Avicenna and Evidence Based Medicine 2 2 Evidence Based Medicine (EBM) is now generally perceived to be the dominant operating system in con-ventional medicine. It is unsurprising then that some have counseled complementary and alternative medicine practitioners to resist EBM 1,2. The efficacy of medicinal herbs does need to be established and toxicity, contraindications and side effects also need to be investigated, and this is best done with clinical research and trials that at this time are being conducted almost exclusively on efficacy and are limited in number most probably because of funding. Very little to no attention is being given to the more traditional fresh herbal extracts 3,4. Many herbal medicines are now being supported by scientific evidence and have been shown to exert sig-nificant effects in the body, relieve symptoms, treat disease and improve everyday function. Any "expert" who still states there’s no scientific evidence to support the use of herbal medicines hasn’t done their homework. One of interesting example is saffron (Crocus sativus) for Alzheimer’s disease and depression that has been mentioned by Avicenna in his famous book. Avivenna’s famous works is the Canon of Medicine, which was a standard medical text at many medieval universities. The Canon of Medicine was used as a text-book in the universities of Montpellier and Leuven as late as 1650. Avicenna Canon of Medicine provides a complete system of medicine according to EBM. Saffron is the world’s most expensive spice, derived from the flower of Crocus sativus. Each saffron crocus grows to 20–30 cm and bears up to four flowers, each with three vivid crimson stigmas 3,4. Indeed, it is a Persian herb with a history as long as the Persian Empire itself. Iran, the world's largest producer of saffron has been investing in research into saffron's potential medicinal uses 3,4. To date, five published randomized controlled trials have been published about effects of saffron on depression. The first evidence-based study on this subject was published in 2004 showing that saffron was as efficacious as imipramine in the short-term treatment of mild to moderate depression in adults 5. Importantly, saffron was more tolerable than imipramine (which often causes anticholinergic side effects). Subsequently, saffron was compared to placebo in a six-week randomized controlled trial of 40 adult patients with mild to moderate depression. Saffron resulted in about 12-point reduction on Hamilton Depression Rating Scale (HDRS) compared with only five points seen with the placebo. Tolerability profile of saffron was similar to the placebo 5. Later, several studies provided evidence for antidepressant effects of different Crocus sativus L. constituents compared with both placebo and fluoxetine. Both petal and stigma of Crocus sativus L. have shown beneficial effects for treatment of depression 6,7. Crocus sativus L. is increasingly being studied as a memory enhancer. Saffron can attenuate the deleterious effect of ethanol on memory registration and retrieval, and prevent ethanol-induced inhibition of hippocampal long-term potentiation 3,4. Crocin seems to be involved in spatial memory and recognition and blocked scopolamine-induced performance deficits in the step-through passive avoidance and radial water maze tests 3,4. Saffron showed similar protective effects on recognition and spatial memory in chronic stress and hypoperfusion models of memory impairment 3,4. In an animal model of Alzheimer’s Disease (AD) induced by intraventricular injection of streptozocin, Khalili et al showed that administration of crocin resulted in significantly better results in passive avoidance test 3,4. In a 16-week placebo-controlled study, 46 patients with mild to moderate AD were assigned to saffron 15 mg twice daily or placebo. At the end of the trial, saffron was associated with a significantly better outcome on cognitive function than placebo. Importantly, tolerability of saffron was similar to placebo 8. In a 22-week donepezil-controlled study, saffron 15 mg twice daily was compared to donepezil 5 mg twice daily. Saffron was as efficacious as donepezil, but was associated with lower frequency of side effects than donepezil 9. Now if we read again the monograph regarding saffron in the Avicenna’s book we will find an evidence based medicine approach. 2 2 Shahin Akhondzadeh Tehran University of Medical Sciences Tehran University of Medical Sciences Iran Editorial 173.pdf Guyatt G. Evidence-based medicine. ACP J Club 1991;114 (Suppl2):A16.##Evidence-Based Medicine Working Group. Evidence-based medicine: a new approach to teaching the practice of medicine. JAMA 1992;268(17):2420-2425.##Akhondzadeh S. Herbal medicine in the treatment of psychiatric and neurological disorders. In: L’Abate L, (eds). Low cost approaches to promote physical and mental health: Theory research and practice. New York: Springer; 2007.##Akhondzadeh S, Abbasi SH. Herbal medicine in the treatment of Alzheimer's disease. Am J Alzheimers Dis Other Demen 2006;21(2):113-118.##Akhondzadeh S, Fallah-Pour H, Afkham K, Jamshidi AH, Khalighi-Cigaroudi F. Comparison of Crocus sativus L. and imipramine in the treatment of mild to moderate depression: a pilot double-blind randomized trial [ISRCTN45683816]. BMC Comp Alt Med 2004;4:12.##Akhondzadeh S, Tahmacebi-Pour N, Noorbala AA, Amini H, Fallah-Pour H, Jamshidi AH, et al. Crocus sativus L. in the treatment of mild to moderate depression: A double-blind, randomized and placebo controlled trial. Phytother Res 2005;19(2):148-151.##Akhondzadeh Basti A, Moshiri E, Noorbala AA, Jamshidi AH, Abbasi SH, Akhondzadeh S. Comparison of petal of Crocus sativus L. and fluoxetine in the treatment of depressed outpatients: a pilot double-blind randomized trial. Prog Neuropsychopharmacol Biol Psychiatry2007;31(2):439-442.##Akhondzadeh S, Sabet MS, Harirchian MH, Togha M, Cheraghmakani H, Razeghi S, et al. Saffron in the treatment of patients with mild to moderate Alzheimer's disease: a 16-week, randomized and placebo-controlled trial. J Clin Pharm Ther 2010;35(5):581-588.##Akhondzadeh S, Shafiee Sabet M, Harirchian MH, Togha M, Cheraghmakani H, Razeghi S, et al. A 22-week, multicenter, randomized, double blind controlled trial of Crocus sativus in the treatment of mild-to-moderate Alzheimer's disease. Psychopharmacology (Berl) 2010;207(4):637-643.##

