The therapeutic potential of angiotensin-converting enzyme inhibitor enalapril to ameliorate muscle atrophy in a murine model.

Muscle atrophy due to limb immobilization and inactivity is a common consequence of many diseases and treatment processes. One of the systems activated in inflammatory conditions is the renin-angiotensin system (RAS). The present study was conducted with the aim of investigating the effects of one of the angiotensin-converting enzyme (ACE) inhibitors, enalapril, on improving muscle atrophy caused by immobility. The study was conducted in three groups: a control, an atrophy, and an atrophy group treated with enalapril on Balb/c mice. After tying a splint to cause atrophy in one of the legs, daily treatment with enalapril intraperitoneally (dissolved in DMSO) at a dose of 10 mg/kg/day was done for 7 days. On the eighth day, the splint was opened and half of the mice were evaluated. Then, in the recovery phase, treatment with enalapril was continued in the remaining mice for 10 days without a splint. At the end of each phase, the mice were examined for the muscle strength of the lower limb muscles, and histological and biochemical analyses were subsequently carried out. The tissue level of the oxidative stress index MDA was evaluated, which showed a significantly lower level in the enalapril group compared to the atrophy group (*P<0.1). Also, inflammatory factors in the enalapril group showed a decrease compared to the atrophy group. The strength of four limbs in the mice of the treatment group (-18.36 ± 1.70 %) was significantly higher than that of the atrophy group (-30.33 ± 3 %) at the end of the atrophy phase and also after 10 days of recovery. The results suggest that the use of enalapril that reduces the activation of angiotensin II-dependent pro-oxidant and pro-inflammatory pathways may improve the functional disorder and muscle necrosis in the murine model of muscle atrophy.

Age and Sex-related Chromogranin A Gene Polymorphisms and its Association with Metabolic Syndrome Components.

Introduction: The purpose of this study was to determine the possible differences in genetic polymorphisms and serum levels of chromogranin A (CgA), according to age and sex, in subjects with and without metabolic syndrome (MetS). Methodology: The genotyping and serum level of CgA and biochemical parameters were measured by the T-ARMS-PCR and PCR-RFLP and ELISA and spectrophotometer methods, respectively. Results: A comparison of males with and without MetS showed significantly lower high-density lipoprotein-cholesterol (HDL-C) levels than those of females.At ages 30-70 years, both sexes showed significant differences in triglycerides (TG), fasting blood sugar (FBS), CgA levels and waist circumference (WC) when compared to the two groups. Both sexes with MetS indicated significant differences in systolic blood pressure (SBP) at ages 40-70 years, while at ages 40-59 years, there was a significant difference in HDL-C level in males.There was a significant correlation between serum levels of FBS, TG, SBP and WC (in both sexes), and CgA in subjects with MetS. Significant correlation was found between HDL-C level and diastolic blood pressure (DBP), and CgA level in males and females, respectively. CgA genotype frequency (T-415C and C+87T polymorphisms) showed no significant differences between males and females with and without MetS, while there was only a significant difference in frequency of the genotypes T-415C when compared to males with and without MetS. Conclusion: The CgA appears to be strongly associated with MetS components in both sexes. Variation in CgA gene expression may affect the T-415C polymorphism in males. This may mean that the structure of CgA genetics differs in different ethnic groups. Differences in the serum level and expression of CgA gene may show valuable study results that it may be expected a relationship between these variables and the MetS.

The association of paraoxonase I gene polymorphisms Q192R (rs662) and L55M (rs854560) and its activity with metabolic syndrome components in fars ethnic group.

