en 2008-2835 2008-4625 276 5669 11181 gregorian >2014 >January-March 6 1 online 1 fulltext

en 24523951 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 https://www.ajmb.org/En/Article.aspx?id=173 https://www.ajmb.org/PDF/En/FullText/173.pdf ShahinAkhondzadehTehran University of Medical Sciences, Tehran, Iran739

en 24523952 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. Hepatitis B virus, Hep G2 cell line, p53 gene, X gene 3 9 https://www.ajmb.org/En/Article.aspx?id=136 https://www.ajmb.org/PDF/En/FullText/136.pdf RoghyehKordestaniDepartment of Microbiology, School of Biological Science, Shahid Beheshti University, Tehran, Iran637HamidehMirshafieeDepartment of Microbiology, School of Biological Science, Shahid Beheshti University, Tehran, Iran638Seyed MasoudHosseiniDepartment of Microbiology, School of Biological Science, Shahid Beheshti University, Tehran, Iran639ZohrehSharifiBlood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran640

en 24523953 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. Cell proliferation, Epithelial cells, Immunophenotyping, Placenta, Stem cells, Trypsin 10 20 https://www.ajmb.org/En/Article.aspx?id=137 https://www.ajmb.org/PDF/En/FullText/137.pdf MerajTabatabaeiDepartment of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran642NarimanMosaffaDepartment of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran643ShohrehNikooReproductive Immunology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran644MahmoodBozorgmehrNanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran645RoyaGhodsMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran8SomaiehKazemnejadReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran64SiminRezaniaInstitute of Biophysics, Medical University of Graz, , Austria187BaharehKeshavarziDepartment of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran649SoheilaArefiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran78FahimehRamezani-TehraniReproductive Endocrinology Research Center, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran652EbrahimMirzadeganReproductive Immunology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran653Amir-HassanZarnaniImmunology Research Center, Iran University of Medical Sciences, Tehran, Iran7

en 24551431 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. Cell differentiation, Neurotrophin, Real-time PCR, Survival 21 26 https://www.ajmb.org/En/Article.aspx?id=138 https://www.ajmb.org/PDF/En/FullText/138.pdf AbolghasemEsmaeiliCell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan, Isfahan, Iran654SedighehAlifarjaCell and Molecular Biology Division, Department of Biology, School of Sciences, University of Isfahan, Isfahan, Iran655NosratNourbakhshTorabinejad Dental Research Center, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran656ArdeshirTalebiDepartment of Pathology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran658

en 24551432 <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&times;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> Adipose derived stem cells, IP-10, SDF-1, Transfection, Tumor immunotherapy 27 37 https://www.ajmb.org/En/Article.aspx?id=139 https://www.ajmb.org/PDF/En/FullText/139.pdf MahboobehRazmkhahShiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran662MansoorehJaberipourShiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran444AbbasGhaderiDepartment of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran424

en 24523954 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. 18S rRNA, Flow cytometry, HeLa cells 38 46 https://www.ajmb.org/En/Article.aspx?id=140 https://www.ajmb.org/PDF/En/FullText/140.pdf ZahraAmidzadehStudent Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran663AbbasBehzad-BehbahaniDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran664NasrollahErfaniShiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran665SedighehSharifzadehDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran666RezaRanjbaranDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran667LeiliMoezziDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran668FarzanehAboualizadehDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran669Mohammad AliOkhovatDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran670ParniyanAlaviDiagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran671NegarAzarpiraShiraz Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran672

en 24523955 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. Azoospermia, Fourier transform infrared spectroscopy, Seminal plasma, Metabolome 47 52 https://www.ajmb.org/En/Article.aspx?id=141 https://www.ajmb.org/PDF/En/FullText/141.pdf KambizGilanyReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran673Roudabeh SadatMoazeni PouraciDepartment of Chemistry, Sharif University of Technology, Tehran, Iran674Mohammad RezaSadeghiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran40

en 24523956 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. K-562, MicroRNAs, MiR-26b 53 56 https://www.ajmb.org/En/Article.aspx?id=142 https://www.ajmb.org/PDF/En/FullText/142.pdf SadeghAlijaniDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran675ShabanAlizadehDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran219AhmadKazemiDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran677ZahraKashani KhatibStudents Scientific Research Center (SSRC), Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran678MasoudSoleimaniDepartment of Hematology, Tarbiat Modares University, Tehran, Iran221MohamadrezaRezvaniDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran680NedaMinayiDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran681FarshidKaramiDepartment of Hematology, Allied Medical School, Tehran University of Medical Sciences, Tehran, Iran682BehnooshTayebiDepartment of Hematology, Qaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran683

en 24551433 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. Camel, IGF-I, Oocyte, Steroids, Ultrastructure 57 61 https://www.ajmb.org/En/Article.aspx?id=143 https://www.ajmb.org/PDF/En/FullText/143.pdf MojtabaKafiDepartment of Animal Reproduction, School of Veterinary Medicine, Shiraz University, Shiraz, Iran685Seyed FakhroddinMesbahDepartment of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran686NajmehDavoodianDepartment of Animal Reproduction, School of Veterinary Medicine, Shiraz University, Shiraz, Iran687AliKadivarResearch Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran688