فصلنامه خون
سال نهم شماره 4 (پیاپی 38، زمستان 1391)

Arsenic trioxide (As2O3) has been shown to have anti-cancer effects on a wide range of cancers. As2O3 has proved to be the most efficient in treatment of acute promyelocytic leukemia (APL) patients. It primarily acts by triggering apoptosis in cancer cells. To better understand the molecular mechanisms involved in As2O3 -induced apoptosis، in the present study we examined the effects of As2O3 both on the expression of Bax and Bcl-2 (the major regulators of apoptosis)، and p53 genes in HL-60 cell line. In this experimental study، the MTT colorimetric assay was used to assess the growth-inhibitory effect of As2O3 on HL-60 cells. The mRNA levels of p53، Bax and Bcl-2 were analyzed by using Real-time PCR. Data were analyzed by using SPSS18، t-test and ANOVA tests. As2O3 inhibits the growth of HL-60 cells in a dose dependent manner at 24h. Significant decrease (89% ± 3%) in cell viability was observed at 4μM and the IC50 value was about 16 μM. In HL-60 cell، no expression of p53 was detected. Real-time PCR results showed while Bax mRNA expression was not significantly affected in As2O3 treated HL-60، the Bcl-2/Bax ratio expression level significantly decreased in a dose dependent manner. As2O3 exerts part of its antitumor effect by inducing apoptosis through decreasing Bcl-2/Bax mRNA ratio in p53-null leukemic cell line HL-60. More research on the protein level is required for these results to be reinforced.

Having a better understanding about blood use in trauma care helps us plan to ensure adequate blood and design new protocols to avoid blood loss. In this cross-sectional study، 362 of traumatic patients having received blood transfusion in Rajaie Trauma Center in 2011 were randomly selected. Through a questionnaire eliciting information about demographic characteristics، the amount of administered packed RBCs، platelets، and fresh frozen plasma (FFP)، the prognosis in patients who received more than 10 packed RBC units with patients who received less than 10 was compared. We used t-test and Chi-squared test for data analysis. The average units of RBC، platelet and FFP administered per trauma case were 3. 31 ± 3. 74 (1-25)، 0. 92 ± 5. 15 (0-76)، and 2. 59 ± 6. 6 (0-55)، respectively. The mean number of RBC units transfused did not correlate with sex، type of injury، age، and patient transport time (p> 0. 05). The mean number of blood units used was higher for patients with lower Glasgow scores and for those who died after their injury (p< 0. 05). The mortality rate، the mean number of FFP and platelet units transfused، and the need for surgery were higher in patients with massive transfusion (p< 0. 05). Given the positive correlation of morbidity and mortality rates in traumatic patients with the amount of blood transfusion، it seems imperative for blood to be used appropriately and judiciously. Designing massive transfusion protocols can lead to better blood usage، decline complications، and ensure blood adequacy.

Biologival features of adenoviruses have made them vectors of choice for gene transfer in basic research as well as in clinical treatment and vaccination. The key steps in the trasnsfer of adenovirus vector gene are transfection monitoring and viral titer determination. We investigated the transfection of non-reporter adenoviruses as well as viral titration using molecular methods. In this experimental study، after the recombinant adenovirus vector construction، the presence of transgenes in gene construct was evaluated by PCR with transgene specific primers and sequencing. After calcium-phosphate transfection of the packaging cell line (HEK 293A)، transfection was monitored by PCR using transgene specific primers (diagnostic and compound primers) to analyze DNA extracted from the transfected packaging cell line. The viral titer was determined by Real Time PCR (RT-PCR) using diagnostic primer to analyze DNA extracted from viral lysates. The presence of transgene in gene construct was confirmed by transgene specific primers and sequencing. Evaluation of DNA extracted from transfected packaging cells confirmed the efficiency of monitoring of transfection. The viral titer was estimated to be about 4 × 107 viral particles/ml by RT-PCR. Careful primer design is the key to the efficient monitoring of non-reporter adenovirus vector transfection. The appropriate genomic purification and the use of RT-PCR technique would also avoid the disadvantages of functional titration of adenoviruses.

