Tehran University Medical Journal

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Tumor angiogenesis shown by Microvessel Count (MVC) or Microvessel Density (MVD), is assessed by several studies as prognostic factor in some types of tumors, and also in colorectal carcinoma. This article is payed to correlation between clincopathologic factors and tumor angiogenesis. In this study, immunohistochemical techniques are used for vascular evaluation in specimens from twenty-nine colorectal carcinoma, and stained for Factor VIII-Related Antigen (F8RA) by using monoclonal antibody. Uni and multivariate analysis disclosed that total MVC was higher in tumor [76.3±33 (×100=2.5 mm²/field) and 29.8±11 (×200=0.785 mm²/field)] than in normal tissue [37.7±15.8 (×100) and 17.6±7.8 (×200)], (P=0.022, P=0.000009). Microvessel quantification was significantly higher in stage D (115±36.6, ×100 and 26.7±6.4, ×200, P=0.002 and P=0.04). In this study MVD has correlation with vascular invasion (P=0.024, ×100 and P=0.007, ×200), the mean tumor vessel count although was increased with clinicophatologic findings such as age<60 years, male, right colon involvement, infiltrating type, mucinous carcinoma, transmural penetration, grade III, lymphatic and perineural invasion, but was not statistically significant. Lymph node and hematogenous metastasis and size of tumor also, was not important. As a conclusion, MVD was increased in tumor and has shown correlation with metastasis, and vascular invasion. Resulting angiogenesis increase risk of metastasis.

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Background: The important of angiogenesis for the progressive growth and viability of solid tumors is well established. Only few data are available for hematologic neoplasms.

Materials and Methods: To investigate the role of angiogenesis in the acute myloid leukemia (AML) bone marrow biopsies from 30 adults with newly diagnosed, untreated AML(day 0) were evaluated. Further studies were done after completion on remission induction of treatment (day 35 of 7×3 regimen n=13, complete remission in AML (m3) treat with arsenic trioxide n=17). Micro-vessels were scored in at least 3 areas of highest micro-vessel density in representative section of each bone marrow specimen using immunohistochemistry for Von Willbrand factor.

Results: Median micro-vascular density (MVD) were in AMLM3 patients before treatment, %6.81±3.58 and after treatetment %3.48±3.06 (p<0.0001). In other AML patients MVD were befor treatment %3.38 and after treatment %3.6.

Conclusion: In conclusion, there is evidence of increased micro-vessel density in the bone marrow of patients with AML, which supports the hypothesis of an important role of angiogenesis in AML. MVD was reduced with chemotherapy and arsenic. Furthermore , these finding suggest that antiangiogenesis therapy might constitute a novel strategy for the treatment of AML.

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Background: This study was designed to investigate the process of bone formation caused by implantation of octa calcium phosphate (OCP) in rat tibiae.

Methods: We used 25 young male Sprague-Dawley rats. A full thickness standardized trephine defect, 3-mm in diameter, was surgically created on the superior end of right and left tibia. Amount of 6-mg synthetic Octa calcium Phosphate was implanted into a bony defect on the right tibia as a experimental group. No OCP particles were implanted in the left tibia as a control group that was otherwise treated identically. Bone formation was examined histologically on 7th, 10th, 14th, 21st, 28th days after implantation.

Results: In the experimental group, on the 7th day after implantation, a few clusters of cartilage cells were observed between the OCP particles near the defects margin. Osteogenesis was initiated locally between the OCP particles in central position of the defects on 10th day after implantation. By 14th day after implantation, Alcian blue staining showed hypertrophic chondrocytes that replaced by new bone. In addition to bone formation locally around the OCP particles, more apposition of new bone was observed near the defects margin on 14th and 21st days after implantation. At the end of study implanted OCP was surrounded by newly formed bone. In the control group, at the end of study, bone formation was observed only along and near the defects margin.

Conclusion: These results demonstrate that octa calcium phosphate could be used in the repair of the long bone defects.

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Background: Tumor cells need food and oxygen supply for growth and division. Therefore one of the most promising areas of cancer therapy focuses on using agents that inhibit tumor angiogenesis. Inhibition of angiogenesis prevents cell growth, division and metastasis. Previous studies showed that plasminogen related Protein-B has an anti-tumor activity in mice. This protein has a high level of homology with preactivation Peptide (PAP) of human plasminogen. According to this high homology, antiangiogeneic activity of PAP was investigated in an in vitro angiogenesis model.

