per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
245
256
article
Cloning and Study of Expression of Helicobacter Pylori FlaAGene in Eukaryotic System
Motaram Sadeghi
1
Abbas Doosti
abbasdoosti@yahoo.com
2
Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
Background: Helicobacter pylori is the most common bacterium causing chronic infections worldwide. Expression of flagella and bacterial motility are essential in colonization and virulence. FlaA gene is one of the flagellin-encoding genes that play the key role in the colonization and bacterial motility, and it has a significant impact on immunization. This study aimed to design, construct and evaluate the Helicobacter pylori flaA gene expression in eukaryotic cells.
Material and Methods: 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 using the specific primers. Then, this gene was cloned into pTZ vector by T/A cloning technique. To express flaA gene and generate the final construct, the flaA gene was removed from pTZ plasmid and subcloned 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 using the SDS-PAGE method.
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 could express the flaA product in a eukaryotic system, successfully.
Conclusion: Given that the pcDNA3.1 (-)-flaA as a final construct can express the flaA protein of the Helicobacter pylori in animal cells. Flagellin protein is one of the essential antigens of the bacterium So we can appropriately say that gene pcDNA3.1(-)-flaA construct is a suitable candidate for usage in the field of gene vaccines against Helicobacter pylori and it can be used to check the immunization in the laboratory animals in the future research.
http://ismj.bpums.ac.ir/article-1-876-en.pdf
Helicobacter pylori
flaA gene
Cloning
Electroporation
per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
257
266
article
Investigation of Association Between KISS1R rs397515615 Polymorphism with Idiopathic Male Infertility
Azadeh Poursharif
1
Hamidreza Vaziri
vaziri@guilan.ac.ir
2
Toba Mirzapour
3
Department of Biology, School of Science, University of Guilan, Guilan, Iran
Department of Biology, School of Science, University of Guilan, Guilan, Iran
Department of Biology, School of Science, University of Mohaghegh Ardabili, Ardabil, Iran
Background: Infertility refers 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 the gonadotropin-releasing hormone (GnRH). Kisspeptin (product of Kiss1 gene) and its receptor (KISS1R) are as vital upstream regulators in integrating of central and peripheral signals with GnRH release. In this paper, we studied an association between polymorphism rs 397515615 in the KISS1R gene and idiopathic male infertility.
Material and Methods: In this research, we evaluated a total of 50 men with idiopathic infertility and 50 healthy men (control group). The AS-PCR method was applied for the determination of the codon polymorphism. Also, we used chi-square (χ2) 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 control group (P=0.02), while the allele distribution (G/-) between case and control groups showed no significant difference (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 sample size and diverse geographical distribution are required for more conclusive results.
http://ismj.bpums.ac.ir/article-1-877-en.pdf
rs 397515615
KISS1R gene
Male-factor Infertility
Polymorphism
per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
267
277
article
Identification of E. coli O157:H7 by Using Specific Primers for rfbE and stx2b Genes
Mostafa Bakhshi
1
Firouz Ebrahimi
2
Shahram Nazarian
kpnazari@ihu.ac.ir
3
Yousef Tarverdizade
4
Davood Sadeghi
5
Department of Biology, School of Basic Science, Imam Hossein University, Tehran, Iran
Department of Biology, School of Basic Science, Imam Hossein University, Tehran, Iran
Department of Biology, School of Basic Science, Imam Hossein University, Tehran, Iran
Department of Biology, School of Basic Science, Imam Hossein University, Tehran, Iran
Department of Biology, School of Basic Science, Imam Hossein University, Tehran, Iran
Background: E. coli O157:H7 is one of the intestinal pathogens which can cause severe lesions in the gastrointestinal system. These bacteria can produce toxins and are the main cause of hospital infections. Their detection is usually done by culture on sorbitol-MacConkey agar which is a time-consuming test. The aim of this study was to develop a rapid, yet accurate method to identify this bacterium using a PCR based technique.
Material and Methods: rfbE and stx2b genes were selected for identification of specific E. coli O157:H7 strain. Then amplification was performed by PCR following designing specific primers. Sorbitol-MacConkey agar was used to verification of growth ability of selected colonies during PCR.
Results: By the appearance of the bonds belong to rfbE and stx2B genes on an agarose gel, the ability of designed primers for gene detection in samples of E .coli O157:H7 was verified. A sorbitol-MacConkey agar medium was used to evaluate the growing potency of colonies selected during PCR.
Conclusion: In this study we demonstrated that we could develop a fast, accurate, and relatively comfortable method for detection of E. coli O157:H7 strain by using PCR technique and specific primers instead of using current methods.
http://ismj.bpums.ac.ir/article-1-878-en.pdf
Identification of E. coli O157:H7 strain
polymerase chain reaction
rfbE gene
stx2b gene
per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
278
286
article
P63 marker Expression in Usual Skin Cancers Compared With Non Tumoral Skin Lesions
Abdolhamid Esmaili
hamidsporteduction@gmail.com
1
Mohsen Dehghani Zahedani
2
Fatemeh Nili
3
Department of Pathology, School of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran
Department of Pathology, School of Medicine, Hormozgan University of Medical Sciences, Hormozgan, Iran
Department of Pathology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Background: Non-melanoma skin cancers including basal cell carcinoma and squamous cell carcinoma are the most common malignant diseases in human. The aim of this study was to determine the expression of the P63 marker in common skin cancers and non-tumoral skin lesions and compared the difference between them.
