The Importance of Ethics in Medical Biotechnology 2 2 The rapid progress of modern biotechnology has presented a number of new and unique ethical and social challenges within the context of human medical science. Research in  medical biotechnology has led to increased knowledge of disease, acceleration of the treatment process, improved pharmacotherapy for infectious diseases and hope for the struggle against incurable diseases such as ALS, MS and Alzheimer’s 1. Medical biotechnology promises major advances in human health and therefore, any limitations on the right to freedom of scientific research should be for significant reasons only, and as least restrictive as possible, so as not to impede scientific wisdom and prevent damage to the scientific undertaking. At the same time a duty exists to ensure that research in this area of biotechnology is conducted in ethically acceptable ways. A balance needs to be struck between recognizing the potential benefits which biotechnology research offers to individuals and the community as a whole, and the duty to ensure that research in this area is conducted ethically 2. Researchers in the biotechnology industry face challenges unlike researchers in other sectors. Unlike most other industries, advances and research in the biotechnology industry are often front page news and has to face intense scrutiny by press, academics, government and the public. As biotechnology is a newly emerging field, a further challenge facing the industry is the lack of historical precedence in the sector to provide guidance for the safe and ethical development of the technology. In biotechnology research, the usual ethical principles applicable to health research involving animals and human participants must be observed and such research must be scientifically sound. 89 89 Shahin Akhondzadeh Editorial 209.pdf Akhondzadeh S. Hippocampal synaptic plasticity and cognition. J Clin Pharm Ther 1999;24(4):241-248.##Akhondzadeh S. Ethical issues in medical biotechnology. Avicenna J Med Biotechnol 2014;6(3):129.##

The Immunosuppressive Activity of Amniotic Membrane Mesenchymal Stem Cells on T Lymphocytes 2 2 Background: Mesenchymal Stem Cells (MSCs) are isolated from different sources like placenta. The placenta and its membranes like Amniotic Membrane (AM) are readily available and easy to work with. There is only limited knowledge on the immunomodulatory properties of human Amniotic Membrane-derived Mesenchymal Stem Cells (hAM-MSCs). The aim of this study was to survey the suppressive activity of hAM-MSCs on T lymphocytes in vitro. Methods: Human AMs were obtained after caesarean section births from healthy women. After enzymatic digestion, cells were cultured and hAM-MSCs were obtained. In addition, human T lymphocytes were isolated and co-cultured with hAM-MSCs for 72 hr in the presence or absence of phytohemagglutinin (PHA). Subsequently, proliferation of T cells was analyzed using BrdU and subsequently flow cytometry technique. Besides, the production of IL-4 and IFN-γ was examined by ELISA method. Additionally, the expression of activation markers (CD38, HLA-DR) was studied on T lymphocytes by flow cytometry technique. Results: It was revealed that hAM-MSCs could significantly suppress the proliferation of T lymphocytes (p≤0.01) and significantly decrease the production of IFN-γ by T cells (p<0.05). hAM-MSCs also down regulated the expression of activation markers on the surface of T lymphocytes, CD38 and HLA-DR. The difference was significant between the case and control samples (p<0.05). All the comparisons were carried out between the case (Tcell+PHA+hAM-MSCs) and control (Tcell+PHA) groups. Conclusion: In conclusion, hAM-MSCs could inhibit the (mitogen-activated) T cells even in the absence of blood monocytes. Besides, hAM-MSCs-mediated inhibition of T lymphocytes was combined with down regulation of activation markers. 90 96 Fatemeh Alikarami Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Fatemeh Yari Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Naser Amirizadeh Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Mahin Nikougoftar Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Mohammad Ali Jalili Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran AmnionMesenchymal stem cellsT-lymphocyte 210.pdf Yoo KH, Jang IK, Lee MW, Kim HE, Yang MS, Eom Y, et al. Comparison of immunomodulatory properties of mesenchymal stem cells derived from adult human tissues. Cell Immunol 2009;259(2):150-156.##Sensebé L, Krampera M, Schrezenmeier H, Bourin P, Giordano R. Mesenchymal stem cells for clinical application. Vox Sang 2010;98(2):93-107.##Gieseke F, Böhringer J, Bussolari R, Dominici M, Handgretinger R, Müller I. Human multipotent mesenchymal stromal cells use galectin-1 to inhibit immune effector cells. Blood 2010;116(19):3770-3779.##Hass R, Kasper C, Böhm S, Jacobs R. Different populations and sources of human mesenchymal stem cells (MSC): A comparison of adult and neonatal tissue-derived MSC. Cell Commun Signal 2011;9:12.##Vija L, Farge D, Gautier JF, Vexiau P, Dumitrache C, Bourgarit A, et al. Mesenchymal stem cells: Stem cell therapy perspectives for type 1 diabetes. Diabetes Metab 2009;35(2):85-93.##Le Blanc K, Tammik L, Sundberg B, Haynesworth SE, Ringdén O. Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex. Scand J Immunol 2003;57(1):11-20.##Bacigalupo A, Valle M, Podestà M, Pitto A, Zocchi E, De Flora A, et al. T-cell suppression mediated by mesenchymal stem cells is deficient in patients with severe aplastic anemia. Exp Hematol 2005;33(7):819-827.##Groh ME, Maitra B, Szekely E, Koç ON. Human mesenchymal stem cells require monocyte-mediated activation to suppress alloreactive T cells. Exp Hematol 2005;33(8):928-934.