Methods: PAP encoding region of human plasminogen gene was isolated by Polymerase Chain Reaction and ‎cloned in pGEX-2T vector. This plasmid was expressed in Escherichia coli as a fusion protein (GST-PAP). ‎GST-PAP was expressed as inclusion body and purified by affinity chromatography on GSH-sepharose ‎resin after refolding. antiangiogenic effects of purified protein were surveyed with Matrigel assay‏.‏‎ ‎

Results: The GST-PAP was expressed and purified and its accuracy was confirmed by SDS-PAGE analysis ‎and immunoblotting. Microscopic studies showed that GST-PAP inhibited angiogenesis in Matrigel system ‎which is shown by shrinking the length of capillary like structures and a decrease in the number of tubule. ‎While applying concentarations of 25μg/ml of GST-PAP and concentrations above that, antiangiogenic ‎activity of GST-PAP was significant comparing to the controls. ‎

Conclusion: Finding shows that GST-PAP can inhibit network formation in Matrigel system. This findings ‎support the theory that PAP is a potent angiogenesis inhibitor.‎

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Background: Plasminogen has a central role in fibrinolyrtic system can activate through various activators (PAs) to its active form plasmin and perfoem its vital function that is fibrin clot lysis. Furthermore the fibrinolyrtic system plays a major role in angiogenesis. The fibrinolyrtic system activation control cell migration and invasion. In addition to this, plasmin regulates tumor growth. Monoclonal antibodies, as biological tools, play an important role in basic researches.

Methods: In the first step the effects of antibodies on the activation of fibrinolyrtic system with PAs were evaluated with several methods including macroscopic observation, quantitative measurement of DD/E fragments by D-dimer assay and activation of plasminogen by S-2251 synthetic substrate (ELISA method), subsequently we studied the effect of antibodies on angiogenesis process in an in- vitro model.

Results: Results showed that MC2B8 that is an inhibitor of plasminogen activation in presence of plasminogen activators can inhibit angiogenesis process: A1D12 that is against N-terminal domain of Glu-plasminogen, in addition to activation of fibrinolyrtic system in presence of plasminogen activators, can activate in vitro angiogenesis process.

Conclusion: Plasmin formation is a critical step in invasion and migration of endothelial cells to form new vessels. Plasmin directly participates in angiogenesis by direct fibrin and other matrix components degradation, and indirectly by activating matrix degrading metalloproteinase and angiogenic growth factors. According to the in- vitro results, MC2B8 and A1D12 monoclonal antibodies play roles in this process in a dose dependent manner.

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Background: Despite advances in cancer diagnosis and treatment, survival rate of patients suffering from glioblastoma multiform (GBM) has not been significantly improved. Therefore, novel therapeutic adjuncts to routine therapies have been suggested over time. Inhibition of angiogenesis by antiangiogenic drugs is one of the new approaches to inhibit the growth of malignant cells. Microvessel density (MVD) assay is a technique performed by counting immunohistochemically-stained blood vessels. Nowadays, athymic nude mice are widely used for the establishment of xenograft tumor models in cancer research. The aim of this study was to evaluate the MVD of autochthonous xenograft models of GBM isolated from Iranian patients for use in pharmaceutical research on antiangiogenic drugs.Methods: Fresh tumor samples of GBM were obtained from three patients in Cancer Institute of Tehran University of Medical Sciences in Fall of 2010 and Winter of 2011. After preliminary processing, minced tumor samples were implanted heterotopically on flanks of athymic nude mice. Two months later, the animals were sacrificed and the xenograft tumor samples were sent to the pathology laboratory. After establishing the proof of the xenograft tumor type, MVD-CD34, an endothelial cell marker, was assessed by counting hot spot areas in 22 samples.Results: The mean number of microvessels in these xenograft tumor models was 30±2.1. Conclusion: These autochthonous xenograft models of GBM can be used in preclinical settings for research on antiangiogenic drugs regarding a pharmacogenomics-based treatment regimen for the Iranian population. Moreover, such models can be used in future studies for determining the sensitivity or resistance to antiangiogenic drugs in individualized cancer therapy.

