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Background and Objective: Alpha thalassemia is one of the most common hemoglobin disorders. Some combination of alpha globin gene mutations may cause HbH disease with severe anemia or intermediate thalassemia. genotype common deletions are routinely tested for suspicious alpha thalassemia couples but because of lack of information about the nature and frequency of point mutations and higher expenosor of sequencing, less attention was paid to them. This study was done to determine the prevalence of common point mutations of alpha globin gene in Babol, Iran.

Materials and Methods: This descriptive study was carried out on DNA of 153 adult suspected to α-thalasemia with deleted α- golobolin gene referred to genetic laboratory in Babol, Iran during 2005-09. a1 and a2 genes were amplified by using specific biotinilated primers by PCR method. PCR products were assayed using 11 specific probs corresponding to common point mutations in alpha gene (C19, IVSI (-5nt), C59, Hb constant spring, Hb Icaria, Hb seal Rock, IVSI (148), C14, poly A (-2bp), poly A2, Poly A1) and fixed on byodine C membrabe. Hybridization between the probes and PCR products was visualized after a colorimetric reaction using of conjugated streptavidin peroxidase and TMB (tetra methyle Benzidine) and H²O².

Results: The prevalence of point mutations in poly A2, 5nt, Hb constant spring and poly A1 were 28.75%, 14.38%, 7.84% and 2.61%, respectively.

Conclusion: Point mutation in alpha globin genes was detected in %53.60 out of 153 adults suspected with alpha thalassemia without common deletion mutations.

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Background and Objective: According to world health organization statistics, at least 5.2% of world population is carrier for a main hemoglobin disorder. Previous reports showed that more than 10% of people are carrier for beta-thalassemia Northern Iran. This study was done to determine the prevalance of hemoglobinopathies in premarriage individuals referred to Babolsar, Iran.

Materials and Methods: This descriptive study was carried out on 8500 individuals  (4200 women and 4300 men) whome were attended the thalassemia counseling program in Babolsar, North of Iran during 2006-09. After performing the CBC test, for those MCV and MCH were less than 80 and 27 respectively, Hemoglobin A2 was evaluated. Subjects whome were volunteers for more comprehensive tests, basic and acidic electrophoresis and genetic tests were applied, subsequently.

Results: 1200 (14.11%) subjects had low hematological indexes. 474 (5.57%) subjects had high HbA2 and were classified as beta-thalassemia carriers and 726 (8.54%) had normal HbA2 level and were classified as alpha-thalassemia carriers. 6 (1.2%) subjects were identified with HbF level more than 10 and were identified as carriers for beta-gene cluster deletion carrier. Also, 16 (3.2%) individuals had HbE, 16 (3.2%) had HbS, 4 had HbD and 4 had HbH (0.33% in 1200 and 0.047% in 8500 subjects). Genetic study of 317 individuals for beta carriers and 145 subjects for alpha-carriers showed IVSII-1G>A (74.5%) in beta-globin and single gene deletion of 3.7 (47.5%) in alpha-globin genes were the most frequent mutations.

Conclusion: This study showed that carriers for alpha - thalassemia (8.5%) are more frequent compared with beta- thalassemia (5.57%). Also other hemoglobin variants included HbS, HbE, HbD or different beta-gene cluster deletions in the region are considerable and should be screened.

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Background and Objective: Down syndrome is a genetic disorder due to trisomy of 21 chromosome, which mainly is manifested by moderate to severe mental problems, including physical, sensory and motor symptoms. This study was done to determine the related factors associated with the birth of children with Down syndrome. Materials and Methods: This cross-sectional study was carried out on 913 mothers with children or fetuses of Down syndrome who were referred to genetic counseling clinics in Tehran, Iran during 2011. Diagnosis of Down syndrome confirmed by the chromosomes culture. Maternal age, maternal age at menarche, parents interfamilial marriage, maternal blood group, the number of abortions and children, collected through the questionnaires. Results: Out of 913 affected children and fetuses with Down syndrome, 420 (46%) and 493 (54%) were female and males respectively. 1.17 considered to be The male/female ratio. 32.4% of mothers were in 25-30 years of age and 42% of parants had interfamilial marriages. The abortion in mothers varied from lack of ahortion (96.7%) up to 5 repetitive abortion (0.1%). The number of children in families with affected subject consisted of 1 (15.1%), 4 (16.5%) and 11-13 (1.3%). Conclusion: This study showed that Down syndrome infants are mostly born with the mothers of 25-30 years age.

