Journal of Advances in Medical and Biomedical Research
Volume:22 Issue: 94, 2014

The important role of co-stimulatory molecules in immunological responses has been clearly demonstrated. With the development of new methods in molecular immunology, preventing the co-stimulatory signals in order to induce tolerance has emerged as a novel strategy. RNA interference is a new method that can specifically and effectively reduce target gene expression. In this study, we used a lentiviral system to target the expression of CD40 gene in dendritic cells. ShRNA sequence is able to reduce the expression of CD40 gene on murine dendritic cells. Our results indicate 64-fold decreased expression of CD40 mRNA in the group receiving lentiviral shRNACD40 as well as 31% decrease in expression of CD40 protein on the surface of dentridic cells (p<0.0001). Also, dendritic cells cannot stimulate T cells upon contact with them. This study indicates that a decrease in expression of CD40 molecule induces tolerogenic dendritic cells and prevents Th1 immunological response. Prevention of Th1 response would shift the immunological responses to Th2 response, and this type of response can be effective in autoimmune diseases including rheumatoid arthritis and type I diabetes. References 1- Groux H, Fournier N, Cottrez F. Role of dendritic cells in the generation of regulatory T cells. Sem in Immunol. 2004; 16: 99-106. 2- Kanako L, Reizis L, Reizis B. Dendritic Cells: Arbiters of Immunity and Immunological Tolerance. Cold Spring Harb Perspect Biol. 2012; 4: a007401. Avalable from: www.cshperspectives.org. 3- Morel PA. Dendritic cell subsets in type1diabetes: friend or foe? Frontiers in Immunol. 2013; 4. Avalable from: http://dx.doi.org/ 10.1155/ 2013/972865. 4- Machen J, Harnaha JO, Lakomy R, Styche A, Trucco M, Giannoukakis N. Antisense oligonucleotides down-regulating costimulation confer diabetes-preventive properties to nonobese diabetic mouse dendritic cells. The J Immunol. 2004; 173: 4331-41. 5- Maldonado R, Andrian UH. How tolerogenic dendritic cells induce regulatory T cells. Adv Immunol. 2010; 108: 111-165. 6- Peters AL, Stunz LL, Bishop GA. CD40 and autoimmunity: the dark side of a great activator. Sem Immunol. 2009; 21: 293-300. 7- Li L, Wang H, Wang B. Anergic cells generated by blocking CD28 and CD40 costimulatory pathways in vitro ameliorate collagen induced arthritis. Cell Immunol. 2008; 254: 39-45. 8- Zheng X, Suzuki M, Zhang Z, et al. RNAi-mediated CD40-CD154 interruption promotes tolerance in autoimmune arthritis. Arth Res & Ther. 2010; 12: R13. 9- Pletinckx K, Dohler A, Pavlovic V, Lutz M.B. Role of dendritic cells maturity/ costimulation for generation, homeaostasis and suppressive activity of regulatory T cells. Fronties in Immunol. 2011; 2: 7-22. 10- Gavrilov K, Saltzman WM. Therapeutic siRNA: Principles, challenges and strategies. Yale J Biol Med. 2012; 85(2): 187-200. 11- Manjunath N, Wu H, Subramanya S, Shankar P. Lentiviral delivery of short hairpin RNAs. Adv Drug Delivery Rev. 2009; 61: 732-45. 12- Inaba K, Inaba M, Romani M, et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with GM-CSF. J Exp Med. 1992; 176: 1693-702. 13- Virus packaging protocol. Stem Cell Biotechnology Research Center protocol, Iran. 2013. Avalable from: www.stemcellstech. com/index.php/fa/. 14- Muller G, Muller A, Jonuleit H. Fetal calf serum –free generation of functionally active murine dendritic cells suitable for invivo therapeutic approaches. J Inv Dermatol. 2000; 114: 142-8. 15- Marin-Gallen S, Clemente-Casares X, Planas R, et al. Dendritic cells pulsed with antigen-specific apoptotic bodies prevent experimental type 1 diabetes. Clin and Exp Immunol. 2009; 160: 207-14. 16- Hasse C, Ejrnaes M, Juedes AE, Wolf T. Immunomodulatory dendritic cells require autologous serum to circumvent nonspecific immunosuuressive activity in vivo. Blood. 2005; 106: 4225-33. 17- Feili-Hariri M, Dong X, Alber SM, Watkins SC, Salter RD, Morel PA. Immunotherapy of NOD mice with bone marrow–derived dendritic cells. Diabetes. 1999; 48: 2300-08. 18- Karimi MH, Ebadi P, Pourfathollah AA, et al. Immune modulation through RNA interference-mediated silencing of CD40 in dendritic cells. Cellular Immunol. 2009; 259: 74-81. 19- Mei S, Jin-Dong LI, Rui J, et al. Construction and characterization of lentiviral shRNA expression vector targeting rat CD40 gene in dendritic cells. Chem Res. 2009; 25(5): 666-72. 20- Jantsch J, Turza N, Volke M, et al. Small interfering RNA (siRNA) delivery into murine bone marrow-derived dendritic cells by electroporation. J of Immunological Methods. 2008; 337: 71-7. 21- Giannoukakis N, Trucco M. Dendritic cell therapy for type 1 diabetes suppression. Immunother. 2012; 4(10): 1-12. 22- Giannoukakis N. Tolerogenic dendritic cells for type 1 diabetes. Immunother. 2013; 5(6): 1-3. 23- Lee CN, Lew AM, Wu L. The potential role of dendritic cells in the therapy of Type1 diabetes. Immunother. 2013; 5(6): 591-606.

