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Systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) are multisystemic autoimmune diseases with multifactorial nature. Considering limitations of the current conventional serological tests for diagnosis of these diseases, researchers strive to find new and more valid biomarkers. The methylation level of interferon-induced protein 44-like (IFI44L) promoter was evaluated in 69 patients with SLE, 61 patients with RA, and 71 healthy subjects. Quantitative methylation of the promoter region of IFI44L gene was measured in DNA extracted from peripheral blood mononuclear cells (PBMCs) with methylation-quantification endonuclease-resistant DNA (MethyQESD) method. Our findings revealed a substantial hypomethylation of IFI44L promoter in SLE and RA patients compared with healthy volunteers (mean: 60.36%±64.54%, 47.59%±30.34%, and 89.17%±76.96%, respectively; PSLE= 0.018, PRAIFI44L promoter methylation had a sensitivity of 84/06% and 93/65% and specificity of 32/39% and 29/58, respectively. The promoter methylation level was not meaningfully different between SLE and RA patients (P > 0.05). Moreover, our analysis revealed that the methylation level of IFI44L promoter was not statistically significantly different between SLE disease activity and renal involvements (P > 0.05). While RA patients with a higher concentration of CRP had a lower DNA methylation level (P = 0.012). The methylation level of IFI44L promoter was lower in PBMCs of Iranian patients with SLE and RA than that control group. Furthermore, DNA methylation level of IFI44L promoter had a negative correlation with RA disease activity. However, there was not a significant association between clinical characteristics of SLE.

مقدمه: سرطان‌های بدخیم کولورکتال، چه در صورت تکی و چه در صورت خانوادگی، اگر با افزایش میزان ناپایداری در میکروستلایت همراه باشند، پیش‌آگهی بهتری دارند، راهکار متفاوتی برای جراحی در آن‌ها اتخاذ می‌شود و پاسخ متفاوتی نسبت به داروی شیمی‌درمانی 5FU (Fluorouracil) خواهند داد. بنا بر این، امروزه برای بیماران مبتلا به سرطان کولورکتال ارثی و غیر ارثی، آزمایش سنجش وضعیت ناپایداری میکروستلایت انجام می‌شود. هدف این مطالعه، سنجش کارایی نشانگر تک نوکلئوتیدی MT1XT20 در جمعیت بیماران مبتلا به سرطان کولون ارثی غیر پولیپی (HNPCC یا Hereditary nonpolyposis colorectal cancer) کشور ایران بود. علاوه بر نشانگر پیش‌گفته، از دو روش بررسی وضعیت میکروستلایت با پنل 5 نشانگری و استاندارد Promega و نیز آزمایش ایمنوهیستوشیمیایی استفاده شد. روش‌ها: در این مطالعه، نشانگر تک نوکلئوتیدی MT1XT20 و کیت استاندارد Promega از طریق روش PCR (Polymerase chain reaction) بر روی 20 خانواده‌ی مبتلا به HNPCC ایرانی مورد سنجش قرار گرفت و آزمایش ایمنوهیستوشیمیایی نیز برای بررسی وضعیت 4 پروتئین مهم دخیل در مسیر ترمیم ناجور جفت شدگی مورد استفاده قرار گرفت. یافته‌ها: در روش سنجش ناپایداری میکروستلایت از طریق پنل Promega و نشانگر تک نوکلئوتیدی MT1XT20 به ترتیب 8 (40 درصد) و 5 نمونه (25 درصد) از 20 نمونه وضعیت ناپایداری نشان دادند. در روش ایمنوهیستوشیمیایی نیز، 7 نمونه از 20 نمونه (35 درصد) وضعیت کمبود پروتئین مسیر ترمیم ناجور جفت شدگی را نشان دادند. در میان نشانگرهای مورد استفاده در کیت Promega، نشانگر BAT26 ناپایدارترین نشانگر بود و در هر 8 نمونه، Microsatellite instability-high (MSI-H) تشخیص داده شده توسط کیت Promega ناپایداری نشان داد. نشانگرهای NR21 و NR24 در 7 نمونه (5/87 درصد) ناپایداری نشان دادند و نشانگرهای BAT25 و MONO27 به ترتیب با نشان دادن ناپایداری در 6 (0/75 درصد) و 5 نمونه (5/62 درصد)، پایدارترین نشانگرها بودند. نتیجه‌گیری: MT1XT20 در جمعیت ایتالیایی، به عنوان نشانگر تک نوکلئوتیدی مورد اعتماد ارزیابی شده است، اما این مسأله در رابطه با جمعیت ایرانی صدق نمی‌کند. در عوض، BAT26 با نشان دادن ناپایداری در تمام نمونه‌های MSI-H ارزیابی شده توسط کیت Promega، می‌تواند به عنوان نشانگری موثق و قابل مقایسه با پنل 5 نشانگری Promega در جمعیت ایرانی در نظر گرفته شود.