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Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: Breast cancer is the most common form of hereditary cancer worldwide and is an important cause of morbidity and mortality. Approximately 5-10% of breast and ovarian cancers are due to the highly penetrating germline mutations in cancer predisposing genes. Two genes, BRCA1 and BRCA2, account for at least half of these cases. The demand for BRCA1 and BRCA2 mutation screening is rapidly increasing as their identification will affect the medical management of people at increased risk for the disease. Therefore, the aim of this study was to investigate BRCA1/2 mutations in 100 high risk Iranian families.
Methods:  One hundred families who met the minimal risk factors for breast/ovarian cancer were screened among the families referred to Kawsar Human Genetics Research Center for the diseases in 2009-2011. The entire coding sequences and each intron/exon boundaries of BRCA1/2 genes were screened for by direct sequencing and MLPA in both patients and the controls.
Results:  In the present study, we could detect the following novel mutations: p.Gly1140Ser, p.Ile26Val, p.Leu1418X, p.Glu23Gln, p.Leu3X, p.Asn1403His, p.Asn1403Asp, p.Lys581X, p.Pro938Arg, p.Thr77Arg, p.Leu6Val, p.Arg7Cys, p.Leu15Ile, p.Ser177Thr, IVS7+83(-TT), IVS8 -70(-CATT), IVS2+9(G>C), IVS1-20(G>A), IVS1-8(A>G), p.Met1Ile, IVS2+24(A>G), IVS5-8 (A>G), IVS2(35-39)TTcctatGAT, IVS13+9 G>C in BRCA1 and p.Glu1391Gly, p. Val1852Ile, IVS6-70(T>G), 1994-1995 (InsA) in BRCA2.
Conclusion: Ten mutations seemed to be pathogenic and the disease-causing mutations were seen in 16% of the families. In addition, from the total number of substitutions and reassortments (42), 80% related to BRCA1 and 20% to mutations in BRCA2 genes.

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Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: Gastric cancer is one of the most common diseases of digestive system with a low 5-year survival rate and metastasis is the main cause of death. Multi-factors, such as changes in molecular pathways and deregulation of cells are involved in the disease development. Epidermal growth factor receptor pathway (EGFR) which is associated with cell proliferation and survival can influence cancer development. EGFR function is governed by its genetic polymorphism thus, we aimed to study the tyrosine kinase domain gene mutations of the receptor in patients with gastric cancer.
Methods : In this experimental study, 123 subjects (83 patients with gastric cancer and 40 normal subjects) were investigated in north of Iran for EGFR gene polymorphisms during 1 year. Genomic DNA was extracted by DNA extraction kit according to the manufacture's protocol. Polymerase chain reaction single-stranded conformation polymorphism (PCR-SSCP) and silver staining were performed for investigating EGFR gene polymorphisms.
Results : The participants included 72 men and 44 women. Gene polymorphism in exon 18 was present in 10% of the study population but SSCP pattern in exon 19 did not show different migrate bands neither in patients nor in normal subjects.
Conclusion: It seems that screening for tyrosine kinas gene polymorphism of epidermal growth factor receptor in patients with gastric cancer and use of tyrosine kinas inhibitors could be useful in the prevention of disease progress and improvement of treatment process for a better quality of life in these patients.

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Background: Achillea millefolium or yarrow is a native plant in Europe and Iran. Yarrow has been used as a medicine historically, mainly because of its astringent effects. It is reported to be associated with the treatment of several ailments. Nowadays use of plants for medical purpose has become very common. Achillea millefolium L, Yarrow, is being used in traditional and modern medicine due to various chemical compounds. Considering the importance of birth control, finding a drug with less side effects inhibiting spermatogenesis seems to be necessary. The aim of the present study was to investigate the effect of ethanol extract of Achillea millefolium L. on spermato-genesis of male wistar rats.
Methods: In this study, 32 adult male wistar rats were used. The animals were divided to four groups of eight rats. The first group, received 200 mg/kg Achillea millefolium L. interaperitoneally, the second and third groups received 400, 800 mg/kg Achillea millefolium L. interaperitoneally, respectively. In the fourth group (control) distilled water was administered. After 20 days, the rats were sacrificed and testis tissues were histologically evaluated.
Results: Comparing to control group, in the experimental groups received the high doses of the extract, thickening of seminiferous tubules basement membrane, loss of germinal epithelium and testicular hyperemia were demonstrated (P<0.001).
Conclusion: Based on the results, high concentrations of Achillea millefolium L. leaded to structural and spermatogenesis changes in testis tissue.

