Hematological phenotype analysis of fetuses and patients with hemoglobin H disease / 中华地方病学杂志

Objective:To analyze the relationship between hematological and genotype characteristics of fetuses and patients with hemoglobin (Hb) H disease and their natural disease progression.Methods:From 2010 to 2022, a total of 1 252 blood samples from fetuses and patients with Hb H disease who visited the Guangxi Zhuang Autonomous Regional Maternal and Child Health Hospital were collected (including 174 umbilical cord blood samples, 1 062 peripheral blood samples from patients over 2 years old, and 16 peripheral blood samples from patients with rare cases of genotype Hb H). Additionally, 278 peripheral blood samples were collected from patients aged 0 - 2 years old with Hb H 3.7, Hb H 4.2, Hb H CS, and Hb H WS disease for the study of trends in red blood cell development. Multiple probe hybridization and microarray comparative genomic hybridization technology combined with first-generation Sanger sequencing were used for rare mutation detection.Results:Among the 1 062 Hb H disease patients over 2 years old, 62.34% (662/1 062) had gene deletion (--/-α), of which Hb H 3.7 (-- SEA/-α 3.7) and Hb H 4.2 (-- SEA/-α 4.2) were the most common, accounting for 42.28% (449/1 062) and 19.11% (203/1 062) of the total, respectively. Among the non-deletion genotypes (--/αα T or α Tα/αα T), Hb H CS (-- SEA/α CS), Hb H WS (-- SEA/α WS) and α CSα/α CSα accounted for 16.85% (179/1 062), 16.48% (175/1 062) and 1.98% (21/1 062), respectively. The 81.12% (537/662) of patients with deletional Hb H disease showed mild to moderate anemia, with Hb H detection rates ranging from 75% to 80%. Among non-deletional Hb H disease, Hb H WS disease showed the mild (blood Hb concentration > 95 g/L in 90% of patients) phenotype while Hb H CS and Hb H QS (-- SEA/αα QS) patients had moderate to severe anemia, with Hb H detected in peripheral blood at higher levels than in other types of Hb H disease patients. Except for Hb H CS and Hb H QS, which did not show a significant increase in Hb A2 levels when complicated with β-thalassemia, Hb A2 levels were increased (> 3.5%) in all other types of Hb H disease patients. When Hb H disease was complicated with β-thalassemia, Hb H peaks were not detected in either type of Hb H disease. The results of red blood cell development trend detection showed that erythrocyte counts were elevated in patients with Hb H disease compared to their normal counterparts; whereas, blood Hb, mean erythrocyte volume (MCV) and mean erythrocyte hemoglobin content (MCH) were lower than in their normal counterparts ( P < 0.05) and decreased to the minimum at 6 months to 1 year of age. Patients with Hb H CS disease, as the most severe form of anemia, had the highest MCV values ( P < 0.001). The results of fetal cord blood with Hb H disease showed that α CSα/α CSα caused severe intrauterine anemia, followed by Hb H QS and Hb H CS. The content of Hb Bart's in umbilical cord blood was negatively correlated with the severity of anemia ( rs = - 0.58, P < 0.001). When Hb H disease was complicated with β-thalassemia, there was no significant improvement in fetal anemia, and the Hb Bart's content did not change significantly ( P > 0.05). In addition, Hb H 21.9 (-α 21.9kb/-- SEA) and Hb H 2.4 (-α 2.4/-- SEA) were common in patients with deletion rare Hb H. In patients with non-deletion rare Hb H, αα Amsterdam-A1/-- SEA and αα Hb G-Georgia/-- SEA were both first reported. Conclusions:There is heterogeneity in clinical manifestations of patients with different types of Hb H disease or same type of Hb H disease at different developmental stages. When patients with Hb H are complicated with β-thalassemia, the phenotype of patients with the deletion type is improved, while that of patients with the non-deletion type is not. Compared to normal individuals, patients with Hb H disease have lower blood Hb concentration, MCV and MCH, and more rapid physiological changes in red blood cells.

