Molecular genetic characteristics of a family which coinheritance of rare-88 C>G (HBB:c.-138 C>G) β-thalassemia mutation with α-thalassemia and review of the literature / 中华预防医学杂志

The molecular genetic characteristics of a family with rare -88 C>G (HBB: c.-138 C>G) β-thalassemia gene mutation were studied using cohort study. The cohort study was conducted from June to August 2022 by Prenatal Diagnosis Center of Sanya Women and Children's Hospital Managed by Shanghai Children's Medical Center. The phenotype and genotype were analyzed by hematological cytoanalyzer, automatic electrophoretic analysis system, and next-generation sequencing (NGS). And then, Sanger sequencing was used to verify the rare gene results. The results showed that the proband, her father, her uncle and her younger male cousin had discrete microcytosis (MCV 70.1 fl, 71.9 fl, 73.1 fl and 76.6 fl, respectively) and hypochromia (MCH 21.5 pg,22.0 pg,22.6 pg and 23.5 pg, respectively), elevated hemoglobin A2 level (5.3%, 5.4%, 5.4% and 5.5%, respectively), slightly elevated or normal fetal hemoglobin (Hb F), but no anemia. The proband was identified to have co-inherited ɑ-thalassemia (Hb Westmead gene heterozygous mutation, ɑwsɑ/ɑɑ) and β-thalassemia with a rare -88 C>G (HBB: c.-138 C>G) heterozygous mutation (β-88 C>G/βN). Her mother had the same α-thalassemia as the proband. Her father, her uncle and her younger male cousin had the same rare -88 C>G heterozygous mutations as the proband. While her grandmother and younger brother were not carrier of thalassemia. In conclusion, 4 cases of rare -88 C>G(HBB:c.-138 C>G) heterozygous mutation had been detected in a Chinese family. Carriers of this beta-thalassemia are clinically asymptomatic. This study enriches the knowledge of the thalassemia mutation spectrum in Chinese people and provides valuable information for genetic counseling, prenatal diagnosis, and prevention of thalassemia, providing a scientific basis for improving the quality of birth population and preventing birth defects.

Bilateral maculopathy in a thalassemia patient on iron chelation therapy: a case report / Maculopatia bilateral em paciente com talassemia em terapia quelante de ferro: relato de caso

ABSTRACT A 10-year-old Malay girl with underlying HbE/beta-thalassemia, on regular blood transfusion and deferoxamine iron chelation therapy, presented with two-month history of bilateral blurring of vision. On examination, her vision was 6/36 both eyes. Other optic nerve functions were normal. Anterior segment examination of both eyes was unremarkable. Fundus examination of both eyes revealed dull foveal reflex. Optical coherence tomography of both maculae showed increased central subfield thickness. Fundus fluorescence angiography showed patchy hypofluorescence over macular region for both eyes and late staining, indicating retinal pigment epithelium anomalies. A diagnosis of iron-chelation-therapy-related bilateral maculopathy was made. Patient was co-managed with pediatric hematology team to adjust the dose of deferoxamine, and was given three monthly appointments to monitor the progression of maculopathy at the ophthalmology clinic. However patient defaulted ophthalmology follow-up after the first visit.

RESUMO Uma menina malaia de 10 anos de idade com doença de base- B/beta-talassemia, em transfusão de sangue regular e terapia quelante de ferro deferoxamina, apresentou história de dois meses de visão turva bilateral. Ao exame, sua visão era de 6/36 em ambos os olhos. Outras funções do nervo óptico estavam normais. O exame do segmento anterior de ambos os olhos foi normal. Exame do fundo de ambos os olhos revelou reflexo foveal opaco. A tomografia de coerência óptica de ambas as máculas mostrou aumento da espessura do subcampo central. A angiografia de fluorescência do fundo mostrou hipofluorescência irregular sobre a região macular de ambos os olhos e coloração tardia, indicando anomalias de epitélio pigmentar da retina. Um diagnóstico de maculopatia bilateral relacionada à terapia quelante de ferro foi feito. A paciente foi avaliada em conjunto com a equipe de hematologia pediátrica para ajustar a dose de deferoxamina, e foram oferecidas três consultas mensais na clínica oftalmológica, para monitorar a progressão da maculopatia. No entanto, ela não compareceu para acompanhamento oftalmológico após a primeira visita.