OBJECTIVES: Metabolic syndrome (MetS) may cause premature development of some diseases. PON1 genes may be involved in the pathogenesis of MetS. The aim of study was to evaluate the association between Q192R and L55M gene polymorphisms and its enzyme activity with the MetS components in subjects with and without MetS. METHODS: Polymerase Chain Reaction and Restriction Fragment Length Polymorphism analysis were performed to determine polymorphisms of the paraoxonase1 gene in subjects with and without metabolic syndrome. Biochemical parameters were measured by spectrophotometer. RESULTS: The MM, LM, and LL genotype frequencies of the PON1 L55M polymorphism were 10.5, 43.4, and 46.1%, and 22.4, 46.6, and 31% and; the QQ, QR, and RR genotype frequencies of the PON1 Q192R polymorphism were 55.4, 38.6 and 6%; and 56.5, 34.8 and 8.7% in subjects with and without MetS, respectively. The L and M allele frequencies were 68 and 53%; and 32 and 47% for PON1 L55M in subjects with and without MetS, respectively. The Q and R allele frequencies for PON1 Q192R were 74 and 26% in both groups. There were significant differences in HDL-cholesterol level and PON1 activity in the genotypes QQ, QR, and RR of the PON1 Q192R polymorphism in subjects with MetS. CONCLUSIONS: The PON1 Q192R genotypes had only effected on PON1 activity and HDL-cholesterol level in subjects with MetS. Different genotypes of the PON1 Q192R seem to be important candidates to make the subjects susceptible to MetS in the Fars ethnic group.

Genetic polymorphisms -137 (G > C) (rs187238) and -607 (C > A) (rs1946518) and serum level of interleukin 18 in Fars ethnic groups with metabolic syndrome in Northern Iran.

INTRODUCTION: We aimed to determine the genetic polymorphisms and serum level of interleukin 18 in Fars ethnic groups. MATERIAL AND METHODS: 226 Fars ethnic groups were participated. The ATP III criteria were used to assess MS components. The SNPs of the IL-18 gene were determined with ARMS-PCR. RESULTS: The GG, GC, and CC genotypes of -137 were 50%, 40%, and 10%. The CC, CA, and AA genotypes of -607 were 45%, 37%, and 18%. The GG, GC, and CC genotypes of -137 were 44.20%, 43.40%, and 12.40%, and were 55.75%, 36.28%, and 7.97% in subjects with and without MS, respectively. The CC, CA, and AA genotypes of -607 were 48.70%, 37.20%, and 14.20% and were 41.60%, 37.20%, and 21.20% in both groups, respectively. CONCLUSION: IL-18 gene may different in specific populations, different ethnic groups and geographic regions. The IL-18 polymorphisms might not be used as a marker of metabolic syndrome.

The effect of simvastatin on gene expression of low-density lipoprotein receptor, sterol regulatory element-binding proteins, stearoyl-CoA desaturase 1 mRNA in rat hepatic tissues.

The study aimed to assess the effect of simvastatin on gene expression of LDLR, SREBPs, and SCD1 in rat hepatic tissues fed with high-fat diets (HFD) and its association with some biochemical parameters. Thirty-two male Wister albino rats were divided into four equal groups (three test and one control groups). The biochemical parameters were determined by using spectrophotometer techniques and the Elisa method. Low-density lipoprotein receptor, sterol regulatory element-binding proteins, stearoyl-CoA desaturase1, Beta-actin were analysed by real-time quantitative polymerase chain reaction (RT-PCR) method. At the end of study, the livers of the rats were separated and changes of hepatic tissue were determined. LDLR, SREBP2, and SCD1 expression increased significantly when compared G1 versus G4 and G2 versus G4. The expression of LDLR, SREBP2, and SCD1 also increased significantly when compared G2 versus G3, G1versus G3 and G1 versus G3 and G2 versus G3. The serum level of cholesterol, triglyceride, glucose, LDL, and HDL increased significantly when compared G1 versus G3. LDL showed significantly decreased when compared G1 versus G2. Cholesterol, glucose and HDL and triglyceride levels were increased significantly when compared G1 versus G4 and G2. Treatment of rats with HFD and simvastatin 20 mg/kg, triglyceride and LDL were almost the same as a control group and LDLR expression increased 98% in liver tissue. Gene expressions may be up-regulated in liver tissue and they showed different effects on biochemical parameters.

Age- and Gender-Related Serum Level of Interleukin 18 in Iranian Fars Ethnic Group with Metabolic Syndrome.

