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Background: Heterogeneous mutations in the human coagulation factor IX gene lead to an X-linked recessive bleeding disorder known as hemophilia B. The disease is distributed worldwide with no ethnic or geographical priority.

Materials and Methods: The aim of this study was to characterize the factor IX gene mutations in 28 unrelated Iranian hemophilia B patients. Polymerase chain reaction (PCR) and direct sequencing was performed for all functionally important regions of the gene. Haplotype analysis was performed using three markers.

Results: We identified 24 point mutations and four small deletions (one novel mutation). Overall, 20 different mutations were found and patients with common mutations had identical haplotype.

Conclusion: These data confirm high molecular heterogeneity of the mutations causing hemophilia B and will enable carrier testing and prenatal diagnosis for family members.

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Background: The frequency of pyruvate kinase (PK) deficiency, an autosomal recessive defect, is approximately 3 per 10,000 individuals in Shiraz and surrounding areas, and is increased due to high consanguinity marriage frequency. The purpose of this study is to obtain data on the frequency and spectrum of gene mutation of PK in newborns, from Shiraz and surrounding areas.

Materials and Methods: Two hundred eleven neonates with neonatal jaundice were studied for erythrocyte pyruvate kinase activity using the method recommended by International Committee for Standardization in Hematology (ICSH).

Results: In 22 the PK enzyme activity was below 60%, where the erythrocyte PK activity from 35 healthy cord bloods ranged from 3.9 – 9.8 IU/g Hb. Genomic DNA analysis for PK-R gene mutation was examined in 12 out of 22 cases (heterozygote 3.8%, 95% CI=0.012-0.064 homozygous 1.8%, 95% CI=0.001-0.036). All mutations in four homozygote and 8 heterozygote neonates in this cohort have been reported from the region previously, except for mutation G1675C that has not been found before. Therefore, the rate of recurrence of PK-R gene defect is 4 times more frequent in those with neonatal jaundice in comparison with the regional frequency (0.038).

Conclusion: Erythrocyte enzymopathies, especially PK deficiency, should be considered in the differential diagnosis of non-immune hemolytic anemia as well as for genetic counseling in our area.

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FMS-like tyrosine kinase-3 is a receptor tyrosine kinase expressed by immature hematopoietic cells and is important for the normal development of stem cells and the immune system. Mutations of FMS-like tyrosine kinase-3 have been detected in about 30% of patients with acute myelogenous leukemia and a small number of patients with acute lymphoblastic leukemia. The FMS-like tyrosine kinase-3 mutations most often involve small tandem duplications of amino acids within the juxtamembrane domain of the receptor and are called internal tandem duplications. The other type of mutations in FMS-like tyrosine kinase-3 is located in the activation loop of the second tyrosine kinase domain. Multiple studies have shown that activating mutations of FMS-like tyrosine kinase-3 are common in blasts from patients with AML but are rarely found in adult patients with acute lymphoblastic leukemia. In addition, activating FLT3 mutations occur only rarely in T-ALL cases. Here we report a 3-year-old girl with ALL who had a mutation in FMS-like tyrosine kinase-3 / internal tandem duplications. Keywords: FMS-like tyrosine kinase-3, mutation, acute lymphoblastic leukemia, childhood.

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Background: Beta thalassemia gene mutations are among common mutations in southwest Iran. However, Hemoglobin E (Hb E) and Hb E/β⁰ thalassemia account for a small number of hemoglobinopathies in Iran. This is the first study to directly address the existence of Hb E and consequently Hb E/β⁰ thalassemia in southwest Iran. Methods: This retrospective study discovered seven cases of Hb E/β⁰ thalassemia among 700 patients with hemoglobinopathies referring to Health Institute and Research Center for Thalassemia and Hemoglobinopathy in southwest Iran. EDTA and clot blood samples were obtained and analyzed for complete blood counts, hemoglobin electrophoresis, LDH, bilirubin, ferritin and amplification refractory mutation system (ARMS) technique by polymerase chain reaction (PCR) and DNA sequencing. Results: Out of 700 cases, seven patients with Hb E/β⁰ thalassemia were detected (1%). Four patients were classified into non-transfused dependent Hb E/β⁰ thalassemia and three cases were classified into transfusion dependent Hb E/β⁰ thalassemia group. Alpha thalassemia (deletional and non-deletional) and XmnI gene polymorphism were not found in either of cases. Conclusion: Hb E/β Thalassemia is not a common hemoglobin disorder in southwest Iran. Phenotype heterogeneity is common in Iranian patients from a mild asymptomatic anemia to severe anemia that can be presented in the early years of life. This was the first report of Hb E/β⁰ thalassemia from Iran. Keywords: Hb E/β⁰ Thalassemia, Southwest Iran, Transfusion dependent, Non-transfusion dependent, Hb E mutation.