Diagnostic Value of HBA / 中国实验血液学杂志

OBJECTIVE@#To explore the diagnostic value of HBA@*METHODS@#1 178 couples in the department of women's health of Chongqing maternal and child health hospital were selected for pregnancy examination. Peripheral venous blood was extracted and analyzed for parallel blood routine test, hemoglobin capillary electrophoresis and thalassemia gene detection.@*RESULTS@#A total of 265 cases of thalassemia gene carriers were screened out in 1 178 couples； 91.3% β@*CONCLUSION@#HBA

The Clinical Value of Combined Detection of RBC, Ret-He and HbA2 for Thalassemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the distribution of Ret-He and RBC in thalassemia and the value of combining HbA2 in the detection of thalassemia among patients with microcytic or hypochromic.@*METHODS@#145 patients with microcytic or hypochromic outpatient or hospitalization in our hospital from May 2018 to December 2019 were selected and were divided into the thalassemia group(68 cases) and the non-thalassemia group (77 cases), and at the same time, the patients were divided into four groups of the non-anemia, mild anemia, moderate anemia and severe anemia group according to the degree of anemia. The Ret-He, RBC, RDW-CV and HbA2 in patients were detected, and the distribution of these parameters were compared, and the joint detection of Ret-He, RBC and HbA2 about its sensitivity, specific and other indicators of auxiliary diagnosis of thalassemia were analyzed.@*RESULTS@#Among patients with microcytic or hypochromic, according to the anemia grade Ret-He gradually decreased from the non-anemia group to the severe anemia group (P<0.05); while RDW-CV was increased gradually from the mild anemia group to the severe anemia group (P<0.05); both RBC and Ret-He were increased in the thalassemia group as compared with the non- thalassemia group (P<0.05); while RDW-CV was decreased in the thalassemia group as compared with the non-thalassemia group (P<0.05); meanwhile Ret-He in the α-thalassemia group was higher than that in the β-thalassemia group. ROC curve analysis showed that combined with HbA2, the specificity was 93.51%, the sensitivity was 66.18%, the positive predictive value was 90% and the negative predictive value was 75.189% when Ret-He was truncated with 19.25 pg and RBC was truncated with 4.95×10@*CONCLUSION@#Among patients with microcytic or hypochromic, the distribution of RBC, Ret-He and RDW-CV was different in the thalassemia group and the non-thalassemia group, and was also affected by the degree of anemia. Combined Ret-He and RBC could improve the diagnostic specificity for thalassemia, which were screened by HbA2 in patients with microcytic or hypochromic.

Discriminant indexes to simplify the differential diagnosis between iron deficiency anemia and thalassemia minor in individuals with microcytic anemia

SUMMARY INTRODUCTION: Microcytic anemias are very common in clinical practice, with iron deficiency anemia (IDA) and thalassemia minor (TT) being the most prevalent. Diagnostic confirmation of these clinical entities requires tests involving iron metabolism profile, hemoglobin electrophoresis, and molecular analysis. In this context, several discriminant indices have been proposed to simplify the differential diagnosis between IDA and TM. OBJECTIVE: The aim of this paper was to demonstrate the clinical relevance of the use of discriminant indices in individuals with microcytic anemia to simplify the differential diagnosis between iron deficiency anemia and minor thalassemia. METHODS: A bibliographic and cross-sectional search was performed in the PubMed, SciELO and LILACS databases, using the following descriptors: iron deficiency anemia, thalassemia minor, and differential diagnosis. RESULTS: More than 40 mathematical indices based on erythrocyte parameters have been proposed in the hematological literature in individuals with microcytosis. Green & King indexes (IGK), Ehsani index, and erythrocyte count (RBC) had excellent performances, especially when their efficacy was observed in adults and children. CONCLUSIONS: Confirmatory tests for differential diagnosis between IDA and TM require time-consuming and costly methods. Despite the excellent performances of IGK, Ehsani index, and RBC, none of them presented sufficient sensitivity and specificity to establish a diagnosis. However, they can provide a powerful additional tool for diagnostic simplification between IDA and TM.