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Background and Objective: Sphingosine-1-phosphate (S1P) is involved in regulation of proliferation, differentiation, hypertrophy and anti-apoptosis and activation of satellite cells. This study was done to evaluated the effect of 8 weeks resistance training on sphingosine-1-phosphate level and gene expression of SK1 enzyme, isoforms of MHCs in skeletal muscles of male Wistar rats. Materials and Methods: This experimental study was done on Twenty four 8-week-old 190-250 gr male Wistar rats. The rats were allocated randomly into control (N=12) and training (N=12) groups. Resistance training was done using a 1 meter height ladder with 2 cm grid with an 85 degree incline, and weights attached to rat's tails. The content of S1P present in the chloroform layer was determined by means of high performance liquid chromatography (HPLC). Determination of relative mRNA expression was performed by Real-time PCR. Data were analyzed using SPSS-17, Kolmogorov-Smirnov and independent t-test. Results: Resistance exercise training increased the total content of S1P in FHL (fast-twitch) and soleus (slow-twitch) muscles in comparison with control group (P<0.05). Resistance exercise training changed the gene expression of FHL SK1, SOL SK1, FHL MHC I, Sol MHC I, FHL MHC IIa, Sol MHC IIa, FHL MHC IIb, Sol MHC IIb, FHL MHC IIx, Sol MHC IIx in comparison with control group (P<0.05). Conclusion: This study showed that S1P level and gene expression of SK1, MHCs increased at skeletal muscles after training.

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Background and Objective: Toll-like receptors (TLRs) have been discovered as the most important receptors in innate immunity. One of the most important TLRs is TLR4, the key receptor for the LPS component of gram-negative bacteria. Two polymorphisms, D299G (rs4986790) and T399I (rs4986791), in TLR4 gene are associated with a decreased response to LPS. This study was done to estimate the expression of different polymorphisms of TLR4 gene in colorectal cancer cell line by flowcytometery. Materials and Methods: In this laboratory study, the HCT116 cells were transfected with plasmids containing different variants of TLR4 gene including Flag-tagged-TLR4 wild type, flag-tagged D299G and T399I Using TurboFect transfection reagent. Transfection efficiency was evaluated by GFP plasmid. Expression of different variants of TLR4 was assessed in transfected cells by flowcytometery. Data were analyzed using SPSS-11.5 and chi-square test. Results: TLR4 was detected on HT29 and CaCo2 cell lines at low levels. HCT116 cells did not express detectable amounts of TLR4 by flowcytometery prior to transfection. Gene transfer efficiency for GFP plasmid was about 80% in HCT116 cells by flowcytometery and microscopic analysis. TLR4 expression and LPS responsiveness significantly was higher in HCT116 cells which were transfected with wild type TLR4 gene compared to non-transfected and mutant transfected cells (P<0.05). Conclusion: Lower expression of TLR4 on cells with mutant TLR4 showed that these polymorphisms affect on expression patterns of TLR4 on colon cancer cells.

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Background and Objective: Hemoglobinopathies are among the most prevalent genetic disorders worldwide, and occur as a result of mutations in the gene involved in synthesizing hemoglobin chains. By now more than 1000 defects in hemoglobin chains are discovered. Hemoglobin D (Hb D) is one of these disorders, identified by a single nucleotide mutation on codon 121 of beta globin chain. This study was carried out to evaluate Hb D mutations through molecular methods in Mazandaran province of Iran. Materials and Methods: This descriptive laboratory study was done on 70 patients with an electrophoresis band in hemoglobin-S zone in Mazandaran province of Iran during 2010-11. Capillary zone electrophoresis was done to find out Hb D in 51 patients. Subsequently, PCR-RFLP was performed to evaluate the samples at molecular level. Results: Molecular investigation revealed all cases are carriers of hemoglobin D-Punjab. Two patients were shown to be homozygote carriers of the abnormal gene. Conclusion: This study showed all Hb D affected patients were carriers of Hb D Punjab.

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Papillon-Lefevre Syndrome is a rare autosomal recessive disorder. This syndrome accompanied by palmoplantar hyperkeratosis and severe periodontal destruction of primary and permanent teeth. The teeth erupt normally but due to the severe alveolar bone loss both in deciduous as well as permanent dentitions, these teeth are exfoliated within two or three years after eruptions and by the age of 15 or 17. Patients are usually edentulous. Due to periodontal disease, the dentists are often the first ones who diagnose the syndrome. A 15-year-old girl was referred to dental clinic complaining of permanent teeth mobility. All patient’s teeth except 13, 14, 17, 23, 27, 37, 43, 44 and 47 had been extracted. The third molars were impacted. Patient has advanced periodontal disease and all teeth have mobility. There was hyperkeratosis at the palms and soles. The teeth were extracted and treated with complete denture. Early diagnosis of the papillon-lefevre syndrome can help to preserve teeth. Dental treatment included extraction of all deciduous teeth, professional prophylaxis, conventional periodontal therapy, systemic antibiotics, oral retinoid, complete dentures and implants.