Visfatin is a secreted protein from visceral adipose tissue that has been linked to obesity and associated with health risk factors. The purpose of this study was to examine the effects of 8 weeks of interval endurance rope training on plasma visfatin and insulin resistance in overweight nonathletic adolescents In this semi-experimental study, 36 healthy overweight and obese male adolescents were randomly assigned to experimental (n=18) and control (n=18) groups. The experimental group underwent 8 weeks of interval endurance rope training (45 min/d, 4 d/wk). Blood samples were taken before and after the completion of exercise training to assess levels of visfatin, insulin resistance and lipid profiles. Independent T-test and Pearson correlation coefficient were used to analysis the data. The results showed that 8 weeks of interval endurance rope training decreased the plasma visfatin, body fat percent, BMI, insulin resistance, waist circumference and triglycerides levels in in the experimental group (P<0.05). Also, there was a significant and positive relationship between visfatin, body fat percent and plasma triglyceride levels (P<0.05). 8 weeks of interval endurance rope training with decreased triglyceride, insulin resistance and obesity induced a significant reduction of plasma visfatin in overweight and obese adolescents.

Pomalidomide - a combination of Lenalidomide and Thalidomide drugs- is one of the newest anticancer drugs. Pomalidomide induces apoptosis in cancer cells. Furthermore, few studies indicating its relatively low cytotoxic effects on normal peripheral blood cells have been carried out. However, there is yet no information about the effects of Pomalidomide on bone marrow cells that contain important immune cells. In this study, the effect of Pomalidomide on normal bone marrow mononuclear cells was studied. The samples obtained from individuals who clinically needed bone marrow examination, but finally were diagnosed with no serious pathologic conditions in their leukocytes. Remained bone marrow aspirates were obtained and mononuclear cells were isolated. Half of the mononuclear cells were cultured with Pomalidomide (final concentration 10ìM) and the remaining half(control group) were cultured with media alone. After 48h incubation, vital activity (MTT) and cell apoptosis induction rates were assessed. The results showed that the vital activity of bone marrow mononuclear cells cultured in the presence of Pomalidomide increased significantly compared to the control group (P<0.05). However, Pomalidomide had no apoptosis induction and the rate of apoptotic cells had no significant difference with control group. Pomalidomide moderately stimulates the viability (or activity) of normal bone marrow mononuclear cells to improve and maintain them. Also, unlike most anticancer drugs it has no observable toxic effect or apoptosis induction on these cells. So, it may be concluded that Pomalidomide may be more favorable in preventing the side effects of anticancer drugs on normal cells.

