فصلنامه کومش
سال سیزدهم شماره 2 (پیاپی 42، زمستان 1390)

Anterior pituitary glycoprotein hormones include thyroid stimulating hormone, lutinizing hormone, follicule stimulating hormone, and gonadotropine hormone. Each of them contains alpha and beta subunits. The alpha subunit gene is the same in all of these hormones and contains 4 exones and 3 intrones. The beta subunit is responsible for specific function of each hormone. The aim of this study was to clone alpha chain cDNA of glycoprotein hormones in a proper vector for eukaryotic system. To clone cDNA, alpha subunit of glycoprotein hormones was amplified by using one pair primers and T.vector as template and cloned in Not I and Bam HI sites of pcDNA3.1 plasmid. The recombinant plasmid transformed to E.coli Top10F΄ cell and colonies that contain plasmid were selected by Colony PCR. The accuracy of extracted plasmid of these clones was approved by enzyme digestion and sequencing. Enzyme analysis showed that pcDNA3.1-F351 a had correct structure and sequencing confirmed by 100% homology of the gene with reported alpha gene in Gene Bank. Because of its proper structure, this plasmid is able to transform to Eukaryotic system and translation.

Chitin, a linear polysaccharide and is one of the most abundant natural polysaccharides is the main component in numerous of organisms. Chitinase enzymes are essential enzymes catalyzing the conversion of chitin to its monomeric or oligomeric components, therefore, chitinase can used as a natural antifungal agent. The purpose of this study is cloning and expression of recombinant chitinse gene in Bacillus subtilis in the treatment offungal infection effects and replacement with carcinogenic and synthetic products. Chitinase gene of Bacillus.pumilus was cloned to integrate vector Pdh5 Through designed the appropriate pimers.built construct using dual crossing over for transferred into the amylase gene (amyE) of Bacillus subtilis168; and its expression was induced by IPTG and its function was evaluated. Recombinant construct containing chitinase gene was produced. Gene expression by creating halo around the recombinant bacteria colonies in minimal media including chitin as carbon source was observed. Create a halo containing chitin in the minimal media represents the performance of recombinant chitinase enzyme to catalyze the chitin. Considering antifungal property can be effectively treated as a matter of human fungal infection and safe substance to excretion biological pest and pollutants.

Peroxisome proliferator-activated receptor gamma (PPAR g) is a ligand-dependent transcription factor involved in various disease processes including inflammation and carcinogenesis. This study aimed to determine polymorphism of PPAR g gene and its association with Helicobacter pylori infection and gastrointestinal diseases in patients. Two hundred patients with helicobacter pylori infection were examined. H. pylori infection was diagnosed by histology, rapid urease test (RUT), culture, ELISA and PCR. PPAR g polymorphism was analyzed by PCR-based restriction fragment length polymorphism. In total 200 patients (4 gastric cancer, 141gastritis, 35 peptic ulcer, 18 duodenal ulcer, 2 peptic ulcer and duodenal ulcer) with Helicobacter pylori infection were enrolled. The frequency of PPAR g G (Ala12) allele (5%) showed a significant association with gastric cancer (P=0.004) or gastritis (P=0.007). PPAR g GC (Pro12Ala) allele (35%) showed a significant association with duodenal ulcer (P=0.03) or gastritis (P=0.002). Pro12Ala PPAR g polymorphism is associated with gastric cancer and gastritis and is a potential marker for genetic susceptibility to these two diseases in the presence of H. pylori infection. Finally, our study suggests the potential association between PPAR g polymorphism and H. pylori infection in the development of gastric cancer and peptic ulcer and gastritis.

Immune response to recombinant L7/L12, in addition to protective role, may show its importance in detection Brucellosis tests. The aim of this study was to examine antigenicity of recombinant L7/L12 from Brucella abortus by Brucellosis human sera. We amplified L7/L12 gene by polymerase chain reaction (PCR) method and sub- cloned to prokaryotic expression vector pET28a. Escherichia coli BL21-DE3-plySs was transformed with pET28a-L7/L12 and gene expression was induced by IPTG. Recombinant L7/L12 was further analyzed by Western Blot. Sera reactivity of five infected individual were further analyzed against the recombinant L7/L12 protein. The sequencing result was confirmed by Sanger method and it was the same as L7/L12 gene. Escherichia coli BL21 (DE3) pLysS was transformed with pET28a-L7/L12 and gene expression was induced by IPTG. The expressed protein was purified by affinity chromatography by Ni-NTA resin. The data also indicated that L7/L12 protein from Brucella abortus recognized by patient sera. Our data showed that recombinant L7/L12 protein can be produced by pET28a in Escherichia coli. This protein was recognized by sera in infected human as an antigen. Therefore, recombinant L7/L12 has same epitopes with natural form of this antigen. Recombinant L7/L12 also seems to be a promising antigen for protection and serologic diagnosis of Human brucellosis.