##Asari S, Itakura S, Ferreri K, Liu CP, Kuroda Y, Kandeel F, et al. Mesenchymal stem cells suppress B-cell terminal differentiation. Exp Hematol 2009;37(5):604-615.##Spaggiari GM, Capobianco A, Abdelrazik H, Becchetti F, Mingari MC, Moretta L. Mesenchymal stem cells inhibit natural killer-cell proliferation, cytotoxicity, and cytokine production: role of indoleamine 2,3-dioxygenase and prostaglandin E2. Blood 2008;111(3):1327-1333.##Veryasov VN, Savilova AM, Buyanovskaya OA, Chulkina MM, Pavlovich SV, Sukhikh GT. Isolation of mesenchymal stromal cells from extraembryonic tissues and their characteristics. Bull Exp Biol Med 2014;157(1):119-124.##Fibbe WE, Noort WA. Mesenchymal stem cells and hematopoietic stem cell transplantation. Ann NY Acad Sci 2003;996:235-244.##Bensidhoum M, Chapel A, Francois S, Demarquay C, Mazurier C, Fouillard L, et al. Homing of in vitro expanded Stro-1- or Stro-1+ human mesenchymal stem cells into the NOD/SCID mouse and their role in supporting human CD34 cell engraftment. Blood 2004;103(9):3313-3319.##Bartholomew A, Patil S, Mackay A, Nelson M, Buyaner D, Hardy W, et al. Baboon mesenchymal stem cells can be genetically modified to secrete human erythropoietin in vivo. Hum Gene Ther 2001;12(12):1527-1541.##Díaz-Prado S, Muiños-López E, Hermida-Gómez T, Rendal-Vázquez ME, Fuentes-Boquete I, de Toro FJ, et al. Isolation and characterization of mesenchymal stem cells from human amniotic membrane. Tissue Eng Part C Methods 2011;17(1):49-59.##Mihu CM, Rus Ciucă D, Soritău O, Suşman S, Mihu D. Isolation and characterization of mesenchymal stem cells from the amniotic membrane. Rom J Morphol Embryol 2009;50(1):73-77.##Parolini O, Caruso M. Review: Preclinical studies on placenta-derived cells and amniotic membrane: an update. Placenta 2011;32 Suppl 2:S186-195.##Kang JW, Koo HC, Hwang SY, Kang SK, Ra JC, Lee MH, et al. Immunomodulatory effects of human amniotic membrane-derived mesenchymal stem cells. J Vet Sci 2012;13(1):23-31.##Aggarwal S, Pittenger MF. Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood 2005;105(4):1815-1822.##Le Blanc K, Rasmusson I, Sundberg B, Götherström C, Hassan M, Uzunel M, et al. Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells. Lancet 2004;363(9419):1439-1441.##Ghannam S, Bouffi C, Djouad F, Jorgensen C, Noël D. Immunosuppression by mesenchymal stem cells: mechanisms and clinical applications. Stem Cell Res Ther 2010;1(1):2.##Uccelli A, Moretta L, Pistoia V. Immunoregulatory function of mesenchymal stem cells. Eur J Immunol 2006;36(10):2566-2573.##Sato K, Ozaki K, Oh I, Meguro A, Hatanaka K, Nagai T, et al. Nitric oxide plays a critical role in suppression of T-cell proliferation by mesenchymal stem cells. Blood 2007;109(1):228-234.##Si ZG, Bai H, Wang CB, Xue ZW, Wang Q, Wu T, et al. [Effect of human bone marrow mesenchymal stem cells on T lymphocyte killing K562 cells]. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2007;15(6):1216-1219. Chinese.##Ueta M, Kweon MN, Sano Y, Sotozono C, Yamada J, Koizumi N, et al. Immunosuppressive properties of human amniotic membrane for mixed lymphocyte reaction. Clin Exp Immunol 2002;129(3):464-470.##Dazzi F, Krampera M. Mesenchymal stem cells and autoimmune diseases. Best Pract Res Clin Haematol 2011;24(1):49-57.##Klyushnenkova E, Mosca JD, Zernetkina V, Majumdar MK, Beggs KJ, Simonetti DW, et al. T cell responses to allogeneic human mesenchymal stem cells: immunogenicity, tolerance, and suppression. J Biomed Sci 2005;12(1):47-57.##

High Prevalence of Y Chromosome Partial Microdeletions in Overweight Men 2 2 Background: Microdeletions of the Y chromosome are one of the most frequent genetic causes of spermatogenic failure in infertile men. But their role in gaining weight is unclear. The present study investigated the possible association of these partial microdeletions and obesity. Methods: In a case-control study, 180 males were selected. The prevalence of microdeletions was assessed using PCR in AZFc area of Y chromosome and statistical analysis was done using the Fisher exact test and Pearson correlation. Results: In our study, inverse relationship was observed between body mass index and testosterone level (p-value: 0.005). Fisher exact tests showed that there was a significant association between gr/gr mutation and BMI (p-value: 0.044). Conclusion: Our study revealed that Y chromosome microdeletions are more common in obese men. Furthermore, microdeletions such as gr/gr, which were observed in normal men, could cause decreased testosterone level. So, they may contribute to gaining weight. 97 100 Atefeh Biabangard zak Department of Biology, School of Basic Sciences, Golestan University Department of Biology, School of Basic Sciences, Golestan University Iran Masoud Golalipour Department of Biology, School of Basic Sciences, Golestan University Department of Biology, School of Basic Sciences, Golestan University Iran Gholamreza Hadadchi Department of Biology, School of Basic Sciences, Golestan University Department of Biology, School of Basic Sciences, Golestan University Iran ObesityTestosteroneY chromosome microdeletions 211.pdf Blakemore AI, Froguel P. Is obesity our genetic legacy? J Clin Endocrinol Metab 2008;93(11 Suppl 1):S51-56.##Tarnoki AD, Tarnoki DL, Medda E, Cotichini R, Stazi MA, Fagnani C, et al. Bioimpedance analysis of body composition in an international twin cohort. Obes Res Clin Pract 2014;8(3):e201-298.##Rankinen T, Zuberi A, Chagnon YC, Weisnagel SJ, Argyropoulos G, Walts B, et al. The human obesity gene map: the 2005 update. Obesity (Silver Spring) 2006;14(4):529-644.##Vogt PH, Edelmann A, Kirsch S, Henegariu O, Hirschmann P, Kiesewetter F, et al. Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet 1996;5(7):933-943.