Caracterización clínica y epidemiológica de población pediátrica costarricense con alfa-talasemia

Objetivo: Describir las características clínicas y epidemiológicas de pacientes con alfatalasemia atendidos en un hospital nacional pediátrico. Métodos: Estudio observacional descriptivo de corte transversal. Se estudia a 60 pacientes del Servicio de Hematología del Hospital Nacional de Niños "Dr. Carlos Sáenz Herrera", Caja Costarricense de Seguro Social, del 1° de enero de 2018 al 31 de enero de 2019, cuyas edades están comprendidas entre los 0 meses y 12 años 11 meses de edad, con índices hematimétricos sugestivos de alfa-talasemia y con electroforesis de hemoglobina patrón AA con HbA2 normal o disminuida. Análisis molecular: identificación de 21 mutaciones y deleciones más frecuentes para el gen de alfa-globina: 3.7, 4.2, 20.5, MED, FIL, SEA, THAI, anti-3.7 triplicación, HbConstant Spring, HbQuonSze, Hb Adana, HbKoya Dora, HbIcara, HbPakse, a2 poli A-1/2, a2-cd142, a1-cd14, a2-init-cd, a2-cd19, a2-IVS1, a2-cd59. Se utiliza el método de amplificación por reacción en cadena de la polimerasa e hibridación inversa del ácido desoxirribonucleico genómico en leucocitos de sangre periférica de los pacientes. Resultados: Se confirma la enfermedad en 44/60 casos (73%). La edad media al diagnóstico para estos casos es de 4.9 años (desviación estándar 3.0), predominó el sexo femenino en 52.3% de los casos. La provincia de Guanacaste reportó la mayor prevalencia de la enfermedad. El defecto genético delecional -3.7 Kb es el genotipo más frecuente. El fenotipo en el 77.2% de los casos indicó portador silente de alfa-talasemia. El 84.1% de los sujetos positivos para alfa-talasemia correlacionó con hipocromía, microcitosis y eritrocitosis en el hemograma inicial. El 9% de los casos evidenció la coexistencia de alfa-talasemia y anemia por deficiencia de hierro. Conclusiones: Este estudio demuestra que los hallazgos de índices eritrocitarios que indiquen hipocromía y microcitosis con aumento del cómputo de eritrocitos, índices férricos normales y una electroforesis de hemoglobina con patrón normal (AA) sugieren ser estudiados molecularmente por alfa-talasemia. La electroforesis de hemoglobina reportada como normal no excluye la condición de alfa-talasemia y debe realizarse el estudio molecular.

Objective: Describe the clinical and epidemiological characteristics of patients with alpha thalassemia in the Hematology Service of a national pediatric hospital. Methods: Cross-sectional descriptive observational study. 60 patients from the Hematology Service of the National Children's Hospital "Dr. Carlos Sáenz Herrera", Costa Rican Social Security Fund, from January 1, 2018 to January 31, 2019, with hematometric indices suggestive of alphaThalassemia, with AA standard hemoglobin electrophoresis with normal or decreased HbA2 with ages between 0 months and 12 years 11 months old. Molecular analysis: Identification of 21 mutations and deletions that includes the detection of the most frequent deletions/mutations for the alpha globin gene: 3.7, 4.2, 20.5, MED, FIL, SEA, THAI, anti-3.7 tripling, HbConstant Spring, HbQuonSze, Hb Adana, HbKoya Dora, HbIcara, HbPakse, a2 poli A-1/2, a2-cd142, a1-cd14, a2-init-cd, a2-cd19, a2-IVS1, a2-cd59. The reverse hybridization PCR amplification method of genomic DNA in peripheral blood leukocytes of patients isused. Results: Of the 60 cases studied, in 44/60 (73%) cases the disease is confirmed. The average age at diagnosis for these cases is 4.9 years (SD 3.0), the female sex predominated in 52.3% of the cases. Guanacaste reported the highest prevalence of the disease. The deletional genetic defect -3.7 Kb was the most frequent genotype and the phenotype in 77.2% of the cases was he silent carrier of alpha thalassemia. In 84.1% of subjects positive for alpha thalassemia, it correlated with hypochromia, microcytosis, and erythrocytosis in the initial blood count. 9% of the cases showed the coexistence of alpha thalassemia and iron deficiency anemia. Conclusions: This study demonstrates that the findings of erythrocyte indices that indicate hypochromia and microcytosis with increased erythrocyte count; normal iron indices and a normal hemoglobin (AA) electrophoresis pattern suggest that they should be studied molecularly for alpha thalassemia. Hemoglobin electrophoresis reported as normal does not exclude the condition of alpha thalassemia and the molecular study must be carried out.