RESUMO INTRODUÇÃO: Anemias microcíticas são muito comuns na prática clínica, sendo a anemia ferropriva (AF) e a talassemia menor (TM) as mais prevalentes. A confirmação diagnóstica dessas entidades clínicas requer testes que envolvem o perfil do metabolismo do ferro, eletroforese de hemoglobinas e análises moleculares. Nesse contexto, vários índices discriminantes têm sido propostos para simplificação do diagnóstico diferencial entre AF e TM. OBJETIVO: O objetivo deste artigo foi demonstrar a relevância clínica da utilização de índices discriminantes em indivíduos com anemia microcítica, para simplificação do diagnóstico diferencial entre anemia ferropriva e talassemia menor. MÉTODOS: Foi realizada uma pesquisa bibliográfica e transversal nas bases de dados PubMed, SciELO e Lilacs, utilizando-se os seguintes descritores: anemia ferropriva, talassemia menor e diagnóstico diferencial. RESULTADOS: Mais de 40 índices matemáticos baseados em parâmetros eritrocitários foram propostos na literatura hematológica em indivíduos com microcitose. Os índices de Green & King (IGK), o índice de Ehsani e a contagem de eritrócitos (RBC) obtiveram excelentes desempenhos, especialmente quando sua eficácia foi observada em adultos e crianças. CONCLUSÕES: Testes confirmatórios para o diagnóstico diferencial entre AF e TM demandam métodos que consomem bastante tempo e alto custo. Apesar dos excelentes desempenhos do IGK, do índice de Ehsani e do RBC, nenhum deles possui sensibilidade e especificidade suficientes para firmar diagnóstico. No entanto, podem fornecer uma poderosa ferramenta adicional para simplificação diagnóstica entre AF e TM.

Síndrome drepanocítico: Asociación de hemoglobina S Y &ß talasemia / Sickle cell syndrome. Association between hemoglobin S and ß thalassemia

El síndrome drepanocítico HbS/β talasemia responde a la herencia de tipo mendeliana en simultáneo de un alelo βs de la hemoglobina S (HbS) y un alelo de β talasemia. Vinculado fundamentalmente a individuos que comparten ascendencia africana y de países del Mediterráneo. La mutación responsable de la HbS es puntual, mientras que para la β talasemia existen más de 200 mutaciones que causan diferentes grados de deficiencia de síntesis de la cadena de β globina, lo cual justifica la heterogeneidad clínica y genética de este síndrome. Se presenta el caso clínico de un adulto joven de escasos recursos que consulta por dolores óseos de larga data. Registra hemogramas con anemia y marcada microcitosis. Se le realizó electroforesis de Hb detectándose un pico anómalo en posición de HbS y elevada fracción de HbA2. El resultado de la electroforesis de hemoglobina indica dos posibles alteraciones moleculares en simultáneo, por tal motivo se realizó el estudio molecular de las mutaciones más frecuentes en nuestra población de β talasemia y de la mutación puntual responsable de la hemoglobinopatía S. A partir de la clínica y datos del laboratorio bioquímico se diagnosticó el síndrome drepanocítico y se confirmó por biología molecular la portación de las mutaciones IVS-Int 110 G > A (β talasemia) y del codón 6 A > T (GAG→GTG: Glu→Val) responsable de la hemoglobinopatía S. Dado que es una enfermedad de alto impacto sanitario, es importante un adecuado asesoramiento genético a toda la familia.