##Imken L, El Houate B, Chafik A, Nahili H, Boulouiz R, Abidi O, et al. AZF microdeletions and partial deletions of AZFc region on the Y chromosome in Moroccan men. Asian J Androl 2007;9(5):674-678.##Hucklenbroich K, Gromoll J, Heinrich M, Hohoff C, Nieschlag E, Simoni M. Partial deletions in the AZFc region of the Y chromosome occur in men with impaired as well as normal spermatogenesis. Hum Reprod 2005; 20(1):191-197.##Lardone MC, Parodi DA, Ebensperger M, Peñaloza P, Cornejo V, Valdevenito R, et al. AZFc partial deletions in Chilean men with severe spermatogenic failure. Fertil Steril 2007;88(5):1318-1326.##Ferlin A, Tessari A, Ganz F, Marchina E, Barlati S, Garolla A, et al. Association of partial AZFc region deletions with spermatogenic impairment and male infertility. J Med Genet 2005;42(3):209-213.##Banegas JR, Gutiérrez-Fisac JL, Rodríguez-Artalejo F, Cruz JJ, Guallar P, Herruzo R. Obesity and body mass index in Spain: the 'single population' theory revisited. Eur J Clin Nutr 2001;55(9):782-785.##Ardern CI, Janssen I, Ross R, Katzmarzyk PT. Development of health-related waist circumference thresholds within BMI categories. Obes Res 2004;12(7):1094-1103.##Wang C, Jackson G, Jones TH, Matsumoto AM, Nehra A, Perelman MA, et al. Low testosterone associated with obesity and the metabolic syndrome contributes to sexual dysfunction and cardiovascular disease risk in men with type 2 diabetes. Diabetes Care 2011;34(7):1669-1675.##Stewart TM, Liu DY, Garrett C, Jørgensen N, Brown EH, Baker HW. Associations between andrological measures, hormones and semen quality in fertile Australian men: inverse relationship between obesity and sperm output. Hum Reprod 2009;24(7):1561-1568.##MacDonald AA, Herbison GP, Showell M, Farquhar CM. The impact of body mass index on semen parameters and reproductive hormones in human males: a systematic review with meta-analysis. Hum Reprod Update 2010;16(3):293-311.##Mårin P, Holmäng S, Gustafsson C, Jönsson L, Kvist H, Elander A, et al. Androgen treatment of abdominally obese men. Obes Res 1993;1(4):245-251.##Jones TH, Saad F. The effects of testosterone on risk factors for, and the mediators of, the atherosclerotic process. Atherosclerosis 2009;207(2):318-327.##Tsui S, Dai T, Warren ST, Salido EC, Yen PH. Association of the mouse infertility factor DAZL1 with actively translating polyribosomes. Biol Reprod 2000;62(6):1655-1660.##Leder BZ, Rohrer JL, Rubin SD, Gallo J, Longcope C. Effects of aromatase inhibition in elderly men with low or borderline-low serum testosterone levels. J Clin Endocrinol Metab 2004;89(3):1174-1180.##

Expression of Recombinant Human Insulin-like Growth Factor Type 1 (rhIGF-1) in Escherichia coli 2 2 Background: Human insulin-like growth factor type 1 (hIGF-1) is a protein consisting of 70 amino acids (MW=7.6 kDa) and mainly synthesized by liver. Mecasermin (Trade name INCRELEX) is the synthetic form of the protein which is used as an effective treatment for particular disorders such as short stature, type 1 and 2 diabetes, and wound healing. Current study was aimed to investigate the expression of human insulin-like growth factor type1 in Escherichia coli (E. coli) BL21 (DE3) expression system in order to produce an active recombinant form of the protein. Methods: For the purpose of the study, firstly codon optimization was done for hIGF-1 gene, using bioinformatics databases. Then, the gene was synthesized and inserted in pET-24a vector by a cutting strategy included NdeI and BamHI-HF enzymes. In the next step, gene was run in agarose gel and purified. The constructed expression cassette was transformed into E. coli BL21 (DE3) cells through CaCl2 heat shock method. Identification and confirmation of the transformed colonies were performed using screening PCR method. Synthesis of hIGF-1 was induced by IPTG. The expression in induced strains was analyzed by SDS-PAGE and western blotting techniques. Confirmation of cloning and IGF-1 expression cassette was carried out through genetic engineering procedures. Results: Analysis of transformed E. coli strain with SDS-PAGE and western blotting techniques confirmed that gene was expressed in host cells. Molecular weight of the expressed protein was estimated to be 7.6 kDa. Conclusion: hIGF-1 expression cassette for cloning and expression in E. coli was designed and the protein of interest was successfully induced and identified. In addition, E. coli BL21 (DE3) can be used as a suitable host for production of recombinant hIGF-1 and this technology has a potential to be localized. 101 105 Hamidreza Iranpoor Department of Medical Biotechnology, Faculty of Advanced Technologies in Medicine, Golestan University of Medical Science Department of Medical Biotechnology, Faculty of Advanced Technologies in Medicine, Golestan University of Medical Science Iran Eskandar Omidinia Genetics and Metabolism Research Group, Pasteur Institute of Iran Genetics and Metabolism Research Group, Pasteur Institute of Iran Iran Venus Vatankhah Faculty of Medical Sciences, Shahrekord University of Medical Sciences Faculty of Medical Sciences, Shahrekord University of Medical Sciences Iran Vahid Gharanjik Islamic Azad University Gorgan Branch Islamic Azad University Gorgan Branch Iran Majid Shahbazi Cellular and Molecular Research Center, Taleghani Children Hospital, Gorgan University of Medical Sciences Cellular and Molecular Research Center, Taleghani Children Hospital, Gorgan University of Medical Sciences Iran CloningEscherichia coliGene expressionInsulin-like growth factor type 1 212.pdf Schulz MF, Buell G, Schmid E, Movva R, Selzer G. Increased expression in Escherichia coli of a synthetic gene encoding human somatomedin C after gene duplication and fusion. J Bacteriol 1987;169(12):5385-5392.