فهرست مطالب nooshin shabab

Resveratrol (RezV) which is found in several plants including grapes and types of berries has a vital role in inducing apoptosis and suppressing cell proliferation. Although the role of Bcl-2 in the apoptosis has been known in several pathways, the role and mechanism of miR-21 in the regulation of apoptosis in colorectal cancer (CRC) cells are unclear.

The main aim of this study was to evaluate the effects of RezV on the expression level of miR21, Bax, and Bcl2 in colorectal tumor cells.

In this study, the effect of RezV on the viability of CRC cells was evaluated by MTT assay. Then, the expression level of miR-21 was evaluated by real-time polymerase chain reaction (PCR) method. For evaluating HCT-116 cells apoptosis, the expression level of Bax and Bcl2 that are involved in the apoptosis pathway was investigated by the same method.

RezV inhibits the viability of HCT-116 cells. MiR-21 gene expression was decreased after 24 hours of treatment with RezV. The reduction of miR-21 expression leads to the reduction of the Bcl2 gene expression level. Moreover, increasing the Bax/Bcl2 ratio enhances HCT-116 cells apoptosis.

In summary, RezV might be used as a co-treatment agent for CRC. On the other hand, conducting the in vivo study to evaluate the effects of RezV was critical.

Usually, chemoradiotherapy can be used for the treatment of locally advanced colorectal cancer (CRC) before surgery. On the other hand, some studies have shown that fractional radiation of tumor cells leads to chemoresistance. The aim of this study was to evaluate the chemoresistance of radioresistant sub-line (RR sub-line). This study was done in Hamadan University of Medical Sciences in 2017-2018. MTT assay and sub-G1 fraction analysis by flow cytometry were used to evaluate cross-resistance of RR sub-line to gefitinib and regorafenib. Real-time PCR was used to investigate the role of four miRNAs and their target genes in the cross-resistance of RR sub-line. The t test and repeated measures test were used for the assessment of statistical significance between groups. The IC50 of gefitinib and regorafenib for RR sub-line were significantly higher than those of the parental cell line. On the other hand, the resistance index of RR sub-line for gefitinib and regorafenib were 1.92 and 1.44, respectively. The sub-G1 fraction of RR sub-line following treatment with gefitinib and regorafenib was significantly lower than that of the parental cell line (P=0.012 and P=0.038, respectively). The expression of miR-9, Let-7e, and Let-7b in RRsub-line was significantly lower than that of the parental cell line. However, NRAS, IGF1R, NFKB1, and CCND1 found to be upregulated in RR sub-line in comparison with the parental cell line. We can conclude that the acquired RR sub-line was cross-resistance to gefitinib and regorafenib. Furthermore, miR-9/NFKB1, let-7b/CCND1, let-7e/NRAS, and IGF1R played essential roles in the chemoradioresistance of CRC.

Induced oxidative stress in diabetes mellitus (DM) plays a critical role in insulin resistance. Fork head-related transcription factor (FOXO) proteins are important transcriptional factors involved in oxidative stress and insulin resistance. Resveratrol (RSV) is a polyphenol with hypoglycemic and antioxidant properties. The aims of the present study were to examine the effects of RSV on FOXO gene expression, serum superoxide dismutase (SOD) activity, insulin level, and insulin resistance in type 2 diabetic (T2DM) rats. Thirty male Wistar rats were used in this study. DM was induced in rats (n=24) using streptozotocin (STZ) and nicotinamide; then, they were divided into 4 groups of 6 rats each. Six untreated normal rats were used as normal control group; diabetic rats in groups 2 to 5 were treated with 0, 1, 5 and 10 mg /kg body weight of RSV, respectively for 30 days. At the end of the experimental period, the rats were sacrificed, their sera were separated, and adipose tissues were obtained and stored at −80 °C. Serum glucose and SOD activity levels were determined biochemically, and serum insulin level was determined by ELISA method. Gere expression in FOXO1 and FOXO3a in adipose tissue was evaluated using real‐time PCR. Results indicated that RSV significantly reduced blood glucose level, increased insulin level and improved insulin sensitivity. RSV resulted in an increased serum SOD activity and caused decreased FOXO1 and FOXO3a expression in adipose tissue of rats with T2DM. Therefore, by attenuation of FOXO expression in adipose tissue of T2DM rats, RSV showed a hypoglycemic potential and antioxidant properties, and consequently ameliorated insulin resistance.