Sickle cell syndrome HbS/β thalassemia is an inheritable mendelian type disease where two affected alleles are simultaneously present, one from HbS (βS) and the other from β thalassemia. That situation is mainly linked to individuals who share African and Mediterranean ancestors. The mutation responsible for HbS is a point mutation, whereas for β thalassemia, there are more than 200 mutations that cause different degrees of deficiency synthesis of β globin chain, which justifies the clinical and genetic heterogeneity of this syndrome. It is presented a clinical case of a young adult man with limited resources that consulted by longstanding bone pain. The patient presented anemia with a marked microcytosis. Hemoglobin electrophoresis was performed, an abnormal peak in position of HbS and high HbA2 fraction were detected. These last results indicated two possible molecular alterations simultaneously, for this reason the molecular study was performed looking for the most common β thalassemia mutations in our population and, the point mutation responsible for S hemoglobinopathy. Clinical data and biochemical laboratory allowed the diagnosis of sickle cell syndrome. The molecular study confirmed the syndrome carrying mutations IVS-I nt 110 G > A, responsible for β thalassemia and, codon 6 A > T (GAG → GTG: Glu → Val) responsible for S hemoglobinophaty. Since it is a disease of high health impact, it is important to provide genetic counseling to the whole family.

Análise funcional de microRNAs e fatores de transcrição durante a diferenciação eritroide in vitro em pacientes com talassemia beta intermediária / Functional analysis of microRNAs and transcription factors during in vitro erythroid differentiation in beta thalassemia intermedia patients

As talassemias beta são caracterizadas pela redução parcial ou completa da síntese de cadeias da globina beta. Estudos mostraram que vários fatores de transcrição estão envolvidos na regulação da expressão de genes eritroides, incluindo o complexo transcricional de GATA1. Além disso, os microRNAs podem atuar como reguladores pós-transcricionais durante o desenvolvimento eritroide. Neste trabalho, foram avaliadas as possíveis relações dos microRNAs tanto com fatores de transcrição eritroide-específicos quanto com os genes das globinas. Para isso, foi realizada a quantificação da expressão dos microRNAs e dos fatores de transcrição durante a diferenciação eritroide in vitro de pacientes com talassemia beta intermediária. Posteriormente, selecionamos o miR-210-3p para os estudos subsequentes e vetores lentivirais foram utilizados para inibir a expressão desse miR em células K562 e KU812. Diminuição da expressão da globina gama e aumento do nível de KLF1 foram observados nas células inibidas. Análise in silico demonstrou que o miR-210-3p possui dois sítios de ligação no RNAm de KLF1 e propõem-se que a expressão aumentada de KLF1 possa colaborar para a diminuição da expressão do gene da globina beta via miR-210-3p. Pela primeira vez é proposto um potencial RNAm alvo para o miR-210-3p que esteja relacionado ao switching da HbF e um possível mecanismo modulador da expressão dessa molécula. (AU)

The beta thalassemia is characterized by partial or complete reduction of the synthesis of beta globin chains. Studies have shown that many transcription factors are involved in regulating the expression of erythroid genes, including the GATA1 complex. Moreover, microRNAs can act as post-transcriptional regulators during erythroid development. In this study, we evaluated the possible relationship of microRNAs both erythroid-specific transcription factors and with the globin genes. Thus, the expression of microRNAs and transcription factors during erythroid differentiation in vitro patients with thalassemia intermediate was realized. Subsequently, miR- 210-3p was selected for subsequent studies and lentiviral vectors were used to inhibit the expression of this miR in K562 and KU812 cells. Down-regulation in the gamma globin and high levels of KLF1 were observed in the inhibited cells. Moreover, in silico analysis showed that miR-210- 3p could target two regions of mKLF1 and we suggest miRNA mediates actions to induce ?-globin expression through KLF1. Our results show, for the first time, a potential target of miR-210-3p and it is related to the hemoglobin switching.(AU)