Background: The purpose of the present study was to investigate the age- and gender-related serum level of interleukin 18 (IL18) in male and female Iranian Fars ethnic group with metabolic syndrome components. Methods: The study included 226 native Iranian Fars ethnic groups. One hundred sixteen females and 110 men were selected. There were 60 females and 50 males with metabolic syndrome and 56 females and 60 males without metabolic syndrome. The serum fasting blood glucose (FBS), lipid profiles, and IL18 were measured. The National Cholesterol Education Program Adult treatment Panel III criteria were used to determine metabolic syndrome components. Results: There were significant differences between the males and females [except high-density lipoprotein (HDL)] with and without metabolic syndrome for the mean body mass index, FBS, HDL-cholesterol, waist circumference (WC), triglyceride (TG), and IL18 levels in all age groups. Serum IL18 was the highest in males and females in age groups 61-70 and 41-50 years with metabolic syndrome, respectively. Serum IL18 levels significantly correlated with TG and waist WC in males (and also correlated with HDL) and females with the metabolic syndrome. There were significant correlations between IL18 and TG and WC in males (and also correlated with HDL) in ages 61-70 years and females in ages 41-50 years with the metabolic syndrome. Conclusions: The increased IL18 level in both gender and different ages may have an important role in the alteration of some metabolic syndrome components. These alterations may be made to happen in different related metabolic diseases. IL18 seems to be a useful biomarker for the management of metabolic syndrome components and the risk factors of cardiovascular disease.

Gender and Body Mass Index-Related Serum Level of Adipokines and Metabolic Syndrome Components in Bipolar Patients Who Received Lithium and Valproic Acid.

Background: This is the study to assess alterations on adiponectin, leptin, and metabolic syndrome components in women and men bipolar disorder (BD) patients with normal weight and obesity who received valproic acid (VPA) and lithium (Li). Methods: Thirty-six women and 51 men were included. Commercial kits were used to determine all parameters. Metabolic syndrome components were determined according to the NCEP ATP III criteria. Results: Patients who received Li and VPA significantly differ in waist circumference (WC) and triglyceride (TG) levels (in women and men). Normal weight patients received both drugs, significant differences were considered in high-density lipoprotein-cholesterol (HDL-C), WC, and TG levels compared to healthy controls, but there were significant differences in TG, leptin, and adiponectin levels in obese patients who received VPA. There were significant negative and positive correlation between leptin and adiponectin and WC and TG in women and men BD patients treated with VPA and Li. There were significant positive correlation between leptin and adiponectin and WC and TG and significant negative correlation with HDL-C in normal weight BD patients treated with VPA and Li, respectively, while there was only a significant positive correlation between leptin and adiponectin, and TG in obese BD patients treated with VPA. Conclusions: It looks like that patients treated with both drugs for our suggested time may increase leptin and adiponectin levels. Correlation differences between leptin and adiponectin, and metabolic syndrome components may be important parameters in women, men, normal weight, and obese BD patients. Monitoring of body composition and adipokines may benefit in medical care of these patients.

Effect of Spirulina on Lipid Profile, Glucose and Malondialdehyde Levels in Type 2 Diabetic Patients

Abstract The study aimed to assess possible spirulina effects on lipid profile, glucose, and malondialdehyde levels in new cases of type 2 diabetes. The subjects consisted of 30 new cases of types 2 diabetes that divided into two groups; each consisted of 15 diabetic patients. Group I did not take any functional food or supplement and received no spirulina supplementation. Group II or experimental group also did not take any functional food or supplement but received spirulina supplementation. Analysis of data was done using SPSS 16.0. The Kolmogorov-Smirnov test, paired t-test, Wilcoxon test, and Spearman correlation analysis were used to analyze the data. After eight weeks of spirulina supplementation, significant differences were shown in the serum levels of total cholesterol, low-density lipoprotein (LDL)-cholesterol, triglyceride, and malondialdehyde. The serum fasting blood glucose, lipid profiles, and malondialdehyde levels at baseline were negatively and positively correlated with changes in these parameters. Spirulina supplementation may have a beneficial effect on lipid profile and malondialdehyde (MDA) levels through an interventional 8 weeks. This effect may protect subjects against free radicals and the development of some diseases such as atherosclerosis. The spirulina supplementation also showed a potential lipid-lowering effect on new case type 2 diabetic patients which may help the diabetics to have control on lipid levels. In addition, spirulina may be used as a functional food for the management of lipid profiles and MDA levels.