Background: Integral membrane proteins involve in a variety of valuable cellular processes including signal transduction, transport and cell-to-cell communication. Thus, due to their importance, are targeted to treatment of diseases or disorders. To achieve this goal, it is necessary to obtain adequate membrane proteins. A variety of expression systems have been used to overexpress these proteins but mammalian cells can produce membrane proteins structurally close to natural form of them and are favourable for this purpose. TOSO/FAIM3 (Fas Apoptosis Inhibitory Molecule), highly conserved membrane protein, playing important role in cell survival, immune surveillance, and homeostasis and in this study was overexpressed on HEK-293T membrane. Methods: Eukaryotic expression vector pEZ-M67-TOSO was transformed into Escherichia coli top 10F’ strain to multiply. Then, the plasmid was extracted and digested with restriction enzyme Eco31I to make a linear plasmid. Subsequently, HEK-293T cells were transfected with linear plasmid and treated with hygromycin to select TOSO expressing stable cells. After three weeks, resistant colons were expanded, Chromosomal DNA and total RNA were extracted and the presence of TOSO-cDNA in HEK-293T genome and the expression level of TOSO were evaluated using polymerase chain reaction (PCR) and Real-time PCR methods, respectively. Findings: The integration of TOSO expressing cassette in HEK-293T genome was confirmed. HEK-293 cells expressed 1700 mRNA TOSO molecule/cell which was in the range of protein with abundant expression. Conclusion: Mammalian cells represent membrane proteins in desirable form versus other expression systems. For this reason, we used HEK293T cells to overexpress transmembrane TOSO protein. In this investigation, we successfully constructed the HEK293 cell line with stable TOSO overexpression, which will facilitate further researches to characterize biophysical and biochemical structure and function of TOSO and pharmaceutical biological researches about it.

BACKGROUND: The objective of this clinical trial was to assess the effects of probiotic soy milk and soy milk on anthropometric measures and blood pressure (BP) in type 2 diabetic (T2D) patients. METHODS: A total of 40 patients with T2D, 35-68 years old, were assigned to two groups in this randomized, double-blind, controlled clinical trial. The patients in the intervention group consumed 200 ml/day of probiotic soy milk containing Lactobacillus planetarium A7 and those in control group consumed 200 ml/day of soy milk for 8 weeks. Anthropometric and BP measurements were performed according to standard protocols. For detecting within-group differences paired-sample t-tests was used and analysis of covariance was used for determining any differences between two groups. (The trial has been registered in the Iranian Registry of Clinical Trials, identifier: IRCT: IRCT201405265062N8). RESULTS: In this study, we failed to find any significant changes between probiotic soy milk and soy milk in term of body mass index (26.65 ± 0.68 vs. 26.33 ± 0.74, P = 0.300) and waist to hip ratio (1.49 ± 0.08 vs. 1.54 ± 0.1, P = 0.170). Although soy milk did not have any effect on BP, probiotic soymilk significantly decreased systolic (14.7 ± 0.48 vs. 13.05 ± 0.16, P = 0.001) and diastolic BP (10 ± 0.7 vs. 9.1 ± 1, P = 0.031). CONCLUSION: In our study, probiotic soy milk in comparing with soy milk did not have any beneficial effects on anthropometric measures in these patients. We need more clinical trial for confirming the effect of probiotic foods on anthropometric measure in diabetic patients. However, probiotic soy milk decreased systolic and diastolic BP significantly. &nbsp

