IRANIAN SOUTH MEDICAL JOURNAL (ISMJ)
Year:2017 | Volume:20 | Issue:3 |Papers Count:9

Background: Helicobacter pylori is the most common bacterium causing chronic infections worldwide. Expression of flagella and bacterial motility are very important incolonization and virulence. FlaA geneis one of theflagellin-encoding genes that play the key role in the colonization and bacterial motility and it has a significant impact in immunization. The aim of this study wasto design, construction and the evaluation of the Helicobacter pylori flaA gene expression in eukaryotic cells. MaterialandMethods: In this experimental study, genomic DNA was purified from the Helicobacter pylori standard strain and flaA gene was amplified and isolated by PCR method with use of the specific primers. Then, this gene was cloned into pTZ vector by T/A cloning technique. In order to flaA gene expression and generation of final construct, the flaA gene was removed from pTZ plasmid and sub-cloned into the pcDNA3. 1 (-) expression vector. The pcDNA3. 1 (-)-flaA construct was transformed into CHO cells by electroporation, and flaA eukaryotic gene expression was studied on SDS-PAGE. Results: The results showed that flaA gene PCR product was cloned into pTZ vector and amplified in Escherichia coli TOP10F strain. Also the enzymatic digestion and sequencing showed that the pcDNA3. 1 (-)-flaA was performed. Finally, the evaluation of the Helicobacter pyloriflaA gene expression in CHO cells showed that the generated gene construct can expressed the flaA product in eukaryotic system, successfully. Conclusion: Given that the pcDNA3. 1 (-)-flaA as a final construct is able to express the flaA protein of the Helicobacterpylori in animal cells. Flagellin protein is one of the important antigens of the bacteriumSo we can properly say that gene pcDNA3. 1(-)-flaA consteuct is an appropriate candidate for use in the field of gene vaccines against Helicobacter pylori and in future researches can be used to check the immunization in laboratory animals.

Background: Infertility referes to the inability of a couple to conceive over an average period of one year with unprotected sexual intercourse. In mammals, Fertility is initiated at puberty by the pulsatile secretion of gonadotrophin releasing hormone (GnRH). Kisspeptin (product of Kiss1 gene) and its receptor (KISS1R) are as vital upstream regulatorsin integrating of central and peripheral signals with GnRH release. In this paper, we studied an association between polymorphism rs 397515615 in the KISS1R gene with idiopathic male infertility. Material and Methods: In this research, we were evaluated a total of 50 men with idiopathic infertility and 50 healthy men (control group). AS-PCR method was applied for determination of the codon polymorphism. Also we used chi-square (χ 2) test to study of an association between genotype and allele frequencies in cases and control groups. Results: The results showed that the genotypic frequencies between two groups revealed a significant difference between the patients and healthy (P=0. 02), while Allele distribution (G/-) between case and control groups was not significant (P=0. 20). Conclusion: The results of this study show that the polymorphism of rs 397515615 KISS1R gene probably associated with idiopathic infertility in men. However, further studies with larger and different geographical population are needed to confirm the results.

Background: E. coli O157: H7 is one of the intestinal pathogens which causes serious lesion in gastrointestinal system. Detection of this bacteria that able to produce toxin and is the major responsible for hospital infection, usually done by culture on sorbitol-MacConkey agar which is time-consuming test. The aim of this study was the preparation a fast and precise method in order to identification of this bacterium by molecular method based on PCR technique. Material and Methods: rfbE and stx2b genes were selected for proprietary identification of E. coli O157: H7 and the amplification of them were performed by PCR following designing specific primers. Sorbitol-MacConkey agar was used to verification of growth ability of selected colonies during PCR. Results: By appearance of the bonds belong to rfbE and stx2B genes on agarose gel, the ability of designed primers in gene detection in samples of E. coli O157: H7 was verified. Colonies which selected during PCR have growth potency on sorbitol-MacConkey agar medium. Conclusion: It was revealed that we can prepare a fast, precise and relative comfortable method for detection of E. coli O157: H7 strain by using PCR technique and specific primers than other available methods.

Background: Non-melanoma skin cancers including basal cell carcinoma and squamous cell carcinoma are the most common cancers in human. The aim of this study was to determine the expression of P63 marker in usual skin cancers compared with non-tomoral skin lesions. Materials and Methods: In this cross-sectional study, sampling was performed from archival blocks of Shahid Mohammadi hospital patients during 2010-2011. 60 samples (including 30 samples of non tumoral skin lesions and 30 samples of basal cell carcinoma and squamous cell carcinoma) were studied and evaluation of p63 gene expression was done with Immunohistochemistry method. T-test and Chi-square were used for analysis of data. Results: P63 gene were expressed in 4 cases (13. 33 %) of non tumoral lesions and all tumoral lesions (100 %). In tumoral lesions, 5 cases (16. 66 %) showed 1+ severity experssion, 11 cases (36. 66%) 2 + severity experssion and 14 cases (46. 66 %) 3+severity experssion. All 4 non tumoral lesions shoed 1+ severity experssion of P63gene. Conclusion: The results of this study indicated that the incidence and severity of gene expression of P63 can be use for differentiation between basal cell carcinoma and squamous cell carcinoma as well as nontumoral skin lesions.