von Willebrand Disease (vWD) is the most common inherited haemorrhagic abnormality. The disease is caused by deficiency of von Willebrand Factor (vWF). vWF is encoded by vWF gene and vWD type3 is the severest form of the disease inherited in autosomal recessive pattern. In the present study، the impact of various vWF gene mutations on mRNA expression was investigated in vWD patients. Blood samples were obtained from vWD patients with known mutations to determine the vWF gene expression level. Peripheral blood platletes were isolated and the total RNA was extracted. To evaluate the vWD gene expression of patients and carriers، the quantitative real-time PCR of the platelet cDNAs was performed. Patients in families with the nonsense mutation in exon 35 showed significantly lower levels of vWF mRNA compared to the hetrozygote carriers of the mutation and normal controls (p=0. 005). However، there was no significant difference between vWF mRNA levels in patients and carriers in the other families who had splice site mutation in exon 10 (p=0. 288). The impact of the mutation on the protein synthesis and function has yet to be investigated. Conclusions Each mutation in vWF gene has its particular impact on the gene expression. It has been shown that even mutations of the same type may have different effects on mRNA expression. Evaluation of cDNA is not applicable in the cases in which the mRNA is affected by NMD pathway.

Bacterial contamination of platelet units is an important cause of transfusion-associated morbidity and mortality. Automated bacterial blood culturing system satisfies many of the requirements of an ideal test. In this research we investigated several factors affecting detection by automated culture of bacteria in platelet units. In this study E. coli was selected to model a fast growing organism and Staphylococci epidermidis was used to model a slow growing organism. Staphylococci epidermidis and E. coli were inoculated into freshly prepared platelet units to yield 10 CFU/Ml. At the time of inoculation t=0، t=6، t=24، and t=48 hours، 0. 5، 1. 0 and 2. 0 ml samples of the contaminated platelet units were transferred into culture bottles. E. coli was detected in 100% of experiments when 0. 5، 1. 0 or 2. 0 ml samples were taken at t=0، t=6، t=24، and t=48 hours. For Staphylococci epidermidis 83. 3% of contaminated platelet units was detected when 0. 5 or 1. 0 ml samples were taken at t=0 hours and 91. 6% of units was detected when 2 ml samples were taken at t=0 hours. Staphylococci epidermidis also was detected in 100% of units when 0. 5، 1. 0، or 2 ml samples were taken at t=6، t=24، and t=48 hours. The data from this preliminary evaluation suggest that sampling times of 0 and 6 hours and 0. 5 ml sampling volume are suitable to provide confidence in detection of E. coli and Staphylococci epidermidis in platelet units using this culture method.

Hemoglobinopathies are the most common single-gene disorders in Iran. Sistan and Balouchistan has 2300 hemoglobinopathy patients who judiciously receive blood in the southeast of Iran. In this study، the prevalence rate of hemoglobinopathies was evaluated in the province. This descriptive analytical study was performed on 2129 people at the age range of 5-10 years who were selected by multistage cluster sampling method. The number of samples in each city was proportional to its population size. Hematologic parameters were evaluated by automated cell counter on EDTA whole blood، hemoglobin A2 (HbA2) was determined by column chromatography، and hemoglobins were separated on Acetate Cellulose at pH= 8. 6. The samples with abnormal hemoglobins underwent electrophoresis at pH= 6. 1. The results indicated 9. 7% affected with minor thalassemia، with the frequency being variable between 6. 2% in north cities to 12. 9% in south cities of the province. A total of 0. 9، 1. 7 and 0. 1% were heterozygous for hemoglobin S، D، and C، respectively. HbA2 level of 72. 4% of heterozygous hemoglobinopathies was estimated to be within the range of healthy individuals. The prevalence rate of β-thalassemia gene in the southern region of the province was higher than the average national rate. Current pre-marriage screening protocol is not able to identify all hemoglobinopathies in heterozygous cases. It is recommended where hemoglobinopathies are common، hemoglobin S detection be included in the screening program.

