Iranian Journal of Blood and Cancer
Volume:1 Issue: 2, Winter 2009

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

The capacity of human HbAA erythrocytes of non-malarious blood to withstand osmotic stress in the presence of five antimalarial drugs، Chloroquine phosphate، Quinine، FansidarTM، CoartemTM and HalfanTM was studied in vitro. Aqueous solutions of four increasing concentrations of the drugs used in this investigation were in the order: 0. 2%، 0. 4%، 0. 6% and 0. 8% (w/v). Spectrophotometric method was employed to ascertain the osmotic fragility index of the erythrocytes. The mean corpuscular fragility (MCF) index (X+S. D) of the control sample was 0. 351+0. 06 g/100ml. The MCF values of the control and test samples were statistically compared (p value = 0. 05). At the drug concentration of 0. 8g، MCF values (g) of 0. 401±0. 005، 0. 391±0. 003، 0. 364±0. 02، 0. 344±0. 02 and 0. 338±0. 04 were obtained for halfan، coartem، quinine، chloroquine phosphate and fansidar، respectively. An overview of the results showed that in a concentration dependent manner، Choroquine phosphate and FansidarTM exhibited a diminishing capacity to stabilize red blood cell membrane while HalfanTM CoartemTM and Quinine elicited an increasing propensity to disrupt erythrocyte membrane integrity

Studies have demonstrated that sickle cell trait can be found in an asymptomatic healthy carrier with normal complete blood count (CBC) and red blood cell (RBC) indices. According to Iranian Ministry of Health bulletin instructions, prenuptial Thalassemia Screening Program (TSP) primarily depends on RBC indices which are measured through a routine CBC. Only when these levels are below the standard values in both couples, hemoglobin electrophoresis is performed. Because of normal values of RBC indices in the sickle cell trait, prenuptial TSP might miss it. To approach and prove this hypothesis, we carried out a prospective randomized pilot study of one year duration on 50 known cases of sickle cell trait (the parents of sickle cell and sickle-Thalassemia referral cases to Ahvaz Sickle Cell Center, affiliated to Ahvaz Jondishapour University of Medical Sciences). An informed written consent was obtained from all patients. Complete blood count was done including a blood film, and CBC was done by Helena Automated Cell Counter. Sickle preparation was done by freshly made sodium bisulfate 2%. It was read after one hour for the first time and was then read after 24 hours by macroscopic and microscopic methods. Hemoglobin electrophoresis was done by Helena apparatus. Serum ferritin was measured by Immunoradiometric Assay (IRMA). Iron deficient cases were treated by oral Iron agents Data of the first screening line showed MCV>80 and MCH>27 in more than 94% and HbA2 value of less than %3 in %100 of sickle cell trait cases. Sickle cell trait with normal indices might be skipped through pre-marriage TSP, and their carrier status is not determined without performing hemoglobin electrophoresis. We recommend mandatory hemoglobin electrophoresis and sickle preparation to be added to the first line of prenuptial Thalassemia Screening Program in the sickle target zones such as the southern parts of Iran.

The present study was carried out on the human population of Kashmir valley to evaluate the status of the biochemical parameters of infected population. Blood samples were collected from 514 individuals, 298 (57.97%) males and 216 (42.02%) females, 187 (36.38%) were found sero-positive for human toxocariasis. Alkaline phosphatase level was found higher in infected children and adults than in uninfected population. Serum bilirubin concentration was not affected by Toxocara infection. The mean value of serum creatinine in uninfected persons was similar to that of infected persons. There was no effect of Toxocara infection on blood urea level in infected persons. The mean value of blood urea in uninfected persons and infected persons was normal. The blood glucose level was not affected by Toxocara infection. From above results alkaline phosphatase level was only found to be affected in Toxocara infection.

The frequency of the multi-drug resistance 1 (MDR1) gene C3435T polymorphism differs in various ethnical populations such as Asian, African, and Caucasians populations. A silent C3435T polymorphism in exon 26 of MDR1 has been reported to be associated with a decreased expression of P-gp in TT genotypes carriers compared with CC genotypes carriers. To evaluate the association between MDR1 gene C3435T polymorphism and acute lymphocyte leukemia (ALL), 126 ALL patients (72 males and 54 females) with a mean age of 11.42 ± 6.55 and 139 healthy controls (79 males and 60 females) with a mean age of 12.15 ± 7.5 who were referred to Dr. Sheykh hospital, Mashhad, Iran, between 2005-2007 were enrolled in our study and their C3435T MDR1 polymorphism was investigated using PCR-RFLP. The mutant homozygous TT and TC genotypes were found to be associated with the incidence of ALL (p<0.05). There was no significant difference for T allele frequency between ALL patients and healthy controls (OR=1.08, 95% CI; 0.84-1.66, p=0.33). TT genotypes carriers are at higher risk of developing ALL than carriers of other genotypes.