Effect of Hepatitis B Virus X Gene on the Expression Level of p53 Gene using Hep G2 Cell Line 2 2 Background: The HBV-X (HBX) protein is believed to contribute to the development of HCC. However, the molecular mechanisms involved in HBX-mediated hepatocarcinogenesis remain obscure. In this study, the effect of hepatitis B virus X gene and its protein product HBxAg on expression of p53 gene in Hep G2 cell line was investigated. Methods: Viral DNA extracted from HBV-positive serum and HBX gene region was amplified using polymerase chain reaction (PCR). Then, PCR product was cloned into the pcDNA3 vector. After confirmation of cloning, the recombinant plasmid pcDNA3-X was transfected into HepG2 cell line using lipid-mediated DNA-transfection procedure. SDS-PAGE and western blotting methods were used to identify expression of HBX protein. Relative quantification was used to analyze the p53gene expression using the 2-ΔΔ Ct method. Results: Recombinant plasmid pcDNA3–HBX was confirmed by restriction endonucleases digestion and colony-PCR. The results of SDS-PAGE and western blot assays showed that HBX gene could be expressed in Hep G2 cell line. There was no significant difference between the expression levels of p53 compared with GAPDH gene as housekeeping gene (p<0.05). Conclusion: There was no significant difference in the protein levels between the transfected cells with X gene containing HBX130 and HBX131 double mutations and p53 gene. It is necessary to do more studies on Hepatitis B virus to understand the role of HBX on the development of liver cancer and its function on p53 tumor suppressor protein. 3 9 Roghyeh Kordestani Blood Transfusion Research Center, High Institute for Research and Education in Transfusion MedicineDepartment of Microbiology, School of Biological Science, Shahid Beheshti University Blood Transfusion Research Center, High Institute for Research and Education in Transfusion MedicineDepartment of Microbiology, School of Biological Science, Shahid Beheshti University IranIran Hamideh Mirshafiee Blood Transfusion Research Center, High Institute for Research and Education in Transfusion MedicineDepartment of Microbiology, School of Biological Science, Shahid Beheshti University Blood Transfusion Research Center, High Institute for Research and Education in Transfusion MedicineDepartment of Microbiology, School of Biological Science, Shahid Beheshti University IranIran Seyed Masoud Hosseini Department of Microbiology, School of Biological Science, Shahid Beheshti University Department of Microbiology, School of Biological Science, Shahid Beheshti University Iran Zohreh Sharifi 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 Hepatitis B virusHep G2 cell linep53 geneX gene 136.pdf Neuveut C, Wei Y, Buendia MA. Mechanisms of HBV-related hepatocarcinogenesis. J Hepatol 2010;52(4):594-604.##Dewantoro O, Gani RA, Akbar N. Hepato-carcinogenesis in viral hepatitis B infection: The role of HBx and p53. Acta Med Indones 2006;38(3):154-159.##Kew MC. Epidemiology of hepatitis B virus infection, hepatocellular carcinoma, and hepatitis B virus-induced hepatocellular carcinoma. Pathol Biol 2010;58(4):273-277.##Kew MC. Hepatitis B virus x protein in the pathogenesis of hepatitis B virus-induced hepatocellular carcinoma. J Gastroenterol Hepatol 2011;26(Suppl 1):144-152.##Miller RH, Robinson WS. Common evolutionary origin of hepatitis x virus and retroviruses. Proc Natl Acad Sci USA 1986;83(8):2531-2535.##ian Z, Liu J, Li L, Li X, Clayton M, Wu MC, et al. Enhanced cell survival of Hep3B cells by the hepatitis B x antigen effector, URG11, is associated with upregulation of beta-catenin. Hepatology 2006;43(3):415-424.##Lee JH, Han KH, Lee JM, Park HJ, Kim HS. Impact of hepatitis B virus (HBV) x gene mutations on hepatocellular carcinoma development in chronic HBV infection. Clin Vaccine Immunol 2011;18(6):914-921.##Ghabeshi S, Sharifi Z, Hosseini SM, Mahmoodian Shooshtari M. Correlation between viral load of HBV in chronic hepatitis B patients and precore and basal core promoter mutations. Hepat Mon 2013;13(2):e7415.##Abdel-Hafiz HA. Role of hepatitis B virus x protein in DNA repair during hepatocellular carcinoma development. J Carcinogene Mutagene 2011;S3:001.##Qu ZL, Zou SQ, Cui NQ, Wu XZ, Qin MF, Kong D, et al. Upregulation of human telomerase reverse transcriptase mRNA expression by in vitro transfection of hepatitis B virus X gene into human hepatocarcinoma and cholangiocarcinoma cells. World J Gastroenterol 2005;2811(36):5627-5632.##Chen HY, Tang NH, Li XJ, Zhang SJ, Chen ZX, Wang XZ. Transfection and expression of hepatitis B virus x gene and its effect on apoptosis in HL-7702 cells. World J Gastroenterol 2004;10(7):959-964.##Ou DP, Tao YM, Tang FQ, Yang LY. The hepatitis B virus X protein promotes hepatocellular carcinoma metastasis by upregulation of matrix metalloproteinases. Int J Cancer 2007;120(6):1208-1214.##Lin N, Chen HY, Li D, Zhang SJ, Cheng ZX, Wang XZ. Apoptosis and its pathway in X gene-transfected HepG2 cells. World J Gastroenterol 2005;11(28):4326-4331.##Vitvitski-Trépo L, Kay A, Pichoud C, Chevallier P, de Dinechin S, Shamoon BM, et al. Early and frequent detection of HBxAg and/or anti-HBx in hepatitis B virus infection. Hepatology 1990;12(6):1278-1283.