Background: Cystinuria, one of the first diagnosed inborn errors of metabolism, recognized by hyperexcretion of cystine, lysine, ornithine and arginine into the urine. So far, two genes associated with cystinuria have been identified: SLC3A1 (2p16.3) that encodes the heavy subunit rBAT of the renal b0,+ transporter and SLC7A9 (19q13.1), which encodes the light subunit b0,+AT. Patients with type A cystinuria have two SLC3A1 mutations, whereas patients with type B have two SLC7A9 mutations and finally patients with type AB have one mutation in each gene. Considering the population-specific distribution of mutations in disease, limited studies on the genetic bases of the cystinuria have been done in Middle East. This research presents the results of mutation analysis on patients with cystinuria in Iran. Methods: Thirty unrelated patients with cystinuria operated to remove kidney stones were screened by urologist .The patients were analyzed for mutation using amplification refractory mutation system (ARMS) and polymerase chain reaction-Restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) methods. Findings: We found some variations including missense mutations, polymorphism and intron variant, but the most frequent mutations, M467T and also T216M and R333W, were not detected in our patients. Conclusion: Our research may confirm the ethnic distribution of mutations in cystinuria and this study can expand our concept of the genetic basis of cystinuria in Iranian patients.

Milk thistle [Silybum marianum (L.) Gaertn.] is an important medicinal-industrial plant. The medicinal compounds of milk thistle are derived from its seeds. The plant is commercially cultivated for the production of silymarin. Cultivation offers the opportunity to optimise achene yield and silymarin content. Furthermore, efforts should be made to develop new cultivars. The main objective of milk thistle breeding is to develop high yielding cultivars with elevated silymarin content. There are few developed cultivars of milk thistle (e.g. Argintiu, Budakalaszi, Szibilla, Khoreslo, Babak Castle, Mirel, Silma and Silyb). Different genotypes of milk thistle have variable amount of silymarin e.g. a ‘Royston’ genotype is rich in silymarin (6-10%). Further progress can be made by using the genetic diversity available in abundance in different genotypes. The main concern of the breeder should be to increase the yield and silymarin contents of the seeds. Asynchronous flowering and seed shedding are also major problems in milk thistle cultivation. At the time of harvest, the plants have flower heads at all stages of development resulting non-uniform maturation of seeds. Therefore, a breeding effort in milk thistle should be planned to obtain plants with simultaneous flowering and reduced crop losses. The developed cultivar Argintiu is characterised by simultaneous seed maturation in flower heads. The wild populations, as valuable gene pools, could be exploited for the improvement of milk thistle crop.

Background: Micro ribonucleic acids (miRNA) are a type of ribonucleic acids with 20-24 nucleotides. They seem to have an important regulating role in cells. MiR-211 is an miRNA whose expression decreases in cancer cells, especially human melanoma cells. It is of high importance in suppression of tumor growth and invasion. Up to now, no study has evaluated miR-211 expression in murine melanoma cells for in vivo and in vitro conditions. We evaluated and compared the expression of miR-211 in murine melanoma cell line and melanoma tumors using real-time polymerase chain reaction (PCR). Methods: Melanoma B16F10 cell line was cultured. Subcutaneous melanoma tumors were induced by injecting this line to C57 mice. Melanoma cell line and melanoma tumors were lysed and miRNAs were extracted. After complementary deoxyribonucleic acid (cDNA) synthesis, real-time PCR of samples was performed using miR-211 primers. Data was statistically analyzed by Student’s t-test. Findings: Average expression of miR-211 in melanoma cell line was significantly higher than melanoma tumors. Conclusion: Due to the lower expression of miR-211 in murine melanoma tumors in comparison to murine melanoma cells, miRNA may be involved in invasive properties of melanoma cells in the mouse model of melanoma. Keywords: MiRNA, Murine melanoma cells, Real time polymerase chain reaction