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Background and Objective: Breast cancer is one of the most frequent malignancies among women. This study was done to determine the BRCA1 gene expression in 7,12-Dimethylbenz(a)anthracene (DMBA) induced breast cancer in rats. Materials and Methods: In this experimental study, the breast cancer was induced by DMBA in Sprague dawley rats. After tumors arise, cell cultures were prepared and G-banding staining was performed on metaphase chromosomal smear. According to databases, genes in the affected area were collected and after comparing genome of the rats and human in changed chromosomal segments, a gene list was prepared. FISH technique was performed on BRCA1 gene to prove accuracy of chromosomal banding results. Results: Structural changes such as deletion occurred in chromosomes 10, which BRCA1 is located on. 24.7% of cells showed evidence of physical deletion in both copy of BRCA1 gene and 23.8% of cells showed deletion in one copy. Conclusion: Induced DMBA Breast cancer cells showed deletion in BRCA1 copy numbers. This gene may be involved in animal breast tumor model.

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Background and Objective: Mayer Rokitansky Kuster Hauser (MRKH) syndrome is characterized by Mullerian duct aplasia in an XX individual with female phenotype presenting primary amenorrhea at adolescence. This study was done to determine the mutations of cystic fibrosis transmembrane conductance regulator (CFTR) gene including DF508, G542X, N1303K, W1282X in patients with MRKH syndrome. Methods: This case-control study was performed on 25 females with MRKH syndrome and 25 healthy females. Blood sample was taken from each subject. DNA genomic was isolated by standard methods and common mutations of CFTR gene analyzed by ARMS-PCR. Results: DF508 gene was found in 3 in case and one individual in control group. G542X, N1303K and W1282X gene was not detected. Conclusion: DF508 gene was found in 12% of patients with MRKH syndrome.

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Background and Objective: SLC26A4 gene mutations after GJB2 mutations are the second currently identifiable genetic cause of autosomal recessive non syndromic hearing loss (ARNSHL) which currently is used in molecular diagnosis of ARNSHL. Several potential STR markers related to this region have been reported .This study was carried out to identity the informativeness of D7S2456 CA repeat STR marker in SLC26A4 gene region in five ethnic groups of the Iranian population. Methods: In this descriptive study, The locus was genotyped in 165 unrelated healthy individuals of five different ethnics including Fars, Azari, Turkmen, Gilaki and Arabs ethnic groups using polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis (PAGE) and fluorescent capillary electrophoresis. Data was analyzed by Gene Marker HID Human STR Identity software, Gene Pop program and Microsatellite Tools software. Results: Analysis of the allelic frequency revealed the presence of 9 alleles for D7S2456 marker in the Iranian population, which allele 5 at the D7S2456 locus with 55% frequency was the most frequent. The most frequent heterozygosity with rate of 81.8% belongs to Azari ethnic group. Analysis of deviations from Hardy-Weinberg equilibrium demonstrated that all the ethnics except Fars were in equilibrium for D7S2456 locus. D7S2456 marker is a moderately informative marker in Iranian ethnic population (PIC value within 0.44 and 0.7). Conclusion: D7S2456 is a moderately informative marker in diagnosis of SLC26A4 based autosomal recessive non syndromic hearing loss in Iranian population by linkage analysis.

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Background and Objective: Human erythropoietin (EPO) is a glycosylated hormone with molecular weight of about 40 KDa which is synthesized in kidneys and plays an important role in proliferation and differentiation of erythrocytes.This study was done to assess and analyze the expression of recombinant EPO in Leishmania tarentolae host. Method: In this descriptive study, the EPO gene was codoned , optimized with bioinformatics database prior to be synthesized. It was cleaved by KpnIandXbaI enzymes and cloned into pLEXSY expression vector. The constructed expression cassette was transfected into Leishmania tarentolae through electroporaton method. Identification and confirmation of transfected colonies was performed using PCR expression diagnostic primers and EPO specific primers. Induction of the cloned gene was done with tetracycline. The expression in induced strains was analyzed by SDS-PAGE and western blotting techniques. The amount of recombinant protein was quantified by ELISA method. Confirmation of cloning and EPO expression cassette was carried out through genetic engineering procedures. Results: Expression analysis of transfected parasitic strain with SDS-PAGE and western blotting confirmed gene integration into chromosomal of host as well as expression. The optimal conditions for expression were found to be 10μg of tetracycline and 72h induction time. Molecular weight of expressed protein estimated to be 40 KDa and expression level was determined to be 12.4 mg/l which was equal to 1% of total protein mass. Conclusion: EPO expression cassette for cloning and expression in Leishmania tarentolae was designed and protein of interest was successfully induced and identified Leishmania tarentolae can be used as a suitable host for production of recombinant EPO and this technology has a potential for localization.

