Mazandaran University of Medical Sciences
Research in Molecular Medicine
2322-1348
6
2
2018
5
1
CAR-T Cells: an Innovative Therapeutic Strategy against Pediatric Acute Lymphoblastic Leukemia
1
4
EN
Mohsen
Sheykhhasan
Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran; Department of Mesenchymal Stem Cell, the Academic Center for Education, Culture and Research, Qom Branch, Qom, Iran
Hamed
Manoochehri
Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Leila
Naserpour
Department of Reproductive Biology, The Academic Center for Education, Culture and Research, Qom Branch, Qom, Iran
Naser
Kalhor
Department of Mesenchymal Stem Cell, The Academic Center for Education, Culture and Research, Qom Branch, Qom, Iran
Mazandaran University of Medical Sciences
Research in Molecular Medicine
2322-1348
6
2
2018
5
1
Effect of UBE2Q on BECN and CEA Protein in Colorectal Cancer Cells
5
11
EN
Niloofar
Borumand
Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran.
Background: Expression of UBE2Q1, UBE2Q2, and members of the ubiquitin-conjugating enzyme family (E2) are affected in colorectal cancer (CRC). The BECN gene plays a key role in CRC cells. In gastrointestinal carcinoma therapy, tumor-associated antigens such as CEA are typically used.To investigate the association between UBE2Q1 and Beclin1 autophagy marker and CEA protein expression in LS180 CRC cell line.
Materials and methods: In this study, changes in the expression of BECN marker in LS180 cell lines with the vector containing UBE2Q1 were investigated using real-time PCR. The expression of CEA protein was also evaluated by western blotting. Statistical analyses were performed with Graph Pad Prism software.
Results: The results indicated reduced expression of BECN autophagy marker (P=0). Therefore, in the presence of UBE2Q1, cancer cells have less ability to induce autophagy. However, CEA protein levels in LS180 transfected cells with a UBE2Q1-ORF-containing plasmid decreased when compared to non-transfected cells.
Conclusion: The use of pharmacologic inhibitors related to the autophagy mechanism can be a novel approach in cancer therapy.
Mazandaran University of Medical Sciences
Research in Molecular Medicine
2322-1348
6
2
2018
5
1
Prevalence of bla VIM, bla IMP, and bla KPC Genes among Carbapenem-Resistant Klebsiella pneumoniae (CRKP) Isolated from Kurdistan and Isfahan Hospitals, Iran
12
20
EN
Leila
Gheitani
1 Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
Hossein
Fazeli
1 Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
Background: Carbapenem resistance among Klebsiella pneumoniae is an emerging problem worldwide. One of the main mechanisms of resistance to carbapenems is the potential of Klebsiella pneumoniae to produce carbapenemase enzyme.This study was conducted to determine the frequency of blaVIM, blaIMP, and blaKPC among carbapenem-resistant K. pneumoniae (CRKP) isolated from Kurdistan and Isfahan hospitals.
Materials and methods: This study was carried out in Iran using 183 samples from the Besat and Alzahra hospitals in 2017. Antibiotic susceptibility tests were performed by Kirby-Bauer disc diffusion. The modified Hodge test (MHT) was used to investigate the presence of carbapenemase. The β -lactamases genes were detected by PCR.
Results: The highest and lowest rates of resistance were observed against cefotaxime (98.2%) and gentamicin (43.6%), respectively. Among the 183 isolates, 134 (73.2 %) were positive by the MHT. The prevalence rates of blaVIM, blaIMP, and blaKPC were 4 (2.18%), 1 (0.5%), and 0%, respectively.
Conclusion: The prevalence of CRKP strains is a major concern and infection control processes are needed. These gene showed a low prevalence in our country, likely because other mechanisms of resistance to carbapenems are involved.
Mazandaran University of Medical Sciences
Research in Molecular Medicine
2322-1348
6
2
2018
5
1
Circulating miR-95 is a Potential Biomarker of Chronic Lymphocytic Leukemia
21
28
EN
Behnaz
Nateghi
Department of Biochemistry, Faculty of Science, Nourdanesh Institutions of Higher Learning, Meimeh, Isfahan, Iran
Parisa
Behshood
Department of Microbiology, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
Sima
Fathullahzadeh
Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran.
Omid
Mardanshah
Department of Laboratory Sciences, Sirjan Faculty of Medical Sciences, Sirjan, Iran.
Background: MicroRNAs (miRNAs) have crucial roles in cellular and molecular processes related to different malignancies including chronic lymphocytic leukemia (CLL). Studies revealed altered miR-95 expression in several diseases. Long non-coding RNAs (lncRNAs) are a heterogeneous group of non-coding and regulatory RNAs. The present study was conducted to investigate the association of miR-95 expression with CLL by quantitative real-time PCR.