salvia officinalis L. consists of, glycosides, flavonoids, and anthocyanins. Due to the increasing use of medicinal plants in treatment of diseases and their side effects on various organs, this study was conducted to evaluate the effect of salvia officinalis on biochemical blood parameters in male rats. In this experimental study, 24 male Wistar rats were randomly assigned to four groups of 6 animals each. The control group received normal saline and treatment groups received 75, 150 and 300 mg/kg body weight of salvia extract for 14 days. Body weight was measured in days 0, 7, and 14. At the end of the experiment, serum levels of liver function enzymes such as AST and ALT, total proteins, albumin, creatinine, cholesterol and triglyceride were assessed. Results were analyzed using one- way ANOVA and Tukey test. According to the results, plasma concentrations of protein, albumin and creatinine showed a significant increase (P<0.05) but, a significant decrease was observed in liver enzymes. No significant changes were observed in serum cholesterol, triglyceride levels and body weight in the control and salvia groups (P>0.05). The results indicated that Salvia Officinalis probably contains flavonoidic components with antioxidant effect leading to the increase in albumin and decrease in liver enzymes.

Virgin Olive oil contains Oleocanthal as an anti-inflammatory agent. The main purpose of this study was to compare the effects of ultrasound therapy and phonophoresis with virgin olive oil on female athlete's chronic Low Back Pain (LBP). Thirty female athletes with chronic LBP voluntarily participated in this pilot study. Patients were randomly assigned to Phonophoresis (n=15) and Ultrasound (n=15) groups and were treated with virgin olive oil or ultrasound gel for 10 sessions. Frequency and duration of treatment by ultrasound were 1 MHz and 10-min, respectively. Subjects completed Mc Gill, Modified Oswestry and Ronald-Morris standard questionnaires before and after 10 sessions of physiotherapy. There was a significant reduction in symptoms of LBP based on Mc Gill, Ronald Morris, and Modified Oswestry questionnaires at the end of the therapy in both groups compared to pre-treatment status (P<0.05). Although the mean post-treatment scores were lower than the mean pre treatment scores in all questionnaires in phonophoresis compared with ultrasound group, there was no significant difference in symptoms of LBP between two groups (P>0.05). It seems both ultrasound and phonophoresis with virgin olive oil are effective methods for treatment of chronic LBP in female athletes.

Some evidence about the relationship between heme oxygenase and many cancers is available. Heme oxygenase has anti-apoptotic effects and contributes to tumor growth. The aim of this study was to evaluate the effect of heme oxygenase on melanoma tumor cells mitosis and tumor size in C57BL/6 mice. B16F10 melanoma cells were injected subcutaneously to 18 C57B 6 mice with 8 weeks of age. Mice were randomly divided into three groups: the first group received Zinc protoporphyrin (Znpp), heme oxygenase inhibitor (n = 6), the second group, hemin (heme oxygenase activator) (n = 6) and the third group((control group) (n = 6)) received diluent injection. They received their injection every other day from the first day of study for 16 days. Tumors were extracted in the 16th day of study. Length and width of tumors were measured and mitotic cell activities were evaluated using immunohistochemistry staining and counting Ki-67 posetive cells. Our study results showed that size and weight of tumor and mitotic index in treated mice by Hemin were higher than two other groups. (P≤0.05). But mitotic index, tumor size and tumor weight in the recipient Znpp group and the control group were not statistically different. Stimulation of Hemeoxygenase-1 pathway by hemin injection can increase melanoma tumor growth. However, inhibition of this pathway by Hemeoxygenase -1 inhibitors may not be effective in reducing tumor growth and size.