##Wong EY, Seetharam R, Kotts CE, Heeren RA, Klein BK, Braford SR, et al. Expression of secreted insulin-like growth factor-1 in Escherichia coli. Gene 1988;68(2): 193-203.##Buell G, Schulz MF, Selzer G, Chollet A, Movva NR, Semon D, et al. Optimizing the expression in E. coli of a synthetic gene encoding somatomedin-C (IGF-I). Nucleic Acids Res 1985;13(6):1923-1938.##Dalboge H, Skriver L. Method of producing IGF-1. Google Patents; 1995.##Laron Z. Insulin-like growth factor 1 (IGF-1): a growth hormone. Mol Pathol 2001;54(5):311-316.##Yilmaz A, Davis ME, Simmen RC. Reproductive performance of bulls divergently selected on the basis of blood serum insulin-like growth factor I concentration. J Anim Sci 1999;77(4):835-839.##Chung BH, Choi YJ, Yoon SH, Lee SY, Lee YI. Process development for production of recombinant human insulin-like growth factor-I in Escherichia coli. J Ind Microbiol Biotechnol 2000;24(2):94-9.##Choi JH, Lee SJ, Lee SJ, Lee SY. Enhanced production of insulin-like growth factor I fusion protein in Escherichia coli by coexpression of the down-regulated genes identified by transcriptome profiling. Appl Environ Microbiol 2003;69(8):4737-4742.##Yamada H, Saito Y, Fujimoto T, Noguchi Y, Mori T, Miura T, et al. Large scale purification of recombinant insulin-like growth factor I (IGF-I, mecasermin) from a fused protein produced in Escherichia coli. J Ferment Bioeng 1996;82(2):134-139.##Seyedi E, Rahaie M, Mofid M, Haddad L, Namvaran M, Fallah J, et al. Recombinant production of mecasermin in E. coli expression system. Res Pharm Sci 2014;9(6):453-461.##Chen Z, Chen H, Wang X, Ma X, Huang B. Expression, purification, and characterization of secreted recombinant human insulin-like growth factor-binding protein-6 in methylotrophic yeast Pichia pastoris. Protein Expr Purif 2007;52(2):239-248.##Kim SO, Lee YI. High-level expression and simple purification of recombinant human insulin-like growth factor I. J Biotechnol 1996;48(1):97-105.##Steube K, Chaudhuri B, Märki W, Merryweather JP, Heim J. α‐Factor‐leader‐directed secretion of recombinant human‐insulin‐like growth factor I from Saccharomyces cerevisiae. Eur J Biochem 1991;198(3): 651-657.##Elliott S, Fagin KD, Narhi LO, Miller JA, Jones M, Koski R, et al. Yeast-derived recombinant human insulin-like growth factor I: production, purification, and structural characterization. J Prot Chem 1990;9(1):95-104.##Studier FW, Moffatt BA. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 1986;189(1):113-130.##Yamada H, Saito Y, Fujimoto T, Noguchi Y, Mori T, Miura T, et al. Large scale purification of recombinant insulin-like growth factor I (IGF-I, mecasermin) from a fused protein produced in Escherichia coli. J Ferment Bioeng 1996;82(2):134-139.##Georgiou G, Segatori L. Preparative expression of secreted proteins in bacteria: status report and future prospects. Curr Opin Biotechnol 2005;16(5):538-545.##Upadhyay AK, Murmu A, Singh A, Panda AK. Kinetics of inclusion body formation and its correlation with the characteristics of protein aggregates in Escherichia coli. PLoS One 2012;7(3):e33951.##Han Y, Zhang X-H, Chen Y, Li Y, Chen H, Fang L. High-level expression, purification, polyclonal antibody preparation against recombinant OprD from Pseudomonas aeruginosa. African J Biotechnol 2011;10 (7):1246-1251.##

Retinoic Acid as the Stimulating Factor for Differentiation of Wharton’s Jelly- Mesenchymal Stem Cells into Hepatocyte-like Cells 2 2 Background: Wharton’s Jelly-Mesenchymal Stem Cells (WJ-MSCs) are pluripotent cells with differentiation capability into most cell lineages. The aim of the current work was to examine the role of Retinoic Acid (RA) in differentiation process of these cells into hepatocyte-like cells and determine the morphological and functional patterns. Methods: Human WJ-MSCs were extracted, cultured and expanded; after approximately 95% of confluence, the cells were treated with hepatogenic media containing RA. The cells were subsequently analyzed for morphological changes, glycogen storage, albumin production, and specific gene expression. Results: WJ-MSCs expressed high levels of CD90 (93.6%) and CD105 (90.7%), but low levels of CD34 (0.3%) and CD45 (0.8%). Albumin production had significant difference in the two groups (p≤0.05). The data showed specific characteristics in favor of considering the differentiated cells as hepatocyte-like cells such as obtaining morphologic, functional, and αFP and HNF1-α expression patterns which in turn were higher in cells exposed to RA. Conclusion: Based on the data of present study, RA is an effective molecule in inducing differentiation of WJ-MSCs into hepatocyte-like cells; therefore, it may be considered as a promising factor for targeting therapy of liver disorders. 106 112 Keywan Mortezaee Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Bagher Minaii Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Fatemeh Sabbaghziarani Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Iraj Ragerdi Kashani Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Gholamreza Hassanzadeh Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Parichehr Pasbakhsh Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran Mohammad Barbarestani Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Department of Anatomy, School of Medicine, Tehran University of Medical Sciences Iran GlycogenPluripotent stem cellsRetinoic acidWharton’s jelly-mesenchymal stem cell 213.pdf Friedman SL. Liver fibrosis-from bench to bedside. J Hepatol 2003;38 Suppl 1:S38-53.##Tsai PC, Fu TW, Chen YM, Ko TL, Chen TH, Shih YH, et al. The therapeutic potential of human umbilical mesenchymal stem cells from Wharton's jelly in the treatment of rat liver fibrosis. Liver Transpl 2009;15(5): 484-495.