Some studies have determined that polymorphism in insulin gene are associated with increased insulin level and resistant to insulin and also cause to increase risk of colorectal cancer (CRC). The goal of this study was to evaluate incidence of the insulin gene polymorphism (rs689) in an Iranian population and to investigate the role of this polymorphism in increased risk of CRC.

Genotyping of the insulin gene were determined in a series of 110 colorectal cancer patients and 110 controls by using polymerase chain reaction and restriction fragment length polymorphism genotyping assays (PCR-RFLP).

P value for genotype AT compared with AA, was 0.052 (OR=1.88, CI=0.99-3.5) and TT versus AA was 0.57 (OR=1.33 CI=0.48- 3.6). The results showed that the insulin gene polymorphism (rs689) is not a predisposing factor to increased risk to CRC (P=0.14). Incidence of mutant allele between patients and controls had no significant differences (OR=1.53 95% CI=0.98- 2.39, Pe=0.057).

These findings suggest that the insulin gene polymorphism (rs689) is not associated with increased risk of CRC.

Toxoplasmosis is a shared human-animal disease with worldwide distribution caused by Toxoplasma gondii. More than half of the world's population is seropositive for toxoplasmosis. The possibility of reactivation of the old infection or acquisition of infection from donor’s tissue increases in the transplant recipient patients who receive immunosuppressive therapy. In this study, IgM and IgG anti-toxoplasma immunoglobulins seroconversion in renal transplant recipients was evaluated before and after transplantation. This was a prospective cohort study on a total of 102 recipients. Two serum samples were obtained from each patient. The first sample was taken before administration of any immunosuppressive drugs and second sample was taken 3 months after transplantation. The IgM and IgG anti-toxoplasma antibodies were assayed by ELFA and ELISA techniques. IgM/ISAGA method was also used. ELFA identified 65 (63.7%) pre-transplantation samples as IgG+ and did not detect any positive IgM samples. However, IgM was detected in 3 (2.9%) post-transplantation samples by this method. Forty nine (48%) pre-transplantation samples were reported IgG+ by ELISA and no IgM positive sample was identified by this method. ELISA method detected 2 (1.9%) IgM positive reactions in post-transplantation samples. By IgM/ISAGA method, we detected no IgM positive reactions in pre-transplantation samples whereas 3 months later (second sampling) IgM antibody was detected in 3 (2.9%) cases. Secondary toxoplasmosis infection was observed in 30 cases per 1000 recipients, which indicates that screening for toxoplasmosis infection should be performed in developed countries for these patients. On the other hand, as the risk of re-active toxoplasmosis infection exists in developing nations, they should consider the necessary preventive measures to control this condition. In addition, we suggest that ELFA is the best method because of the most valuable results.

Clinical studies indicate that females have higher blood leptin levels when compared with the male gender, however to date there is no available report on the impact of offspring gender on maternal milk leptin (ML) levels. Therefore, this study was designed to investigate the effect of child gender on ML levels. A cross-sectional study was carried out across Urmia city health centers, Iran. 115 mothers were selected across a wide range of population. A questioner was designed with respect to infant gender, age and weight. Questioners were filled by all applicants and a 10 ml breast milk sample was taken for further analysis of ML levels and specific gravity (SG) determination. Mothers were divided into 5 different groups based on offspring's age. Maternal ML levels were significantly different according to offspring gender. ML levels were significantly higher in mothers with girls than mothers who have given birth to baby boy. However, there was no significant difference between ML levels with respect to infant age. SG level was found to be different among all groups and no significant correlation was found between SG and ML levels of all applicants. It was concluded that maternal ML levels varies with respect to the offspring gender. ML levels in infant girls are higher than that of the boy; however the mechanism of action is still unclear. A normal reduction in salt and water retention that occurred postpartum may cause slight elevation in SG of the breast milk.