The Rh antigens are expressed as parts of a protein complex in the RBC membrane. This complex is a tetramer, consisting of two molecules of RhAG and two molecules of Rh proteins. To express RhD in RBC membrane, expression of RhAG is essential. This co-expression only occurs in the erythroid lineage. K562 cell line has an erythroid lineage. Cord blood was used as a source of RHD gene in which nucleated RBCs are rich. Mononuclear cells were isolated using Ficol method. RNA was extracted by trizol followed by cDNA synthesis. RHD gene was isolated with specific primers. The RHD cDNA was ligated to pcDNA3.1 vector and cloned into E. coli. The recombinant pcDNA-RHD construct was transfected to K562 cell line. Stable cells expressing RhD were selected in the presence of geneticin. RT-PCR and western blot analysis were performed to detect recombinant RhD. Stable cells expressing recombinant RhD were established. RT-PCR results showed exogenous expression of recombinant RhD which was further confirmed by western blot analysis. Overall, our results revealed that K562 is suitable for expression of RhD. The recombinant RhD may be helpful to further investigate the molecular basis of RhD protein.

Although rare, synovial sarcoma is the most common malignant non-rhabdomyosarcomatous soft tissue sarcoma in children and adolescents. Synovial sarcoma typically involves the soft tissues of the extremities, especially near large joints, but it can occur anywhere in the body in locations far from joint spaces. Although this tumor typically affects adults in their fourth decade of life, nearly half of the reported cases have been children and adolescents. We report a rare case of head and neck synovial sarcoma presented with tonsilar hemorrhage and painful facial contracture. Cervical computed tomography (CT) scan with contrast injection showed an asymmetrical respiratory tract image. Significant thickening of right (anterior) parapharyngeal soft tissue was revealed indicating a soft tissue mass. The mass was completely resected in a surgical procedure. In pathological examination of the mass, biphasic synovial sarcoma was reported. At first, we treated the patient with VIE (Vincristin, Ifosfamide, Etoposide),but because of the severe neutropenia and hemorrhagic cystitis, we changed the protocol to VAC (Vincristin, Actinomycine, Cyclophosphamide). The girl is 6 years old now and in a good condition without any pain or bleeding. Also, she has a normal cervical CT scan after more than two years. She is followed up every week and receives monthly Vincristin.

Dermatomyositis (DM) is a rare idiopathic inflammatory myopathy with characteristic skin lesions. Case series have shown an association between dermatomyositis and malignancy. Malignancy has been found in 15-25% of the adult patients with dermatomyositis. A 50-year-old new case of breast cancer, was admitted with muscle weakness and inability to walk. Physical examination revealed fever, periorbital edema, a heliotrope rash on the upper eyelids, a large right-sided breast mass and erythematous plaques on the upper chest, abdomen and legs. During hospitalization, she developed a nasal speech, dysphasia, and nasal regurgitation of food and water. A clinical diagnosis of dermatomyositis secondary to breast adenocarcinoma was made according to the skin lesions, muscle weakness, elevated ESR, and increased CPK and LDH levels. Wide spectrum antibiotics and Dexamethasone were administered but fever persisted. Although chemotherapy was recommended, she developed respiratory failure and aspiration pneumonia and died. Dermatomyositis is one of the paraneoplastic syndromes which are associated with breast cancer. Although treatment of dermatomyositis generally includes corticosteroids with or without immunosuppressants, cancerdirected specific therapy including surgery and/or chemotherapy would be more effective. Dermatomyositis should be considered in breast cancer patients with skin lesions and muscle weakness and cancer-specific therapy should be started as soon as possible.

Hemophilia B is a bleeding disorder with a recessive X-linked inheritance pattern, in which the infected individuals have low levels of factor IX in their plasma. Affected individuals may have bleeding episodes after trauma or spontaneously considering the plasma level of factor IX. In order to prevent these episodes and to control bleeding, they should use coagulation factor concentrates that may be associated with the formation of inhibitors. This study was conducted in the northeast of Iran in 2006. Among 48 individuals who agreed to participate in our survey, 3 individuals (6.25%) had used FFP, 38 s (79.16%) factor IX concentrate and 7 (14.58 both FFP and factor IX concentrate in the 6 months prior to the study. Of them, three participants (6.3%) had factor IX inhibitor which was assayed using Bethesda method. Three hemophilia B (6.3%) patients had factor IX inhibitor, but no correlation was found between the existence of the inhibitor and the type of coagulation therapy. Our findings did not show any correlation between factor IX inhibitor and type of coagulation therapy used in the 6 month period (p=0.65). None of the hemophiliacs had used coagulation factor as a prophylaxis regimen and most of them (83.33%) had injected coagulation factor on demand.