فصلنامه خون
سال ششم شماره 1 (پیاپی 22، بهار 1388)

. The aim of this study was to clone and express human recombinant factor VII in CHO cell line as an eukaryotic host cell. Materials and Methods In this descriptive study, FVII cDNA was isolated from HepG2 cell line and cloned to pcDNA 30.1(+) vector. The constructs were transfected to CHO cell line. A cell line that permanently expressed recombinant factor VII was established. The expression of recombinant FVII was determined by RT-PCR, ELISA, SDS-PAGE and western blot analysis. Biological activity of recombinant factor VII was determined by prothrombin time assay in factor FVII-depleted plasma. Results The results showed that FVII was successfully cloned and expressed. After 3 weeks stable cell lines were generated in the culture of the CHO cell line in the presence of geneticin. RT-PCR, ELISA, SDS-PAGE and western blot analysis results indicate the expression of FVII in the stable clones. A three- to four-fold decrease of the specific coagulant activity of rFVII was observed that indicates of rFVII being biologically active. Conclusions Four thousands to Six thousands vials of rFVII were imported to our country having high cost for the government. Therefore, production of rFVII through recombinant DNA technology within lab scale is the first step to overcome the problems.

Existence of leukocytes in all blood products causes a wide variety of side effects after transfusion. As a consequence, the use of filter technology for leukoreduction has been widely practiced. In this study, absolute leukocyte count in three types of home-made bedside filtered packed cell units is evaluated. Materials and Methods Ninety three packed cell units from blood donors were prepared in Tehran Regional Educational Blood Transfusion Center, stored at 4ºC, and filtered by two types of home-made filters within 1 hour at room temperature. The first type of filters was made prior to 2007 and the second type underwent optimization after 2007. Eight samples were filtered by control group filters with CE certificate. The results were analyzed with SPSS 11.5 and chi-square test. Results The mean values of leukocyte count/unit by CD45 and True Count Method were respectively 9×106 and 10×106 in 55 bags filtered by the first type filters; and these values were 4.2×106 and 4.8×106 in 30 bags filtered by the second type filters, whereas the mean value of leukocyte count/bag in 8 bags filtered by control filters was 2.3×106

Background and Objectives Acute Promyelocytic Leukemia (APL) is one subtype of acute myeloblastic leukemia; it responds to differentiation using All-Trans Retinoic Acid (ATRA). Recently, arsenic trioxide (As2O3) has been added to this method. The cellular mechanism of differentiation therapy by arsenic is not yet clear. We decided to study the relationship between cell differentiation using arsenic trioxide and nucleostemin gene as a proliferation marker. Materials and MethodsIn this descriptive study, we treated NB4 cell (a cell line in APL) with 0.5, 1, and 2 µM of arsenic trioxide in 6 well plates for five days. Then, cellular differentiation was assessed by flowcytometry for CD11b. Nucleostemin gene expression was also assessed by Real Time PCR. ResultsAccording to the results, cell proliferation has occurred by 0.5 µM arsenic trioxide and no differentiation was observed during 10 days of culture. With 1 µM concentration of arsenic, CD11b has raised from 5.2% to 13.6% during five days of culture (p < 0.001). Morever, 1 µM of arsenic caused decrease in nucleostemin gene copy number from 130 to 70. ConclusionsAccording to the results, 1 µM of arsenic has increased CD11b in the cells and caused a partial differentiation during five days of culture. Increase in CD11b marker has also been associated with decrease in nucleostemin gene expression as a proliferation marker.

Background and ObjectivesIt is suggested that HA-1 mismatching among hematopoietic stem cell recipients-donors be associated with acute graft-versus-host disease (aGVHD). So the aim of this study was to evaluate HA-1 frequency and examine the correlation between HA-1 disparity and GVHD patients who received transplantation from HLA-A2 identical siblings. Materials and MethodsExtracted DNA samples were collected from 55 HLA-A2-positive donor-recipient pairs. All the patients received peripheral blood stem cell transplant (PSCT) from HLA-identical siblings. HA-1 was detected by SSP-PCR method. The HA-1 typing was performed using SSP method. Data were analyzed using Chi-square, Man withney and Z test with SPSS 11.5. ResultsThirty patients showed to be GVHD I-IV and 25 pairs were without any GVHD signs. The frequency rates of HA-1R and HA-1H alleles in patients were 0.55 and 0.45, respectively; it showed no significant difference with the frequency rates (0.53 and 0.47) of this allels in donors (p>0.05). HA-1 disparity was detected in 8 out of the 55 donor/recipient pairs (14.5%). aGVHD (grades I-IV) was occurred in 6 of patients. Two patients with HA-1 disparity did not show any GVHD signs. X2 test showed there was not any relationship between the incidence of acute graft-versus-host-disease (aGVHD) and HA-1 incompatibility in the patiens. ConclusionsIn spite of higher frequency of HA-1 disparity in GVHD+ group, our data did not reflect any significant association between HA-1 disparity and risk of acute GVHD.