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Background: The aim of this study was compared the efficacy of  the designed primers and already published primers for detection of the exoA, oprL and algD genes by PCR assay  for finding a rapid, accurate and highly sensitive and specific procedure to detect the Pseudomonas aeruginosa in the serious and fatal infections such as cystic fibrosis disease, burned individual.
Methods: A total of 150 clinical specimens were inoculated in to routine and selective culture media for Pseudomonas aeruginosa isolation. Specific primers were designed by bioinformatics analysis for detection of the virulence genes exoA, oprL and algD. The available sequences of these three genes were obtained from NCBI and multiple alignments were performed to find the conserved sequences of each gene for primer designing. Both multiple alignment and primer designing steps were carried out by AlleleID software, version 7.0.
Results: Microbiological culture methods were showed that 70 Pseudomonas aeruginosa strains isolated from the 150 clinical specimens. PCR assay performed by using the designed primers shown 68, 70 and 69 positive results from 70 direct specimens for exoA, oprL and algD respectively that shown 97.2%, 100% and 98.6% sensitivity for above genes. PCR assay performed by using the already published primers shown 57, 49 and 28 positive results for above genes respectively that shown 81.5%, 70% and 40% sensitivity.
Conclusion: The present study shows that by using the high specific primers for detection of the mentioned genes of the Pseudomonas aeruginosa. The conventional PCR assay detected the early colonization of the organism in Cystic Fibrosis patients with more sensitivity and specificity before several mounts to obtain positive culture. Indeed PCR assay with high specific primers has more sensitivity and specificity as a rapid and accurate diagnosis of the organism in other deadly infections by using the direct clinical specimens.

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Background: Nasal polyp (NP) is a benign mucosal mass located in both sinuses and nares which is mostly seen in association with cystic fibrosis, asthma or oversensitivity to aspirin. The prominent histological feature of NP is inflammatory cell infiltration with eosinophil predominance. Superantigens role in causing NP complications is already proven. Superantigens, which are mostly originated from Streptococci and Staphylococci, activate T cells strongly and increase the process of production and release of cytokines, and secretion of IgE from B cells, which in turn directly affects proinflammatory cells such as eosinophils, both in their tissues infiltration and functions.
Methods: The samples are collected from patients referring to ENT clinic in Rasoul Akram training Hospital in Tehran after thorough clinical and paraclinical examinations. For control group the samples collected from patients undergoing rhinoplasty. All the samples kept frozen and sent to immunology lab. The DNA of the excised tissues extracted and amplified by using the superantigens specific primers and PCR product detected by gel electrophoresis. The date analyzed by using mean and SD and χ2 analytical tools. 
Results: Fifteen healthy individuals, 25 patients with rhinosinusitis and 24 with polyposis entered this trial. Group A Streptococcus toxin detection was significantly more frequent in those with nasal polyp and rhinosinusitis compared to healthy individuals (P=0.001 and 0.005, respectively), but the results were almost the same for those with nasal polyp and rhinosinusitis (P=0.4).
Conclusion: Streptococci may play an important role in induction or clinical exacerbation of polyposis and group A Streptococcus pyogenes exotoxin (SPEs) with superantigenic effects may have a crucial role in etiology and pathogenesis of polyps with or without rhinosinusitis. It is postulated that, T cells polyclonal activation by SPEs may cause recruitment of inflammatory cells in nasal mucosa. These inflammatory cells include IgE producing B cells laeding to allergic and inflammatory reactions in NP.