Effects of Onosma dichroanthum Boiss. root extract on AGS human gastric cancer cell-line.

One of the cancer-related deaths is gastric cancer in this area. Onosma dichroanthum Boiss. roots have been used as an antiseptic and anti-inflammatory for wound healing treatment. The aim of this study was to assess the in vitro cytotoxic and anticancer effects of O. dichroanthum Boiss. roots from the Golestan province of Iran. After identification of the taxonomical effect of O. dichroanthum Boiss., different concentrations of the hydroalcoholic root extract were used. Three different time periods (24, 48, and 72 h) were used to treat AGS gastric cancer and L-929 normal fibroblasts cell lines. The evaluation of different concentrations of root extract was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The 48 h treatment affected cell survival, while the concentration of 64 µg/mL was determined as IC50 concentrations at 48 h incubation time. The 48 h incubation time with 64 µg/mL showed the best effectiveness on cancerous cell-line while being safe for normal cell-line. Our results show that O. dichroanthum Boiss. roots extract may have cytotoxic and safe effects on gastric cancer cell-line and normal cells in 48 h treatment periods, respectively. The results indicated the O. dichroanthum Boiss. may be as an effective anticancer agent (gastric cancer).

Melatonin and doxorubicin co-delivered via a functionalized graphene-dendrimeric system enhances apoptosis of osteosarcoma cells.

A functionalized graphene-dendrimeric system was designed via Fe3O4 nanoparticle (NP) as a magnetic nanocarrier for co-delivery of doxorubicin (DOX) and melatonin (MLT). Accordingly, ß-Cyclodextrin (ß-CD) was modified by creating amine functional groups. The modified ß-CD was grafted with Graphene oxide (GO), and the resulting platform gain many functional groups, including the hydroxyl (-OH), carboxylic acid (-COOH), and amine functional groups (-NH2). Finally, magnetic NPs were synthesized on the prepared platform to efficiently controlling and targeting drugs to tumor sites. The human osteosarcoma cell lines including Saos-2 and MG-63, as well as Human Bone Marrow Mesenchymal Stem Cells (hBM-MSC) line, were used to determine the in vitro biological effects of the functionalized graphene-dendrimeric system. The magnetic nanocarrier has encapsulation efficiency (EE) values of 99.92% for DOX and 21.5% for MLT. The biocompatibility tests of the nanocarrier revealed that the magnetic nanocarrier was appropriate as a drug carrier. Co-delivery of DOX and MLT with an efficiently anticancer performance was also was confirmed by cellular uptake, 4',6-diamidino-2-phenylindole (DAPI) staining, and apoptosis analysis in comparison with free DOX and MLT. Moreover, there was a synergy in the antitumor effect when MLT was combined with DOX, especially in the nano-formulation form, which may be due to the down-regulation of X-linked Inhibitor of Apoptosis (XIAP), survivin, and human telomerase catalytic subunit (hTERT) (p < 0.0001). Overall, the result of our study suggests that the designed carrier is a promising nanocarrier for targeted co-delivery of DOX and MLT with improved anticancer efficacy in cancer cells and thus reduced toxicity in normal cells.

The association between leptin and adiponectin, and metabolic syndrome components and serum levels of lipid peroxidation in bipolar disorder patients treated with lithium and valproic acid.