Gestational diabetes and hypothyroidism in pregnancy are the most common endocrine disorders which are considered as insulin resistant conditions. Maternal thyroid hormones play an important role in embryogenesis, fetal maturity, and child’s IQ level. It seems that subclinical hypothyroidism in women with gestational diabetes has a higher prevalence. Since thyroid function tests during pregnancy are not considered as part of a routine testing, therefore, we aimed to study thyroid function associated with insulin resistance in pregnant women. In this descriptive-analytic study, 142 pregnant women were randomly entered in GDM and normal groups. After obtaining demographic data and measuring all patients’ BMI, Glucose Tolerance Test (GTT) was performed. Serum insulin, thyroid function test and anti –TPO antibody were measured on fasting blood samples. Data were analyzed using descriptive statistics, T test, Chi-square test and multivariate regression analysis. 68 pregnant women with GDM and 74 normal pregnant women participated in this study. Patients with GDM had higher age and BMI (P< 0.0001). After eliminating the confounding variables, no significant correlation was found between TSH and insulin or HOMA-IR. Elevated anti-TPO was seen in 14.7% of GDM and 6.8% of normal women (P= 0.171). Significant correlation of anti TPO level was found between both insulin level (P= 0.01) and HOMA-IR (P=0.03). This study did not show any association between thyroid dysfunction and GDM. However, a higher anti-TPO level was seen in GDM patients. Further investigations with more sample sizes are recommended.

The aim of this study was to detect bacteria associated with biliary tract diseases and relationship between their class 1, 2 and 3 integrons and antibiotic resistance patterns. Bile samples of biliary patients undergoing cholangiopancreatography were examined for the presence of bacteria and antibiotic resistance patterns using biochemical tests and disk diffusion method. PCR was used for detection of the presence of integrase genes intI1, intI2, and intI3 in total DNA and plasmid extracts of these bacterial isolates. Out of 102 bile samples, 41.2% were positive by culture The isolated bacteria belonged to E. coli (35.5%), Enterococcus spp. (19.4%), Klebsiella pneumoniae (17.74%), Pseudomonas aeruginosa (17.74%), Acinetobacter spp. (6.45%) and Staphylococcus epidermidis (1.6%). All isolates were resistant to amoxiclav and 87.5% of the isolates showed multidrug resistance (MDR) pattern. intI1and intI2 were found in E. coli (59.09%, 50%), P. aeruginosa (81.81%, 45.45%), K. pneumonia (63.6%, 27.27%) and Acinetobacter spp. (100%, 100 %), respectively. These results showed the presence of class 1 and 2 integrons on the extracted plasmids and indicated a significant association between class 1 integron and resistance to gentamicin and ciprofloxacin. High frequency of class 1 and 2 integrons among the bile isolates and their association with increased antimicrobial resistance phenotypes suggests bile components as selective agents for bacterial strains encoding these elements.

Breast cancer is the most common cancer among women worldwide. Saffron is one of the most well-known plants as an antioxidant and anticancer. The aim of this study was to assess the impact of saffron on the viability of breast cancer cells and simultaneous interaction effect of Paclitaxel drug. In this study, saffron extract (Crocus sativus L.) was prepared by Soxhlet method. Breast cancer cells (4T1) were prepared from the Pasteur cell bank then passaged several times until the required number of cells obtained. Cytotoxicity of Paclitaxel and the extracts of saffron were assessed separately and simultaneously using colorimetric MTT assay and trypan Blue test with certain concentrations of drug and extracts in the treatment period of 48 and 72 h. Then, the rate of apoptogenic changes in each compound was evaluated by Hoechst and PI methods. Data from 48 and 72 h treatment of extract of Crocus sativus showed antitumor effects on breast cancer cells. These effects were dose and time dependent. The synergistic effects in 48 hour treatment indicated a significant increase in cytotoxicity of saffron/Paclitaxel (P <0.05). The results showed that ethanol and aqueous extract of saffron and Paclitaxel have significant anticancer properties against breast cancer cells. It was also found that the aqueous and ethanol extracts of saffron can considerably increase cytotoxicity in cancer cells induced by Paclitaxel while the aqueous extract of saffron showed better results.