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Background: Squamous cell carcinoma (SCC) is the most frequent oral cancer. Protec-tive effects of the consumption of vegetables and fruits on various forms of cancer in-cluding oral cancer have been determined. Tomato (Solanum lycopersicum L.) because of its lycopene and bioflavonoids contents possesses anti-carcinogenic properties. The aim of this study was to evaluate the preventive effects of tomato pulp on pre-neoplastic changes induced by 4-Nitroquinoline-1-oxid (4-NQO) in epithelial cells of lingual mucosa in the rats. Methods: Forty-eight male Wistar rats were randomly allocated into four equal groups. Group 1 served as control. Groups 2 to 4 assigned to receive 30 ppm 4-NQO in drinking water for 12 consecutive weeks. When the feeding of 4-NQO was started to the rats of groups 3 and 4, they received tomato pulp (20 and 40 ml/kg bw) daily through the oral gavage. Finally, histological evaluations for carcinogenesis were performed for tongues epithelial tissue. Results: There were no pathological alterations in epithelial tissue of lingual mucosa in control rats. In the epithelial cells of lingual mucosa of 4-NQO treated rats, premalig-nant alterations appeared after 12 weeks of the last application of the drug. Administration of tomato pulp at both doses (20 and 40 ml/kg bw) during the experiment reduced the severity of the lesions, as well as caused a significant reduction in the frequency of pre-neoplastic lesions of tongue epithelial cells (P= 0.024 and P= 0.008). The incidence of severe epithelial cells dysplasia of lingual mucosa in the high dose treatment group was significantly smaller than of low dose treatment group (P= 0.037). Conclusion: The results obtained showed that tomato pulp is effective in inhibiting the development of neoplasms in epithelial cells of lingual mucosa induced by 4-NQO in the rat.

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Background: Colorectal cancer is a major cause of morbidity and mortality throughout the world, and its treatments include surgery, chemo-radiotherapy. Despite improvements in clinical outcomes of patients with this tumor over the past decades, prognosis remains poor with a 5-year survival rate of <10%. Angiogenesis inhibitor agents have been recently added to the treatment regimen of this disease. In the past two decades, it has been recognized that selective inhibitors of the cyclooxygenase -2 (Cox-2) enzyme result in the regression in the size of colorectal tumor, and one of its reasons is attributed to angiogenesis inhibition. The present study aimed at identifying the molecular pathways of angiogenesis inhibition by celecoxib. Methods: HCT-116, which is one of the cell lines of Colorectal cancer (separated from human colorectal adenocarcinoma) was provided by the National Cell bank of Iran (NCBI) affiliated to Pasteur Institute. It was then cultured in DMEM (high glucose) culture medium containing 10% FBS, and then treated in the active substance of celecoxib at pharmacological concentrations of 50 mM (C50) and 100 mM (C100). Afterwards, RNA was extracted and cDNA was prepared. The oligonucleotide of HIF-1 Alpha gene (angiogenesis initiator) was prepared and the level of HIF-1 alpha gene expression was assessed with a real-time PCR device in three control, C50 and C100 groups. Results: HIF-1 alpha gene expression significantly decreased in the celecoxib treatment group (compared with control group) with the concentration of C100 (P< 0.001), but no change was observed in the concentration of C50. Conclusion: Angiogenesis is a key factor in the carcinogenesis process and FDA today approved bevacizumab as a first-line treatment for patients with metastatic colorectal cancer. The results of this study showed one of the causes of angiogenesis reduction in celecoxib-treated colorectal cancer. According to clinical findings and basic studies, celecoxib will be hopefully used as a first-line therapy along with chemotherapy in the near future in colorectal cancer. The advantages of this treatment method include its low cost and low side effects.