##Zhang H, Wu LY, Zhang S, Qiu LY, Li N, Zhang X, et al. Anti-hepatitis B virus x protein in sera is one of the markers of development of liver cirrhosis and liver cancer mediated by HBV. J Biomed Biotechnol 2009;2009:289068.##Hwang GY, Lin CY, Huang LM, Wang YH, Wang JC, Hsu CT, et al. Detection of the hepatitis B virus x protein (HBx) antigen and anti-HBx antibodies in cases of human hepatocellular carcinoma. J Clin Microbiol 2003;41(12):5598-5603.##Yun C, Lee JH, Park H, Jin YM, Park S, Park K, et al. Chemotherapeutic drug, adriamycin, restores the function of p53 protein in hepatitis B virus X (HBx) protein-expressing liver cells. Oncogene 2000;19(45):5163-5172.##Neuveut Ch, Wei Y, Buendia MA. Mechanisms of HBV-related hepatocarcinogenesis. J Hepatol 2010;52(4):594-604.##Teodoro JG, Branton PE. Regulation of apoptosis by viral gene products. J Virol 1997;71(3):1739-1746.##Thomas M, Matlashewski G, Pim D, Banks L. Induction of apoptosis by p53 is independent of its oligomeric state and can be abolished by HPV-18 E6 through ubiquitin mediated degradation. Oncogene 1996;13(2):265-273.##Rubenwolf S, Schutt H, Nevels M, Wolf H, Dobner T. Structural analysis of the adenovirus type 5 E1B 55-kilodalton-E4orf6 protein complex. J Virol 1997;71(2):1115-1123.##Lin Y, Nomura T, Yamashita T, Dorjsuren D, Tang H, Murakami S. The transactivation and p53-interacting functions of hepatitis B virus X protein are mutually interfering but distinct. Cancer Res 1997;57(22):5137-5142.##Truant R, Antunovic J, Greenblatt J, Prives C, Chromlish JA. Direct interaction of the hepatitis B virus HBx protein with p53 leads to inhibition by HBx of p53 response element-directed transactivation. J Virol 1995;69(3):1851-1859.##Chung TW, Lee YC, Ko JH, Kim CH. Hepatitis B virus x protein modulates the expression of PTEN by inhibiting the function of p53, a transcriptional activator in liver cells. Cancer Res 2003;63(13):3453-3458.##Lee AT, Ren J, Wong ET, Ban KH, Lee LA, Lee CG. The hepatitis B virus x protein sensitizes HepG2 cells to UV light-induced DNA damage. J Biol Chem 2005;280(39):33525-33535.##

Isolation and Partial Characterization of Human Amniotic Epithelial Cells: The Effect of Trypsin 2 2 Background: Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells (hAECs) remains controversial. The aim of the present study was to characterize immuno-phenotypic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of trypsin used for tissue digestion. Methods: Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four trypsin types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor (EGF). To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining. Results: Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of trypsin used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies. Conclusion: Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC. 10 20 Meraj Tabatabaei Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Nariman Mosaffa Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Shohreh Nikoo Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Iran Mahmood Bozorgmehr Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Roya Ghods Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Somaieh Kazemnejad Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Simin Rezania Institute of Biophysics, Medical University of Graz Institute of Biophysics, Medical University of Graz Austria Bahareh Keshavarzi Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Soheila Arefi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Fahimeh Ramezani-Tehrani Reproductive Endocrinology Research Center, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences Reproductive Endocrinology Research Center, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences Iran Ebrahim Mirzadegan Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Reproductive Immunology Research Center, Avicenna Research Institute, ACECR Iran Amir-Hassan Zarnani Nanobiotechnology Research Center, Avicenna Research Institute, ACECRImmunology Research Center, Iran University of Medical Sciences Nanobiotechnology Research Center, Avicenna Research Institute, ACECRImmunology Research Center, Iran University of Medical Sciences IranIran Cell proliferationEpithelial cellsImmunophenotypingPlacentaStem cellsTrypsin 137.pdf Diwan S, Stevens LC. Development of teratomas from the ectoderm of mouse egg cylinders. J Natl Cancer Inst 1976;57:937-942.##Akle CA, Adinolfi M, Welsh KI, Leibowitz S, McColl I. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers. Lancet 1981;2(8254):1003-1005.##Toshio M, Thomas L, Hongbo C, Donna BS. Stem cell characteristics of amniotic epithelial cells. Stem Cells 2005;23:1549-1559.##Sakuragawa N, Kakinuma K, Kikuchi A, Okano H, Uchida S, Kamo I, et al. Human amnion mesenchyme cells express phenotypes of neuroglial progenitor cells. J Neurosci Res 2004;78(2):208-214.##Alviano F, Fossati V, Marchionni C, Arpinati M, Bonsi L, Franchina M, et al. Amniotic membrane is a high throughput source for multipotent mesenchymal stem cells with the ability to differentiate into endothelial cells in vitro. BMC Dev Biol 2007;7:11-20.##Zheng YB, Gao ZL, Xie C, Zhu HP, Peng L, Chen JH, Chong YT. Characterization and hepatogenic differentiation of mesenchymal stem cells from human amniotic fluid and human bone marrow: a comparative study. Cell Biol Int 2008;32(11):1439-1448.##Liu ZS, Xu YF, Feng SW, Li Y, Yao XL, Lu XL, Zhang C. Baculovirus-transduced mouse amniotic fluid-derived stem cells maintain differentiation potential. Ann Hematol 2009;88(6):565-572.##Perin L, Giuliani S, Jin D, Sedrakyan S, Carraro G, Habibian R, et al. Renal differentiation of amniotic fluid stem cells. Cell Prolif 2007;40(6):936-948.