BACKGROUND: The aim of study is to assess a relation between the adiponectin and leptin levels, and metabolic syndrome components and lipid peroxidation treated with Li and VPA in bipolar disorder patients and compared with controls. MATERIALS AND METHODS: 56 patients and 31 healthy controls were enrolled. The ATP III criteria were used to determine metabolic syndrome components. Leptin, adiponectin, lipid peroxidation and lipid profiles were measured. RESULTS: Malondialdehyde in Li patients was higher than VPA patients. BMI, waist circumference (WC), triglyceride, malondialdehyde and adiponectin levels were increased, whereas HDL-cholesterol (VPA treated patients) and leptin were decreased in patients compared with controls. Leptin and adiponectin were correlated with WC, triglyceride and malondialdehyde in both groups. Adiponectin was correlated with HDL-cholesterol in VPA patients. CONCLUSION: Patients should be checked metabolic syndrome components, serum leptin and adiponectin level occasionally to prevent possible deficiency or pathologic increase of these parameters.

Effect of Rubus anatolicus Leaf Extract on Glucose Metabolism in HepG2, CRI-D2 and C2C12 Cell Lines.

INTRODUCTION: The aim of this study was to assess the effects of Rubus anatolicus on glucose metabolism in HepG2, CRI-D2 and C2C12 cell lines. MATERIALS AND METHODS: R. anatolicus was collected in Golestan province, Iran. Three different cell lines HepG2 (human liver cell), CRI-D2 (mice pancreatic cell) and C2C12 (rat myoblast) were used for cell culture experiments. Cell viability was measured using MTT assay. Cells were treated with various concentrations of the extract (6.25-400 µg/mL) and then the extracellular glucose level and intracellular glycogen content were measured using colorimetric methods. The insulin level of the culture medium was measured using the ELISA method. RESULTS: Our findings showed that R. anatolicus extract enhances glucose uptake and consumption by all three cell lines. The R. anatolicus extract exposure also elevated cellular glycogen content in HepG2 and C2C12 cells (for 200 and 100 µg/mL) significantly. We found a significant increase in glucose uptake and consequently higher stimulation of insulin secretion in CRI-D2 cell pancreatic cells treated with R. anatolicus extract. CONCLUSION: The R. anatolicus appears to activate glucose uptake and cellular glycogen synthesis probably by activating the glycogenesis or inhibition of glycogenolysis pathways. The extract enhances insulin secretion in the pancreatic cells by increased glucose uptake.

Evaluation of Relationship Between Arylesterase-Based Activity and Genetic Variants of Paraoxonase1 in T2DM Patients within Golestan Province.

Arylesterase activity of Paraoxonase-1 (PON1) enzyme may be play important role in initiation and progression of several diseases. Activity or serum level of Arylesterase can be affected by many genetic alterations such as SNPs. The reduction in the activity and serum level of Arylesterase could be involved in Type2 diabetes mellitus (T2DM). The aim of this investigation is to examine the association between Arylesterase activity and promoter polymorphism (- 108C > T) of PON1gene in patients with T2DM in Golestan Province, northern area of Iran. Achievement of this purpose was due to DNA obtaining from blood then SNP determination using PCR-RFLP and Arylesterase activity measurement in the serum of 90 normal individuals and 90patients suffering diabetes. Data was processed by SPSS software version 16. The significant association was observed between the Arylesterase activity and three genotypes of PON1 gene such as CC, CT, and TT in subjects with T2DM. In the present study, it has been shown level of Arylestrase activity of PON1 in patients (117.33 ± 63.96) is lower than it in control group (289.33 ± 68.38); P < 0.05. Our results declared that activity of Arylesterase in diabetic patients was significantly lower than the healthy individuals.

MicroRNA-146a as a Prognostic Biomarker for Esophageal Squamous Cell Carcinoma.