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Helicobacter pylori (H. pylori) is the causative agent in development of gastroduode-nal diseases, such as chronic atrophic gastritis, peptic ulcers, mucosa associated lym-phoid tissue (MALT) lymphoma, and gastric cancer. H. pylori has been associated with inflammation in cardia, showing the fact that infection with this bacterium could also be a risk factor for gastric cardia cancer. Gastric cancer is the fourth most common cancer worldwide. This is the second leading cause of cancer-related deaths, and ap-proximately 700,000 people succumb each year to gastric adenocarcinoma. It has been estimated that 69% of the Iranian population currently harbor H. pylori infection. The prevalence of duodenal ulcer and gastric cancer is high in Iranian populations. However, this has been largely influenced by geographic and/or ethnic origin. Epidemi-ology studies have shown that host, environmental, and bacterial factors determine the outcome of H. pylori infection. The bacterium contains allelic diversity and high genet-ic variability into core- and virulence-genes and that this diversity is geographically and ethnically structured. The genetic diversity within H. pylori is greater than within most other bacteria, and its diversity is more than 50-fold higher than that of human DNA. The maintenance of high diversification makes this bacterium to cope with particular challenges in individual hosts. It has been reported that the recombination contributed to the creation of new genes and gene family. Furthermore, the microevolution in cagA and vacA genes is a common event, leading to a change in the virulence phenotype. These factors contribute to the bacterial survival in acidic conditions in stomach and protect it from host immune system, causing tissue damage and clinical disease. In this review article, we discussed the correlation between H. pylori virulence factors and clin-ical outcomes, microevolution of H. pylori virulence genes in a single host, microevolu-tion of H. pylori during primary infection and progression of atrophic gastritis to ade-nocarcinoma, and H. pylori infection status in Iran. Finally, we put forward the hy-pothesis that if the pattern of nucleotide sequence evolution shifts from recombination (r) to mutation (m) and the r/m ratio is reduced, bacterial pathogenicity may be re-duced while maintaining the bacterial life. However, this hypothesis should be further studied with future experiments.

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Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, involves almost all organs such as skin, heart, kidneys and central nervous system. The disease is characterized by vascular and connective tissue inflammation in a recurring pattern of remission and flare. Although the exact pathophysiology of disease has not been fully understood yet, the fundamental defect in SLE is attributed to dysfunction of T lymphocytes in controlling of B-cell that leads to polyclonal activation of B lymphocytes and production a large quantity of autoantibodies against nuclear and cytoplasmic components. These autoantibodies can damage tissues either directly or as a result of immune complex deposits. Several factors are involved in pathogenesis of SLE which can be divided into three major groups, environmental factors, genetic components, and immunological disturbances. They could breakdown body tolerance towards endogenous antigens and cause abnormal immunologic response to the healthy tissue, resulting in tissue damage. SLE occurs more frequently in female than male. It seems that immunological factors have important role in SLE. Inflammation and vascular endothelium irregularities are a number of main pathologies seen in SLE. Cytokines are protein mediators that play an essential role as regulator of innate and adaptive immune response against microbial agents or self-antigens. Influences of cytokines in autoimmune diseases such as SLE are poorly understood. Studies in both experimental animal models of lupus and patients with SLE have revealed a number of cytokine pathways that are important in the disease process. These studies showed that overexpression of inflammatory cytokines increases the proliferation of auto reactive B-cells and results in higher production of autoantibodies. Among them, the role of B-cell activating factor (BAFF), a proliferation-inducing ligand (APRIL), TNF-α, IFN-α, IL-6, IFN-γ, IL-23/IL-17, IL-10, IL-21 are prominent, which is associated with the generation of pathogenic autoantibodies and formation of immune complexes. In this paper, the role of cytokines and their encoding genes are described, while therapeutic applications are also briefly presented.

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Background: Cancer and obesity are two major public health concerns. More than 12 million cases of cancer are reported annually. Many reports confirmed obesity as a risk factor for cancer. The molecular relationship between obesity and breast cancer has not been clear yet. The purpose of this study was to investigate priorities of effective genes in the molecular relationship between obesity and breast cancer. Methods: In this study, computer simulation method was used for prioritizing the genes that involved in the molecular links between obesity and breast cancer in laboratory of systems biology and bioinformatics (LBB), Tehran University, Tehran, Iran, from March to July 2014. In this study, ENDEAVOUR software was used for prioritizing the genes and integrating multiple data sources was used for data analysis. Training genes were selected from effective genes in obesity and/or breast cancer. Two groups of candidate genes were selected. The first group was included the existential genes in 5 common region chromosomes (between obesity and breast cancer) and the second group was included the results of genes microarray data analysis of research Creighton, et al (In 2012 on patients with breast cancer). The microarray data were analyzed with GER2 software (R online software on GEO website). Finally, both training and candidate genes were entered in ENDEAVOUR software package. Results: The candidate genes were prioritized to four style and five genes in ten of the first priorities were repeated twice. In other word, the outcome of prioritizing of 72 genes (Product of microarray data analysis) and genes of 5 common chromosome regions (Between obesity and breast cancer) showed, 5 genes (TNFRSF10B, F2, IGFALS, NTRK3 and HSP90B1) were the priorities in the molecular connection between obesity and breast cancer. Conclusion: There are some common genes between breast cancer and obesity. So, molecular relationship is confirmed. In this study the possible effect of gene F2 polymorphism in making breast cancer associated with obesity risk factor was confirmed, the fact that past studies have not been reported.