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Background and Objective: Hemoglobin D-Punjab is one of the variant of hemoglobin caused by a mutation on position 121 of beta globin gene which is frequent in India, Pakistan and Iran. Heterozygote form of this variant is mainly asymptomatic while in combination with hemoglobin S, severe form of anemia occure. This study was carried out to determine the beta globin gene haplotypes associated with hemoglobin D-Punjab in Northern Iran. Methods: This descriptive study was carried out on families of 18 individuals whom were carriers of hemoglobin D-Punjab in Sari in Northern Iran. Genomic DNA was extracted from peripheral blood samples using Phenol-chloroform standard protocol. In order to identify different haplotypes associated with hemoglobin D-Punjab, PCR-RFLP method and family linkage analysis were used. Results: In 17 subjects hemoglobin D-Punjab was linked to [+ - - - - + +] haplotype and in one case association with [- + + - + + +] haplotype was observed. Conclusion: The hemoglobin D-Punjab alleles have mainly unicentric origin and [- + + - + + +] rare haplotype may have different genetic origin or is created as a result of gene recombination.

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Background and Objective: Mirtazapine is a norepinephrine and serotonergic antidepressant that is used in the theraphy of major depressive disorders. This study was carried out to determine the genotoxic and cytotoxic effect of mirtazapine using chromosome aberration and mitotic index tests in human peripheral blood lymphocytes. Methods: In this descriptive -analytic study genotoxic and cytotoxic effect of mirtazapine at 24 and 48 hours treatment periods on four concentration (10, 25, 40, and 55µg/ml) was performed on peripheral blood lymphocyte of four subjects. Results: Mirtazapine significantly reduced the mitotic index in the all concentrations but it non-significantly increased the chromosome aberration at 24-hours and 48-hours treatment periods. Conclusion: Mirtazapine has cytotoxic effect but it has no genotoxic effect on human lymphocyte.

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Background and Objective: The interleukin-18 (IL-18) gene on chromosome 11 has been suggested as a susceptibility factor for allergies. It is a member of the IL-1 family that was originally described as interferon (IFN-γ)–inducing factor. IL-18 might initiate Th2 responses with production of IgE via the stimulation of IL-4 and IL-13 synthesis in mast cells and in basophil and eosinophil recruitment, such as allergic inflammation. This study was done to assess the association of Interleukin-18 gene polymorphism -137G/C with allergic rhinitis. Methods: This case-control study was performed on 293 allergic rhinitis patients and 218 healthy control volunteers .The IL-18 polymorphism was analyzed by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. Results: The frequency of the GG, GC and CC genotypes were 64.2%, 32.1% and 3.7% in healthy controls and 60.1%, 35.5% and 4.4% in allergic rhinitis patients, respectively. This diference was not significant. Conclusion: This study suggests that IL-18 polymorphism gene -137G/C may not be participated as a risk factor in the pathogenesis of allergic rhinitis.

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Background and Objective: Despite enormous progress in the understanding of human reproductive physiology, the underlying cause of male infertility remains undefined in about 50.0% of cases, which are referred to as idiopathic infertility and affects about 5.0-7.0% of the general male population. Human apurinic/apyrimidinic endonuclease (ApE1) is a multifunctional protein that has an important role in the base excision repair (BER) pathway. ApE1 SNP T>G found in exon 5 led to substitution of Asp>Glu at codon 148. This study was done to evaluate the association of ApE1 Asp148Glu polymorphism and the risk of idiopathic male infertility. Methods: In this case-control study, blood samples were collected from 90 patients diagnosed with idiopathic male infertility and 90 healthy men, genotyped by Allele-Specific PCR (AS-PCR) method by using specific primers that were designed and the association between genotype and allele frequencies in cases and controls were estimated. Results: There was no significant association between ApE1 gene polymorphism at codon 148 in case and control groups. Conclusion: No significant association was found between the Asp148Glu polymorphism and idiopathic male infertility.