Background: Breast cancer is the most common form of hereditary cancer worldwide and is an important cause of morbidity and mortality. Approximately 5-10% of breast and ovarian cancers are due to the highly penetrating germline mutations in cancer predisposing genes. Two genes, BRCA1 and BRCA2, account for at least half of these cases. The demand for BRCA1 and BRCA2 mutation screening is rapidly increasing as their identification will affect the medical management of people at increased risk for the disease. Therefore, the aim of this study was to investigate BRCA1/2 mutations in 100 high risk Iranian families.Methods: One hundred families who met the minimal risk factors for breast/ovarian cancer were screened among the families referred to Kawsar Human Genetics Research Center for the diseases in 2009-2011. The entire coding sequences and each intron/exon boundaries of BRCA1/2 genes were screened for by direct sequencing and MLPA in both patients and the controls.Results: In the present study, we could detect the following novel mutations: p.Gly1140Ser, p.Ile26Val, p.Leu1418X, p.Glu23Gln, p.Leu3X, p.Asn1403His, p.Asn1403Asp, p.Lys581X, p.Pro938Arg, p.Thr77Arg, p.Leu6Val, p.Arg7Cys, p.Leu15Ile, p.Ser177Thr, IVS7+83(-TT), IVS8 -70(-CATT), IVS2+9(G>C), IVS1-20(G>A), IVS1-8(A>G), p.Met1Ile, IVS2+24(A>G), IVS5-8 (A>G), IVS2(35-39)TTcctatGAT, IVS13+9 G>C in BRCA1 and p.Glu1391Gly, p. Val1852Ile, IVS6-70(T>G), 1994-1995 (InsA) in BRCA2.Conclusion: Ten mutations seemed to be pathogenic and the disease-causing mutations were seen in 16% of the families. In addition, from the total number of substitutions and reassortments (42), 80% related to BRCA1 and 20% to mutations in BRCA2 genes.

Background: Severe mitral stenosis is occasionally associated with significant tricuspid regurgitation (TR) and this association has an adverse impact on morbidity and mortality in patients undergoing mitral valve intervention. However, the effect of successful mitral balloon valvotomy (MBV) on significant TR is not fully elucidated. The aim of this study was to investigate the course of TR after MBV in patients with severe mitral stenosis with TR.Methods: The present study was performed in Tabriz Madani heart center from March 2007 to February 2008. Among 110 patients with mitral stenosis who were candidates of MBV, 68 cases with more than mild TR were selected and the fate of TR after MBV and its predictors were evaluated. Results: Among 68 patients who were enrolled in this study, 58 individuals (85.3%) were female with mean age of 36.85± 14.32 years. Before intervention, 48 patients (70.6%) had severe TR and 20 (29.4%) cases had moderate TR. After intervention, 14 patients suffered from mild TR, 22 from moderate TR and 32 patients from severe TR (P

Background Helicobacter pylori (H. pylori) is a gram negative curved bacilli colonized in the human stomach. It causes duodenal ulcer, gastritis and is associated with adenocarinoma. There might be a possible relationship between cagA, vacA and ice a genes and clinical outcomes. The aim of this study was to identify the frequency of vacA genotypes of H. pylori isolated from the upper gastrointestinal. Methods One hundred H. pylori strains were isolated from patients with different gastrointestinal disease who admitted to Isfahan Al-zahra hospital. Vac A alleles were typed using PCR with specific primers. Findings There were four Vac A mosaicsms, including 28 for s1a/m1 (28%), 23 for s1b/m1 (23%), 26 for s1a/m2 (26%) and 23 for s1b/m2 (23%), s2 form was not found. Conclusion The results showed there is no significant relationship between different genotypes of vacA and the related diseases. Key words: Helicobacter pylori, vac A, Adenocarinoma, Duodenal ulcer, Normal /Gastritis, PCR

Soluble forms of nonclassical human leukocyte antigen (HLA)-G have recently been suggested as immunomodulatory factors in multiple sclerosis (MS). HLA-G inhibits the effecter function of T cells and natural killer (NK) cells. Also regulatory T cells (Treg) are considered as pivotal players in MS pathogenesis. Thus, we aimed to evaluate the presence of HLA-G molecules and Treg cells in Relapsing-Remitting Multiple Sclerosis (RRMS) patients and compare it to healthy controls.Patients with RRMS (n=205, mean age=31.32±8.53) and healthy subjects (n=205, mean age=32.2±7.48) were studied. The patients subgrouped to untreated and treated with Interferon beta. Then sHLA-G levels (sHLA-G1 and sHLA-G5) were measured using ELISA method. Treg (CD4+CD25+ Foxp3+) cells in patients who had sHLA-G>10 U/ml were characterized by using flow cytometry.Our data showed that there was no significant differences between RRMS patients and healthy controls in sHLA-G concentration (p>0.05). Treg cell frequencies were higher in the patients who had sHLA-G >10 U/ml compared to healthy subjects (p