BACKGROUND AND AIMS: MicroRNAs including miR146a have a regulatory role on the expression of genes and act with binding to 3'-UTR region of the genes. Cyclooxygenase-2 (COX-2) is involved in carcinogenesis as an inflammatory marker, and microRNA-146a (miR-146a) as a negative regulatory factor. We aimed to evaluate miR146a expression as a prognostic or diagnostic biomarker for esophageal squamous cell carcinoma (ESCC) and also an association between miR146a and COX2 expression. MATERIALS AND METHODS: We quantified the level of miR-146a and COX-2 expression in cancerous and adjacent normal tissue samples obtained from 34 patients with ESCC, using real-time-PCR. Statistical analyses were conducted using one-sample t-test. Receiver-operating characteristic (ROC) curve and Kaplan-Meier analysis were applied to assay miR146a as a diagnostic and prognostic marker, respectively, during 4 years of the study. Furthermore, the Cox regression model was performed to assay the hazard ratio (HR). The association between miR-146a and COX2 expression level in ESCC patients was evaluated by nonparametric Spearman's rho analysis. RESULTS: The results revealed a reduction of miR-146a expression in 50% of cancerous tissue when compared with adjacent normal regions (P-value=0.127). COX-2 expression in 80% of ESCC patients was higher than in the controls (P-value=0.001). Overall, in 60% of cases, direct association was seen between microRNA-146a and COX-2 expression level (correlation coefficient= 0.438, P-value=0.011). COX2 can be considered as a diagnostic biomarker (AUC=0.834, sensitivity=72%, specificity =83%, P-value<0.0001) but miR146a cannot be considered as a diagnostic biomarker (AUC=0.553, sensitivity=88%, specificity =28%, P-value=0.453). Survival analysis by Kaplan-Meier method showed miR146a and COX2 expression can be probably considered as prognostic biomarkers for ESCC because patients with high expression of miR146a had 7 months shorter life span and patients with low expression of COX2 had 8 months shorter life span. CONCLUSION: COX2 expression is a diagnostic biomarker. MiR-146a and COX2 expression can probably be considered as prognostic biomarkers for survival in ESCC.

The most effective polymorphisms of paraoxonase-1 gene on enzyme activity and concentration of paraoxonase-1 protein in type 2 diabetes mellitus patients and non-diabetic individuals: A systematic review and meta-analysis.

INTRODUCTION: Many studies have evaluated the association of paraoxonase-1 (PON1) gene polymorphisms with enzyme activity and concentration in type 2 diabetes mellitus (T2DM). However, the exact impact of these polymorphisms is not still obvious. Hence, we conducted a systematic review and meta-analysis to clarify the association of PON1 polymorphisms with its enzyme characteristics in T2DM patients and non-diabetic individuals. METHODS: We searched electronic databases including PubMed, Web of Science, Embase and Scopus for publications by April 2018. The pooled response ratio (rr) for the association and their corresponding 95% confidence intervals (CIs) were calculated using a fixed-effect model. RESULTS: Fifteen relevant studies fulfilled our inclusion criteria. The results showed a 1.25-fold increase in total PON1 activity in non-diabetic group against T2DM patients (p-value = 0.024). Also, only Q192R and L55M polymorphisms had sufficient studies to be included in the meta-analysis. All three genotypes of Q192R polymorphism showed significantly different activities between the study groups with the highest pooled effect size for RR genotype (rrQQ < rrQR < rrRR) while this difference was seen only in LL genotype of L55M polymorphism. Therefore, Q192R polymorphism was more correlated with type 2 diabetes mellitus. In case of concentration, there was no significant differences between two groups (p-value = 0.897). CONCLUSION: Current meta-analysis suggested that the observed difference of total PON1 activity was due to the different activity of various genotypes of PON1 enzyme in case of L55M and Q192R polymorphisms so that LL and RR genotypes had the most important role in the establishment of mentioned difference.

Polymorphism of Secretary PLA2G2A Gene Associated with Its Serum Level in Type2 Diabetes Mellitus Patients in Northern Iran.