Enfermedad por hemoglobina H: primer caso de dobles heterocigotos hemoglobina Constant Spring / Sudeste Asiático en Costa Rica / Hemoglobin H disease: first case of double heterozygous hemoglobin Constant Spring / Southeast Asian in Costa Rica

Resumen La enfermedad por hemoglobina H es un cuadro clínico que se presenta en las alfa talasemias, las cuales son enfermedades que cursan con anemia microcítica hipocrómica, debidas principalmente a deleciones en el gen de alfaglobina, lo que disminuye la producción de la cadena de alfa globina y promueve la formación de variantes de hemoglobina. Cuando se detectan variantes de hemoglobina en las alfa talasemias, por lo general, se debe a genotipos homocigotas o dobles heterocigotas para mutaciones y deleciones del gen de alfa globina coheredadas. En este artículo se describe el primer caso en Costa Rica, de dos hermanos con enfermedad por hemoglobina H, que fenotípicamente presentaron las variantes de hemoglobina H y hemoglobina Constant Spring en el análisis electroforético de la hemoglobina, y cuyo análisis molecular del gen de alfa globina detectó tanto la deleción sudeste asiático como la mutación para hemoglobina Constant Spring, siendo diagnosticados como dobles heterocigotos por alfa talasemia (genotipo --SEA/ααCS).

Abstract Hemoglobin H disease occurs in patients with alpha thalassemia, diseases associated with hypochromic microcytic anemia, mainly due to deletions in the alpha globin gene, which decreases the production of the alpha globin chain and promotes the formation of hemoglobin variants. When hemoglobin variants are detected in alpha thalassemias it is usually due to homozygoys or doublé heterozygous genotypes, for mutations and deletions of the alpha globin gene. This article describes the first case in Costa Rica of two siblings with hemoglobin H disease, who phenotypically presented the hemoglobin H and Constant Spring hemoglobin variants in the electrophoretic analysis of the hemoglobin, and whose molecular DNA analysis of the alpha globin gene detected both, the Southeast Asian deletion and the mutation for Constant Spring Hemoglobin, being diagnosed as compound heterozygous for alpha thalassemia (genotipe --SEA/ααCS).

Significance of hemoglobin H detection in the diagnosis of α-thalassemia and MDS / 国际检验医学杂志

Objective To evaluate the significance of hemoglobin H in the diagnosis of alpha thalassemia and Myelodysplastic syndrome. Methods A retrospective analysis of the complete clinical data of hemoglobin H in our hospital from January 2007 to October 2017 was performed. The hemoglobin H content in 34 cases of alpha thalassemia and the corresponding blood routine, reticulocyte, Bilirubin, and plasma free hemoglobin levels were analyzed by Pearson correlation. Results 39 cases of hemoglobin H positive cases, of which 34 cases diagnosed as alpha thalassemia,5 cases diagnosed as myelodysplastic syndrome; for alpha thalassemia, hemoglobin H content and reticulocyte, plasma free hemoglobin and Indirect bilirubin test results are related(r =0. 453 ,0. 398,0. 412,P<0. 05). Conclusion Hemoglobin H is not only found in alpha thalassemia but detected in other types of hematologic disorders.