Beta talasemia intermedia: características clínicas y estudio molecular. Serie de casos clínicos / Beta thalassemia intermedia: clinical characteristics and molecular analysis. Case series

La beta talasemia intermedia es una hemoglobinopatía de amplio espectro clínico, que surge de la presencia de una o dos mutaciones en el gen HBB, asociada a modificadores genéticos secundarios y/o terciarios. Analizamos las características clínicas y de laboratorio de 29 pacientes con beta talasemia intermedia, evaluados en un período de 23 años. La edad mediana fue de 10,8 años (rango: 0,34-60,4). El 100% de los pacientes mostró anemia microcítica hipocrómica, y solo el 17,2% presentó esplenomegalia y requerimiento transfusional esporádico. El análisis molecular de los pacientes detectó 3 con los dos genes HBB afectados; 2 con un gen HBB afectado y genes alfa cuadriplicados/triplicados; 23 con un gen HBB afectado y genes alfα triplicados; y 1 con dos genes HBB afectados y polimorfismos de genes gama. La correcta identificación de estos pacientes aseguró un adecuado consejo genético y la implementación de controles clínicos regulares.

Beta thalassemiaintermediaisaquantitative haemoglobinopathy covering a broad clinical spectrum, that results from the presence of one or two HBB gene mutations associated with secondary and/or tertiary genetic modifiers. We analyze the clinical and laboratory features of 29 patients with beta thalassemia intermedia, assessed over a period of 23 years. Median age was 10.8 years (range: 0.34-60.4). Hypochromic microcytic anemia was seen in 100% of the patients, while only 17.2% had splenomegaly and occasional transfusion requirement. The molecular analysis of patients detected: 3 with two HBB affected genes; 2 with one HBB affected gene and alpha quadruplicate/triplicate genes; 23 with one HBBaffected gene and alpha triplicate genes and 1 with two HBB affected genes and polymorphisms of gamma genes. The adequate identification of these patients enables us to give appropriate genetic counseling and implementation of regular clinical follow up

Talassemia beta maior e gestação na adolescência: relato de dois casos / Beta thalassemia major and pregnancy during adolescence: report of two cases

A talassemia beta maior é uma doença hematológica hereditária rara em que deficiência na síntese de cadeias globínicas beta causa anemia grave. O tratamento consiste de transfusão sanguínea e quelação de ferro. Descrevemos dois casos de adolescentes com talassemia beta maior, com gestação não planejada e início tardio de pré-natal. Uma delas apresentou piora da anemia, necessidade transfusional aumentada, restrição de crescimento fetal e senescência placentária. A outra apresentava também hipotireoidismo e baixo peso materno, e foi internada por duas ocasiões durante a gestação, por choque hemorrágico do dengue e por infecção respiratória associada a vírus influenza H1N1. Uma delas apresentou restrição de crescimento fetal e teve parto vaginal no termo complicado com hipotonia uterina. Ambas necessitaram de transfusão sanguínea no pós-parto e optaram por medroxiprogesterona como método contraceptivo subsequentemente. Esse relato ressalta a importância de orientação contraceptiva para essas mulheres e o papel do cuidado pré-natal especializado em conjunto com hematologista.

Beta thalassemia major is a rare hereditary blood disease in which impaired synthesis of beta globin chains causes severe anemia. Medical treatment consists of chronic blood transfusions and iron chelation. We describe two cases of adolescents with beta thalassemia major with unplanned pregnancies and late onset of prenatal care. One had worsening of anemia with increased transfusional requirement, fetal growth restriction, and placental senescence. The other was also diagnosed with hypothyroidism and low maternal weight, and was admitted twice during pregnancy due to dengue shock syndrome and influenza H1N1-associated respiratory infection. She also developed fetal growth restriction and underwent vaginal delivery at term complicated by uterine hypotonia. Both patients required blood transfusions after birth and chose medroxyprogesterone as a contraceptive method afterwards. This report highlights the importance of medical advice on contraceptive methods for these women and the role of a specialized prenatal follow-up in association with a hematologist.