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Background: Breast cancer is one of the most common malignancies among Iranian women; however, its clinicopathological feature is uncertain. We pioneered a genetic counseling program among patients with breast cancer and their families in Isfahan. This is the first report of this program.
Methods: This was a descriptive cross-sectional study on women with breast cancer registered in Ala Cancer Control and Prevention Center (ACCPC)during 2014. The women and/or their first/second relatives were enrolled for genetic counseling, then their demographic and clinicopathological data were analyzed using SPSSsoftware. 
Results: The records of 258 patients with breast cancer and their families were studied. The mean age of the patients at diagnosis was 44.2 years (range: 25-71 years). Of these, 88 (34.1%) patients had ≤40 years at diagnosis. Only 2 (0.8%) patients were men. Also, 21 (8.1%) out of the 258 patients had died at the time of genetic counseling. Distant metastasis was found in 40 (15.5%) patients at diagnosis. The most common pathological feature of breast tumor was invasive ductal carcinoma (68.2%) and the rarest were sarcoma (0.4%) and papillary carcinoma (0.4%). Triple-negative molecular phenotype breast cancer was reported in 25 (9.7%) patients. Estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) were negative in 32.2%, 27.1%, and 44.2% of the patients’ tumors, respectively. P53 had been checked in 41.5% of the patients of which about 70.1% were mutant. Overall, 895 cases of cancer were reported  among the patients and their families (3.5 patients per family: range=1-9) of which breast, gastric, and colorectal cancers with an incidence of 43.9%, 8.3% and 5.5%, were the most common malignancies, respectively. 
Conclusion: Early-onset breast cancer and positive family history for cancer were seen in a significant proportion of the patients in our center, indicating the importance of genetic counseling among the patients and their families.

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Background: Mutation in NPM1 gene has been reported to be the most common genetic mutation in de novo acute myeloid leukemia (AML). AML with NPM1 gene mutation usually presents with higher initial leukocyte and blast cell counts and negative CD34 expression. We aimed to investigate the difference of initial leukocyte counts, bone marrow blast cell counts and expression of CD34 among patients with AML with and without NPM1 mutation.
Methods: In this study, 25 de novo patients with AML were investigated for NPM1 exon 12 gene mutation using ASO-RT-PCR. Initial leukocyte counts, bone marrow blast cell counts and expression of CD34 on blasts were examined in all patients.
Results: 13 of 25 de novo patients with AML (52%) had NPM1 gene mutation. Initial leukocyte counts in AML patients with NPM1 gene mutation was not significantly higher than patients without this mutation (23.400 /µL versus 16.000 /µL, P=0.53). Blast cell counts were not significantly higher in AML patients with NPM1 gene mutation than patients without mutation. (41% versus 19%, P=0,18). Expression of CD34 was not significantly different between AML patients with and without NPM1 gene mutation (P=0.48).
Conclusion: There were no difference in initial leukocyte count, blast cell count and CD34 expression among patients with AML with and without NPM1 exon 12 type A gene mutation.