The First Case of X-linked Alpha-thalassemia/Mental Retardation (ATR-X) Syndrome in Korea

Mutation of the ATRX gene leads to X-linked alpha-thalassemia/mental retardation (ATR-X) syndrome and several other X-linked mental retardation syndromes. We report the first case of ATR-X syndrome documented here in Korea. A 32-month-old boy came in with irritability and fever. He showed dysmorphic features, mental retardation and epilepsy, so ATR-X syndrome was considered. Hemoglobin H inclusions in red blood cells supported the diagnosis and genetic studies confirmed it. Mutation analysis for our patient showed a point mutation of thymine to cytosine on the 9th exon in the ATRX gene, indicating that Trp(C), the 220th amino acid, was replaced by Ser(R). Furthermore, we investigated the same mutation in family members, and his mother and two sisters were found to be carriers.

Enfermedad por hemoglobina H: primer caso de la variante de hemoglobina H tipo, menos alfa elevado a potencia 3.7 / SEA, en Costa Rica / Hemoglobin H disease: first case of hemoglobin H variant type (-a 3.7 / SEA) in Costa Rica

La enfermedad por Hemoglobina H es la forma más común de talasemia intermedia y posee muchas características que requieren cuidadosa consideración en su manejo clínico. En lamayoría de los casos, la enfermedad por Hemoglobina H resulta de un estado doble heterocigoto producido por unadeleción tipo α0 que remueve ambos genes de α-globina en uno de los cromosoma 16 y de una deleción tipo α+ en uno de los genes de α-globina en el otro cromosoma 16, resultando enuna condición tipo (--/-α). El exceso de cadenas β de globina precipita y forma una hemoglobina anormal característica; la hemoglobina H (Hb H), un tetrámero de β globina (β4). Lospacientes con hemoglobina H que se encuentran en estado compensado pueden tener niveles de hemoglobina entre 9 y 10 g/dL, sin embargo durante las crisis hemolíticas, que se desarrollan durante o después de infecciones agudas con fiebres altas, la hemoglobina puede llegar a disminuirsignificativamente y los pacientes pueden desarrollar shock y fallo renal. Aún cuando la esplenectomía eleva la hemoglobina significativamente, no se recomienda porque la mayoría delos pacientes tienen un nivel aceptable de hemoglobina mientras se encuentren compensados. Se presenta el primercaso descrito en Costa Rica de enfermedad por hemoglobina H variante del sudeste asiático (-α3.7/ --SEA).

Hemoglobin H (Hb H) disease is the most common form of thalassemia intermedia and has many features that require careful consideration in its management. In the majority of cases, the disease results from double heterozygosity for α0- thalassemia due to deletions that remove both linked α-globin genes on one chromosome 16, and deletional α+ from single α-globin gene deletions on the other chromosome 16 resulting in a (--/-α) condition. The excess β globin chainprecipitates and forms a characteristic abnormal hemoglobin: hemoglobin H a β globin tetramer (β4). In a steady state,patients with Hb H disease have hemoglobin levels around 9 to 10 g/dL however, during a hemolytic crisis, which frequently occur in or after acute infections causing high fever, the hemoglobin may drop significantly and the patients can develop shock or renal shutdown. Even though splenectomy leads to significant elevation of hemoglobin levels, it is not recommended because the majority of patients do well with said steady-state hemoglobin levels. We present here the first case of hemoglobin H (-α3.7/ --SEA) southeast Asia variant described in Costa Rica.