##Elwan M, Sakuragawa N. Evidence for synthesis and release of catecholamines by human amniotic epithelial cells. Neuroreport 1997;8(16):3435-3438.##Kakishita K, Elwan MA, Nakao N, Itakura T, Sakuragawa N. Human amniotic epithelial cells produce dopamine and survive after implantation into the striatum of a rat model of Parkinson’s disease: a potential source of donor for transplantation therapy. Exp Neurol 2000;165(1):27-34.##Dobreva MP, Pereira PN, Deprest J, Zwijsen A. On the origin of amniotic stem cells: of mice and men. Int J Dev Biol 2010;54(5):761-777.##Hunt JS, Wood GW. Interferon-gamma induces class I HLA and beta 2-microglobulin expression by human amnion cells. J Immunol 1986;136(2):364-367.##Parolini O, Alviano F, Bagnara GP, Bilic G, Bühring HJ, Evangelista M, et al. Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells. Stem Cells 2008;26(2):300-311.##Banas RA, Trumpower C, Bentlejewski C, Marshall V, Sing G, Zeevi A. Immunogenicity and immunomodulatory effects of amnion-derived multipotent progenitor cells. Hum Immunol 2008;69(6):321-328.##Murphy S, Rosli S, Acharya R, Mathias L, Lim R, Wallace E, Jenkin G. Amnion epithelial cell isolation and characterization for clinical use. Curr Protoc Stem Cell Biol 2010;1E.6.1-1E.6.25.##Nikoo S, Ebtekar M, Jeddi-Tehrani M, Shervin A, Bozorgmehr M, Kazemnejad S, Zarnani AH. Effect of menstrual blood-derived stromal stem cells on proliferative capacity of peripheral blood mononuclear cells in allogeneic mixed lymphocyte reaction. J Obstet Gynaecol Res 2012;38(5):804-809.##Darzi S, Zarnani AH, Jeddi-Tehrani M, Entezami K, Mirzadegan E, Akhondi MM, et al. Osteogenic differentiation of stem cells derived from menstrual blood versus bone marrow in the presence of human platelet releasate. Tissue Eng Part A 2012;18(15-16):1720-1728.##Tabatabaei-Panah AS, Jeddi-Tehrani M, Ghods R, Akhondi MM, Mojtabavi N, Mahmoudi AR, et al. Accurate sensitivity of quantum dots for detection of HER2 expression in breast cancer cells and tissues. J Fluoresc 2013;23(2):293-302.##Phelan MC. Basic techniques for mammalian cell tissue culture. Curr Protoc Cell Biol 1998; Chapter 1:Unit 1.1.##Dobreva MP, Pereira PN, Deprest J, Zwijsen A. On the origin of amniotic stem cells: of mice and men. Int J Dev Biol 2010;54(5):761-777.##Ilancheran S, Michalska A, Peh G, Wallace EM, Pera M, Manuelpillai U. Stem cells derived from human fetal membranes display multilineage differentiation potential. Biol Reprod 2007;77(3):577-588.##Liu YH, Vaghjiani V, Tee JY, To K, Cui P, Oh DY, et al. Amniotic epithelial cells from the human placenta potently suppress a mouse model of multiple sclerosis. PLoS One 2012;7(4):1-8.##Moll R, Franke WW, Schiller DL, Geiger B, Krepler R. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 1982;31(1):11-24.##Huang HL, Hsing HW, Lai TC, Chen YW, Lee TR, Chan HT, et al. Trypsin-induced proteome alteration during cell subculture in mammalian cells. J Biomed Sci 2010;17:36.##Hori J, Wang M, Kamiya K, Takahashi H, Sakuragawa N. Immunological characteristics of amniotic epithelium. Cornea 2006;25(10 Suppl 1):S53-58.##Li H, Niederkorn JY, Neelam S, Mayhew E, Word RA, McCulley JP, et al. 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Messenger RNA Expression Patterns of Neurotrophins during Transdifferentiation of Stem Cells from Human-Exfoliated Deciduous Teeth into Neural-like Cells 2 2 Background: Stem cells from Human Exfoliated Deciduous teeth (SHED) have the capability to differentiate into neural cells. Neurotrophins including Nerve Growth Factor (NGF), Brain-Derived Neurotrophic Factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4) have neurogenesis, neurotrophic, or neuroprotective effects and are expressed in developing teeth. The aim of this study was to measure quantitative changes in mRNA expression levels of neurotrophins in neural-like cells differentiated from dental pulp stem cells. Methods: Isolated total RNA from SHED, dental pulp and neural-like cells (n=3) were transcribed into cDNA. Then real time PCR was done. Expression levels of mRNA for NGF, BDNF, NT-3, and NT-4 genes were compared in these three cells. Results: In neural like cells, BDNF mRNA increased (372.1113.5) significantly (p<0.01) after differentiation. NGF mRNA increased to more than 266 times the dental pulp level after differentiation. A similar pattern was seen for the expression of NT3 after differentiation. NT4 mRNA enhancement was 1344630.8 and 30.77.9 fold in neural like cells and SHED cells, respectively. Results show alterations with different degrees and direction in neurotrophins mRNA expression levels in these cells. Conclusion: Our results suggest that neurotrophins dental pulp cells, SHED cells and neural like cells derived from SHED cells produce neurotrophic factors. Since the large amounts of neurotrophins are expressed in SHED and neural like cells they may have important role in survival and differentiation of dental pulp stem cells and obtained information may lead to a novel method for tooth regeneration. 21 26 Abolghasem Esmaeili Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan Iran Sedigheh Alifarja Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan Cell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan Iran Nosrat Nourbakhsh School of Dentistry, Isfahan University of Medical SciencesTorabinejad Dental Research Center, School of Dentistry, Isfahan University of Medical Sciences School of Dentistry, Isfahan University of Medical SciencesTorabinejad Dental Research Center, School of Dentistry, Isfahan University of Medical Sciences IranIran Ardeshir Talebi Department of Pathology, School of Medicine, Isfahan University of Medical Sciences Department of Pathology, School of Medicine, Isfahan University of Medical Sciences Iran Cell differentiationNeurotrophinReal-time PCRSurvival 138.pdf Lindvall O, Kokaia Z. 