Isolation and cloning of Helicobacter Pylori ureE gene into pIRES2-DSRed expression vector to generate a gene vaccine

Background and Aim: As one of the factors of gastric ulcers and cancer, Helicobacter pylori can live in the acidic environment of stomach for many years due to having urease enzyme. This enzyme requires Ni2+ and a group of auxiliary proteins such as ureE for its catalytic activity. Urease is not only a requisite factor to colonize the Helicobacter pylori but it is also pathogenic with different mechanisms. Regarding the high prevalence of these bacteria finding a way to prevent infection with them is necessary. The present research aimed at homogenizing . cloning of Helicobacter Pylori ureE gene into pIRES2-DS Red expression vector in order to create a DNA [gene] vaccine

Materials and Methods: In this experimental study, the ureE gene fragment was amplified through PCR method and it was cloned using T/A cloning in the pTZ vector. Sub-cloning of the gene was done in pIRES2-DS Red vector using T4-ligase enzyme and it was transformed into E. coli TOP10F strain; and, then, amplified. The gene construct, which is a DNA vaccine candidate, was transferred to CHO cells using electroporation method to investigate the gene expression in eukaryotic systems. UreE gene expression was assessed in eukaryotic cells by means of SDS-PAGE

Results: UreE gene cloning was confirmed in two vectors including pTZ as a replicative vector and pIRES2- DS Red expression vector by PCR, enzyme digestion, and sequencing methods. SDS-PAGE results confirmed the successful expression of the ureE gene in the eukaryotic system of CHO cells

Conclusion: The recombinant pIRES2-DSRed-ureE construct is capable of successfully generating of polypeptides derived from Helicobacter Pylori ureE gene expression in bestial cells. Given that the protein product of the ureE gene is one of the most important proteins of the mentioned bacterium, . the created recombinant DNA in this research can be used as a DNA vaccine candidate against Helicobacter pylori in future

Cytotoxic effect of rosemary extract on gastric adenocarcinoma [AGS] and esophageal squamous cell carcinoma [KYSE30] cell lines

Aim: The present study was conducted to survey the potential cytotoxic influence of freeze-dried aqueous extract of its fruits on gastrointestinal cell lines, namely AGS [human gastric carcinoma] and KYSE30 [human esophageal squamous cell carcinoma]

Background: Rosemary [Rosmarinus officinalis] is a wild medicinal plant shown to have anticancer activity. Carnosic and rosmarinic acids are compounds, obtained from it through several extraction methods

Methods: The aqueous extract of the fruits of R.officinalis was freeze-dried, and KYSE30 and AGS cancer cell lines were treated with crude extract. Cytotoxic effect of the extracts on the cell lines was examined using 3-[4, 5-Dimethylthiazol-2-yl]-2, 5- diphenyltetrazolium bromide [MTT] and neutral red assay. Apoptotic cells were detected with ethidium bromide/acridine orange [EB/AO]. Cell-cycle distributions were evaluated by flow cytometry

Results: IC50 values were 4.1, 1.8 and 1.3 mg/mL for AGS cell lines after 24, 48 and 72 hours by MTT assay, respectively, and 4.4, 2.1 and 1.1 mg/mL by neutral red assay, respectively. IC50 values for KYSE30 cell lines were 600, 180 and 150 mg/mL after 24, 48 and 72 hours by MTT assay, and 860, 270 and 230 mg/mL by neutral red. EB/AO staining increased in apoptotic cells. After 24 h of treatment at different concentrations, significant increases and decreases in population were shown at G2/M and G1 phases, respectively

Conclusion: The aqueous extract of the fruits of R.officinalis was freeze-dried, and KYSE30 and AGS cancer cell lines were treated with crude extract. Cytotoxic effect of the extracts on the cell lines was examined using 3-[4, 5-Dimethylthiazol-2-yl]-2, 5- diphenyltetrazolium bromide [MTT] and neutral red assay. Apoptotic cells were detected with ethidium bromide/acridine orange [EB/AO]. Cell-cycle distributions were evaluated by flow cytometry

[ effectiveness of acceptance and commitment therapy on stress coping strategies in women with ulcerative colitis]

The main purpose of the present paper is to examine the effectiveness of acceptance and commitment therapy on perceived stress, stress coping strategies and illness perception in women with Ulcerative Colitis. This is an experimental study performed as pretest and post-test. The population consists of woman suffering from Ulcerative Colitis in Isfahan. The sample includes 37 Colitis selected by multi-stage random sampling and randomly assigned to experiment and control groups [control=20, experimental=17]. The acceptance and commitment therapy was performed in the experimental group within two months once a week. The tools applied by this study include Perceived Stress Scale, Brief Illness Perception Scale Stress Coping Strategies Scale and Brief Illness Perception Scale. The results of this research showed that the means of coping strategies in experimental group has changed significantly in comparison with control group [p<0.05]. Thus, the acceptance and commitment therapy could be used along with other methods of the treatment for the women with Ulcerative Colitis

[ case report of the rupture of a 12 weeks pregnant uterus following the previous traumatic abortion]

Rupture of uterus is a life-threatening complication with high fetal and maternal morbidity and mortality. Hysteroscopy, traumatic abortion, inappropriate use of oxytocin, manipulations, hydramnios, multifetal pregnancy, etc. are causes of rupture of uterus. A31-year-oldwoman G4P2A1L2, with 12-week pregnancy and with history of two cesarean deliveries and one traumatic abortion two months before the current pregnancy, presented to hospital with abdominal pain and bleeding. In clinical examination, whole abdomen had tenderness and patient complained of difficulty in breathing, nausea, and vomiting. In vaginal examination, the cervix was closed and the patient's bleeding was a little more than spotting. Vital sign decreased and the patient's dyspnea deteriorated. Hemoglobin level decreased from 13 to 4/5mg/dl. During vaginal sonography, the sonologistreported that there was no possibility of evaluation due to the fullness of abdominal cavity with fluid and blood, and there was a probability of ruptured ectopic pregnancy along with intrauterine pregnancy .The patient was transferred to operating room after the sonography. In the operating room, the fetus with gestational sac was seen in abdomen after laparotomy. The diagnosis of the rupture of uterus was from the previous site of incision, which was repaired by gynecologist