Background and ObjectivesMultiple myeloma (MM) is a clonal disorder of hematopoietic stem cell. We have been doing in-patient stem cell transplant (SCT) in Iran since 1991; however, this is the first time we have decided to embark on out-patient SCT. Materials and MethodsIn this cohort study, the selected Multiple Myeloma patients received outpatient stem cell transplantation. They were then discharged and followed by an out-patient SCT team including a general physician, a staff nurse and a care giver during the neutropenic period. All data were collected and analyzed using ANOVA test, Caplan mayer curve and SPSS 11.5. ResultsForty four patients received in-patient or out-patient autologus SCT. The rates of median hospital stay were 28 (19-54) and 6.5 days (1-8) in in-patient and out-patient groups, respectively. Median home visit by team was 10.5 days. There were not significant differences (p<0.001) between these groups as far as apheresis days, granulocyte colony stimulatig factor (GCSF) dosage as mobilization, number of mono nuclear cell (MNC) or cluster of differentiation (CD)34+ cell parameters are concerned. There was also a significant decrease in total cost of SCT in out-patient group by 70% (p<0.017) including visit cost with 80% decrease (p<0.01), drug cost with 50% (p<0.004), laboratory cost with 70% (p<0.02), and hospital cost with 70% (p<0.04). ConclusionsOur results show that out-patient autolgous SCT in multiple myeloma patients is feasible and its complications are manageable. Significant reduction in cost and bed requirement is also inevitable.

Background and ObjectivesHepatitis A virus (HAV) is a hepatotropic virus and a member of the Hepatovirus genus within the Picorna-viridae family. Among HAV-infected individuals, children under 6 years old are typically asymptomatic, whereas older children and adults develop jaundice. Studies carried out in different regions of the world have demonstrated a reduction of HAV infection in children under five and a gradual increase among adolescents and adults. The frequency of acute hepatitis A infection was investigated among different age groups by molecular and serological procedures in this study. Materials and MethodsThis research was a cross sectional study. During the period of 12 months, from October 2005 to 2006, a total of 308 patients with a diagnosis of Hepatitis A were admitted and analyzed at Tehran Regional Educational Blood Transfusion Center in Tehran. The serum samples were tested for viral hepatitis (HAV, HBV, and HCV). Of these samples, 62 were positive for HBV or HCV and were excluded from this study. The 246 remaining serum samples were included. Serum specimens were tested for IgM anti-HAV antibody using a commercial enzyme immunoassay. Differences in the prevalence of IgM anti-HAV among different age groups were analyzed using the chi-square test. HAV RNA was detected by nested Reverse Transcription-Polymerase Chain Reaction (RT-PCR). ResultsIgM anti-HAV antibodies were detected in 29 (11.8%, CI 95%= 7.77-15.83) of 246 patients. The frequency rates of IgM anti-HAV antibodies in the studied population were 17.8% within the age range of 0-10 years, 23.1% within the range of 11-20 years, 14.7% within the range of 21-30 years, 3.2% within the range of 31-40 years, and 0% within the age range of above 40 years. Significant difference was observed between different age groups. The frequency of IgM anti-HAV was higher within the age range of 11-20 years (23.1%) as compared to other age groups. HAV RNA was detected in 25 of 29 (86.2%) serum samples. ConclusionsWith improvement in sanitary conditions, it is necessary to perform new studies on HAV infection prevalence and incidence in different age groups of the country. These data will be useful for future vaccination planning.

Background and ObjectivesThalassemia is the most common hereditary anemia in Iran. Despite improved hematologic care, multi endocrine dysfunction is a common complication in these patients. The main objective of this study was to estimate the prevalence of thyroid dysfunction in patients including both thalassemia major and thalassemia intermedia. Materials and MethodsIn this descriptive cross sectional study, a questionnaire was designed for 195 patients who were reffered to Iranian Blood Transfusion Organization Clinical Laboratory of Tehran from Thalassemia Clinic in Winter 2007. The following items were brought up in the questionnaire: sex, age, height, weight, splenectomy time, amount of transfused blood, blood transfusion interval, desferoxamine dosage, type of thalassemia (major or intermedia), serum thyroid hormones, and ferritin levels. Then, the correlation of thyroid functional status with age, serum ferritin level, type of thalassemia, splenectomy and desferoxamine dosage was evaluated in both β thalassemia major and thalassemia intermedia groups. ResultsWe had 178(91.3%) β thalassemia major (50.6% male, 49.4% female with the mean age of 17.2 ± 8 years) and 17(8.7%) thalassemia intermedia (23.5% male, 76.5% female with the mean age of 23.2 ± 8.8 years). One hundred sixty two (83.1%) patients were euthyroid, 27 (13.8%) had subclinical hypothyroidism (CI 95%= 9-18.6), and 6(3.1%) were primary hypothyroid (CI 95% = 0.7-5.5). Mean ferritin levels for euthyroid group were 1923 ± 1470 ng/ml, for subclinical hypothyroidism group 1723 ± 1346 ng/ml, and for primary hypothyroidism 1569 ± 734 ng/ml, respectively. No significant correlation was found between abnormal thyroid function (subclinical and primary hypothyroidism) and serum ferritin levels (p=0.55), age (p=0.11), type of thalassemia (p=0.68), splenectomy (p=0.62) and desferoxamine dosage (p=0.33). ConclusionsBased on our results, thalassemia patients present a sort of thyroid dysfunction such as hypothyroidism. So, more effective treatment with desferoxamine and regular follow up of patients for evaluation of thyroid function satus are recommended.

Background and ObjectivesHBV infection is one of the most common infections with side effects. Clinical personnel are at risk for HBV infection; so immunization with vaccine and detection of HBs Ab level are very important. Materials and MethodsThis study is a descriptive analytic research on 501 clinical personnel that had received HBV vaccine. HBs Ab titres were estimated with standard kits and the final analysis was made by using X2, simple scatter methods and SPSS 16. ResultsOut of 501 patients, 293 (58.5%) were female and 207 (41.5%) male. The age average was 34.38 ± 8.65 years and the average BMI was 24.13 ± 3.17. The average HBs Ab titre was 58.13± 12.3 mIU/ml and the average HBs Ab titre was higher in women than men (p<0.05). ConclusionsImmunization with HBV vaccine is very necessary for clinical personnel especially hospital care workers. Moreover, HBs Ab titre of such cases should be regularly evaluated.