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Background: Staphylococcus aureus is one the most common pathogens causing community-acquired infections and a major concern for public health, and the other hands antibiotic resistance is also of great concern for public health authorities also Staphylococcus aureus produce a lot of virulence factors such as variety of exoproteins included toxic shock syndrome and exfoliative toxin which causes colonization and different infections in their host. The aims of current study were to evaluate the prevalence of Toxic shock syndrome toxin 1 (TSST-1) and ETs genes in isolated S. aureus strains using polymerase chain reaction (PCR) assay. Methods: This cross-sectional study was performed on 100 methicillin-resistant staphylococcal aureus (MRSA) and 100 methicillin-sensitive staphylococcal aureus (MSSA) isolated from clinical specimens of inpatients, outpatients hospitals and nasal carriers in Hamadan University from October 2013 to August 2014. Identified species by biochemical methods were confirmed by the PCR method. Antibiotic resistance was performance by disk diffusion and the presence of TSST-1 and ETs genes was investigated using PCR. Results: Of the 100 isolates MRSA examined, the most frequent resistance was observed to ciprofloxacin (95%), followed by tetracycline (91%), erythromycin (92%), Gentamicin (90%), Rifampin (85%), trimethoprim-sulfamethoxazole (85%), clindamycin (80%) and cefoxitin (100%). Of the 100 isolates MSSA examined, the most frequent resistance was observed to erythromycin (68%), ciprofloxacin (66%), followed by tetracycline (52%), gentamicin (25%), clindamycin (46%), rifampin (45%), trimethoprim-sulfamethoxazole (66%) and cefoxitin (0%). Prevalence of TSST-1 and ETs genes were determined 13% (n=26) isolates, totally. Also the prevalence of TSST-1 was 11% (n=22) and ETs genes was 2% (n=4) isolates and none of the investigated isolates carried eta gene. Conclusion: The increasingly prevalence of MRSA and emerging its antibiotic resistance in clinical isolates can be considered a serious problem for public health. Detection of the high rate prevalence of TSST genes in current study is considered as a serious problem and existing and circle of these strains in according to colonization in community especially old people and immunocompromised patients is very serious.

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Angiogenesis, as the process of new vessel formation from pre-existing vessels is dependent on a delicate equilibrium between endogenous angiogenic and antiangiogenic factors. However, under pathological conditions, this tight regulation becomes lost which can result in the formation of the different diseases such as cancer. Angiogenesis is a complex process that includes many gene products that are produced by different cells. Each of the processes influenced by specific genes that their expression can be regulated by hypoxi inducible factor-1 (HIF-1). Hypoxia, the imbalance between the oxygen in need and the oxygen available, usually occurs in tumors and ischemic cardiovascular diseases. In order to overcome this challenge, tumors regulate and control the expression of genes related to angiogenesis, cell cycle and metabolism using hypoxia-inducible factor 1 (HIF-1). HIF-1 was first recognized as a transcription factor involved in hypoxia-induced erythropoietin expression. As angiogenesis pathway molecules are being described, this factor has been characterized as a key transcription regulator for these molecules. In this review article, after discussing HIF-1 structure and characterization, the role of this important factor in angiogenesis and cancer as a pathological case and finally, the clinical applications has been evaluated. Articles related to the key words of hypoxia, HIF-1 and angiogenesis were searched from valid databases such as Springer Link, google scholar, Pubmed and Sciencedirect. Then, the articles related to the role of hypoxia and HIF-1 in activation of genes that are involved in angiogenesis and cancer were searched and selected for this study. Studies show that, HIF-1 activation of genes including vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2), etc., induced angiogenesis in the tumor cells. Furthermore, the activation of genes such as insulin-like growth factor 2 (IGF2), transforming growth factor &alpha; (TGF-&alpha;) and MAPK and PI3K signaling pathway will also enable the survival and proliferation of tumor cells. HIF-1 by activating genes involved in angiogenesis and also activates signaling pathways associated with cell survival and proliferation plays an important role in the stability and growth of tumors. Therefore, better understanding of molecular mechanisms associated with this factor can be effective in the treatment of cancer.