##Kern S, Eichler H, Stoeve J, Klüter H, Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells 2006;24(5):1294-1301.##Lennon DP, Edmison JM, Caplan AI. Cultivation of rat marrow-derived mesenchymal stem cells in reduced oxygen tension: effects on in vitro and in vivo osteochondrogenesis. J Cell Physiol 2001;187(3):345-355.##Yang B, Guo H, Zhang Y, Chen L, Ying D, Dong S. MicroRNA-145 regulates chondrogenic differentiation of mesenchymal stem cells by targeting Sox9. PLoS One 2011;6(7):e21679.##Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006;8(4):315-317.##Silva GV, Litovsky S, Assad JA, Sousa AL, Martin BJ, Vela D, et al. Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model. Circulation 2005;111(2):150-156.##Roobrouck VD, Clavel C, Jacobs SA, Ulloa-Montoya F, Crippa S, Sohni A, et al. Differentiation potential of human postnatal mesenchymal stem cells, mesoangioblasts, and multipotent adult progenitor cells reflected in their transcriptome and partially influenced by the culture conditions. Stem Cells 2011;29(5):871-882.##Lei Z, Yongda L, Jun M, Yingyu S, Shaoju Z, Xinwen Z, et al. Culture and neural differentiation of rat bone marrow mesenchymal stem cells in vitro. Cell Biol Int 2007;31(9):916-923.##Wang Y, Nan X, Li Y, Zhang R, Yue W, Yan F, et al. Induction of umbilical cord blood-derived beta2m-c-Met+ cells into hepatocyte-like cells by coculture with CFSC/HGF cells. Liver Transpl 2005;11(6):635-643.##Cho PS, Messina DJ, Hirsh EL, Chi N, Goldman SN, Lo DP, et al. Immunogenicity of umbilical cord tissue derived cells. Blood 2008;111(1):430-438.##Taghizadeh RR, Cetrulo KJ, Cetrulo CL. Wharton's Jelly stem cells: future clinical applications. Placenta 2011;32 Suppl 4:S311-315.##Turnovcova K, Ruzickova K, Vanecek V, Sykova E, Jendelova P. Properties and growth of human bone marrow mesenchymal stromal cells cultivated in different media. Cytotherapy 2009;11(7):874-885.##Weiss ML, Medicetty S, Bledsoe AR, Rachakatla RS, Choi M, Merchav S, et al. Human umbilical cord matrix stem cells: preliminary characterization and effect of transplantation in a rodent model of Parkinson's disease. Stem Cells 2006;24(3):781-792.##Mitchell KE, Weiss ML, Mitchell BM, Martin P, Davis D, Morales L, et al. Matrix cells from Wharton's jelly form neurons and glia. Stem Cells 2003;21(1):50-60.##Huang P, Lin LM, Wu XY, Tang QL, Feng XY, Lin GY, et al. Differentiation of human umbilical cord Wharton's jelly-derived mesenchymal stem cells into germ-like cells in vitro. J Cell Biochem 2010;109(4):747-754.##Chen MY, Lie PC, Li ZL, Wei X. Endothelial differentiation of Wharton's jelly-derived mesenchymal stem cells in comparison with bone marrow-derived mesenchymal stem cells. Exp Hematol 2009;37(5):629-640.##Wu LF, Wang NN, Liu YS, Wei X. Differentiation of Wharton's jelly primitive stromal cells into insulin-producing cells in comparison with bone marrow mesenchymal stem cells. Tissue Eng Part A 2009;15(10): 2865-2873.##Wells JM, Melton DA. Early mouse endoderm is patterned by soluble factors from adjacent germ layers. Development 2000;127(8):1563-1572.##Oh SH, Miyazaki M, Kouchi H, Inoue Y, Sakaguchi M, Tsuji T, et al. Hepatocyte growth factor induces differentiation of adult rat bone marrow cells into a hepatocyte lineage in vitro. Biochem Biophys Res Commun 2000;279(2):500-504.##Ishikawa T, Factor VM, Marquardt JU, Raggi C, Seo D, Kitade M, et al. Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice. Hepatology 2012;55(4):1215-1226.##Ying M, Wang S, Sang Y, Sun P, Lal B, Goodwin CR, et al. Regulation of glioblastoma stem cells by retinoic acid: role for Notch pathway inhibition. Oncogene 2011;30(31):3454-3467.##Hong SH, Gang EJ, Jeong JA, Ahn C, Hwang SH, Yang IH, et al. In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells. Biochem Biophys Res Commun 2005;330(4): 1153-1161.##Seo MJ, Suh SY, Bae YC, Jung JS. Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun 2005; 328(1):258-264.##Wang X, Cui J, Zhang BQ, Zhang H, Bi Y, Kang Q, et al. Decellularized liver scaffolds effectively support the proliferation and differentiation of mouse fetal hepatic progenitors. J Biomed Mater Res A 2014;102(4):1017-1025.##Angelucci S, Marchisio M, Di Giuseppe F, Pierdomenico L, Sulpizio M, Eleuterio E, et al. Proteome analysis of human Wharton's jelly cells during in vitro expansion. Proteome Sci 2010;8:18.##Forte G, Minieri M, Cossa P, Antenucci D, Sala M, Gnocchi V, et al. Hepatocyte growth factor effects on mesenchymal stem cells: proliferation, migration, and differentiation. Stem Cells 2006;24(1):23-33.##Troyer DL, Weiss ML. Wharton's jelly-derived cells are a primitive stromal cell population. Stem Cells 2008;26(3):591-599.##Can A, Karahuseyinoglu S. Concise review: human umbilical cord stroma with regard to the source of fetus-derived stem cells. Stem Cells 2007;25(11):2886-2895.##Lee KD, Kuo TK, Whang-Peng J, Chung YF, Lin CT, Chou SH, et al. In vitro hepatic differentiation of human mesenchymal stem cells. Hepatology 2004;40(6):1275-1284.##Zhang YN, Lie PC, Wei X. Differentiation of mesenchymal stromal cells derived from umbilical cord Wharton's jelly into hepatocyte-like cells. Cytotherapy 2009;11(5):548-558.##Schwartz RE, Reyes M, Koodie L, Jiang Y, Blackstad M, Lund T, et al. Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. J Clin Invest 2002;109(10):1291-1302.##Bonora-Centelles A, Jover R, Mirabet V, Lahoz A, Carbonell F, Castell JV, et al. Sequential hepatogenic transdifferentiation of adipose tissue-derived stem cells: relevance of different extracellular signaling molecules, transcription factors involved, and expression of new key marker genes. Cell Transplant 2009;18(12):1319-1340.