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SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA 2003;100(10):5807-5812.##Sakai Y, Yoshiura Y, Nakazawa K. Embryoid body culture of mouse embryonic stem cells using microwell and micropatterned chips. J Biosci Bioeng 2011;111(1):85-91. ##Nosrat CA, Ebendal T, Olson L. Differential expression of brain-derived neurotrophic factor and neurotrophin 3 mRNA in lingual papillae and taste buds indicates roles in gustatory and somatosensory innervation. J Comp Neurol 1996;376(4):587-602. ##Nosrat CA, Tomac A, Lindqvist E, Lindskog S, Humpel C, Stromberg I, et al. Cellular expression of GDNF mRNA suggests multiple functions inside and outside the nervous system. Cell Tissue Res 1996;286(2):191-207.##Nosrat CA, Fried K, Lindskog S, Olson L. Cellular expression of neurotrophin mRNAs during tooth development. Cell Tissue Res 1997;290(3):569-580.##Nosrat CA, Fried K, Ebendal T, Olson L. NGF, BDNF, NT3, NT4 and GDNF in tooth development. 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Downregulation of MMP2 and Bcl-2 in Adipose Derived Stem Cells (ASCs) following Transfection with IP-10 Gene 2 2 <p>Background: Mesenchymal Stem Cells (MSCs) are recently introduced as novel immunological gene carriers for treatment of cancer. It is believed that balance between the expression of angiogenic and anti-angiogenic factors, such as SDF-1 and IP-10, may regulate neovascularization within the tumor. Methods: In this study, we compared the expression of important tumor promoting mediators in IP-10-transfected Adipose Derived Stem Cells (ASCs) to those transfected with SDF-1. ASCs were isolated from adipose tissue of a normal subject undergoing cosmetic mamoplasty surgery using collagenase. ASCs were transfected with IP-10 or SDF-1 propagated plasmids by electroporation method and Lipofectamin 2000. Expressions of SDF-1, CXCR4, IP-10, Bcl-2, MMP2, IL-10, IGF-1, and VEGF were detected in transfected ASCs using quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Results: Results showed that the expressions of SDF-1, CXCR4, Bcl-2, MMP2, IL-10, IGF-1, and VEGF were upregulated in SDF-1-transfected ASCs. In contrast, Bcl-2 and MMP2 transcripts showed 45×103 and 10 fold lower expression in ASCs transfected with IP-10 compared to non-transfected cells. Conclusion: Anti-angiogenic chemokines such as IP-10 may modulate tumor promoting properties of ASCs and would be introduced as novel candidates for tumor immunotherapy; however, further studies are needed to be conducted.</p> 27 37 Mahboobeh Razmkhah Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences Iran Mansooreh Jaberipour Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences Iran Abbas Ghaderi Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical SciencesDepartment of Immunology, School of Medicine, Shiraz University of Medical Sciences Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical SciencesDepartment of Immunology, School of Medicine, Shiraz University of Medical Sciences IranIran Adipose derived stem cellsIP-10SDF-1TransfectionTumor immunotherapy 139.pdf Uccelli A, Pistoia V, Moretta L. 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J Biol Chem 2008;283(7):4283-4294.##Kato M, Kitayama J, Kazama S, Nagawa H. Expression pattern of CXC chemokine receptor-4 is correlated with lymph node in human invasive ductal carcinoma. Breast Cancer Res 2003;5(5):R144-150.##Sei S, O’Neill DP, Stewart SK, Yang Q, Kumagai M, Boler AM, et al. Increased level of stromal cell-derived factor-1 mRNA in peripheral blood mononuclear cells from children with AIDS-related lymphoma. Cancer Res 2001;61(13):5028-5037.##Lazarini F, Tham TN, Casanova Ph, Arenzana-Seisdedos F, Dubois-Dalcq M. Role of the α-chemokine stromal cell derived factor (SDF-1) in the development and mature central nervous system. Glia 2003;42(2):139-148.##Li M, Ransohoff RM. The roles of chemokine CXCL12 in embryonic and brain tumor angiogenesis. Semin Cancer Biol 2009;19(2):111-115.##Sun X, Cheng G, Hao M, Zheng J, Zhou X, Zhang J, et al. CXCL12 / CXCR4 / CXCR7 chemokine axis and cancer progression. 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Future Oncol 2009;5(8):1145-1168.##Gimble JM, Katz AJ, Bunnell BA. Adipose-derived stem cells for regenerative medicine. Circ Res 2007;100(9):1249-1260.##Taléns-Visconti R, Bonora A, Jover R, Mirabet V, Carbonell F, Castell JV, et al. Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells. World J Gastroenterol 2006;12(36):5834-5845.##Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002;13(12):4279-4295.##Razmkhah M, Jaberipour M, Erfani N, Habibagahi M, Talei AR, Ghaderi A. Adipose derived stem cells (ASCs) isolated from breast cancer tissue express IL-4, IL-10 and TGF-β1 and upregulate expression of regulatory molecules on T cells: do they protect breast cancer cells from the immune response? Cell Immunol 2011;266(2):116-122.##Razmkhah M, Jaberipour M, Ghaderi A. 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Stem Cells 2008;26(1):151-162.##Rasmusson I, Ringdén O, Sundberg B, Le Blanc K. Mesenchymal stem cells inhibit the formation of cytotoxic T lymphocytes, but not activated cytotoxic T lymphocytes or natural killer cells. Transplantation 2003;76(8):1208-1213.##Najar M, Raicevic G, Boufker HI, Fayyad-Kazan H, De Bruyn C, Meuleman N, et al. Adipose-tissue-derived and Wharton's jelly-derived mesenchymal stromal cells suppress lymphocyte responses by secreting leukemia inhibitory factor. Tissue Eng Part A 2010;16(11):3537-3546.##Giuliani M, Fleury M, Vernochet A, Ketroussi F, Clay D, Azzarone B, et al. Long-lasting inhibitory effects of fetal liver mesenchymal stem cells on T-lymphocyte proliferation. PLoS One 2011;6 (5):e19988.