nL/mL DW from all of the 20 persons. The reagent for Hb F reacted with dilutes from WB 1microL to 1 nL/mL DW from the ten neonates but did not react with those from any pregnant women. CONCLUSIONS: The latex agglutination test can be applied to the specimen with no color detectable to the naked eyes after dilution. The specimen reacted with both the Hb A and Hb F reagents could be determined as fetal blood and the one that reacted with the reagent only for Hb A could be determined as maternal blood.
Agglutination ; Female ; Fetal Blood ; Fetal Hemoglobin ; Hemoglobin A ; Humans ; Indicators and Reagents ; Infant, Newborn ; Laboratories, Hospital ; Latex Fixation Tests* ; Latex* ; Meconium* ; Pregnant Women ; Water

Massive fetomaternal hemorrhage (FMH) is a major cause of unexplained fetal death and neonatal anemia. FMH can be diagnosed using the Kleihauer-Betke test or flow cytometry by identifying the presence of fetal red cells in the maternal blood. However, timely diagnosis is a challenge because many hospitals lack the equipment needed to perform such tests. The authors experienced a case of FMH diagnosed via high-performance liquid chromatography (HPLC) which is generally used in measuring glycated hemoglobin (HbA1c) in a patient with unexplained neonatal anemia. A girl aged 2 days was transferred to our hospital for showing pallor and a hemoglobin level of 5.0 g/dL. HPLC revealed 3% fetal hemoglobin (HbF) in the maternal blood. HPLC is a quick test for quantifying HbF that is readily available in many hospitals and could serve as a promising alternative for diagnosing FMH.
Anemia, Neonatal ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Diagnosis ; Female ; Fetal Death ; Fetal Hemoglobin ; Fetomaternal Transfusion ; Flow Cytometry ; Hemoglobin A, Glycosylated ; Humans ; Infant, Newborn ; Pallor ; Pregnancy ; Prothrombin Time

The fetal hemoglobin, which is structurally different from adult hemoglobin, has higher affinity for oxygen and higher resistance to denaturation with alkali than adult hemoglobin. Intrauterine growth retarded neonates have higher mortality and morbidity than normal neonates. In this article, to determine the changes of fetal hemoglobin by gestational age in infants appropriate in weight for gestational age (AGA)and to explain the mechanism underlying the increased fetal hemoglobin synthesis in intrauterine growth retarded newborn infants, the proportion of fetal hemoglobin and adult hemoglobin at birth and 1 month of age was checked in 25term infants small for gestational age(TSGA). The results were compared with 50 preterm infants appropriate in weight for gestational age (paga) and 40 term infants appropriate in weight for gestational age (TAGA). The results were as forllows. 1)The decrease of fetal hemoglobin by gestational age in infants of AGA at birth was statistically significant (p<0.01). 2) The fetal hemoglobin at birth was 74.47+/-2.4%n the TSGA group, 78.01+/-5.05% in the PAGA group and 68.32+/-4.84% in the TAGA group. The differnce between each group was statistically significant (p<0.01). 3) The fetal hemoglobin at one month of age was 55.68+/-3.76% in the TSGA group, 35.74 (13.33%in the PAGA group and 59.96+/-5.53% in the TAGA group. The difference between TSGA and TAGA infants was not significant (p>0.05). 4) The decrese rate of fetal hemoglobin between first day and one month of postantal age was 54.2% in the PAGA infants, 25.2% in the TSGA infants and 12.2% in the TAGA infants. 5) The fetal hemoglobin at forty weeks of postconceptional age was 13.20+/-5.09%in the transfused PAGA group, 62.34+/-3.01% in the nontransfused PAGA group, 64.08+/-2.08% in the TSGA group and 68.32+/-4.12% in the TAGA infants. The difference between transfused PAGA group and other groups was statistically significant (p<0.05).
Adult ; Alkalies ; Fetal Hemoglobin* ; Gestational Age* ; Humans ; Infant ; Infant, Newborn* ; Infant, Premature ; Mortality ; Oxygen ; Parturition

OBJECTIVE: To evaluate the intrauterine hypoxic effect in term small-for-date (SGA) neonates by comparing the umbilical venous erythropoietin (EPO) concentration between appropriately-grown (AGA) and SGA neonates at delivery and to determine the variables that correlate with the umbilical venous EPO concentration by multiple regression analysis. METHODS: 183 term singleton neonates (gestational weeks > OR =37) were enrolled and divided into 136 cases of AGA (10th-90th percentile of birth weight for the gestational age) and 47 cases of SGA (< 10th percentile of birth weight for the gestational age. At each delivery, blood gas values, concentration of EPO by radioimmunoassay and the number of nucleated erythrocytes (NRBC) per 100 white blood cells in smear of umbilical venous blood were obtained. The placentas were examined microscopically for presence of pathological infarct. Statistical analysis was done by Mann-Whitney U test, x2 test, and univariate and multiple regression analysis using SPSS statistical package (version 10). RESULTS: The median umbilical venous EPO concentration, fetal hemoglobin level were significantly higher in SGA neonates than those in AGA neonates. There was no difference in number of NRBC between AGA and SGA neonates. Multiple regression analysis model with level of NRBC, presence of placental infarct and SGA provided prediction of EPO level in umbilical venous blood at delivery (regression equation: EPO=-103.94+4.75NRBC+68.07placental infarct+36.40SGA F=15.57. r2=0.47). CONCLUSION: Term SGA neonates was considered to have compensatory, not pathological intrauterine hypoxic effect by showing increased level of EPO and normal level of NRBC in umbilical venous blood at delivery, compared with thoses of AGA. In the suspected cases of SGA antenatally, measurement of NRBC level and placental pathologic examination for infarct can be informative for estimating the extent and duration of intrauterine hypoxia.
Anoxia ; Birth Weight ; Erythroblasts ; Erythropoietin* ; Fetal Hemoglobin ; Gestational Age ; Humans ; Infant, Newborn* ; Leukocytes ; Placenta ; Pregnancy* ; Radioimmunoassay