Materials and Methods: In this cross-sectional study, sampling was performed from the archive of sample blocks from patients admitted in Shahid Mohammadi Hospital during 2010-2011. Sixty samples (including 30 samples of non-tumoral skin lesions and 30 samples including basal cell carcinoma and squamous cell carcinoma) were studied. The evaluation of p63 gene expression was performed using immunohistochemistry method. Student’s T-test and Chi-square test were used for analysis of the 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 expression, 11 cases (36.66%) 2+severity expression and 14 cases (46.66 %) 3+severity expression. All 4 non-tumoral lesions showed 1+severity expression of P63gene.
Conclusion: The results of this study indicated that the frequency and severity of gene expression of P63 could be used for differentiation between basal cell carcinoma and squamous cell carcinoma as well as non-tumoral skin lesions.
http://ismj.bpums.ac.ir/article-1-879-en.pdf
Non-tumoral skin lesions
P63
basal cell carcinoma
squamous cell carcinoma
per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
287
300
article
Phospholipase A2 activity of the Persian Gulf upside-down jellyfish venom (Cassiopea andromeda)
Gholamhossean Mohebbi
1
Hossean Vatanpour
2
Amir Vazirizadeh
3
Ammar Maryamabadi
4
Alireza Hasaninejad
5
Samad Akbarzadeh
6
Maryam Farrokhnia
7
Afshar Bargahi
8
Iraj Nabipour
inabipour@gmail.com
9
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
Department of Pharmacology and Toxicology, School of Pharmacy, Shaheed Beheshti Medical Sciences University, Tehran, Iran
The Marine Biology and Fishery Science Department, Persian Gulf Institute, Persian Gulf University, Bushehr, Iran
Department of Chemistry, school of Sciences, Persian Gulf University, Bushehr, Iran
Department of Chemistry, school of Sciences, Persian Gulf University, Bushehr, Iran
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
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 µmol/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.
http://ismj.bpums.ac.ir/article-1-880-en.pdf
Cassiopea andromeda
crude venom
Phospholipase A2
Persian Gulf
per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
301
307
article
The Study the Effect of Vitamin D on Hypothyroidism
Afsaneh Talaei
afsanehtalaeii@yahoo.com
1
Fariba Ghorbani
2
Parisa Naseri
3
Ali Chehrea
4
Endocrine and Metabolism Research Center, Arak University of Medical Science, Arak, Iran
Department of Internal Medicine, School of Medicine, Arak University of Medical Science, Arak, Iran
Department of Statistic, School of Medicine, Arak University of Medical Science, Arak, Iran
Endocrine and Metabolism Research Center, Arak University of Medical Science, Arak, Iran
Background: Vitamin D deficiency or insufficiency is a common worldwide concern. 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 referred 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 start of the study. Both intra-groups and inter-groups differences were analyzed by student t-test and paired t-test analysis.
Results: Male/Female ratio in case and control groups were 0.24 and 0.15 respectively (P=0.1). The prevalence of vitamin D deficiency and insufficiency was 68.7 % (138) and 93.5% (188) and after taking vitamin D was 70% (34.8) and 51.2% (103) respectively.Student t-test showed that TSH level in people who received vitamin D had a significant reduction in comparison with the placebo group(P<0.05). There was a significant difference in the change of TSH level between two groups at vitamin D level 10-30 ng/ml.
Conclusion: Most of the hypothyroid patients had vitamin D deficiency and receiving vitamin D improved thyroid function by TSH suppression in these patients. We recommend the screening for vitamin D deficiency in hypothyroid patients. However, more research is required to explain this hypothesis at the molecular level.
http://ismj.bpums.ac.ir/article-1-881-en.pdf
Vitamin D
Hypothyroid
effect
TSH
per
Bushehr University of Medical Sciences
Iranian South Medical Journal
1735-4374
1735-6954
2017-07
20
3
308
316
article
Association of Helicobacter Pylori Infection with ABO and Rh Blood Groups in Military Students and Soldiers
Shahrokh Iravani
1
Adel Naimi
2
Tohed Jafari koshki
3
Pedram Azimzadeh
4
Kazem Nejati kashki
5
Saead Solali
ssolali@gmail.com
6
AJA Cancer Research Center (ACRC), Aja University of Medical Sciences, Tehran, Iran
Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
Department of Biostatistics, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Gastroenterology and Liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Student research committee, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
Hematology and Oncology research center, Tabriz University of Medical Sciences, Tabriz, Iran
Background: Helicobacter pylori (H. Pylori) is one of the most common infectious bacteria cause diseases such as chronic gastritis, peptic ulcer, adenocarcinoma. Epidemiological studies have demonstrated that individuals who had O blood group were more likely to develop peptic ulcers. The aim of this study was to investigate the associ ation between the prevalence of H. Pylori infection in soldiers and military students and their ABO, Rh blood group in Tehran city.
Materials and Methods: In this descriptive study 417 individuals aged 18-27 years who were selected from military students of AJAUMS (AJA University of Medical Sciences) University of Afsari Imam Ali and other soldiers. Personal, social and health information of individuals were collected through questionnaires. The phenotype of ABO blood groups and Rh in all participants were studied by a standard hem-agglutination test. Antibody levels of Anti- H. pylori IgG in serum of all participants were determined by ELISA test. Collected data analyzed by using SPSS software version 16 and Chi-square test.
Results: Overall 183 (43.9%) of 417 subjects were seropositive, and 234 (56.1%) subjects were seronegative for anti- H. pylori antibody. Prevalence of infection in AJAUMS students compared to other two groups was significantly lower. However, the prevalence of infection in the group of individuals 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 and H. Pylori infection.
http://ismj.bpums.ac.ir/article-1-882-en.pdf
Helicobacter Pylori
ABO blood group
Rh blood group
military students