##Jotzu C, Alt E, Welte G, Li J, Hennessy BT, Devarajan E, et al. Adipose tissue derived stem cells differentiate into carcinoma-associated fibroblast-like cells under the influence of tumor derived factors. 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Assessment of Different Permeabilization Methods of Minimizing Damage to the Adherent Cells for Detection of Intracellular RNA by Flow Cytometry 2 2 Background: Various fixation and permeabilization techniques have been developed for detection of intracellular antigens by flow cytometry; however, there are few studies using flow cytometry to detect the frequency of intracellular nucleic acids, particularly RNA. We tested six different permeabilization methods in order to gain access to a high quality method with minimal damage to intracellular components focusing on 18S rRNA in HeLa cells. Methods: HeLa cells were fixed in 2% paraformaldehyde. A variety of detergents and enzymes including saponin, TritonX-100, Tween-20, NP40, Proteinase K, and streptolysin O were used to optimize a protocol of permeabilization for the flow cytometric enumeration of intracellular 18S rRNA. Treated cells were subjected to standard protocol of flow cytometric in situ hybridization in the presence of FITC-labeled sense and antisense probes to detect 18S ribosomal RNAs. Samples were then analyzed on a FACSCalibur flow cytometer. To evaluate cell morphology, following hybridization the cells were fixed on glass slide, covered with DAPI, and evaluated on a fluorescent microscope with appropriate filter sets. Results: In comparison with other methods, maximum cell frequency in percentage and fluorescent intensity (M1=2.1%, M2=97.9%) were obtained when the cells were treated with 0.2% Tween-20 and incubated for 30 min (p=0.001). Conclusion: Our study indicated that the highest levels of mean fluorescence could be obtained when the cells were treated with Tween-20. However, it should be taken into consideration that for a successful flow cytometric result, other interfering factors such as hybridization conditions should also be optimized. 38 46 Zahra Amidzadeh Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical SciencesStudent Research Committee, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical SciencesStudent Research Committee, Shiraz University of Medical Sciences Iran Iran Abbas Behzad-Behbahani Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Nasrollah Erfani Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences Iran Sedigheh Sharifzadeh Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Reza Ranjbaran Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Leili Moezzi Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Farzaneh Aboualizadeh Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Mohammad Ali Okhovat Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Parniyan Alavi Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences Iran Negar Azarpira Shiraz Transplant Research Center, Shiraz University of Medical Sciences Shiraz Transplant Research Center, Shiraz University of Medical Sciences Iran 18S rRNAFlow cytometryHeLa cells 140.pdf Rieseberg M, Kasper C, Reardon KF, Scheper T. Flow cytometry in biotechnology. Appl Microbiol Biotechnol 2001;56:350-360.##Hanley MB, Lomas W, Mittar D, Maino V, Park E. Detection of low abundance RNA molecules in individual cells by flow cytometry. PLoS One 2013;8(2):e57002. ##Yu H, Ernst L, Wagner M , Waggoner A. Sensitive detection of RNAs in single cells by flow cytometry. Nucleic Acids Res 1992;20(1):83-88.##Robertson KL, Verhoeven AB, Thach DC, Chang EL. Monitoring viral RNA in infected cells with LNA flow-FISH. RNA 2013;16(8):1679-1685.##Ormerod MG. Cell preparation for flow cytometry. Methods Mol Biol 1992;10:359-362.##Holmes K, Lantz LM, Fowlkes BJ, Schmid I, Giorgi JV. Preparation of cells and reagents for flow cytometry. Curr Protoc Immunol 2001;Chapter 5:Unit 5.3.##Verdier M, Jayat C, Ratinaud MH, Troutaud D. Optimization of cell permeabilization for multiparametric flow cytometric analysis with lectin staining. Cytometry 2000;41(1):55-61.##Preobrazhensky SN, Bahler DW. Optimization of flow cytometric measurement of ZAP-70 in chronic lymphocytic leukemia. Cytometry B Clin Cytom 2008;74(2):118-127.##Koley D, Bard AJ. Triton X-100 concentration effects on membrane permeability of a single HeLa cell by scanning electrochemical microscopy (SECM). Proc Natl Acad Sci USA 2010;107(39):16783-16787.##Schimenti KJ, Jacobberger JW. Fixation of mammalian cells for flow cytometric evaluation of DNA content and nuclear immunofluorescence. Cytometry 1992;13(1):48-59.##Fox CH, Johnson FB, Whiting J, Roller PP. Formaldehyde fixation. J Histochem Cytochem 1985;33(8):845-853.##Williams Y, Byrne S, Bashir M, Davies A, Whelan A, Gun’ko Y, et al. Comparison of three cell fixation methods for high content analysis assays utilizing quantum dots. J Microsc 2008;232(1):91-98.##Mercanti V, Cosson p. Resistance of Dictyostelium discoideum membranes to saponin permeabilization. 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Fourier Transform Infrared Spectroscopy: A Potential Technique for Noninvasive Detection of Spermatogenesis 2 2 Background: The seminal plasma is an excellent source for noninvasive detection of spermatogenesis. The seminal plasma of normospermic and azoospermic men has been analyzed for detection of spermatogenesis. Methods: Optical spectroscopy (Attenuated Total Reflectance-Infrared spectroscopy (ATR-IR) and Fourier Transform infrared spectroscopy (FT-IR) has been used to analyze the seminal plasma and the metabolome of seminal plasma for detection of spermatogenesis. Results The seminal plasma of normospermic and azoospermic men has been analyzed by ATR-IR. The results show that there is a pattern variation in the azoospermic men compared to normospermic men. However, the seminal plasma is too complex to show significant pattern variation. Therefore, the metabolome which is a subcomponent of the seminal plasma was analyzed. The seminal plasma metabolome of normospermic and azoospermic men has been analyzed by FT-IR. A significant pattern change was observed. The data combined with chemometrics analysis showed that significant changes are observed at metabolome level. Conclusion: We suggest that FT-IR has the potential as a diagnostic tool instead of testicular biopsy. 