BACKGROUND: Inflammation may occur in Type2 diabetes mellitus. sPLA2 is among the factors that contribute to the activation of pathways involved in inflammation. Several agents affect serum sPLA2 level, one of which is genetic diversity. OBJECTIVE: The current study was performed to determine whether there is a relationship between sPLA2 gene (-763C > G) polymorphism and circulating sPLA2 level in patients with Type 2 diabetes. METHODS: DNA was extracted from blood samples and used for the amplification of sPLA2 gene using ARMS-PCR. RESULTS: A statistical analysis using SPSS (version 16) revealed a significant correlation between -763C > G sPLA2 gene polymorphisms and the disease incidence in patients with T2DM. Among the three possible genotypes (GG, CC, and CG), CG genotype was found to have a higher frequency(53%) in T2DM patients. GG and CC genotypes frequencies were 20 and 27%, respectively. In healthy individuals, the frequencies of CC, GG, and GC genotypes were 77, 9.8% and 13.2%, respectively). Patients with genotype GG had the highest level of sPLA2. We showed that C>G polymorphism at position- 763 is associated with a high level of sPLA2 in both T2DM patients and healthy individuals. The average of sPLA2 circulating level was (170.48± 84.90), (106.62 ± 74.31), in patients and normal individuals, respectively. CONCLUSION: Our findings show that sPLA2 serum level is significantly higher in patients with T2DM disease than that in healthy individuals.

Thiopurine Methyltransferase Genetic Polymorphisms and Activity and Metabolic Products of Azathioprine in Patients with Inflammatory Bowel Disease.

BACKGROUND: Thiopurine S-methyltransferase (TPMT) is a cytoplasmic enzyme that catalyzes thiopurine drugs such as 6-mercaptopurine, 6-thioguanine, and azathioprine. There is a correlation between thiopurine drug metabolism, response, and toxicity and genetic polymorphism of TPMT. The aim of this study is to assess TPMT genetic polymorphisms activity and metabolic products of AZA in patients with IBD. METHODS: Blood samples were obtained from 50 IBD unrelated patients from a private laboratory. We used polymerase chain reaction-restriction length polymorphism (PCR-RFLP) and allele-specific PCRbased assays to determine the TPMT gene for the different variants. A high-performance liquid chromatography system (HPLC) was carried out to determine the whole blood 6-TGN concentration. Determination of serum TMPT activity was done by ELISA kit. RESULTS: In IBD patients, 46/50 (92%) subjects were homozygous for the wild-type allele (TPMT*1/*1). Mutant TPMT*1/*2 and TPMT*1/*3C alleles were found in 4/46 (8%) and 3/47 (6%) of IBD patients, respectively. TPMT*1/*3B variant was not detected in any of the IBD patients. TPMT enzyme activity was higher in wild-type than that mutant variants TPMT*1/*2 and TPMT*1/*3C, suggesting that there are statistically significant differences between 6-TG levels and polymorphisms of TMPT enzyme. 6-TG levels significantly increased in IBD patients mutant variants TPMT*1/*2 and TPMT*1/*3C. CONCLUSIONS: Our results showed that TPMT polymorphisms are associated with 6-TGN levels in patients using AZA. This study suggests that AZA dosage may be determined according to the high or low prevalence of a TPMT genotype. Moreover, the results present the determination of metabolite for assessing possible safe effective dosage of the drug.

Micro-RNAs -106a and -362-3p in Peripheral Blood of Inflammatory Bowel Disease Patients.

OBJECTIVE: MicroRNAs (miRNAs) can regulate various genes after binding to target mRNAs. Studies on Inflammatory Bowel Disease (IBD) in relation with miRNA are much less shown. The aim of the present study was to assess the expression patterns of microRNA 106a and microRNA 362-3p in peripheral blood samples of Inflammatory Bowel Disease (IBD) patients including Crohn's Disease(CD) and Ulcerative Colitis (UC). METHODS: This study consisted of 32 CD, 32 UC patients and 32 controls. The expression level of the micro-RNAs -106a and -362-3p was determined using reverse transcription and real-time RT-PCR. RESULTS: Our findings showed that MiR-106a and miR-362-3p are expressed at significantly higher levels in the peripheral blood from patients with CD and UC compared to controls. MiR-106a and miR-362-3p expression are also different in the peripheral blood of patients regarding the activity score of the disease. There were significant differences of miR362-3p in active UC relative to inactive UC. CONCLUSION: Altogether our findings suggest that miR-106a and miR-363-3p can play an important role in the pathogenesis of IBD. The differences in expression of miR106a and miR362-3p in peripheral blood of the UC and CD patients in an active phase in comparison to inactive disease suggest that these miRNAs may be useful as potential biomarkers for diagnosis and monitoring the disease activity.