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Background: The major issue to address in obesity etiology is to identify the genetic changes in the disease and their occurrence in different populations. Uncovering these genetic changes may be important in developing potential biomarkers for early diagnosis and prognosis of obesity. Among all obesity susceptibility genes studied before, convincing association has been found with variants in the FABP2 gene and this disease; however, the contributions of these genetic variants in different populations and ethnic groups are not similar. Accordingly, this study was carried out to replicate the previous findings to assess whether a missense variation (rs1799883) in this gene is associated with obesity in the Tehran Lipid and Glucose Study (TLGS) population.

Methods: A case–control study was designed to determine the possible association between rs1799883 and occurrence of obesity “in phase IV of the study between the years of 2008 to 2011”. The study group consisted of 217 subjects with body mass index (BMI, kg/m2) greater than 30 as cases and 159 healthy individual as control group (1820). All subjects were recruited among the Tehran Lipid and Glucose Study (TLGS) participants in phase IV of the study between the years of 2008 to 2011. The genomic DNA was extracted from peripheral blood leucocytes using the salting out method and subsequently subjects were genotyped for this marker using The tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Association of risk allele with obesity was assessed using the SPSS software, version 20 (Chicago, IL, USA).

Results: The results showed no significant differences between case and control groups in terms of allele frequency (P=0.61). According to the findings, the presence of T allele as the risk allele was not associated with increased risk of obesity in carriers of this allele compared to individuals carrying the normal allele (OR=1.17; CI%95= 0.62-2.19, P=0.61).

Conclusion: The results did not support the previous findings of an association between genetic polymorphism in the FABP2 gene and risk of obesity. However, a number of replicated studies with other ethnicity are suggested to make a conclusion about the role of this genetic polymorphisms and susceptibility to obesity in Iranian population.

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Background: Spermatogenesis is a complex and highly organized process of proliferation and differentiation of spermatogonial stem cells. Spermatogonial stem cells (SSCs) as a unique stem cell have the potential to self-renewal, differentiation and transmit genetic information to the next generation and play a vital role in maintaining fertility. Sertoli cells as the only somatic cells within the seminiferous epithelium play central roles in the formation of niche and balance between self-renewal and differentiation by secrete many growth factors. Given the importance and widespread use of SSCs, particularly in the treatment of infertility, the aim of this study was to create an optimal environment for the proliferation of SSCs. So we decided to study of undifferentiated (ID4) and differentiated (c-Kit) gene expression in SSCs followed by co-culture with Sertoli cells for a one-month.

Methods: This experimental study was conducted from November 2013 to December 2014 in Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, on immature NMRI mouse (6-3 days old). Initially, Sertoli cells and SSCs were isolated from neonates mouse testes during the two-step enzymatic digestion characteristics Sertoli cells with vimentin marker and SSCs with promyelocytic leukemia zinc-finger (PLZF) marker were confirmed. Then SSCs were cultured in two groups: co-culture with Sertoli and without co-culture (control). Undifferentiated (ID4) and differentiation (c-Kit) gene expression were evaluated by Real-time PCR technique.

Results: Spermatogonial stem cells purity was obtained 66.91% by flow cytometry. The relative expression levels of gene ID4 in co-culture group at the end of each week, compared to the control group showed a significant increase (P<0.05). While the expression of this gene significantly decreased in each group over time (P<0.05). The results of the comparison of the relative expression of c-Kit gene in co-culture group are indicated significant decrease than the control group at the end of each week (P<0.05). In addition, this gene expression was showed significant increase in each group individually over time (P<0.05) ID4 gene expression showed a significant (P<0.05) increase toward the control group, while in the expression of c-Kit was observed a significant (P<0.05) decrease compared with the control group at the end of each week.

Conclusion: According to the results of this study, co-culture with Sertoli cells maintains SSCs in the prolifration stage for long-term, so can be used to optimize the culture medium at the clinic.