Dificuldades na identificação laboratorial da talassemia alfa / Difficulty on laboratory identification of alpha thalassemia

Introdução: Talassemia alfa é uma síndrome associada à redução da síntese de cadeias de globina do tipo alfa. A gravidade das manifestações clínicas está relacionada com a quantidade de globinas produzida e a estabilidade das cadeias beta presentes em excesso. A talassemia alfa mínima resulta da deleção de apenas um dos quatro genes a (-α/αα). Clinicamente apresenta anemia leve com microcitose ou ausência de anemia, sendo o diagnóstico realizado por meio de visualização da hemoglobina (Hb) H por eletroforese alcalina em acetato de celulose ou por identificação de inclusões celulares de Hb H coradas pelo azul de crezil brilhante. Objetivo: Avaliar portadores de talassemia alfa e seus respectivos progenitores, correlacionando perfil hematológico e presença de Hb H, utilizando procedimentos laboratoriais clássicos em três diferentes amostragens. Discussão e conclusão: Os dados obtidos mostram que a presença de Hb H, indicativo de talassemia alfa, pode não ser confirmada em uma análise posterior. Entre os fatores que podem influenciar no não aparecimento de Hb H em pessoa comprovadamente com talassemia alfa está a deficiência de ferro. A talassemia alfa está associada a defeitos envolvendo os genes codificadores da cadeia alfa, mas também pode estar relacionada com desbalanciamento temporário na expressão dos genes globina, diminuição de alfa ou aumento de beta, o que poderia explicar o aparecimento de tetrâmeros de cadeia beta (Hb H), sugerindo diagnóstico de talassemia alfa mínima.

Introduction: Alpha thalassemia is a syndrome with associated with the reduction of alpha globin chain synthesis. The severity of clinical manifestations is related to the amount of globins produced and the stability of beta chains that are present in excess. Alpha thalassemia minor is caused by the deletion of one of the four genes a (-α/αα). Clinically, it presents mild anemia with microcytosis or absence of anemia. The diagnosis is made by the visualization of Hb H through alkaline electrophoresis on cellulose acetate or by the identification of inclusion bodies stained with brilliant cresyl blue. Objective: Evaluate alpha thalassemia carriers and their respective progenitors, correlating their hematology profile and the presence of Hb H by means of standard laboratory procedures in three different samplings. Discussion and conclusion: The results show that the presence of Hb H, which is indicative of alpha thalassemia, may not be confirmed in a subsequent analysis. Iron deficiency in Hb H carriers is among the factors that may influence on the absence of Hb H in alpha thalassemia proven patients. Alpha thalassemia is associated with genetic defects involving alpha chain encoding genes, but may be also associated with a temporary imbalance of globin gene expression, alpha chain reduction or beta increase, which could explain the presence of beta chain tetramer (Hb H) leading to the diagnosis of alpha thalassemia minor.

Molecular characterization of the - -SEA alpha thalassemia allele in Mexican patients with HbH disease / Caracterización molecular del alelo - -SEA de talasemia alfa en pacientes mexicanos con enfermedad por HbH

α-Thalassemia is one of the most prevalent hemoglobin disorders in the world, in South-East Asians, the--SEA allele is widely found in the HbH disease patients. The purpose of this work is to describe the molecular characteristics of Hemoglobin H disease in three patients from two Mexican families, as well to analyze the DNA sequence of the --SEA allele to determine the precise site of the crossover. The -α3.7 and --SEA alleles were identified using an established long-PCR method modified in our laboratory. The crossover site of --SEA mutation was analyzed by DNA sequencing. The three HbH subjects showed the same genotype -α3.7/--SEA. The -α3.7 allele has been observed in almost every racial studied group, whereas the --SEA allele is predominant in South-East Asian countries. DNA analysis through the breakpoint sites of the --SEA allele in both families showed the 5' breakpoint at the third base of codon 28 in the ψα2 gene and the 3' breakpoint within an Alu-Jo sequence, 1,328 nucleotides upstream of the 3'HVR. Therefore the size of the deletion is 19,303 nucleotides. This is the first report in which the flanking deletion sites of the--SEA mutation have been analyzed in Mexican patients, the 5' and 3' ends of the deletion is well determined.