Guidelines for screening, diagnosis and management of hemoglobinopathies.

The b‑thalassemias and sickle cell disorders are a major health burden in India. Diagnosis and management of these disorders both in adults and in newborns using appropriate approaches and uniform technology are important in different regions of a vast and diverse country as India. In view of a National Thalassemia Control Program to be launched soon, a need was felt for guidelines on whom to screen, cost‑effective technologies that are to be used as well as for establishing prenatal diagnosis programs in regional centers. Newborn screening for sickle cell disorders is in its infancy in India and uniform approaches need to be followed. Also, included are guidelines for monitoring and managing patients who are now growing older and need comprehensive care as well as management of complications of the disease.

[Effect of self-management empowering model on the quality of life in adolescents and youths with major thalassemia]

Currently, medical advances ensure life-expectancy of chronic patients such as thalassemia. Improvement in the quality of life of these patients requires that medical treatment be combined with psychosocial support. The present study aimed at investigating the effect of self-management empowerment model on the quality of life in adolescents and youth with major thalassemia. This was randomized clinical trial in two groups. 70 Eligible adolescents and youths with major thalassemia were selected through census in Bushehr and Borazjan thalassemia center. Samples were allocated to case and control groups. Data collection tool was quality of life questionnaire [SF-36]. Intervention was performed in three stages. post-test was performed 1.5 months after pre-test in control group and 1.5 months after intervention in case group. Data were analyzed using SPSS version 18 and chi-square, independent t-test, and mann-witney [P<0.05]. There was no statistically significant difference in terms of demographic data between the two groups [P>0.05]. As a result, mean difference of eight dimensions of SF-36 quality of life were statistically significant between case and control groups before and after the intervention [P<0.0001]. Education based on self-management empowerment model has a great impact on the quality of life in adolescents and youth with major thalassemia

Early diagnosis of co-existent ß-thalassemia and alkaptonuria.

Since the aggregate incidence of inborn errors of metabolism is relatively high, a high degree of suspicion is essential to correctly diagnose an inborn error of amino acid metabolism. We report a case of alkaptonuria an autosomal recessive disorder that occurs due to deficiency of homogentisic acid oxidasein a β-thalassemia infant presenting with reddish discoloration of nappies and clothes, breath holding spells, and microcytic hypochromic anemia. Born to consanguineous cousins, to our knowledge, the combination of β-thalassemia and alkaptonuria, which we have described in this baby, has not been reported earlier.

[Screening and identifying of erythrocyte alloantibodies in patients with Thalassemia major referred to Ahvaz Shafa hospital]

Patients with thalassemia major are clinically dependent on red blood cell [RBC] transfusions. Performing multiple transfusions increase the risk of transfusion-related complications including blood-borne infections, iron overload and also RBC alloimmunization. This cross-sectional descriptive study was conducted on 70 thalassemia cases with regular blood transfusions. The serum samples were screened for the detection of the unexpected antibodies and the positive samples were subjected to antibody identification. Among 70 cases, 6 [8.6%] were identified as unexpected alloantibody; three cases as Anti-K, one as Anti-E and another one as Anti-D. Coincidence of Anti D and Anti-E was detected in one case. Eighteen patients [25.7%] were splenectomized. No significant correlation was seen between the presence of alloantibody and age, sex, the time of first transfusion and spleen condition. Considering that the most prevalent unexpected antibodies [8.6%] identified in this study were against the Kell and Rh system antigens, the evaluation of compatibility for antigens found can be recommended before the performing of transfusion. Therefore, this strategy may decrease the possibility of recipient immunization and production of the unexpected antibodies against donor RBCs

Beta-Thalassemia Minor Is Associated with IgA Nephropathy

No abstract available.

