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Background & objectives: KAI1 is a tumor suppressor gene and inhibitor of metastasis in a wide range of malignancies. While it is ubiquitously expressed in normal tissues, KAI1 expression subjects to the down regulation in tumors. The present research aims semi-quantitative evaluation of KAI1 mRNA expression in Iranian patients with colorectal cancer (CRC) and correlation between expression levels of KAI1 and stage oftumorigenesis, especially metastasis and invasion of CRC as well as pathologic factors of patients.

Methods: RT-PCR was done by specific primers for KAI1 and β-actin genes on the 80 tumor tissues and 14 normal tissues as fresh samples which obtained from 80 unrelated patients referred to Imam Khomeini Hospital.  

Results: According the results, 51.2% and 48.8% of the sample were on and off for KAI1 gene expression, respectively. As a detail, 97.3% of samples in the stage 3 and 4 and 94.5% of metastatic phases samples showed no expression of this gene. Statistical analysis showed that there is a significant difference of the KAI1 expression between four groups of samples; normal, stage 1, 2 and 3 (p<0.05). Also a significant difference was observed between semi-quantitative KAI1 expression and degree of spread to regional lymph nodes (p=0.02) as well as semi-quantitative KAI1 expression and metastasis (p=0.000001).

Conclusion: A significant difference between semi-quantitative expression of KAI1 and degree of spread to regional lymph nodes (p=0.02) and metastasis (p=0.000001) was observed.

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Background & objectives: Epithelial ovarian carcinoma seems to be one of the most lethal cancer types among all gynecological malignancies. The conventional course of therapy includes chemotherapy. Actually most cancers respond to chemotherapy but in the long run drug resistance and side effects cause treatment failure. In addition, milk thistle (silibinin), a plant that has been used from ancient time because of its good effects on different organs, determined to have powerful antioxidant activity.  The aim of this study was to examine the effect of silibinin on SKOV-3 cancer cell line after 48 hours of treatment and P21 gene expression which involves in cell cycle progression.

Methods: The human epithelial ovarian cancer cell line SKOV-3 was cultured as monolayer in 25 cm² flask in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS). Then the numbers of live cells were calculated using hemocytometer method and the cells were seeded in 96-well flat-bottomed culture plates and treated with different concentration of Silibinin. MTT assay was carried out to determine cell viability. To study P21 gene expression, RNA extraction and cDNA synthesis were carried out and real-time PCR was done.

Results: Cell growth was inhibited considerably by Silibinin treated groups compared with control after 48 hours. P21 gene expression was increased as well.

Conclusions: According to the results, Silibinin can be used as an effective drug in cancer treatment. More studies on animal models are also suggested.

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Background & Objectives: hcpD gene in Helicobacter pylori is a member of cysteine-rich proteins family which triggers the host's immune system and antibody production. H. pylori is found in human's stomach and causes digestive diseases such as duodenal ulcer, chronic gastritis and stomach cancer. The objectives of this study were to isolate, amplify and clone H. pylori's hcpD gene in pcDNA3.1 (-) vector and to study its expression in eukaryotic system.

Methods: H. pylori genomic DNA was isolated by extraction kit. The hcpD gene was amplified using PCR reaction and then purified from gel, followed by pTZ cloning. Subcloning of hcpD was performed in pcDNA3.1 (-) eukaryotic expression vector. The accuracy of cloning steps was investigated through PCR, enzymatic digestion by BamHI and EcoRV enzymes, and sequencing, respectively. Transfer of expression construct into CHO cells was done by electroporation. The gene expression in these cells was analyzed using RT-PCR and SDS-PAGE.

Results: PCR results showed amplification of a 933bp segment related to hcpD gene. Successful cloning of the gene in pTZ vector and construction of pTZ-hcpD recombinant vector were achieved. Enzymatic digestion and sequencing confirmed the correctness of subcloning and creation of pcDNA3.1 (-)-hcpD construct. hcpD was expressed in eukaryotic system, and its protein product was observed on SDS-PAGE gel.

Conclusion: pTZ-hcpD construct can be used as a source of H. pylori's hcpD gene for future research, like production of recombinant protein and vaccine in different systems. Furthermore, successful expression of the gene using pcDNA3.1 (-)-hcpD in CHO animal cells shows the potential of vector as a gene vaccine against H. pylori.

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Background & objectives: Irisin is a novel myokine that encoded by FNDC5 gene and effects on obesity, metabolism and glucose homeostasis through browning of white adipose tissue and thermogenesis. The main purpose of this study was to investigate the changes of FNDC5 gene expression and Irisin protein level of visceral fat tissue after eight weeks of resistance training in type 2 diabetic rats.
Methods: Eighteen male Wistar rats (8 week old) were used for this study. Diabetes was induced using nicotinamide and streptozotocin . Five days after inducing diabetes, rats with fasting blood glucose levels between 127-600 mg / dl were selected as diabetic subjects. Rats were homogenized according to the body weight and assigned into two groups including control-diabetes (n=9) and resistance training-diabetes (n=9). Training group exercised resistance training for eight weeks (5 days a week). The resistance training protocol consisted of climbing   a one-meter- high ladder, with a weight attached to a tail sleeve. Quantitative Real time RT-PCR and ELISA Kit were used for assessment of expression level of FNDC5 gene and Irisin protein, respectively. Data were analyzed using independent t- test at p≤0.05.
Results: Resistance training significantly increased the expression level of FNDC5 gene and Irisin protein in visceral adipose tissue in type 2 diabetic rats.
Conclusion: It seems that FNDC5 gene and Irisin protein have an important role in metabolic diseases and can be affected by resistance training. Perhaps the changes in the levels of these metabolic indicators is a potential new target for the treatment of metabolic disorders, such as T2DM (type 2 diabetes).

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Background & objectives: The increase of Bnip3 and Parkin plays an important role in maintaining mitochondrial function and inhibiting ROS. A correlation was observed between the mitochondrial respiratory capacity and the subjects' mRNA levels of Bnip3 and Parkin proteins as a result of exercise, which shows the significant role of mitochondrial dynamics on the improvement of respiratory capacity. This study aimed to investigate the effect of eight weeks of high-intensity interval training and curcumin supplementation on Bnip3 and parkin mitochondrial gene expression levels in cardiomyocytes of male heart attack model rats.
Methods: In this experimental study, 32 male Wistar rats were subjected to myocardial infarction using  intraperitoneal injection of isoproterenol (100 mg/kg for two consecutive days) and, after confirmation of infarction (troponin measurement), randomly divided into four groups; Control, high-intensity interval training, curcumin (pure curcumin 15mg and dimethyl sulfoxide with a concentration of 10% per kg of body weight five days a week by gavage) and combination group (supplement+exercise).The exercise and combination groups were subjected to high-intensity interval training (10 bouts of four-minute activity with an intensity of 85-90% VO2max) for eight weeks. Bnip3 and Parkin gene expression levels were obtained using the Real-time PCR method. The data were analyzed using the Kolmogorov-Smirnov test and one-way analysis of variance.
Results: The results showed that there is a significant difference between the mean of the groups in Bnip3 and parkin gene expression levels. The results of Tukey's test showed that the expression level of Bnip3 and Parkin genes was higher in the intense interval training and exercise-supplement groups than in the control and supplement groups. There was no significant difference between the supplement and control groups; in fact the use of curcumin without training did not affect Bnip3 and PARKIN gene expression levels compared to the control group.
Conclusion: This study shows that intermittent exercise and curcumin consumption have a protective effect on mitochondrial quality during infarction.