La Talasemia-α es uno de los desórdenes de la hemoglobina más prevalences en el mundo. En el sureste de Asia, --SEA es el alelo más frecuente en pacientes con enfermedad por HbH (EHbH). En el presente trabajo se describen las características moleculares de tres pacientes con EHbH de dos familias mexicanas, y se analiza la secuencia de DNA del alelo --SEA, para determinar los sitios de ruptura. Los alelos -α3.7y --SEA se identificaron por un método de PCR modificado en nuestro laboratorio y los sitios de ruptura por secuenciación de DNA. Los tres pacientes con EHbH mostraron el genotipo -a3.7/--SEA. El alelo -α3.7 está ampliamente distribuido en el mundo, mientras que el alelo--SEA predomina en los países del sureste de Asia. El análisis de DNA del alelo--SEA mostró en 5' el sitio de ruptura en el codón 28 del pseudogén ψα2 y en 3', dentro de la secuencia Alu-Jo, localizada a 1,328 nucleótidos de la región HVR3', lo que da un segmento delecionado de 19,303 nucleótidos. Éste es el primer reporte en el que se analizan los sitios que flanquean la deleción del alelo --SEA en pacientes mexicanos y se definen con precisión los extremos 5' y 3' de la deleción.

Significance of Serum Transferrin Receptors and Iron Overload in Children with Hemoglobin H Disease / 实用儿科临床杂志

Objective To measure the levels of serum transferrin receptors (sTfR) and serum ferritin (SF) in children with hemoglobin H disease for investigation the iron metabolism(sideropenic or iron overload) and to guide the proper intervention.To get a best index of judging the iron overload in HbH children by analysis of the values of sTfR and sTfR/lgSF.Methods Peripheral blood specimens were obtained from 50 cases of HbH children and 30 cases of healthy normal controls,the levels of sTfR and SF were examined by enzyme linked immunosorbent assays(ELISA).The diagnosis efficiency of sTfR and sTfR/lgSF in HbH children′s iron metabolism was judged by receiver operator cha-racteristic (ROC) curve analysis.Results The level of sTfR in HbH group was (6.74?1.02) mg?L-1,which was significantly lower than that in healthy control group[(8.09?0.67) mg?L-1](P

Hemoglobin H disease combining hemoglobin Q-thailand and hemoglobin E disease:a clinical and laboratory genealogical research / 中华检验医学杂志

Objective To investigate clinically and in laboratory a genealogical tree with hemoglobin H disease Combining Hemoglobin Q-Thailand and Hemoglobin E Disease.Methods Genealogical laboratory studies were carried out with the following methods:hemoglobin electrophoresis, various biochemical determinations,and DNA analysis.Results Father's genotype of ?-THAL:??/?~Q ?~(4.2); genotype of ?-THAL:?E/N;phenotype:minor ?-THAL carrier combining Hb Q and Hb E multiple heterozygote;mother' s genotype of ct-THAL:--~(SEA)/??;genotype of ?-THAL:?n/?n.According to comprehensive analysis,mother's phenotype:minor ?-THAL,complex minor ?-THAL carrier combining Hb F ? Initial sign of ?-THAL genotype:--~(SEA)/?~Q ?~(4.2);phenotype:deletion type Hb H genotype disease;?- THAL genotype:?E/?E;phenotype:? E homozygote.According to comprehensive analysis:deletion type Hb H combining HbE multiple heterozygote.Youger brother's ?-THAL genotype:--~(SEA)/?~Q ?~(4.2);?-THAL genotype:?n/?n;phenotype:deletion type Hb H disease.Both mother and her youngest son have G6PD deficiency.Conclusions Guangdong Province is an area with high morbidity of ?-THAL and ?-THAL,Hb E and Hb Q as well as G6PD deficiency.There may be some correlation between Hb E and Fib Q in term's of the high morbidity of regional Hb,but the two types of Hb combining Hb H disease are rare in China and the world in point of nonhomologous chromosome.Attention should be paid to the problems of double heterozygote of ?-THAL complex ?-THAL,and THAL complex G6PD deficiency.Data from the study have enriched the scientific information of molecular genetics of erythroeyte thalassemia and of molecular pathology with important significance in genetics guidance and clinical treatment for patients.