Clinical Characteristics of Pediatric Thalassemia in Korea: A Single Institute Experience

Few literatures have elaborated on the clinical characteristics of children with thalassemia from low-prevalence areas. A retrospective analysis was conducted on children genetically confirmed with thalassemia at Seoul National University Children's Hospital in Korea. Nine children (1alpha thalassemia trait, 6beta thalassemia minor, 2beta thalassemia intermedia) were diagnosed with thalassemia at median age of 4.3 yr old with median hemoglobin of 9.7 g/dL. Seven (78%) children were incidentally found to be anemic and only 2 with beta thalassemia intermedia had presenting symptoms. Five children (56%) were initially misdiagnosed with iron deficiency anemia. Despite the comorbidities due to alpha thalassemia mental retardation syndrome, the child with alpha thalassemia trait had mild hematologic profile. Children with beta thalassemia intermedia had the worst phenotypes due to dominantly inherited mutations. None of the children was transfusion dependent and most of them had no complications associated with thalassemia. Only 1 child (11%) with codon 60 (T-->A) mutation of the HBB gene needed red blood cell transfusions. He also had splenomegaly, cholelithiasis, and calvarial vault thickening. Pediatricians in Korea must acknowledge thalassemia as a possible diagnosis in children with microcytic hypochromic hemolytic anemia. High level of suspicion will allow timely diagnosis and managements.

Molecular epidemiology of β-thalassemia in Pakistan: Far reaching implications.

Background: β -Thalassaemia, an autosomal recessive hemoglobinopathy, is one of the commonest genetically transmitted disorders throughout the world. Collective measures including carrier identification, genetic counseling and prenatal diagnosis are required for preventing β-thalassemia. Aim: To achieve this objective, Identification of the spectrum of genetic mutations, especially for various ethnic backgrounds in Pakistan. Therefore, we designed a cross sectional prospective study to identify the frequency of various gene mutations in different ethnic groups of Pakistan. Materials and Methods: Over a 5-year period, DNA from 648 blood samples {including specimens of chorionic villus sampling (CVS)} were analyzed for the twelve most common β-thalassemia mutations found in the Pakistani population by a Multiplex amplification refractory mutation system (ARMS). Each sample was analyzed for the mutation as well as the normal gene, appropriate with negative and positive controls, and reagent blanks. Results: Out of 648 samples mutations were identified in 640 (98.75%) samples by multiplex ARMS. 8 common β-thalassemia mutations were identified in 8 different ethnic groups accounting for 93.9% of the β-thalasemia alleles. Conclusions: Based on the outcome of this study a cost effective proposal is formulated for detection of β-thalassemia mutations.

[Investigating of the effect of familiarzation play on the injective procedural anxiety in B-Thalassemic pre-school children]

During Thalassemia, children should be under frequent blood injections to maintain their lives. The use of intra-vein cutters is one of the most prevalent and painful procedures, and children specially young children usually call it the most stressful aspect of their disease, hospitalization and even their outpatient visit. The present research is an investigation of the influence of familiarization play on the anxiety in the case of injection procedures for children before school age. The present study is a clinical experiment. The research sample consists of 60 Thalassemia children of 3-7 years old, possessing files in Thalassemia sections in Gilan Province. The participants were randomly divided into two groups [experimental group and one control group]. In the experimental group, a familiarization play was carried out for 10-15 minutes prior to the injection procedure. The instruments used in the research include: Demographic Information Questionnaire, the Scale of facial self report of anxiety, and the Questionnaire of child clinical fear self report. The analysis of the data was carried out through SPSS and the statistic test: chi-square, t-test, Mann-whiney and Wilcoxon sign. The data indicated that there was not a significant difference between the average scores of anxiety in the two groups prior to the treatment [P>0.05]. However, after the treatment, the statistic test Mann-whiney and Wilcoxon sign showed a significant difference between the experimental group and the control group in term of the average scores anxiety [P<0.05]. According to the research results, the treatment of familiarization play is influential on anxiety decrease resulted from the injection procedures in children. Therefore it is suggested that child nurses use the method of familiarization play in order to comfort of children with acute and chronic diseases who are frequently under treatment measures