The mesolimbic dopaminergic system that projects from the ventral tegmental area (VTA) to the nucleus accumbens (NAc) is critical for initiation of opioid reinforcement and reward-related effects of drugs of abuse. In the present study, the effects of reversible inactivation of VTA on firing rate of nucleus accumbens neurons and on acquisition and expression of morphine-induced conditioned place preference (CPP) were investigated in rats. Adult male Wistar rats were used in these experiments. In behavioral study, the reversible inactivation of VTA was done through bilateral intra-VTA microinjection of 2% lidocaine during the acquisition and expression of morphine (5 mg/kg; s.c.)-induced CPP and in electrophysiology section, it was done through unilateral intra-VTA microinjection of 2% lidocaine during single unit recording from the NAc neurons. Conditioning score and locomotor activity were recorded by Ethovision software. Firing rate of neurons was recorded by single unit recording technique. The results showed that bilateral intra-VTA administration of lidocaine significantly decreases the acquisition (P<0.01) and expression (P<0.05) of morphine-induced CPP compared to their respective saline-microinjected groups. Moreover, intra-VTA administration of lidocaine had no effect on locomotor activity in these experiments. Also, unilateral intra-VTA administration of lidocaine significantly increased the firing rate of nucleus accumbens neurons. Our results further support the idea that VTA may play an important role in the acquisition and expression of morphine-induced CPP.

With respect to antioxidant effect of Otostegia persica (O. persica) extract and the role of antioxidant agents in diabetes improvement, the aim of this study was to investigate the effect of methanolic extract of O. persica on serum level of glucose and morphology of pancreas in type I diabetic male rats. Type I diabetes was induced in male wistar rats (200-250 g) by injection of 65 mg/kg, i.p of streptozotocin. Before this and 5 days post-injection fasting blood samples were collected. Diabetes was confirmed in rats having fasting blood glucose level above 250 mg/dL. Diabetic rats were divided into 5 groups which received 100, 200 and 300 mg/kg/day O.persica extract, glibenclamide (600µg/kg) and distilled water (0.5 mL) for 14 days individually by gavage, respectively. After 14 days, fasting blood samples were collected and serum levels of glucose and insulin were measured using commercial kits by spectrophotometery and ELISA, respectively. Rat's pancreases disected out and used for histological studies after fixation, tissue preparation process and staining with hematoxylin-eosin dyes. The oral administration of O. persica extract in diabetic rats for 14 days significantly decreased glucose serum level, but it only at dose of 100 mg/kg significantly increased insulin serum level (p<0.05). Furthermore, extract improved pancreas tissue as in all doses it increased number of islets and at doses of 200 and 300 mg/kg increased the mean diameter of islets (p<0.05). Our study demonstrates that O. persica extract has a hypoglycemic effect by improving pancreas islets and increasing insulin secretion.

Various researches showed that depression is a key modifiable risk factor in maintaining of low back pain. The purpose of this study was to investigate the effectiveness of cognitive-behavioral group therapy on depression reduction of the patients with chronic low back pain. In an experimental study including pre-test, post-test, and following up the control group, thirty-five patients with diagnosis of low back pain compliance randomly selected in clinical centers of one, two and three regions of Tehran and randomly assigned to two experimental groups and one control group. All subjects completed depression subscale of the depression anxiety stress scales (DASS) in three phases of pre-test, post-test and follow-up. Cognitive-behavioral group therapy carried out on the experimental groups 8 sessions once a week. The results of variance analysis with repeated measurements showed that cognitive-behavioral group therapy significantly reduced the depression mean scores of the experimental groups as compared to the control group (P<0.05). According to the findings, cognitive-behavioral group therapy is an effective method in decreasing depression in patients with chronic low back pain.