OBJECTIVE: To evaluate the intrauterine hypoxic effect in term small-for-date (SGA) neonates by comparing the umbilical venous erythropoietin (EPO) concentration between appropriately-grown (AGA) and SGA neonates at delivery and to determine the variables that correlate with the umbilical venous EPO concentration by multiple regression analysis. METHODS: 183 term singleton neonates (gestational weeks > OR =37) were enrolled and divided into 136 cases of AGA (10th-90th percentile of birth weight for the gestational age) and 47 cases of SGA (< 10th percentile of birth weight for the gestational age. At each delivery, blood gas values, concentration of EPO by radioimmunoassay and the number of nucleated erythrocytes (NRBC) per 100 white blood cells in smear of umbilical venous blood were obtained. The placentas were examined microscopically for presence of pathological infarct. Statistical analysis was done by Mann-Whitney U test, x2 test, and univariate and multiple regression analysis using SPSS statistical package (version 10). RESULTS: The median umbilical venous EPO concentration, fetal hemoglobin level were significantly higher in SGA neonates than those in AGA neonates. There was no difference in number of NRBC between AGA and SGA neonates. Multiple regression analysis model with level of NRBC, presence of placental infarct and SGA provided prediction of EPO level in umbilical venous blood at delivery (regression equation: EPO=-103.94+4.75NRBC+68.07placental infarct+36.40SGA F=15.57. r2=0.47). CONCLUSION: Term SGA neonates was considered to have compensatory, not pathological intrauterine hypoxic effect by showing increased level of EPO and normal level of NRBC in umbilical venous blood at delivery, compared with thoses of AGA. In the suspected cases of SGA antenatally, measurement of NRBC level and placental pathologic examination for infarct can be informative for estimating the extent and duration of intrauterine hypoxia.
Anoxia ; Birth Weight ; Erythroblasts ; Erythropoietin* ; Fetal Hemoglobin ; Gestational Age ; Humans ; Infant, Newborn* ; Leukocytes ; Placenta ; Pregnancy* ; Radioimmunoassay

A study of normal value of fetal hemoglobin concentration at each age in 1213 cases (852 patients, 361 normal children) who visited Chonnam university hospital from June 1977 to November 1978 was performed by alkali denaturation method, and following results were obtained. 1) The mean value of Hgb F at each age in normal children group was 65.5+/-1.8% in newborn infants during 24 hours, 40.5+/-1.6% in 1 month, 24.3+/-1.2% in 2 months, 15.3+/-1.4% in 3 months, 4.5+/-1.2% in 1 year 1.3+/-0.5% in 5 years, 0.9+/-0.7% in 10 years, 0.5+/-0.3% in 15 years. 2) The mean value of Hgb F at each age in patients group was 68.6+/-2.5% in newborn infants during 24 hours, 42.5+/-1.7% in 1 month, 22.9+/-1.1% in 2 months, 15.2+/-0.9% in 3 months, 5.5+/-1.0% in 1 year, 1.4+/-0.4% in 5 years, 0.9+/-0.6% in 10 years, 0.6+/-0.2% in 15 years. 3) The mean value of Hgb F at each age in both group was 67.5+/-2.0% in newborn infants during 24 hours, 41.8+/-2.1% in 1 month, 23.4+/-1.9% in 2 months, 15.2+/-1.0% in 3 months, 5.1+/-1.1% in 1 year, 1.4+/-0.8% in 5 years, 0.9+/-0.2% in 10 years, 0.6+/-0.2% in 15 years. 4) No significant correlation was found in the concentration of Hgb F at each age between normal children group and patients group, and either in the concentration of Hgb F at each sex of two groups.
Alkalies ; Child* ; Fetal Hemoglobin* ; Humans ; Infant, Newborn ; Jeollanam-do ; Reference Values*

OBJECTIVETo study the effects of matrine on gamma-globin synthesis in K562 cells in vitro.

METHODSBenzidine staining was used to study the dose- and time-dependent effects of matrine on hemoglobin synthesis in K562 cells, and Western blotting was performed to determine the level of hemoglobin F(alpha(2)gamma(2)).

RESULTSBenzidine staining showed that K562 cells treated with matrine of 0.1 mg/ml had a positivity rate for benzidine (BZ%) of 15.67% at 96 h after the treatment, and Western blotting indicated increased synthesis of hemoglobin F.

CONCLUSIONMatrine can induce gamma-globin synthesis and increase hemoglobin F level in K562 cells, the effect of which resembles that of sodium butyrate.

Alkaloids ; pharmacology ; Blotting, Western ; Fetal Hemoglobin ; biosynthesis ; Globins ; biosynthesis ; Humans ; K562 Cells ; Quinolizines ; pharmacology