Bladder cancer is the second most common cancer in the genitourinary tract, showing often recurrence and progresse into invasive states. Epigenetic changes, such as microRNA alteration are involved in bladder cancer tumorigenesis through a variety of signaling pathways. The epigenetic state depends on geographic and lifestyle conditions. The aim of this study was to investigate the expression level of microRNA-99a and microRNA-205 in bladder cancer in Iranian populations and to determine the relationship between their expressions with clinicophatological features. 36 patients with bladder cancer were included in the study. The control group was the healthy adjacent tissue of the same patients. Total RNA was extracted from approximately 50 mg tissue using TRIzol reagent. cDNA was synthesized and Real-Time PCR was carried out using specific primers. The Unisp6 rRNA was used as a reference gene. A significant decrease was found in the expression level of miR-99a in tumor samples, compared to healthy adjacent tissues (P

Atherosclerosis has serious role in coronary arteries disease,so it is important to establish effective strategies for prevention or even treatment of atherosclerosis. Adiponectin, as one of the most abundant adipokines, has insulin sensitivity, anti-inflammatory and anti-atherogenic properties. Disturbed adiponectin actions through its receptor, (AdipoR1 and AdipoR2) may be involved in atherosclerosis development. Some adiponectin effects are mediated by AMPK and PPAR-α signaling. AdipoRon is an orally active synthetic molecule which can bind to AdipoR1, Adipo R2 and activate them. AdipoRon can activate AdipoR1-AMPK- PGC-1α pathway and AdipoR2-PPAR-α pathway. Some studies indicated insulin sensitivity, anti-apoptotic and anti-oxidative effect of AdipoRon. We hypothesize that AdipoRon has anti atherosclerotic effect and may suppress atherosclerosis processes. With confirmation the benefit role of AdipoRon on atherosclerosis, it may be used in patients at risk of atherosclerotic development.

Most cancer studies focus on exploring non-invasive biomarkers for cancer detection. In the present study, we sought to investigate the expression level of microRNA-21 (miR-21), as a potential diagnostic marker, in serum and stool samples from 40 patients with colorectal cancer (CRC) and 40 healthy controls.
Quantitative real-time RT-PCR was applied to determine the relative expression level of miR-21 in serum and stool. At the same time, the sensitivity and specificity of this marker was evaluated by receiver operating characteristic (ROC) curve analysis.
miR-21 expression levels of serum and stool were up-regulated 12.1 (P The results of this study indicated that miR-21 expression levels in serum and stool can be considered as a potential diagnostic biomarker for the diagnosis of CRC patients. However, more studies are required to confirm the validity of miR-21 as a valuable non-invasive diagnostic tool for CRC.

Bipolar disorder is a biological brain disorder which is associated with debilitating fluctuation in mood and adverse effects on patients, their families and society. The importance of genetics and its role in bipolar disorder is a controversial issue to discuss. Evidence indicates a relation between the risk of bipolar disorder and specific genes. Amongst the genes whose role has been established in bipolar disorder, the most notable gene is BDNF (Brain-derived neurotrophic factor)..
The study is based on a case-control methodology. During 18 months, the blood samples of patients diagnosed with bipolar mood disorder who were admitted to Farshchian hospital of Hamadan from March 2011 to September 2012 and for the control group, the blood samples of patients admitted to other parts of Farshchian hospital except psychiatric ward were taken and DNA extraction from white blood cells was performed. In general, 84 patients and 85 controls were examined in this study and an expert in vials containing EDTA anticoagulant collected 4ml of blood samples. These samples were sent to the molecular biology lab of Hamadan University of Medical Science to determine their genetic polymorphisms. Genomic DNA was extracted from peripheral blood cells using the real extraction DNA kit (DNP Tm kit, Cat# DN8115C, CinnaGen co., Iran). The allele specific polymerase chain reaction technique was used to determine the frequencies of listed genotype. Considering the different variations for each gene, primers design was carried out using the Allele ID software (Allele ID 6, premier Bio soft Int, USA). For this purpose, 401 nucleotide sequences of targeted gene polymorphisms was chosen as the control sequence and desired primers for this sequence was designed and ordered (Takapouzist Co., Iran). Finally, using the mentioned method the sequences were amplified and examined on 2% agarose gel during electrophoresis. The young mania rating scale (YMRS) was used to evaluate manic symptoms. A written consent was obtained from each individual patient during the study. In addition, all patients in this study were anonymous and ethical considerations were taken into account. Statistical data analysis was performed using SPSS Software and Chi-square test was used to analyze their significance..
The results of this study, which was conducted on 84 patients in the case group and 85 patients in the control group indicated that the frequencies of evaluated alleles in the case and control groups for AA genotype were 4 and 4, for GA genotype were 23 and 28, and for GG genotype were 53 and 53, respectively..
According to the obtained data, there is no significant relationship between genetic and bipolar disorder. Some studies in this field have also confirmed this issue..

