Research/Original Article- Immunology

The Correlation Between SMG1 Promoter Methylation and Its Expression in Acute Lymphoblastic Leukemia Patient

hoda pourkarim, MEHDI AZAD, Mohammad Taghi Haghi Ashtiani, samaneh keshavarz, Fatemeh Nadali

Archives of Medical Laboratory Sciences, Vol. 2 No. 4 (2016), 16 Farvardin 2017,

Background: Acute lymphoblastic leukemia (ALL) comprises a heterogeneous group of disorders which originate from various important genetic lesions in B and T progenitor cells, including mutations that lead to stage-specific developmental arrest and those that impart the capacity for unlimited self-renewal, resulting in clonal expansion of immature progenitor cells. Acute lymphoblastic leukaemia occurs in both children and adults but its incidence peaks between 2 and 5 years of age. Causation is multifactorial and exogenous or endogenous exposures, genetic susceptibility, and chance have roles. Survival in pediatric acute lymphoblastic leukaemia has improved to roughly 90% in trials with risk stratification by biological features of leukaemic cells and response to treatment, treatment modification based on patients' pharmacodynamics and pharmacogenomics, and improved supportive. The promoter methylation pattern of DNA in cancer cells is different with the normal cells. Suppressor with morphogenetic effect on genitalia family member (SMG1) belongs to a family of phosphoinositide 3-kinase-related kinases and is the main kinase involved in nonsense-mediated mRNA decay.
Materials and Methods: This study was performed to investigate the correlation between SMG1 promotor methylation and its expression levels in acute lymphoblastic leukemia using methylation specific PCR (MSP). Our patients and control samples were collected from Children's Medical Center of children medical center of Imam Khomeini hospital of Tehran. To confirm the MSP results, we used Quantitative Real time-PCR (qRT-PCR ) to measure the expression level of mRNA to find out if there is any relation between pattern of methylation and expression.
Results: After performing MSP, we found that SMG1 promoter was hypermethylated. Hyper methylation of SMG1 was detected in 67/74% (21/31) of ALL samples compared to control group. SMG1 mRNA expression was down- regulated 2.74 fold compared to control group.
Conclusion: The aim of this study was to investigate the effect of methylation pattern on gene expression. Our findings suggest that SMG1 acts as a functional tumor suppressor gene which was down-regulated by CpG islands hypermethylation in ALL patients. It was shown that the methylation of SMG1 was occurred in the 67/74% of samples.

Assessment the prevalence of high-risk human papillomavirus types 16 and 18 in 15 to 45 years old women

Mohammad Reza Rezvani, Maryam Shams, Mahtab Sayaadi, Peyman Beigi, Mahmood Shams

Archives of Medical Laboratory Sciences, Vol. 2 No. 4 (2016), 16 Farvardin 2017,

Background: Cervical cancer is the fourth most common cancer in incidence and the first genital cancer in women around the world, which 95% of them are related to human papillomavirus (HPV) infections. The risk of cervical cancer increases 10-12 time in women with HPV infection. This study aim to evaluate the prevalence of high-risk HPV infections among 15-45 years old women.
Materials and Methods: This cross sectional study was carried out on 92 normal women who admitted at Semnan hygiene center and has 15-45 years old. Cervical samples were collected using Cytobrush cell collector and consequently DNA extraction was performed using commercial DNA extraction kit. Polymerase Chain Reaction (PCR) was done using HPV (GP5, GP6) universal primers accompanied by positive and negative control in each PCR run. In order to extracted DNA template quality control, actin gene used as housekeeping gene.
Results: In this investigation, study subjects age range found to be 15-45 with mean of 30±0.9 years old. HPV infection was not found in patient group. Thus, further approach in order to HPV16 and HPV18 types detection, was not performed. However, other studies represented low to moderate prevalence for HPV in some regions of Iran.
Conclusion: Cervical cancer is one the major health concern and the fourth most common cancer around the world. This cancer is more common in developing countries than developed countries due to lack of screening program. Regard to possible high prevalence rate of HPV virus and its association with cervical cancer, we suggest further determination of the HPV prevalence as well as planning in large-scale vaccination in high risk group.

Assessment of the ratio of cross-matched to transfused blood amounts in pregnant patients in Alzahra Hospital in Tabriz before and after the implementation of the health reform program in 2013-2015

Behzad Bahramzadeh, Monireh Taghizadeh Maleki, Navid Sarakhs Asbaghi, Mohammad Reza Hadigheh Javani, Samaneh Tajfar, Masoomeh Samadi Shams

Archives of Medical Laboratory Sciences, Vol. 2 No. 4 (2016), 16 Farvardin 2017,

Background: Postpartum hemorrhage is the most common cause of mortality in women with vaginal or cesarean delivery. WHO statistics shows that about 500 thousands women have died of complications related to pregnancy or during childbirth in 2013. haemorrhage probability is the main reason to order blood requests in delivery and cesarean units. The purpose of this study is to evaluate the ratio of cross-matched to transfused blood in pregnant women during one year before and after the implementation of health reform program.
Materials and Methods: In this retrospective descriptive study, the requests of blood reserves for pregnant patients with the gravid of 34±7 weeks and the age of 30±16 years old in two periods, before and after the implementation of health reform program were being collected and compared. Blood group antiserums and anti-human globulins with bovine albumins were purchased from LORN company and Baharafshan Company respectively.
Results: The total number of requests for reserved cross-matched blood during the year before the health reform program were 2837 units which 277 of them had been transfused and a year later the total number of requests was for 3029 units, which 270 of them had been transfused. This study showed 6.76 percent increase in requests and 2.53 percent decrease in transfusion in comparison with its past year.
Conclusion: The implementation of health reform program have accompanied by relative reduction of cesarean but considerable increase in vaginal delivery, and to prevent blood loss complications, the requests for reserved cross-matched blood have been increased without noticeable change in the proportion of necessarily transfused amount of them.