Background: The venomous jellyfish Cassiopea andromeda can produce envenomation and different toxicological and biological effects by their nematocysts. The phospholipase A2 enzymes (PLA2) are toxic and induce various pharmacological effects including neurotoxicity, myotoxicity and anticoagulant activities. The main aim of the current project was to screen the in vitro PLA2 activity of the C. andromeda crude venom. To better understand the experimental result; a molecular docking study was also performed. Materials and methods: The live specimens were collected from Nayband lagoon, by a trawl net, and separation of their tentacles was done according to Bloom 's et al., method. The PLA2 activity of crude venom was performed according to the acidimetric method of Tan and Tan. The lyophilized venom was subjected to Gas Chromatography/ Mass Spectroscopy, and the obtained structures were used for docking study against PLA2. The indoxam was considered as standard control. Results: The PLA2 activity of the jellyfish crude venom was 413 ± 0. 08 μ mol/min/mg. Analysis of the crude venom detected seven compounds (i-vii) using GC-MS. Docking data was also confirmed the experimental results. According to the docking results, the highest affinity (-6. 7 (kcal/mol)) was observed in the compound “ Pregn-5-ene-3, 11-dione, 17, 20: 20, 21 bis [methylenebis(oxy)]-, cyclic 3-(1, 2-ethane diyl acetal” . Conclusions: A high PLA2 level was found in the venom of C. andromeda. There was a good correlation between in vitro and in silico studies.

Background: Vitamin D deficiency or insufficiency is a common worldwide problem. The association between hypothyroidism and vitamin D deficiency is controversial. We aimed to study the effect of vitamin D on thyroid function in hypothyroid patients. Material and Methods: In this case-control randomized clinical trial study, 201 hypothyroid patients reffered to endocrinology clinics in Arak, were randomly classified into two groups. All patients were taking levothyroxine. Case group received vitamin D 50000 unit weekly and control group received placebo in addition to levothyroxine. After three months, thyroid function tests were repeated and compared with the results of the beginning of the study both intra groups and inter groups by student t test and paired t test analysis. Results: Male/Female ratio in both case and control groups were 0. 24 and 0. 15 respectively (P=0. 1). The prevalence of vitamin D deficiency and insufficiency were 68. 7 % (138) and 93. 5% (188) and after vitamin D taking were 70% (34. 8) and 51. 2% (103) respectively. Student t test showed that TSH level in people who received vitamin D had a significant decrease in comparison to the people who received placebo (P<0. 05). There were a significant change of TSH level between two groups at vitamin D level 10-30 ng/ml. Conclusion: Most of hypothyroid patients had vitamin D deficiency and vitamin D taking improved thyroid function by TSH suppression in these patients. We recommend the screening for vitamin D deficiency in hypothyroid patients. Although, more researches are needed to clarify molecular explanations of this hypothesis.

Background: Helicobacter pylori (H. Pylori) is one of the most common infectious bacteria cause diseases such as chronic gastritis, peptic ulcer, adenocarcinoma, etc. Epidemiological studies have demonstrated that individuals who had O blood group were more likely to develop peptic ulcers. The aim of present study was to investigate the association between the prevalence of H. Pylori infection in soldiers and military students with ABO, Rh blood group in Tehran city. Materials and Methods: In this descriptive study 417 individuals aged 18-27 years who were selected among military students of AJAUMS (Aja University of Medical Sciences) University of Afsari Imam Ali and soldiers. Personal, social and health information of individuals were collected through questionnaires. Phenotype of ABO blood groups and Rh in all participants were studied by a standard hemagglutination test. Antibody levels of Anti-H. pylori IgG in serum of all participants were determined by ELISA test. Collected data analyzed by using SPSSsoftware version 16 and Chi-square test. Results: Overall 183 (43. 9%) of 417 subjects were seropositive, and 234 (56. 1%) subjects were seronegative. Prevalence of infection in AJAUMS students compared to other two groups showed a significant decrease. However, the prevalence of infection in the group of persons with more than five family members was significantly higher than the group with less than 5. Conclusion: There was no association between ABO, Rh blood groups with H. Pylori infection.