In recent years computerized therapies like computerized cognitive behavioral therapy have helped the therapists. Some advantages of these interventions are reducing the costs and time and spreading the use of psychiatrist interventions as well. Thus, the aim of this study was to explore the efficiency of cognitive behavioral therapy in three methods of web-based intervention, software package and clinical intervention in comparison with untreated in major depression. The sample consisted of 68 students who were selected using availability sampling from students with major depression criterion who were seeking treatment at online and presence consultation center of University of Mohaghegh Ardabili (Iran), and then randomly assigned in three experimental groups and one control group. The instrument consisted of beck depression inventory and structured clinical interview for DSM disorders. The results of this study showed that in comparison to three different methods of cognitive behavioral therapy (web-based intervention, software package and clinical intervention) supported the more effective role of clinical intervention. Computerized therapies present effectiveness learning experiences and result in increasing fast access of therapists and patients to determined therapeutic goals. Results of this research indicate that computerized cognitive behavioral therapy is a feasible and effective intervention in treatment of major depression.

Visfatin, a novel adipokine, was revealed to be associated with obesity, but its role in response to exercise training in human is not known. The purpose of this study was to assess the effects of aerobic and resistance training on plasma visfatin in overweight women. Thirty-eight overweight volunteer women (age 34.84±5.70 y, body mass index 28.82±3.65 kg/m2, body fat percent 36.34±3.39 % and waist-hip ratio 0.85±0.04 (means ± SD)) were randomly divided into 3 groups. 14 subjects participated in an aerobic exercise training (50 min/day, 80-85% heart rate-reserve, 4 days/week), 14 subjects participated in a resistance exercise training (50 min/day, 60-70% 1-RM, 4 days/week), and 10 overweight women were served as control. Fasting blood samples were collected to assess changes in plasma visfatin, insulin and glucose levels before and after 2 months of the training program. Findings indicated a lower and significant (P<0.05) post-training change in circulating levels of visfatin and insulin in the resistance training group as compared with the other groups. Body mass index, waist-hip ratio, and %body fat were significantly decreased (P<0.001) in experimental groups as compared with control group. The resistance exercise training-induced reduction of plasma visfatin is most likely the result of weight loss and body composition changes in overweight women.

Iron deficiency can be followed with major problems in relation to reduced aerobic capacity and power for athletes. The purpose of this research was to study the effect of six weeks aerobic training on young girls’ hemoglobin, red blood cells, hematocrit, iron, serum ferritin and transferrin. 34 female students were selected with an average age of 16.56 years, weight 50.42 kg and height 160.55 cm from 100 volunteers randomly and assigned into three groups: exercise + iron supplementation (group 1), exercise+ placebo (group 2) and control. Before exercise, subject's blood samples were obtained. Experimental groups were trained for three sessions per week for six weeks. The experimental groups 1 and 2 received a tablet containing 27 mg ferrous gluconate and placebo respectively, the night before of each exercise session. After training period, the second stage of blood sampling was performed. Iron supplementation with aerobic exercise increased the amount of iron, serum ferritin and transferring. No significant effects were found on hemoglobin, hematocrit and red blood cells. Placebo intake along with aerobic exercise reduced serum iron, ferritin, hemoglobin, hematocrit and red blood cells significantly. Significant differences between three group's serum iron and ferritin were also observed. The results showed if aerobic exercise performed with iron supplements by young girls, in addition to increasing the amount of iron, it increases iron storage in the form of ferritin and transferrin and possibly their risk of anemia is prevented.

The effect of touch on growth of newborns has been achieved by educated staff, but touch therapy by mothers as the most important care provider for their infants, had not been focused in previous studies. The aim of this work was to study effect of touch therapy by mothers on weight gaining of preterm newborns. In a randomized controlled trail and double blind study, 60 preterm newborns who discharged from neonatal intensive care unit (NICU) to home, randomly allocated to control and touch therapy group (30 newborns in each group). Touch therapy was educated to mother in a direct education, also with offering guide booklet to the mothers. Touch was provided for three 20 minute period per day, until term corrected age. Neonate weight in two groups was measured at the time of discharge and then at the term corrected age by the electronic weighing scale with an accuracy of ± 10 gr. In two groups, mean of age, kind of delivery, level of education, employment and socio-economic status in mothers, also weight, sex and other neonate characteristics had no statistical differences. At the term corrected age, interventional group showed a weight gain of 10 g/day (30%) more than control, which was statistically significant (31 g/day vs 21 g/day) (P=0. 04). An important aspect of this study is that all stimulations were provided by mothers which had positive impact for preterm newborns, So, education of touch therapy to mothers can be recommended before discharge of pre-term newborns.

Cigarette smoking is a known cause of systemic disorders such as bronchogenic carcinoma and cardiovascular diseases. The aim of this cross-sectional study was to estimate prevalence of smoking in 30-70 year old men in Semnan province in Iran.