47 52 Kambiz Gilany Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Roudabeh Sadat Moazeni Pouraci Department of Chemistry, Sharif University of Technology Department of Chemistry, Sharif University of Technology Iran Mohammad Reza Sadeghi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran AzoospermiaFourier transform infrared spectroscopySeminal plasmaMetabolome 141.pdf Owen DH, Katz DF. 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Detection of E. coli O157:H7 from ground beef using Fourier transform infrared (FT-IR) spectroscopy and chemometrics. J Food Sci 2010;75(6):M340-346.##Muller JJ, Neumann M, Scholl P, Hilterhaus L, Eckstein M, Thum O, et al. Online monitoring of biotransformations in high viscous multiphase systems by means of FT-IR and chemometrics. Anal Chem 2010;82(14):6008-6014.##Ioannou-Papayianni E, Kokkinofta RI, Theocharis CR. Authenticity of cypriot sweet wine commandaria using FT-IR and chemometrics. J Food Sci 2011;76(3):C420-427.##Li X, Li QB, Zhang GJ, Xu YZ, Sun XJ, Shi JS, et al. Identification of colitis and cancer in colon biopsies by Fourier Transform Infrared spectroscopy and chemometrics. ScientificWorldJournal 2012;2012:936149. Epub 2012/05/31.##Schmidtke LM, Smith JP, Muller MC, Holzapfel BP. Rapid monitoring of grapevine reserves using ATR-FT-IR and chemometrics. Anal Chim Acta 2012;732:16-25.##Psychogios N, Hau DD, Peng J, Guo AC, Mandal R, Bouatra S, et al. 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Evaluation of the Effect of miR-26b Up-Regulation on Hb-F Expression in Erythroleukemic K-562 Cell Line 2 2 Background: The major hemoglobin in the fetus is hemoglobin F (𝛼2𝛾2), whereas in adult humans, hemoglobin A (𝛼2𝛽2) is predominately expressed. Several studies have indicated that expression of the HbF subunit 𝛾-globin might be regulated post-transcriptionally. This could be done by small non-coding RNAs called microRNAs which target mRNAs in a sequence-specific manner and lead to translational repression or mRNA decay. The aim of this study is to evaluate the effect of miR-26b up-regulation on 𝛾-globin gene expression in K-562 cell line. Methods: These cells were grown in RPMI 1640 and pre miR-26b and were transfected within K-562 cell line using lentiviral vector. After RNA extraction and cDNA synthesis in selected days, miRNA up-regulation was confirmed by miRNA real time PCR and then 𝛾and 𝛽chain and GATA-1 expression were investigated by RT and QRT-PCR. Results: The viability of cells before transfection was 90%. Three and 7 days after transfection, through the use of relative Q-PCR, the 𝛾 chain expression increased 3.7, 6.8 and 3.8 folds and GATA-1 expression increased 2.1, 6.0 and 8.0 in comparison with untransfected cells. Conclusion: The data suggest that miR-26b can be involved in the increase of 𝛾-globin gene expression in K-562 cell line. We suggest that miR-26b may be a significant therapeutic target for increasing HbF levels in patients with sickle cell disease and 𝛽-thalassemia. 53 56 Sadegh Alijani Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Shaban Alizadeh Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Ahmad Kazemi Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Zahra Kashani Khatib Students Scientific Research Center (SSRC), Allied Medical School, Tehran University of Medical Sciences Students Scientific Research Center (SSRC), Allied Medical School, Tehran University of Medical Sciences Iran Masoud Soleimani Department of Hematology, Tarbiat Modares University Department of Hematology, Tarbiat Modares University Iran Mohamadreza Rezvani Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Neda Minayi Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Farshid Karami Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Department of Hematology, Allied Medical School, Tehran University of Medical Sciences Iran Behnoosh Tayebi Department of Hematology, Qaem Hospital, Mashhad University of Medical Sciences Department of Hematology, Qaem Hospital, Mashhad University of Medical Sciences Iran K-562MicroRNAsMiR-26b 142.pdf Bauer DE, Orkin SH. Update on fetal hemoglobin gene regulation in hemoglobinopathies. Curr Opin Pediatr 2011;23(1):1-8.##Sankaran VG, Orkin SH. The switch from fetal to adult hemoglobin. Cold Spring Harb Perspect Med 2013;3(1):a011643.##Platt OS, Thorington BD, Brambilla DJ, Milner PF, Rosse WF, Vichinsky E, et al. N Engl J Med 1991;325(1):11-16.##Serjeant GR. Sickle cell disease. Vol. 2. New York: Oxford university press; 1992.##Labie D, Pagnier J, Lapoumeroulie C, Rouabhi F, Dunda-Belkhodja O, Chardin P, et al. Common haplotype dependency of high G gamma-globin gene expression and high Hb F levels in beta-thalassemia and sickle cell anemia patients. Proc Natl Acad Sci USA 1985;82(7): 2111-2114.##Kim YK, Kim VN. Processing of intronic microRNAs. EMBO J 2007;26(3):775-783.##Bushati N, Cohen SM. microRNA functions. Annu Rev Cell Dev Biol 2007;23:175-205.##Du T, Zamore PD. microPrimer: the biogenesis and function of microRNA. Development 2005; 132(21):4645-4652.##Walker AL, Steward S, Howard TA, Mortier N, Smeltzer M, Wang YD, et al. Epigenetic and molecular profiles of erythroid cells after hydroxyurea treatment in sickle cell anemia. Blood 2011;118(20):5664-5670.##Welch JJ, Watts JA, Vakoc CR, Yao Y, Wang H, Hardison RC, et al. Global regulation of erythroid gene expression by transcription factor GATA-1. Blood 2004;104(10):3136-3147.##