Colorectal cancer remains one of the major cancer- related deaths despite progress in the treatment during past decades. Detection of disease at earlier stages reduces its mortality. The aim of current study was to investigate expression of Cytokeratin 19 (CK19), Cytokeratin 20 (CK20) and Guanylyl Cyclase C (GCC) mRNA in peripheral blood of non- metastatic colorectal cancer patients which may result into introducing of an early detection test. 25 patients with colorectal cancer and 25 healthy controls were recruited. Blood was obtained from all individuals. Expression of CK19 and CK20 and GCC mRNA and 18SrRNA (as reference gene) were determined based on real- time RT-PCR on total RNA from blood. CK19, CK20 and GCC expression had been detected in 68%, 76% & 52% of patient group, respectively, which was higher than healthy group, with 8%, 32% and 0% expression, respectively (p

Early detection is a key to survival for gastric cancer. Molecular markers such as miRNA (microRNA) can have great importance in the early diagnosis of gastric cancer. Expression of miR-21 and miR-221 are deregulated in many types of human cancers. This study aimed to investigate the differences in miRNA expression patterns within the Iranian population. Total RNA was extracted from gastric cancer tissues and adjacent non-cancerous tissues from 32 patients. Expression levels of miR-21 and miR-221 were detected by Real time RT-PCR using a specific primer, with 5s rRNA as the internal reference gene. Our data showed that the expression levels of miR-21 and miR-221 in gastric cancer samples were significantly higher than in paired non-cancerous samples (P < 0.05). The receiver operating characteristic (ROC) analyses yielded the area under the curve (AUC) values of 80.30 for miR-21 and 93.30 for miR-221, and combined ROC analysis revealed the highest AUC value of 96.90 in discriminating GC patients from healthy controls. It seems that miR-21 and miR-221 expression pattern in Iranian patients with gastric cancer are similar to any other population. Considering the increased expression level of two miRNAs in cancerous tissue compared to normal tissue as well as the area under ROC curve, miR-21 and miR-221 can be used for early detection of gastric cancer.

The overall goal of this study was to carry out a set of comparative analyses of arsR gene in plasmid R773 and bacterial chromosome from Escherichia coli BL-21(DE3). PDB and NCBI databases and Chimera, Mega4, CLC main workbench software and 3D-jigsaw and EMBL-EBI servers were applied to perform this study. By using these software and servers, multiple analyses including determination of residue composition, secondary structure and motifs, 3D structure, conserved regions, etc. were done. The results suggest that such high sensitivity to arsenic compounds in ars-containing plasmid R773 may be related to ArsR protein characteristics such as amino acids composition, secondary and tertiary structure, hy-drophobicity, level of interaction with DNA. Bioinformatics studies could be applied to describe the reason of different sensitivities to Arsenic compounds between arsR gene and ArsR protein in plasmid R773 and bacterial chromosome.

Glucose uptake by muscles and fat cells is carried out by the GLUT4 system. Isoforms of the SNAP23, syntaxin-4 and VAMP-2 play an important role in regulating GLUT-4 trafficking and fusion in adipocytes. The changes of SNARE proteins levels and thus impaired GLUT-4 displacement can be one of the etiological causes of type 2 diabetes.Due to changes in the expression of these proteins in diabetes, the aim of this study was to investigate the effect of the natural compound resveratrol with anti-diabetic properties on impaired expression of SNARE proteins in type 2 diabetes. Forty male Wistar rats were used in this study. Type 2 diabetes was induced by administering a single dose of streptozotocin and nicotinamide. The expression of SNAP-23, syntaxin-4 and VAMP-2 proteins were assessed using real-time qRT-PCR. Also, some biochemical parameters were examined, including fasting blood glucose, insulin levels and insulin resistance. The results of this study showed that, resveratrol supplementation increased blood insulin level, reduced the fasting blood glucose, and improved the insulin resistance. In addition, resveratrol supplementation increased the expression of SNAP-23, syntaxin-4 and VAMP-2 proteins that involved in GLUT-4 transport in adipose tissue of diabetic rats. Final results showed that SNARE proteins expression is significantly reduced in diabetic rats and treatment with resveratrol supplementation is associated with the increased expression of these proteins.