Based on the previous researches performed in Iran and worldwide، there is a significant number of subjects carrying alpha-thalassemia gene defect. Therefore، we aimed to assess its frequency amongst a random population selected from premarriage couples who entered the national prevention program of β -thalassemia major at Tehran. Blood samples were collected from 625 randomly selected individuals following obtaining written informed consent through eight cooperating primary health care centers (PHC) in Tehran. Primarily، CBC testing was performed. Subsequently، blood samples underwent DNA extraction and gene-specific amplification using ARMS PCR and Multiplex Gap PCR followed by DNA sequencing of negative samples to identify potentially missed alpha-globin gene mutations. According to CBC results، 147 individuals were considered normal; 479 remaining blood samples underwent molecular assessment which revealed that 95 samples possessed a minimum of one deletional mutation in the alpha-globin genes، resulting in a frequency of 19. 83% in the tested population (CI 95%; 11. 81-27. 85). Furthermore، 50 samples with normal hematological indices (MCH>27، MCV>80) were tested for triplication mutation and revealed to be negative. It has previously been established that Multiplex Gap PCR is capable of detecting 65% of alpha globin gene defects in blood samples with abnormally low hematological indices (MCH<27، MCV<80). Hence، using bio-informatic techniques with generalization of our results to the entire population of Tehran showed the true frequency of alpha-globin gene defect to be actually 29. 90% (CI 95%; 22. 40-37. 40) at the population level.

Major thalassemia is the most common hereditary anemia in Iran. The treatment of the disease is regular blood transfusion which in turn can cause side effects. This study tries to determine the prevalence of diabetes، thyroid and parathyroid diseases in thalassemia patients for the purpose of early diagnosis and treatment. This retrospective cross-sectional study was conducted on 206 thalassemia patients having been referred to Shahid Bahonar Hospital during April 2009 to April 2010. Test results for laboratory parameters of FBS، T4، TSH، Ca، P، PTH and Ferritin were extracted from patients’ medical records. Out of 206 patients، 90 (43. 7%) were male and 116 (56. 3%) female، with a mean ± SD age of 19. 64±7. 06 years. Out of the total number، 186 patients were tested for fast blood glucose; 23 patients (12. 37%) were pre-diabetes and 19 (10. 22%) had diabetes mellitus. Out of 184 patients tested for thyroid function، 38 patients (14 male، 24 female) (20. 65%) had abnormal thyroid test results and 35 were suffering from subclinical hypothyroidism and 3 from clinical hypothyroidism. One (1%) out of 96 patients who were checked for PTH was diagnosed with hypoparathyroidism. Endocrine disease is common in multi-transfused thalassemia patients، and preventive measures such as endocrine tests and other control measures are crucial.

There are different causes for anemia 50 percent of which attributed to iron deficiency. There is considerable evidence، though somehow contradictory، related to the negative effect of iron deficiency on endurance performance. Thus، the present study aims to evaluate the effect of iron supplement intake on aerobic power. In a semi-experimental test، 37 girl students within the age range of 14-17 years randomly selected were divided into three groups of pre-latency، latency with two different iron deficiency levels، and control; difference in iron deficiency level was the main factor for the division of the first two groups. Subjects underwent Bruce maximal aerobic power test before iron supplement intake; then، pre-latency and latency groups received two 50 mg ferrous sulfate tablets as iron supplement on a daily basis. After 45 days، the above tests were conducted again. The mean aerobic power increased from 31. 59 ± 6. 45 to 36. 28 ± 5. 49 in the control group، from 22. 75 ± 7. 76 to 32. 82 ± 4. 58 in the pre-latency group، and from 24. 16 ± 7. 67 to 34. 23 ± 4. 57 in the latency group. The mean serum iron decreased from 119. 50 ± 25. 89 to 97. 83 ± 30. 10 in the control group while it increased from 84. 25 ± 15. 75 to 110. 41 ± 51. 10 in the pre-latency group and from 40. 92 ± 16. 83 to 93. 38 ± 49. 80 in the latency group (p≤ 0. 05). There was shown to be a significant difference between the latency group and the control for the mean differences before and after the supplement intervention. It is shown that those with iron deficiency can reach the normal serum iron level after supplement intake but no significant difference seen on their aerobic power compared to the control group.

Blood bank refrigerators play the main role in blood safety and appropriate blood production. Confirmation of their functions to maintain temperature stability with a PM program is compulsory. The main objective of this study is to identify the level of maintenance and regional climate effects on blood bank refrigerators in different blood centers. In order to study the level of maintenance and possible regional climate effects on each blood bank refrigerator، a survey on 85 local refrigerators in seven regional blood centers was carried out. In this survey، along with the amount of temperature leakage from the body and glass insulators، the quality of ribbons used was also evaluated. The results obtained from this study showed the degree of temperature leakage on bodies of the fridges، metal doors، and glasses to be varied due to different factors like room humidity and temperature changes، air flow around refrigerators، and spaces between refrigerators and other appliances or wall. This study contributes to a higher knowledge level of local manufacturers on how better to produce blood bank refrigerators to be compatible with different climate conditions of Iran.