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Background: Hemoglobinopathies are the commonest single gene disorder in human that affect hemoglobin production and function that occur when mutations alter the amino acid sequence of globin chains. The purpose of the present study was to evaluate the prevalence of hemoglobninopathies detected by capillary electrophoresis method in individuals referred to Masjed-Soleiman health centers by capillary electrophoresis method.
Methods: This study was carried out on 394 individuals referred to Masjed-Soleiman health centers during 2015-2016. Blood samples were collected in EDTA vacutainer tubes, then CBC including blood indexes (MCV, MCH), level of Hemoglobin A, Hb F, Hb A2 and other hemoglobins were evaluated by Sebia minicap (France) and also genetic tests applied for them to confirm results that were aqcuired by capillary electrophoresis method.
Results: 77 (19.5%) subjects had HbA2 ≥3.5%, thus were classified as beta thalassemia carrier and 3.3%, 2.5%, 1.5% and 0.5% of the individuals were heterozygote for Hb S, Hb D, Hb C and Hb Bart, respectively. Results of the genetic analysis showed the mutations in these subjects; cd36-37(-T) was the most frequent mutation in beta thalassemia carriers in this geographic region.
Conclusion: This study showed high frequency of beta thalassemia mutations in the geographic region of Masjed-Soleiman (19.5), and 7.85% of the individuals had hemoglobin variants including Hb S, Hb D and Hb C detected by capillary electrophoresis. Capillary electrophoresis could be a considerable method for detection of hemoglobinopathies.

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Background: Thalassemia is the most common hereditary anemia which has a relatively high prevalence in Iran. In most cases, more than 300 mutations have been identified, which affect genes of alpha and beta globin chains and lead to lack of production or reduction of chains. Iran’s population is composed of different ethnic groups, thus, determining the frequency and distribution of these mutations is essential in different parts of the country. We aimed to assess Thalassemia gene mutations in Kohgiluyeh and Boyer-Ahmad province. Methods: In this cross-sectional study, 656 couples were selected and their Genomic DNA was extracted by DNA extraction kit method and tested using multiplex gap-polymerase chain reaction (gap-PCR), amplification refractory mutation system-PCR (ARMS-PCR), and DNA sequencing. Finally all data were analyzed using the SPSS version 17 software.
Results: More than 13 mutations were found on α-globin genes. Based on gene frequency, the most common mutant allele was –α3.7/αα (rightward) (71.3%) followed by the two gene deletion −α3.7/−α3.7 (2.5%). Other common mutations were polyA2 (2.1%), αcodon 19α/αα (1.7%), –α3.7/αα/–α3.7/αα (1.5%), – (α) 20.5 (0.6%), α−5 nt/αα (0.5%), and other mutations. In this study, more than 21 mutations were identified on beta thalassemia gene. The most common mutation was CD36- /37 (-T) (19.8%). Other common reported mutations included IVSII-1 (G>A) (9.5%), IVS I-110 (G>A) (4.7%), IVSII-745 (C>G) (4.4%), codon 82/83(-G) (3.7%), FSC 8/9 (+G) (1.7%), Codon19(1.5%), 25 bp deletion (beta0) (1.5%), IVS-I-116 (T>G) (1.4%), IVSI-6 (G>C) (1.1%), codon 5 (-CT) (0.9%), codon 88 (-C) (0.5%), and IVSI-1(G>A) (0.3%).
Conclusion: The frequencies of these mutations were different in various parts of the country. Therefore, defining thalassemia mutations is necessary to establish prenatal diagnosis programs leading to lower medical cost in Kohgiluyeh and Boyerahmad province.

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Background: Hemoglobinopathies are inherited blood disorders with an autosomal recessive pattern. We aimed to evaluate the frequency of mutations of thalassemia and hemoglobinopathies among couples referred to health centers of Izeh in Khuzestan Province, Iran.
Methods: This cross-sectional study was performed on 150 couples referred to Izeh Health Centers in 2015-2018. DNA was isolated from peripheral venous blood samples and then the HBB gene was analyzed by using Sanger sequencing. For molecular analysis of α-globin gene, multiplex Gap-PCR and ARMS-PCR was performed to identify mutations of α-thalassemia. 
Results: DNA analysis revealed 13 different mutations for beta thalassemia in studied couples. Three mutations including 36/37 (- T), IVS-II-1 (G>A) and IVS-I-110 (G>A) accounted for 20.7, 19.3 and 13.3% of beta thalassemia mutations, respectively. For alpha thalassemia; α3.7 (49.5%), -- MED (19.1 %) and -α4.2 (3.1%) were identified as the most common mutations. 
Conclusion: Considering common alpha and beta mutations of this geographic area of Iran could be useful concerning genetic counselling in of the population where the rate of consanguineous marriage is high.

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