Materials and methods: Sixty samples, including 30 CLL and 30 healthy controls, were sampled during a period of 4 months. The expression of miR-95 was evaluated by quantitative real-time PCR in peripheral blood mononuclear cells from patients with CLL and in healthy subjects. Additionally, in silico pathway enrichment analysis was performed on validated and predicted targets of miR-95 in several databases, including miRecords and miRTarBase, while the interactions between predicted putative lncRNAs and genes and miRNA expression were examined with miRWalk.
Results: The expression of miR-95 was found to be significantly reduced in patients with CLL compared to that in healthy controls (P < 0.005).
Conclusion: miR-95 showed potential as a biomarker for the early diagnosis of patients with CLL. LncRNAs play a significant role in regulating cellular evolution, differentiation, and other processes and may be important regulators in tumorigenesis.
Mazandaran University of Medical Sciences
Research in Molecular Medicine
2322-1348
6
2
2018
5
1
Expression of Recombinant Factor IX Using the Transient Gene Expression Technique
29
35
EN
Jafar
Vatandoost
Department of Biology, Hakim Sabzevari University, Sabzevar
Mohammad amin
Azimifar
Department of Biotechnology, Sabzevar Branch, Islamic Azad University, Sabzevar, Iran
Background: Pilot and large-scale production of recombinant proteins require the presence of stable clones, but the process of selecting stable clones is time consuming. Moreover, continuous clone culturing in large-scale production may cause loss of incoming plasmid and recombinant genes.
Considering the advancements in Transient Gene Expression (TGE) technology, the large-scale expression of factor IX (FIX) was investigated in HEK cells by the TGE technique.
Materials and methods: HEK cells were seeded in a cell factory, and then transfected by pcDNA-hFIX plasmid using calcium phosphate co-precipitation method. Stable HEK-hFIX cells were also seeded in a cell factory, separately. After adding vitamin K, recombinant FIX was quantified in conditioned media using an ELISA. Moreover, its functional activity was assayed using an aPTT test.
Results: The results showed that the expression and activity of FIX by TGE technology was, respectively, 1.6 and 1.5 times higher than that obtained through stable HEK-FIX cells. Since calculating the specific activity revealed that for all time periods it is 0.2 mU/ng, so the increase in activity is due to the increase in the amount of FIX.
Conclusions: HEK cells with higher transfectability seemed to be an appropriate alternative for transient expression for large-scale protein production. Furthermore, if rapid expression of recombinant proteins is intended, TGE can replace costly and low-yield methods.
Mazandaran University of Medical Sciences
Research in Molecular Medicine
2322-1348
6
2
2018
5
1
Effects of Exercises on IL-6 and GLUT4 Expression in Diabetic Rats
36
45
EN
Ali
Dehghan
Department of Exercise Physiology, Sari Branch, Islamic Azad University, Sari, Iran
Parvin
Farzanegi
Department of Exercise Physiology, Sari Branch, Islamic Azad University, Sari, Iran
Hajar
Abbaszadeh
Department of Exercise Physiology, Sari Branch, Islamic Azad University, Sari, Iran
Background: Diabetes mellitus is one of the most common metabolic disorders worldwide. Empirical evidence has shown exercise to be of value in ameliorating symptoms of diabetes, but the underlying molecular mechanisms involved have not been well-studied. To evaluate the effects of continuous and interval exercises on blood glucose and insulin levels and on IL-6 and GLUT4 expression in diabetic rats.
Materials and methods: Old Wistar rats with streptozotocin-induced diabetes were randomly divided into three groups; namely, the patient (non-exercise control) group, the interval exercise group, and the continuous exercise group. The blood insulin level was measured using a specific ELISA kit, whereas the blood glucose level was determined with an AutoAnalyzer apparatus. Expression of the IL-6 and GLUT4 genes in skeletal muscle was determined using real-time PCR. All data were analyzed using SPSS software.
Results: The expression of IL-6 and GLUT4 was significantly decreased in skeletal muscle of the diabetic rats (p < 0.001). Continuous (9.97-fold; p = 0.000) and interval (7.11-fold; p = 0.000) exercises significantly increased the expression of IL-6 relative to that in the animals of the patient group. Furthermore, continuous (9.36-fold; p = 0.000) and interval (7.65-fold; p = 0.001) exercises also significantly increased the expression of GLUT4 relative to that of the patient group. Both types of exercise training were associated with a significant decrease in the blood glucose and insulin levels compared with the levels in the patient group (p < 0.0001).
Conclusions: Diabetes is significantly associated with IL-6 and GLUT4 downregulation, but this can be reversed through continuous and interval exercises, which also help to lower the blood glucose and insulin levels significantly.