Exercise and physical activity are the most impressive factors in consumption of cellular energy sources which may bring about some changes in key peptides that are effective in adjusting and balancing energy. The purpose of this study was to investigate the probable changes of plasma Neuropeptide Y (NPY) and Ghrelin concentrations after 8 weeks of exercise with different intensities on fat male rats. This experimental study was conducted on 75 adult male rats about 2 months old selected randomly from a laboratory. The rats fattened via stimulating their appetite with lettuce and other vegetables for a month. Their average weight changed from 240±15 gr to 320±20 gr. Then, they were divided into three groups of control (n=25), exercise group with moderate intensity ((n 25) and exercise group with high intensity (n=25). The training program of the study consisted of running three times in a week on Rodent's treadmill for 8 weeks. The results showed that after 8 weeks of high intensity exercises, NPY and Ghrelin levels increased significantly (p<0.05), but moderate intensity exercises did not have meaningful effect on NPY and Ghrelin levels. The results of this research shows that exercise causes negative balance of energy in rats, increase their weight and fatten them. To compensate for this negative balance of energy, NPY hormone and Ghrelin levels are increased.

Pseudomonas aeruginosa is an opportunistic nosocomial pathogen. Evidence suggests that the incidence of enzyme-producing strains of Pseudomonas aeruginosa Metalo Beta Lactamases (MBL) is a major problem in the treatment of infections caused by this organism. The aim of this study was to investigate the frequency of class I and II integrons among metalobetalactamase producing Pseudomonas aeruginosa isolated from clinical samples from Zanjan hospitals. In this cross-sectional study, 120 Pseudomonas aeruginosa isolates from clinical specimens were collected from Zanjan hospitals in 2012-2013. After verifying isolates by biochemical tests, antibiotic susceptibility testing (Kirby-Baur method) was performed according to CLSI guidelines against 7 antibiotics. The DDST was then carried out for detection of MBLs. Finally, the presence of IntI1, IntI2, blaVIM and blaGIM genes was determined by PCR. In our study, the highest and the lowest resistant patterns were related to Cefotaxime and Amikacin with 43.3% and 21.6%, respectively. Of 120 isolates, the metallo- beta-lactamase was detected in 35 (29%) isolates. According to results, the frequency of intI1, intI2, blaVIM genes in MBL producers were 32(91%), 3(9%), and 6(17%), respectively, but no blaGIM positive isolate was detected. Also all of IntI2 positive isolates carried IntI1 and blaVIM,, simultaneously. The results of this study indicate high frequency of class I integron gene among MBL producing Pseudomonas aeruginosa isolates. Due to the importance of integrons in dissemination of antibiotic resistance genes, detection and prevention of these elements are necessary.

Non-specific chronic low back pain is the most common type of chronic low back pain. Rehabilitation to achieve a more rapid improvement in patients with chronic low back pain has always been taken into account. The aim of this study was to evaluate the effect of Pilates exercises on pain in female patients with non-specific chronic low back pain. 27 women with unexplained chronic back pain and hyperlordosis aged 30 to 50 years old were randomly selected and were equally placed in either experimental or control groups. In the pre-test, pain level, performance of patients and the degree of lumbar lordosis were assessed using Quebec, Oswestry questionnaires and Formetric 4D system. Patients in the experimental group participated in an 8- week Pilates exercises while patients in the control group did not participate in any specific program. Then, in the post-test, pain level, performance and the degree of lumbar lordosis of patients in both groups were measured. The patients in the experimental group participated in the modified Pilates exercises for 8 weeks, but those in the control group only received the conventional treatment in the same time period. Data analysis was performed using SPSS 16 and significant level of P”0.05 was considered. The results indicated a significant decrease in pain scores (P”0.0001), a significant improvement in functional disability (P”0.001) and decreased lumbar lordosis (P”0.001) in the experimental group after 8- weeks of Pilates exercises compared to the control group. It can be concluded that Pilates exercises can decrease severity of pain, improve performance level and decrease lumbar lordosis in patients with chronic non- specific LBP.