##Alisi A, Leoni S, Piacentani A, Conti Devirgiliis L. Retinoic acid modulates the cell-cycle in fetal rat hepatocytes and HepG2 cells by regulating cyclin-cdk activities. Liver Int 2003;23(3):179-186.##Tonge PD, Andrews PW. Retinoic acid directs neuronal differentiation of human pluripotent stem cell lines in a non-cell-autonomous manner. Differentiation 2010;80 (1):20-30.##Jian P, Li ZW, Fang TY, Jian W, Zhuan Z, Mei LX, et al. Retinoic acid induces HL-60 cell differentiation via the upregulation of miR-663. J Hematol Oncol 2011;4:20.##Huang J, Bi Y, Zhu GH, He Y, Su Y, He BC, et al. Retinoic acid signalling induces the differentiation of mouse fetal liver-derived hepatic progenitor cells. Liver Int 2009;29(10):1569-1581.##Masaki T, Matsuura T, Ohkawa K, Miyamura T, Okazaki I, Watanabe T, et al. All-trans retinoic acid down-regulates human albumin gene expression through the induction of C/EBPbeta-LIP. Biochem J 2006;397(2): 345-353.##

Inhibition of Coenzyme Qs Accumulation in Engineered Escherichia coli by High Concentration of Farnesyl Diphosphate 2 2 Background: Coenzyme Q10 (CoQ10) is an isoprenoid component used widely in nutraceutical industries. Farnesyl diphosphate synthase (FPPS) is a responsible enzyme for biosynthesis of farnesyl diphosphate (FPP), a key precursor for CoQs production. This research involved investigating the effect of FPPS over-expression on CoQs production in engineered CoQ10-producing Escherichia coli (E. coli). Methods: Two CoQ10-producing strains, as referred to E. coli Ba and E. coli Br, were transformed by the encoding gene for FPPS (ispA) under the control of either the trc or PBAD promoters. Results: Over-expression of ispA under the control of PBAD promoter led to a relative increase in CoQ10 production only in recombinant E. coli Br although induction by arabinose resulted in partial reduction of CoQ10 production in both recombinant E. coli Ba and E. coli Br strains. Over-expression of ispA under the control of stronger trc promoter, however, led to a severe decrease in CoQ10 production in both recombinant E. coli Ba and E. coli Br strains, as reflected by reductions from 629±40 to 30±13 and 564±28 to 80±14 µg/g Dried Cell Weight (DCW), respectively. The results showed high level of FPP reduces endogenous CoQ8 production as well and that CoQs are produced in a complimentary manner, as the increase in production of one decreases the production of the other. Conclusion: The reduction in CoQ10 production can be a result of Dds inhibition by high FPP concentration. Therefore, more effort is needed to verify the role of intermediate metabolite concentration and to optimize production of CoQ10. 113 120 Mojtaba Samoudi Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Iran Negar Omid Yeganeh Department of Biology, Payame-Noor University Department of Biology, Payame-Noor University Iran Hossein Shahbani Zahiri Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Iran Parvin Shariati Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Iran Reza Hajhosseini Department of Biology, Payame-Noor University Department of Biology, Payame-Noor University Iran Coenzyme Q10Decaprenyl diphosphate synthaseFarnesyl diphosphateIsoprenoid 214.pdf 1. 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Multiple strategies for metabolic engineering of escherichia coli for efficient production of coenzyme Q10. Chin J Chem Eng 2011;19 (2):316-326.##Søballe B, Poole RK. Ubiquinone limits oxidative stress in Escherichia coli. Microbiology 2000;146 ( Pt 4):787-796.##Kumar A, Kaur H, Devi P, Mohan V. Role of coenzyme Q10 (CoQ10) in cardiac disease, hypertension and Meniere-like syndrome. Pharmacol Ther 2009;124(3): 259-268.##Damian MS, Ellenberg D, Gildemeister R, Lauermann J, Simonis G, Sauter W, et al. Coenzyme Q10 combined with mild hypothermia after cardiac arrest: a preliminary study. Circulation 2004;110(19):3011-3016.##Quinzii CM, DiMauro S, Hirano M. Human coenzyme Q10 deficiency. Neurochem Res 2007;32(4-5):723-727.##Ochoa JJ, Pamplona R, Ramirez-Tortosa MC, Granados-Principal S, Perez-Lopez P, Naudí A, et al. Age-related changes in brain mitochondrial DNA deletion and oxidative stress are differentially modulated by dietary fat type and coenzyme Q10. Free Radic Biol Med 2011;50(9):1053-1064.##Aguilaniu H, Durieux J, Dillin A. Metabolism, ubiquinone synthesis, and longevity. Genes Dev 2005;19(20):2399-2406.##Ernster L, Dallner G. Biochemical, physiological and medical aspects of ubiquinone function. Biochim Biophys Acta 1995;1271(1):195-204.##Meganathan R. Biosynthesis of the isoprenoid quinones menaquinone (vitamin K2) and ubiquinone (coenzyme Q). Escherichia coli and Salmonella: cellular and molecular biology. 1996;1:642-56.##Okada K, Suzuki K, Kamiya Y, Zhu X, Fujisaki S, Nishimura Y, et al. Polyprenyl diphosphate synthase essentially defines the length of the side chain of ubiquinone. Biochim Biophys Acta 1996;1302(3):217-223.##Zhang D, Shrestha B, Li Z, Tan T. Ubiquinone-10 production using Agrobacterium tumefaciens dps gene in Escherichia coli by coexpression system. Mol Biotechnol 2007;35(1):1-14.##Zahiri HS, Yoon SH, Keasling JD, Lee SH, Won Kim S, Yoon SC, et al. Coenzyme Q10 production in recombinant Escherichia coli strains engineered with a heterologous decaprenyl diphosphate synthase gene and foreign mevalonate pathway. Metab Eng 2006;8(5):406-416.##Huang M, Yunyan C, Jianzhong L. Chromosomal Engineering of Escherichia coli for Efficient Production of Coenzyme Q10. Chin J Chem Eng 2014;22(5):559-569.##Kaur D, Brennan PJ, Crick DC. Decaprenyl diphosphate synthesis in Mycobacterium tuberculosis. J Bacteriol 2004;186(22):7564-7570.##Mekkriengkrai D, Sando T, Hirooka K, Sakdapipanich J, Tanaka Y, Fukusaki E, et al. Cloning and characterization of farnesyl diphosphate synthase from the rubber-producing mushroom Lactarius chrysorrheus. Biosci Biotechnol Biochem 2004;68(11):2360-2368.