Fine Structures of the Oocyte in Relation to Serum, Follicular Fluid Steroid Hormones and IGF-I in the Ovulatory-Sized Follicles in One-Humped Camel (Camelus dromedarius) 2 2 Background: The following study was carried out to determine the ultrastructural features of the oocyte of the ovulatory-sized follicles in relation to concentrations of steroids and IGF-I in the follicular fluid and serum in the dromedary camel. Methods: Camel follicles with a clear and healthy appearance were categorized into three classes: follicles 10 to 13.9, 14-17.9 and 18-30 mm diameter. The Follicular Fluid (FF) and serum samples were assayed for estradiol-17β, progesterone and IGF-I. Recovered Cumulus-Oocyte Complexes (COCs) were prepared for transmission electron microscopy. Results: The mean (±SD) FF concentrations of progesterone and IGF-I was significantly (p<0.05) higher in follicles 18 to 30 mm diameter compared to other groups of follicles. There was no difference in the mean (±SD) serum estradiol-17β, progesterone and IGF-I concentrations between camels with different ovulatory-sized follicles (p>0.05). Oocytes from follicles 18 to 30 mm diameter (group 3) showed more advanced signs of maturation including the disappearance of the nuclear envelope, increased number of microvilli in erect position, the increase in number and size of vesicles and more even distribution of the mitochondria throughout the ooplasm. Conclusion: The final stages of oocyte maturation in dromedary camel is associated with increasing progesterone and IGF-I concentrations and constant high estradiol concentration in the follicular fluid which are paralleled with well-defined ultrastructural changes in oocytes. 57 61 Mojtaba Kafi Department of Animal Reproduction, School of Veterinary Medicine, Shiraz University Department of Animal Reproduction, School of Veterinary Medicine, Shiraz University Iran Seyed Fakhroddin Mesbah Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences Iran Najmeh Davoodian Department of Animal Reproduction, School of Veterinary Medicine, Shiraz University Department of Animal Reproduction, School of Veterinary Medicine, Shiraz University Iran Ali Kadivar Research Institute of Animal Embryo Technology, Shahrekord University Research Institute of Animal Embryo Technology, Shahrekord University Iran CamelIGF-IOocyteSteroidsUltrastructure 143.pdf Skidmore JA. Reproduction in dromedary camels: an update. Anim Reprod 2005;2(3):161-171.##Skidmore JA, Billah M, Allen WR. Investigation of factors affecting pregnancy rate after embryo transfer in the dromedary camel. Reprod Fert Devel 2002;14:109-116.##Badr MR, Abdel-Malak MG. In vitro fertilization and embryo production in dromedary camel using epididymal spermatozoa. Global Vet 2010;4:271-276.##Wani NA, Wernery U, Hassan FAH, Wernery R, Skidmore JA. Production of the first cloned camel by somatic cell nuclear transfer. Biol Reprod 2010;82(2):373-379.##Nili H, Mesbah F, Kafi M, Nasr Esfahani MH. Light and transmission electron microscopy of immature camelus dromedarius oocyte. Anat Histo Embryol 2004;33(4):196-199.##Davoodian N, Mesbah F, Kafi M. Oocyte ultrastructural characteristics in camel (Camelus dromedarius) primordial to large antral follicles. Anat Histo Embryol 2011;40(2):120-127.##Kafi M, Mesbah F, Nili H, Khalili A. Chronological and ultrastructural changes in camel (Camelus dromedarius) oocytes during in vitro maturation. Theriogenology 2005;63(9):2458-2470.##Rahman ZU, Bukhari SA, Ahmad N, Akhtar N, Ijaz A, Yousaf MS, et al. Dynamics of follicular fluid in one-humped camel (Camelus dromedarius). Reprod Dom Anim 2008;43(6):664-671.##Spicer LJ, Santiago CA, Davidson TR, Bridges TS, Chamberlain CS. Follicular fluid concentrations of free insulin-like growth factor (IGF)-I during follicular development in mares. Domest Anim Endocrinol 2005;29(4):573-581.##Spicer LJ. Proteolytic degradation of Insulin-like growth factor binding proteins by ovarian follicles: A control mechanism for selection of dominant follicles. Biol Reprod 2004;70(5):1223-1230.##Skidmore JA. Reproductive physiology in female old world camelids. Anim Reprod Sci 2011;124(3-4):148-154.##Hyttel P, Callesen H, Greve T. Ultrastructural features of preovulatory oocyte maturation in superovulated cattle. J Reprod Fertil 1986;76:615-625.##Grondahl C, Hyttel P, Grondahl ML, Eriksen T, Goteredsen P, Greve T. Structural and endocrine aspects of equine oocytes maturation in vivo. Mol Reprod Devel 1995;42:94-105.##Teissier MP, Chable H, Paulhac S, Aubard Y. Comparison of follicle steroidogenesis from normal and polycystic ovaries in women undergoing IVF: relationship between steroid concentrations, follicle size, oocyte quality and fecundability. Hum Reprod 2000;15(12):2471-2477.##Sirotkin AV, Dukesova J, Makarevich AV, Kubek A, Bulla J. Evidence that growth factors IGF-I, IGF-II and EGF can stimulate nuclear maturation of porcine oocytes via intracellular protein kinase A. Reprod Nutr Dev 2000;40(6):559-569.##Zhao J, Taverne MA, Van der Weijden GC, Bevers MM, Van Den Hurk R. Insulin-like growth factor-1 (IGF-I) stimulates the development of cultured rat pre-antral follicles. Mol Reprod Dev 2001;58(3):287-296.##Revelli A, Delle Piane L, Casano S, Molinari E, Massobrio M, Rinaudo P. Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics: A review. Reprod Biol Endocrinol 2009;7:40-52.##Fair T, Hulshof SCJ, Hyttel P, Greve T, Boland M. Oocyte ultrastructure in bovine primordial to early tertiary follicles. Anat Embryol 1997;195:327-336.##Grado-Ahuir JA, Aad PY, Spicer LJ. New insights into the pathogenesis of cystic follicles in cattle: Microarray analysis of gene expression in granulosa cells. J Anim Sci 2011;89(6):1769-1786.##