Genetic Polymorphisms of Cytochrome p450 (2C9) Enzyme in Patients with Type 2 Diabetes Mellitus in Turkmen and Fars Ethnic Groups.

BACKGROUND: Cytochrome P450 2C9 (CYP450 2C9) has an important role in metabolic processes. Mutations in CYP450 2C9 genes may affect the catalytic activity of this enzyme. The aim of the present study is to assess the genetic polymorphisms of Cytochrome P450 (2C9) enzyme in Turkmen and Fars ethnic groups with type 2 diabetes compared with controls. METHODS: A total of 336 Turkmen and 336 Fars type 2 diabetic patients and 336 healthy Turkmen and Fars individuals were included in this study. Genomic DNA was extracted from whole blood samples and then the CYP2C9 genotyping was done using the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism technique. RESULTS: The CYP2C9*1, CYP2C9*2 and CYP2C9*3 allele frequencies in type 2 diabetic patients were 85.27%, 11.68%, and 3.05%, and in control were 87.13%, 8.56%, and 4.31%, respectively. We found significant differences between allele distribution of 2C9 in type 2 diabetic patients and controls. CYP2C9*2 and CYP2C9*3 allele frequency was significantly different in Turkmen and Fars type 2 diabetic compared to two ethnic controls. The CYP2C9*1/*1 and CYP2C9*1/*2 genotypes frequencies in type 2 diabetic Turkmen showed significant differences compared to Turkmen control. There were significant differences in the genotype frequency of CYP2C9*1/*1, CYP2C9*1/*2, CYP2C9*1/*3, CYP2C9*2/*2, and CYP2C9*2/*3 between type 2 diabetic Fars and Fars controls. Two diabetic ethnic groups showed statistically significant differences in frequencies of CYP2C9*2/*2 and CYP2C9*2/*3 genotypes. CONCLUSION: Our study suggests that diabetic patients with mutant CYP2C9 polymorphism may show different antidiabetic drug metabolism compared to the wild-type allele. In this regard, determination of CYP2C9 alleles and genotypes can be a useful tool for the treatment of diabetic patients with antidiabetic drugs because it may assist physicians' to determine optimal dosage and efficiency of drugs metabolized by this polymorphic enzyme.

Genetic Polymorphism of CYP2C9 Among Sistani Ethnic Group in Gorgan.

Cytochrome P450 2C9 (CYP2C9) is involved in metabolism of many important drugs and its genotype variations is thought to affect drug efficacy and the treatment process. The aim of this study was to assess the distribution of CYP2C9 allele and genotypic variants in Sistani ethnic group, living in Gorgan, South East of Caspian Sea and North East of Iran. This study included 140 Sistani, referred to the health center of Gorgan. CYP2C9 genotyping was carried out by polymerase chain reaction-restriction fragment length polymorphism technique. The allele frequency of CYP2C9*1, CYP2C9*2 and CYP2C9*3 was 76.1, 16.1 and 7.8%, respectively. The frequency of CYP2C9*1/*1, CYP2C9*1/*2, CYP2C9*1/*3, CYP2C9*2/*2, CYP2C9*2/*3 and CYP2C9*3/*3 genotypes was 53.9, 22.1, 11.4, 2.9, 4.3% and nil, respectively. In this study the genotypic variations of the CYP2C9 allele among the Sistani ethnic group was investigated and great differences were observed in comparison to other populations. Our findings suggest that different genotypes of CYP2C9 may influence the pharmacokinetics of some drugs. More studies on the pharmacokinetic effects of CYP2C9 genotypes may help physicians choose optimal dosage of some drugs for treatment and prevention of their side effects. Since different ethnic groups from all over the world use medications, it suggests to investigate the pharmacokinetic effects of CYP2C9 genotypes in different populations.