Bladder cancer (BC) ranks the second most common genitourinary tract malignant tumor with high mortality and 70% recurrence rate worldwide. MiRNAs expression has noticeable role in bladder tumorigenesis. The purpose of this study was to assess miR-200c, miR-30b and miR-141 in tissue samples of patients with BC and healthy adjacent tissue samples and their association with muscle invasion, grade and the size of the tumor. Transurethral resection tissue samples were collected from thirty- five newly diagnosed untreated patients with BC from 2013 to 2014. The control group consisted of adjacent normal urothelium. All samples, observed by two pathologists, were diagnosed transitional cell carcinomas (TCC) with the proportion of tumor cells greater than 80%. Total RNA including miRNAs was extracted from about 50 mg tissue samples by applying TRIzol reagent. 2(-ΔΔ CT) method was used to calculate relative quantification of miRNA expression. Two of 35 patients were females and the other 33 were males. Invasion to bladder muscle was observed in 23 (67%) cases. MiR-141, miR-200-c and miR30-b were up-regulated in 91%, 79% and 64% of malignant tissues, respectively. Down-regulation of miR-141 had a strong association with muscle invasion (P= 0.017). Significant inverse correlation between grading and miRNA-141 level was observed (P= 0.043).

To evaluate the expression of microRNAs in tissue samples from patients with bladder cancer and to compare it with healthy adjacent tissue samples as controls. Thirty five tissue samples from patients with newly diagnosed untreated bladder transitional cell carcinoma and 35 adjacent normal urothelium were collected during 2013 to 2014. TRIzol reagent was used to isolate total RNA including microRNAs. RNA concentration and purity were determined using a nanodrop spectrophotometer. Also 1% agarose gel electrophoresis was used to assess integrity of RNA. Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) method was performed using the PARSGENOME microRNA RT-PCR system. Data was analyzed by STATA 11. A couple of patients were female the remainder were male. Mean age of patients were 71.06 ± 11.43 years. The expression level of miR-30b, miR-141 and miR-200c in case group were significantly higher than that of control normal tissue samples. miR-141 had higher expression rate in malignant tissue than two other miRNAs (P <. 001). There was a more expression rate of miR-200c, miR-141 and miR-30b in bladder cancer tissues than healthy adjacent control tissues. Further studies are needed to draw final conclusion.

One of the limitations in the treatment of common diseases such as cancer chemotherapy is development of multidrug re‌sistance (MDR). Polymorphisms could alter the expression level of MDR1 gene, which plays an important role in MDR. In this research, the frequency of C3435T, C1236T, and G2677T/A polymorphisms of MDR1 gene was investigated in a large group of population from Hamadan city to provide a sample data resource. Peripheral blood (2 ml) was taken, and DNA extraction was carried out. Multiplexed mutagenically separated PCR, which was followed by polyacrylamide gel electrophoresis and silver staining, was applied to detect the mentioned polymorphisms in 935 individuals. Sequencing performed for confirmation of gel electrophoresis resulted in 10 random cases. In total, alleles and genotypes of 933 persons (776 women and 157 men) were determined. The most frequent alleles of the polymorphisms were: 3435T, C1236, and G2677. The most frequent genotypes were: 3435C/T, 1236C/T, and 2677G/A, and their concurrent presence was also found as the most frequent simultaneous genotypes. There was not any meaningful difference among the prevalence of these genotypes in groups of men and women. Our results were close to those of other studies performed in Iran and compared to the other ethnic groups, which showed more similarity to Asian peoples than Europeans. As an aspect of personalized medicine, it could be used by chemotherapists to improve the routine methods of cancer treatment.