This survey was conducted in a sample population of 1695 men 30-70 years old, in October 2005 to February 2006 in both rural and urban areas of Semnan province. A multistage clustered sampling was performed with, first, 760 clusters (households) selected randomly using a systematic sampling technique. Then one or two more households in the neighborhood of each of the former households were also selected. Finally, in each household a 30-70 year old person was randomly selected and asked to complete a questionnaire.

The overall prevalence of current cigarette smoking in men 30-70 years old was 24.1% (95% confidence interval: 22.1-26.1%).18.3% of them were daily and 5.8% occasional smokers. 26.8% of rural and 23.0% city resistant adults were cigarette smokers (P=0.053). Prevalence of cigarette smoking in under 40, 40-49, 50-59 and ≥ 60 years were 21.1%, 32.6%, 25.1% and 11.9%, respectively (all, P<0.001). 19.7% of illiterates, 24.9% of primary, 30.8% of intermediate, 25.4% of high school and 11.8% of university level education were cigarette smokers(P<0.001). 46.6% of them before, and 39.2% between 21-30 years have started cigarette smoking. Mean (±SD) age for starting cigarette smoking was 24.1±8.0 years. Prevalence of cigarette smoking in Semnan, Damghan, Garmsar and Shahrood were 22.5, 23.2, 27.6 and 23.4% (P=0.554), respectively. Only 0.1% and 1.6% of them were pipe and tobacco smokers, respectively. 0.4% tobacco smoking was simultaneously with cigarette smoking. 15.2% non-cigarette smokers were passive smokers. Overall, 39.3% of Semnan province adults, aged 30-70 years, were active or passive cigarette smokers.

Findings showed that prevalence of cigarette smoking in Semnan province in adults aged 30-70 years is high. Therefore, a community-based multiple strategies are required to combat with increasing rate of cigarette smoking and its complications.

Though increased risk of gastric cancer in individuals with family history of the disease has been observed consistently in previous studies, data on the association between gastric cancer and family history of cancer from Iran is scanty. The purpose of this study is to evaluation of gastric cancer risk associated with family history cancer. The present study was designed as unmatched case-control study. Cases were 746 histopathologically confirmed gastric cancer and 746 controls were randomly selected among the healthy participants in a health survey. The family history of cancer was extracted from a standard history form completed by the patients or health care providers. Mantel-Heanszel Odds Ratio was computed for removing the confounding effect of age and sex. A positive family history of cancer was reported by 9.7% and 5.6% of cases and controls, respectively. Gastric cancer risk increased two-fold for subjects reporting any first degree relative with gastric cancer. There is no statistical association among family history of other cancers and gastric cancer (P>0.05). In conclusion, this study showed that family history of gastric cancer, especially in first-degree relatives, increases the risk of gastric cancer. Further studies are needed to better understand the role of genetic factors and environmental factors and their interaction in gastric cancer development in Iranian community.

Leptin and adiponectin, both as adipose tissue-derived adipocytokines, play a substantial role in the pathogenesis and complications of obesity. This study was performed to examine the effects of combined endurance and resistance training on plasma leptin and adiponectin levels and insulin resistance index (HOMA-IR) in sedentary postmenopausal women. 16 sedentary postmenopausal obese women in two experimental and control groups participated in 10 weeks of concurrent training including resistive (40 - 60% of 1 repetition maximum) and swimming endurance training (50-60% of maximal heart rate), three days per week. Plasma leptin and adiponectin levels, HOMA-IR and anthropometric indices were measured before and 48 hours after the last training session. Concurrent training did not significantly change plasma leptin level, but caused a significant increased in adiponectin concentration (P=0.001). Hence, HOMA-IR and all of anthropometric indices improved following 10 weeks of combined training (P<0. 05). Pearson correlation showed a significant correlation between the primary level of adiponectin and primary value of waist circumference (P=0.014), but no significant relationship was found between early level of leptin and anthropometric indices and HOMA-IR (P>0. 05). There were not significant correlation between leptin and adiponectin concentrations and changes of anthropometric indices and HOMA-IR (P>0. 05). Combined endurance and resistance training improves adiponectin concentrations, insulin resistance and anthropometric indices. However, these changes may not be in the line with leptin level changes.