##Suzuki K, Okada K, Kamiya Y, Zhu XF, Nakagawa T, Kawamukai M, et al. Analysis of the decaprenyl diphosphate synthase (dps) gene in fission yeast suggests a role of ubiquinone as an antioxidant. J Biochem 1997;121(3):496-505.##Takahashi S, Nishino T, Koyama T. Isolation and expression of Paracoccus denitrificans decaprenyl diphosphate synthase gene for production of ubiquinone-10 in Escherichia coli. Biochem Eng J 2003;16(2):183-190.##Zahiri HS, Noghabi KA, Shin YC. Biochemical characterization of the decaprenyl diphosphate synthase of Rhodobacter sphaeroides for coenzyme Q10 production. Appl Microbiol Biotechnol 2006;73(4):796-806.##Zahiri HS, Noghabi KA, Samoodi M, Yeganeh NO, Rad SA, Safari A, et al. Effect of concomitant lycopene biosynthesis on CoQ10 accumulation in transformed Escherichia coli strains. Iranian J Biotechnol 2009;7(4):224-232.##Sørensen HP, Mortensen KK. Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol 2005;115(2):113-128.##Fujisaki S, Hara H, Nishimura Y, Horiuchi K, Nishino T. Cloning and nucleotide sequence of the ispA gene responsible for farnesyl diphosphate synthase activity in Escherichia coli. J Biochem 1990;108(6):995-1000.##Casali N, Preston A. E. coli Plasmid Vectors. Methods and Applications. Totowa, N.J.: Humana Press;2003.##Cluis CP, Burja AM, Martin VJ. Current prospects for the production of coenzyme Q10 in microbes. Trends Biotechnol 2007;25(11):514-521.##Huang Q, Roessner CA, Croteau R, Scott AI. Engineering Escherichia coli for the synthesis of taxadiene, a key intermediate in the biosynthesis of taxol. Bioorg Med Chem 2001;9(9):2237-2242.##Kim SW, Keasling JD. Metabolic engineering of the nonmevalonate isopentenyl diphosphate synthesis pathway in Escherichia coli enhances lycopene production. Biotechnol Bioeng 2001;72(4):408-415.##Sandmann G. Combinatorial biosynthesis of carotenoids in a heterologous host: a powerful approach for the biosynthesis of novel structures. Chembiochem 2002;3(7):629-635.##Harker M, Bramley PM. Expression of prokaryotic 1-deoxy-D-xylulose-5-phosphatases in Escherichia coli increases carotenoid and ubiquinone biosynthesis. 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In Silico Evaluation of Nonsynonymous Single Nucleotide Polymorphisms in the ADIPOQ Gene Associated with Diabetes, Obesity, and Inflammation 2 2 Background: The human ADIPOQ gene encodes adiponectin protein hormone, which is involved in regulating glucose levels as well as fatty acid breakdown. It is exclusively produced by adipose tissue and abundantly present in the circulation, with concentration of around 0.01% of total serum proteins, with important effect on metabolism. Methods: Most deleterious nonsynonymous single nucleotide polymorphisms in the coding region of the ADIPOQ gene were investigated using SNP databases, and detected nonsynonymous variants were analyzed in silico from the standpoint of relevant protein function and stability by using SIFT, PolyPhen-2, PROVEAN and MUpro, I-Mutant2.0 tools, respectively. Result: A total of 58 nonsynonymous SNPs consisting of 55 missense variations, 3 nonsense variations were found in the ADIPOQ gene. Next, 14 of the 55 missense variants were predicted to be damaging or deleterious by three different software programs (PolyPhen-2, SIFT, and PROVEAN), and 38 of them were predicted to be less stable (I-Mutant 2.0 and MUpro software). Totally, 10 variants out of 55 missense variants were predicted to be both deleterious and reduce protein stability. Additionally, 3 nonsense variants were predicted to produce a truncated ADIPOQ protein. RMSD and total energy were calculated for 4 nsSNPs out of 10 nsSNPs which were both deleterious and showed a decrease in protein stability. Conclusion: rs144526209 has high root-mean-square deviation (RMSD) and lower total energy value compared to the native modeled structure. It was concluded that this nsSNP, potentially functional and polymorphic in the ADIPOQ gene, might be associated with diabetes, obesity, and inflammation. 121 127 Narayana Swamy A Department of Biotechnology, K L University Department of Biotechnology, K L University India Harika Valasala Department of Biotechnology, K L University Department of Biotechnology, K L University India Sreenivasulu Kamma Department of Biotechnology, K L University Department of Biotechnology, K L University India AdiponectinDiabetes mellitusGestationalNucleotide polymorphism 215.pdf Brochu-Gaudreau K, Rehfeldt C, Blouin R, Bordignon V, Murphy BD, Palin MF. Adiponectin action from head to toe. Endocrine 2010;37(1):11-32.##Meller M, Qiu C, Vadachkoria S, Abetew DF, Luthy DA, Williams MA. Changes in placental adipocytokine gene expression associated with gestational diabetes mellitus. Physiol Res 2006;55(5):501-512.##Nanda S, Savvidou M, Syngelaki A, Akolekar R, Nicolaides KH. Prediction of gestational diabetes mellitus by maternal factors and biomarkers at 11 to 13 weeks. Prenat Diagn 2011;31(2):135-141.##Dastani Z, Hivert MF, Timpson N, Perry JR, Yuan X, Scott RA, et al. Novel loci for adiponectin levels and their influence on type 2 diabetes and metabolic traits: a multi-ethnic meta-analysis of 45,891 individuals. PLoS Genet 2012;8(3):e1002607.##Adzhubei IA, Schmidt S, Peshkin L, Ramensky VE, Gerasimova A, Bork P, et al. A method and server for predicting damaging missense mutations. Nat Methods 2010;7(4):248-249.##Kumar P, Henikoff S, Ng PC. Predicting the effects of coding non-synonymous variants on protein function using the SIFT algorithm. Nat Protoc 2009;4(7):1073-1081.##Choi Y, Sims GE, Murphy S, Miller JR, Chan AP. Predicting the functional effect of amino acid substitutions and indels. PLoS One 2012;7(10):e46688.