Thalassemia represents a group of recessively inherited hemoglobin disorders and is characterized by reduced synthesis of beta globin chain. The homozygous state results in severe anemia which needs regular transfusion every 3-4 weeks. Frequent blood transfusion can lead to iron overload which may result in growth retardation in thalassemic patients. In this descriptive-cross sectional study، we measured weight and height of 100 patients with beta thalassemia major within the age range of 2-18 years old who were under follow up in Dr. Cheikh Hospital. The results were then compared with the control group (340 healthy children). The mean and standard deviation values of hemoglobin and ferritin were 8. 5 ± 1. 5 g/dl and 2183 ± 525 ng، respectively. Short stature was seen in 53. 4% of girls and 56. 4% of boys. Short thrunk was reported in 73. 3% of girls and 74. 5% of boys. Underweight was reported in 48. 9% of girls and 52. 7% of boys. Growth retardation is common in thalassemic patients. Therefore، height and weight should be routinely and frequently assessed and in the case of growth retardation the cause should be investigated. Common causes of growth retardation are chronic anemia، hypoxia، and iron overload.

Studies on the relationship between maternal hemoglobin (Hb) level and adverse pregnancy outcomes have been inconsistent. The aim of this study was to examine the birth outcomes in pregnant adolescents as high risk mothers. In this cross sectional study، 312 healthy pregnant women with GA=37-40 weeks were selected by random sampling. After their medical records were taken and clinical investigation done، Hb tests were conducted. After birth، the correlation between the levels of Hb concentration during pregnancy and birth outcomes was estimated. Statistical analyses were performed with SPSS software using t-test، chi-square، and ANOVA. The prevalence rate of anemia was 23. 2 %. The average birth weight was 3197. 8 ± 398. 25 g and it was higher in mothers with Hb=10. 5-12. 5 g/dL than others (10. 5 > Hb > 12. 5 g/dL) (p< 0. 01). Apgar score less than 8 was higher in the anemic group. Our findings demonstrated that abnormal Hb concentrations increased the risk of adverse birth outcomes in pregnant adolescents. Thus، the intensive care is recommended for them.

Hepatitis B is one of the most dangerous causes of liver infection in the world، and the first known factor responsible for post transfusion infections (PTI). Thus، all donated units and blood products are tested for HBV. HBV genotypes are divided into eight groups with genotype D being dominant in Iran. Occult hepatitis B infection (OBI) is common worldwide and the possibility of transmission of hepatitis B infection is a cause of concern in blood transfusion centers. The present article attempted to review articles published in association with hepatitis B and blood transfusion in Iran to show an image of hepatitis B in Iran compared to the world. Studies done in Iran have shown varied statistics of hepatitis B infections. In thalassemics، hemodialysis patients، and intravenous drug users، 1. 5%، 5. 1% and 5. 8% were HBsAg positive، respectively. OBI rate also varies from 0. 15 to 1. 5 percent. The levels of HBsAg in different reports were shown to be 0. 47%، 0. 41%، 0. 23%، 0. 4% and 1. 08% among blood donors; however، the rate of infection was very low in regular donors. Given the importance of the screening of hepatitis B in blood donors، using sensitive diagnostic tests، especially molecular tests، for reduction in the window period is critical.

In animal models، the results of studies have shown the efficacy of stem cells in repairing tissues and curing different diseases; however، in humans the effectiveness of cell therapy has proven in blood disorders and the immune system. Different phases of clinical trials on the other uses of stem cells are also underway. In this study، the published results of the clinical trials conducted were evaluated. For this purpose، at first، the stages of clinical trials، the guidelines for the clinical translation of stem cells، and the number of clinical trials on the use of stem cells were studied; then، the results of clinical trials using different kinds of stem cells were assessed. The data were obtained by selecting the key word of “stem cells” in the public clinical trials database “http: //www. clinicaltrials. gov” and using several review articles in the field of clinical trials of stem cells. This report demonstrates the safety of stem cell therapy in the primary phases of clinical trials; however، for the efficacy of the therapy to be established the final phases of clinical trials need to be accomplished and evaluated.