Significant Relationship between Salivary and Serum β2-Microglobulin in Prostate Cancer and Benign Prostatic Hyperplasia Patients

Jamal Amri, Heidar Farahani, Mahmoud-Reza Baghinia

Archives of Medical Laboratory Sciences, Vol. 2 No. 4 (2016), 16 Farvardin 2017,

Background: The presence of beta-2microglobulin (β2M) in biological fluids due to lack of binding to the membrane, increases at the time of cell turnover such as cancer and chronic infectious. In recent years, study on the standardization of saliva for clinical diagnosis is increased. The objective of this study was to determine β2M levels and the relationship between salivary and serum β2M in Prostate cancer (PCa) and Benign prostatic hyperplasia (BPH) patients.
Materials and Methods: In a case-control study, forty Subjects including 20 PCa and 20 BPH were enrolled. The concentration of β2M was measured by an enzyme-linked immunosorbent assay. The comparison between β2M levels in the PCa and BPH groups as well as the correlation between salivary and serum β2M were tested using Mann-Whitney U test and spearman correlation coefficient, respectively.
Results: Statistically significant difference was observed between salivary and serum β2M in the PCa and BPH groups (P<0.05). Spearman correlation analysis showed that salivary β2M is correlated positive and significantly with serum β2M in the PCa (r = 0.747, P < 0.05) and BPH (r = 0.513, P < 0.05) groups.
Conclusion: β2M can be a suitable biomarker for the diagnosis of prostate diseases, as well as salivary β2M can be used as an alternative approach to serum β2M for monitoring and diagnosis of prostate diseases.

Designing and construction a DNA vaccine encoding the fusion fragment of cfp10 and Ag85A immunodominant genes of Mycobacterium tuberculosis

Akram Baghani, Masoud Yousefi, Hadi Safdari, Roghayeh Teimourpour, Aida Gholoobi, Zahra Meshkat

Archives of Medical Laboratory Sciences, Vol. 2 No. 4 (2016), 16 Farvardin 2017,

Background: Pathogenic mycobacteria are one of major causes of human morbidity and mortality. Mycobacterium tuberculosis (M. tuberculosis) is an etiological agent of human tuberculosis. Designing new vaccines including DNA vaccines may be considered as new approaches for preventing of TB.
Materials and Methods: M. tuberculosis H37Rv was grown on Lowenstein Jensen medium for 4 weeks at 37ºC and then DNA was extracted. The cfp10 gene was amplified by PCR. After digesting the PCR product and the plasmid, cfp10 fragment was ligated into the vector using T4 DNA ligase. Then, Ag85A was subcloned into pcDNA/cfp10. Escherichia coli strain JM109 bacteria were transformed by the desired construct. Clone confirmations were performed by colony PCR, restriction enzyme digestion and DNA sequencing. Recombinant vector was transfected into HeLa cells and total RNA was extracted, then cDNA was synthesized using oligo-dT. Finally PCR was performed by cfp10 primers.
Results: The cfp10 was amplified by PCR method and the PCR products were visualized by agarose gel electrophoresis. The cfp10 fragments showed 303 bp in length. The cfp10 cloned into pcDNA. Then, Ag85Awas ligated into pcDNA/cfp10 after digestion correctly. Colony-PCR and restriction enzyme digestion and sequencing confirmed the cloning the fusion Ag85A/cfp10 fragment. Finally, after cDNA synthesis, expression of vector was confirmed in eukaryotic system.
Conclusion: Cloning of Ag85A/cfp10 genes of M. tuberculosis were performed correctly. It can use as a DNA vaccine for investigation the immune responses in animal models in future studies.

Review Article

Homing and mobilization of hematopoietic stem cells

Arezou Rahimi, Reza Alimohammadi, Kaveh Tari, Masoud Soleimani, Saeid Abroun, Abbas Hajifathali, Seyed Mahmoud Hashemi, Ali Jalili, Amir Atashi

Archives of Medical Laboratory Sciences, Vol. 2 No. 4 (2016), 16 Farvardin 2017,

Hematopoietic stem and progenitor cells (HSPCs) are non-stop travelers throughout body in both time and space. Understanding the mechanism of HSPCs homing and mobilization is important to enhance the efficacy at bone marrow transplantation and cellular therapy. Mobilized HSPCs has largely replaced than the use of bone marrow as a source of stem cells for both allogeneic and autologous stem cell transplantation. This review describes the specific factors which play a key role in homing and mobilization of HSPCs, includes SDF-1 and its receptor CXCR4, proteases (MMPs and CPM). Moreover, chemokines inducing rapid HPSCs mobilization would be discussed. In this article we showed that many factors such as adhesion molecules and SDF-1/CXCR4 have critical roles in homing hematopoietic stem cells and G.CSF, MMPs, adhesion molecules and ROS involvement in mobilization of stem cells. According to above, we can be rich the peripheral blood of HSPCS using of this factors and antagonist for this receptors on the osteoblastic cells or/and HSPCs to bone marrow transplant.