##Gromiha MM, An J, Kono H, Oobatake M, Uedaira H, Prabakaran P, et al. 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Int J Endocriol Metab 2011;9(1): 253-257.##Zhu N, Pankow JS, Ballantyne CM, Couper D, Hoogeveen RC, Pereira M, et al. High-molecular-weight adiponectin and the risk of type 2 diabetes in the ARIC study. J Clin Endocrinol Metab 2010;95(11):5097-5104.##Rizza S, Gigli F, Galli A, Micchelini B, Lauro D, Lauro R, et al. Adiponectin isoforms in elderly patients with or without coronary artery disease. J Am Geriatr Soc 2010;58(4):702-706.##Kottyan LC, Woo JG, Keddache M, Banach W, Crimmins NA, Dolan LM, et al. Novel variations in the adiponectin gene (ADIPOQ) may affect distribution of oligomeric complexes. Springerplus 2012;1(1):66.##Kadowaki T, Yamauchi T. Adiponectin and adiponectin receptors. Endocr Rev 2005;26(3):439-451.##

An Association Study on IL16 Gene Polymorphisms with the Risk of Sporadic Alzheimer’s Disease 2 2 Background: Interleukin-16 (IL-16) is an important regulator of T cell activation and was reported to act as a chemoattractant agent. There are evidences that IL16 can control the neuroinflammatory processes in Alzheimer’s Disease (AD). This study was performed to investigate the role or association of IL16 polymorphisms, rs11556218 and rs4778889 with the risk of late-onset Alzheimer’s disease (LOAD) in Iranian population. Methods: Totally, 148 AD patients and 137 nondemented and age-matched subjects were recruited in this study. Genotyping of rs11556218 T/G and rs4778889 T/C polymorphisms was performed by PCR-RFLP method using the NdeI and AhdI restriction enzymes, respectively. Results: Statistical analysis of rs11556218 genotypes showed a protective effect against AD in the heterozygote genotype (p=0.001, OR=0.16) as well as rs4778889 (p=0.001, OR=0.23). Frequency of rs11556218 allele T was higher in controls than patients (p=0.001, OR=0.32). However, there was no significant difference in the frequencies of rs4778889 alleles between the AD patients and controls.   Conclusion: Our results indicate that the rs11556218 and rs4778889 polymorphisms have a protective role in the development of sporadic AD in Iranian population. 128 132 Tayyebeh Khoshbakht Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Mohsen Soosanabadi Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Maryam Neishaboury Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Koorosh Kamali Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Masoud Karimlou Department of Biostatistics and Computer, University of Social Welfare and Rehabilitation Sciences Department of Biostatistics and Computer, University of Social Welfare and Rehabilitation Sciences Iran Niloofar Bazazzadegan Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Hamid Reza Khorram Khorshid Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Alzheimer’s diseaseGenetic association studyInterleukin-16Polymorphism 216.pdf Avramopoulos D. Genetics of Alzheimer's disease: recent advances. Genome Med 2009;1(3):34.##Bird TD. Genetic aspects of Alzheimer disease. Genet Med 2008;10(4):231-239.##Bekris LM, Yu CE, Bird TD, Tsuang DW. Genetics of Alzheimer disease. J Geriatr Psychiatry Neurol 2010;23 (4):213-227.##Daw EW, Payami H, Nemens EJ, Nochlin D, Bird TD, Schellenberg GD, et al. The number of trait loci in late-onset Alzheimer disease. Am J Hum Genet 2000;66 (1):196-204.##Rubio-Perez JM, Morillas-Ruiz JM. A review: inflammatory process in Alzheimer's disease, role of cytokines. ScientificWorldJournal 2012;2012:756357.##Akiyama H, Barger S, Barnum S, Bradt B, Bauer J, Cole GM, et al. Inflammation and Alzheimer's disease. Neurobiol Aging 2000;21(3):383-421.##Cruikshank WW, Kornfeld H, Center DM. Interleukin-16. J Leukoc Biol 2000;67(6):757-766.##Di Rosa M, Dell'Ombra N, Zambito AM, Malaguarnera M, Nicoletti F, Malaguarnera L. Chitotriosidase and inflammatory mediator levels in Alzheimer's disease and cerebrovascular dementia. Eur J Neurosci 2006;23(10): 2648-2656.##Manoochehri M, Kamali K, Rahgozar M, Ohadi M, Farrokhi H, Khorshid HR. Lack of Association between Tumor Necrosis Factor-alpha -308 G/A Polymorphism and Risk of Developing Late-Onset Alzheimer's Disease in an Iranian Population. Avicenna J Med Biotechnol 2009;1(3):193-197.##Khorram Khorshid HR, Manoochehri M, Nasehi L, Ohadi M, Rahgozar M, Kamali K. Ccr2-64i and Ccr5 Δ32 Polymorphisms in Patients with Late-Onset Alzheimer's disease; A Study from Iran (Ccr2-64i And Ccr5 Δ32 Polymorphisms in Alzheimer's disease). Iran J Basic Med Sci 2012;15(4):937-944.## Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988;16(3):1215.##Azimzadeh P, Romani S, Mohebbi SR, Kazemian S, Vahedi M, Almasi S, et al. Interleukin-16 (IL-16) gene polymorphisms in Iranian patients with colorectal cancer. J Gastrointestin Liver Dis 2011 ;20(4):371-376.##Streit WJ. Microglial activation and neuroinflammation in Alzheimer's disease: a critical examination of recent history. Front Aging Neurosci 2010;2:22.##Griffin WS. Inflammation and neurodegenerative diseases. Am J Clin Nutr 2006;83(2):470S-474S.##Tong Z, Li Q, Zhang J, Wei Y, Miao G, Yang X. Association between interleukin 6 and interleukin 16 gene polymorphisms and coronary heart disease risk in a Chinese population. J Int Med Res 2013;41(4):1049-1056.##Gao LB, Rao L, Wang YY, Liang WB, Li C, Xue H, et al. The association of interleukin-16 polymorphisms with IL-16 serum levels and risk of colorectal and gastric cancer. Carcinogenesis 2009;30(2):295-299.##Gao LB, Liang WB, Xue H, Rao L, Pan XM, Lv ML, et al. Genetic polymorphism of interleukin-16 and risk of nasopharyngeal carcinoma. 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