Wound infections are a common cause of staphylococcal infections. An ability of S.aureus is to adhere and form biofilm on host surfaces. Biofilm is an exopolysaccharide, a slime matrix around multiple layers of cells and is mediated by expression of the icaADBC operon. The present study evaluated the biofilm forming capacity and the presence of icaAD gene among S.aureus isolated from wound infections. Slime production assay was performed by cultivation on Congo Red Agar plate. In addition, Quantitative biofilm formation determined by microtiter plate assay PCR method used for detection of icaAD gene. Fifty strains were identified, 54% of the isolates produced black colonies on CRA plate, 52% were positive biofilm forming, and all strains carried the icaAD gene. Regarding the ability of S.aureus to form biofilms helps the bacterium to survive hostile environments within the host, suggests that biofilm production is a risk factor for infection. It is important in rapid diagnosis and treatment biofilm forming strains, because biofilm formation may lead to increased antimicrobial resistance and create a significant impediment to wound healing.

The objective of the present study was to evaluate the hormonal responses of buffaloes to GnRH injections given at different moments of the estrous cycle. The estrous cycles of 15 buffaloes were synchronized with 2 im injections of prostaglandin F2α given 11 days apart. The buffalos were randomly assigned to 1 of 3 groups. Buffaloes in the control group received no treatment, whereas GnRH6 buffaloes received a GnRH injection between days 5 and 7 and GnRH16 buffaloes received a GnRH injection between days 15 and 17 of the estrous cycle (estrus = day 0). Up to day 6, the plasma progesterone levels in the three groups were low (1 ± 0.16 ng/ml) but gradually increased until day 12 (4.62 ± 0.41 ng/ml). The amount of plasma progesterone concentrations decreased from day 15 of the estrous cycle in all groups of buffaloes. No significant differences were observed in the plasma level of progesterone and estradiol concentrations of the three groups during the estrous cycle. The peak levels of plasma estradiol were observed on day 20 and the differences were not significant (11.34 ± 1.77, 11.74 ± 1.91 and 12.72 ± 2.49 pg/ml in the control, GnRH6 and GnRH16 groups, respectively (P> 0.05). In conclusion, GnRH given at the beginning or at the end of the estrous cycle did not alter the profile of progesterone and estradiol concentration in buffaloes.

Background: Diabetes mellitus is the most prevalent metabolic disorder worldwide with many subsequent medical complications. An important complication of diabetes is microalbuminuria which is a major indication for development of diabetic nephropathy. The prevalence of diabetic nephropathy in type 2 diabetic patients has been estimated to be 47%. Since the disorder initiates with microalbuminuria, its early detection and prevention are vital. On the other hand, helicobacter pylori infection is more common in diabetic patients due to their low immune tolerance. The vacA positive genotype is reported to be effective in microalbuminuria development in type 2 diabetic patients. In this study, we investigated the relation between H. pylori vacA and microalbuminuria in type 2 diabetic patients. Methods: This study included 88 type 2 diabetic patients among whom 60 had microalbuminuria. Stool samples were collected from all patients and DNA was extracted using QiaAmp Stool DNA Extraction Minikit. In order to detect H. pylori infection, a nested polymerase chain reaction (PCR) protocol was developed and all H. pylori positive samples were genotyped for cagA positivity. Findings: Overall, 16 patients (18.2%) were detected to be H. pylori positive out of whom 12 had microalbuminuria (75%). VacA positivity was detected in one patient without microalbuminuria. Conclusion: We could not establish a relationship between vacA positivity and microalbuminuria in the studied type 2 diabetic patients. Keywords: Helicobacter pylori, VacA, Polymerase chain reaction, Type 2 diabetes, Microalbuminuria