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Background and Objective: Skeletal muscle expresses several neurotrophin and their receptors which providing the basis for neurotrophin signaling within the muscle compartments. This study was done to evaluate the effect of a session of resistance exercise on mRNA expression of NT-3 and TrkC proteins in soleus muscle of Wistar Rats. Methods: In this experimental study, 16 male Wistar rats were randomly allocated into exercise and control groups. The resistance training protocol consisted of climbing a 1-meter–long ladder, with a weight attached to a tail sleeve. Expressions of NT-4/5 and P75, quantitatively were measured using RT-PCR. Results: There was not any significant alteration in NT-3 mRNA in soleus muscle after resistance exercise. However, one session of resistance exercise significantly increased mRNA expression of TrkC (1.7 Folds) in soleus muscle (P<0.05). Conclusion: Resistance exercise increases TrkC expression in soleuse muscle of wistar rats.

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Background and Objective: Breast cancer is a cancer in women with high prevalancy worldwide. Vascular endothelial growth factor (VEGF) is one of the most important Pro-angiogenic factors. +405C/G is one of the common VEGF polymorphism which may have an impact on the level of gene expression and over loading of gene products. This study was done to evaluate the association between VEGF +405C/G gene polymorphism and breast cancer risk in north of Iran.

Methods: This case-control study was carried out on 50 patients with breast cancer and 50 normal aged-matched controls in north of Iran. Genomic DNA was extracted from peripheral blood cells. To determine the genotype of +405 C/G VEGF gene polymorphism, PCR-RFLP method was used.

Results: The prevalence of genotypic frequencies of GG, GC and CC in controls were 42%, 48% and 10%, respectively  and in patients were 22%, 46% and 32%, respectively (P<0.05). The +405C allele was considered as a risk factor in breast cancer (P<0.05).

Conclusion: It seems +405 C/G VEGF gene polymorphism may be associated with the breast cancer in northern Iran.

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Background and Objective: Micro RNAs (or miRNAs) control gene regulation and different biological processes in various tissues and therefore play an important role in various diseases. In some cases, either a single nucleotide polymorphisms (SNPs) in miRNAs or in complementary sequences in their target mRNAs play significant role in human diseases. In this study, the relationship between rs531564 G>C in mir-124-1 with the susceptibility to type 2 diabetes in the Iranian population was examined.

Methods: In this case-control study, 173 individuals affected with type 2 diabetes and 162 healthy individuals were selected. Extracted DNA from peripheral blood was amplified by a pair of relevant primers and then digested by BsmAI restriction enzymes. The obtained electrophoretic patterns were used for genotyping.

Results: The genotype frequencies of GG, GC and CC for rs531564 in the patient group were 0.92, 0.06 and 0.02 respectively, 0.96, 0.04 and 0.00 in the controls. Statistical analysis showed no significant difference between the two groups regarding the genotype frequencies, however the allelic frequencies were significantly different between those groups (P<0.05).

Conclusion: There was no genotype difference between diabetes and healthy individuals, but the allelic C is related with type 2 diabetes among Iranian population.

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Background and Objective: Tumor size results in hypoxic and acidic environment leading to the production of several types of growth factors required for the formation of blood vessels. Afterwards, metastasis of cancerous cells occurs via blood vessels. Therefore angiogenesis inhibition can be a new way of cancer treatment. This study was done to determine the bioinformatics analysis to predict potential Micro-RNAs inhibiting processes of angiogenesis in cancer.

Methods: In this descriptive study, micro-RNAs that are able to connect to MMP genes involved in tumor angiogenesis (MMP1-2-3-8-9-10-11-13) were detected using miRwalk database. Effective Micro-RNAs selection was based on the number of binding sites in 3'UTR genes. MicroRNA data base was used to find sample base pairing sequences.

Results: mir-1302, mir-516a, mir-512, mir-511, mir-516b and mir-548 were determined with the most number of binding sites in genes involved in angiogenesis.

Conclusion: MicroRNAs are worthy options for cell culture and laboratory examination in order to find new ways to prevent the development of cancer by angiogenesis inhibition.

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Neurofibromatosis type1 (NF1) with the incidence of 1 in 3500 births, is the most common disorder which affects skin and peripheral nervous system. NF1 results from mutations in NF1 gene. The NF1 gene spans 350kbp and to date, nearly 2434 mutations in it were reported. The gene with 100 percent penetrance is located on chromosome 17 encoding neurofibromin protein. Recently, many challenges of its genetic analysis have been overcome through the application of new sequencing techniques. In present study patients with neurofibromatosis type 1 have been characterized from clinical symptoms such as presence of café au lait spot, plexiform neurofibroma, optic nerves involvement, presence of several patients in first degree relatives. These patients were in different ages including 73, 63, 44, 20 with different symptoms and severities of disease. In this communication, a NF1 family with 4 cases in 3 generations has been presented.