In fertile women، glycodelin and glutathione peroxidase 3 (GPx3) genes expression rises during the luteal phase، with a peak occurring during the implantation window. The expression of these genes decreases in women with myomas. To determine whether myomectomy would reverse glycodelin and GPx3 expression، we evaluated the transcript levels of these genes in the endometrium of patients before and after myomectomy. Expression of glycodelin and GPx3 genes were examined prospectively during the midluteal phase in the endometrium obtained from infertile women with myoma (n = 12) before and three months after myomectomy. Endometrial expression of these genes was evaluated using quantitative real-time RT-PCR. Endometrial glycodelin mRNA expression levels (normalized to 18S rRNA expression) were increased significantly in endometrium of patients after myomectomy (P = 0. 02). GPx3 mRNA expression was increased insignificantly after myomectomy (P = 0. 43). The results showed that myomectomy increased endometrial glycodelin (significantly) and GPx3 (not significantly) gene expression after 3 months. Study at different times and detecting expression of these genes can reveal more details.

HOXA11 and HOXA10 are expressed in endometrium throughout the menstrual cycle and show a dramatic increase during the mid-luteal phase at the time of implantation. The expression of these genes is decreased in women with myomas. To determine whether myomectomy would reverse HOXA11 and HOXA10 expression, we evaluated the transcript levels of these genes in the endometria of patients before and after myomectomy. Expression of HOXA11 and HOXA10 were examined prospectively during the midluteal phase in endometrium obtained from infertile women (n=12) with myoma before and three months after myomectomy. Endometrial HOXA11 and HOXA10 expression were evaluated using quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Endometrial HOXA11 and HOXA10 mRNAs expression levels (normalized to 18SrRNA) were increased insignificantly in endometrium of patients after myomectomy (p=0.7 and p=0.15 respectively). The results suggest that the alteration in expression pattern of these genes could not account for some aspects of fertility after myomectomy.

Efficient screening for detection of colorectal cancer (CRC) at earlier stages reduces its mortality. The purpose of this study was to investigate expression of carcinoembryonic antigen (CEA) and human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood of CRC patients and to present strategies for early detection screen test.

Twenty seven patients in non-metastatic stage and 27 healthy individuals were studied. Expression of CEA، hTERT mRNA and 18srRNA (18s subunit of ribosomal RNA، as reference gene) were determined based on real-time RT-PCR on 3 µg of total RNA from blood in 3 separate vials (1 µg per vial).

Positive expression rate of CEA mRNA (78%) and hTERT mRNA (81%) were higher in patient group (P<0. 001). These rates were meaningfully higher than the results of individual vials containing only 1 µg of total RNA. Difference between Ct values of markers with 18srRNA (ΔCt) was higher in healthy group than patient one. Therefore، a ΔCt cut-off value was determined for distinguishing between true- and false-positive results. Concurrent expression of both markers was found in 67% of the patients، which was higher than healthy cases (11%). Combination of concurrent marker expression with cut-off point strategy increased specificity to 100%.

These results showed that concurrent evaluation of marker expression and performing the test on 3 µg of samples in 3 separate vials may increase specificity and sensitivity of real-time RT-PCR for early detection of non-metastatic CRC. However، more investigations with larger numbers of samples are needed to verify these results.

Molecular DNA markers are one of the most important tools in molecular biology labs. The size of DNA molecules is determined by comparing them with known bands of markers during gel electrophoresis. There are many different protocols to produce these kinds of molecular markers. In this study we have suggested an efficient strategy to produce molecular weight markers in industrial proportions. To achieve the desired sizes of DNA fragments, a combination of two previously known methods, restriction enzyme digestion and polymerase chain reaction (PCR), were used. The enzymatic digestion process was based on designing and constructing plasmids which equaled in size with the desired length of DNA fragments and produced the desired DNA fragment upon linearization. In the PCR method, the desired length of DNA fragments were cloned in multiple cloning sites of pTZ57R plasmid and in a PCR reaction, the new constructed plasmid was used as a template to produce the final fragment. Upon application of this strategy, 2000 and 3000 bp DNA fragments were produced by enzymatic digestion of plasmids of the same size. Moreover, 100 to 1500 bp fragments were produced during PCR using only a set of forward and reverse primers at the flanking region of pTZ57R multiple cloning site. The highest advantage of this cost-benefit approach is to produce different types of molecular weight markers by using an effective and short protocol.