علی محمد اصغریان

Aim and Among cancers, Lung cancer is considered to be the most important cause of death in worldwide and as the most modern cancer treatments are ineffective with side effects, it seems essential to find impressive alternatives treatment. Nettle plant with the scientific name Urtica dioica L. Of family Urticaceae, is a flowering and Perennial plant that has high nutritional and medicinal value. There fore, the cytotoxic effect of ethanolic extract of nettle leaf (Urtica dioica L.) on A549 cell lines was evaluated.
In this study, for the first time, the cytotoxic effect of ethanolic extract of nettle leaves on A549 cell lines was evaluated. The dioecious nettle leaves in April 2015 of sehezare Tonekabon were collected and studied; Then the antioxidant activity was evaluated by DPPH0 Assay. A549 cell line in DMEM with 20% fetal bovine serum was grown. Different concentrations of ethanolic extract of two legs nettle leaves (5, 2/5, 1/25 and 0/5 mg/ml) for 24, 48 and 72 hours were applied on cancer cells A549. Next the viability of cells using MTT assay was considered.
In this study, the percent of free radical scavenging DPPH0 equals to 67/79 percent and the amount of IC50 was calculated to be 4/30 mg/ml. The results of MTT assay showed that the ethanolic extract of nettle leaves has a cytotoxic effect on the A549 cell line which is dependent to their doses. Also IC50 of the extract concentration for A549 cells was obtained to be 5 mg/ml. MTT results showed no significant differenc in 48 and 72 hours. But the difference was significant at 24 hours.
The ethanolic extract of nettle leaves which is dose-dependent could destroy the cancer cells and showed cytotoxic properties. Thus, this plant can be used as a medicinal plant against lung cancer in wider scope.

Osteoporosis is describesd as a disorder and skeletal disease that decrease bone strength and increases the risk of a bone fractures. Genetic factors have effect role in the progression of the osteoporosis. The aim of this study was to investigate the association between LRP4 gene polymorphism with osteoporosis in a population of postmenopausal women from north of Iran.
In this case-control study, 80 female patients with osteoporosis and 80 healthy females without osteoporosis with average age of 45-60 has been investigated. After DNA extraction from genome samples, polymorphism of LRP4 (rs4752947) gene have been investigated by PCR-RFLP method. Data were analyzed by SPSS software.
Our results showed no significant relationship between polymorphism of LRP4(rs4752947) gene and the risk of osteoporosis disease in two patients and control groups. Also, AT genotype and TT genotype compared with AA genotype increased the chance of disease by 1379 and 3.5, respectively. In addition, TT alleles compared with AA alleles, increased the chance of osteoporosis up to 1.605 times.
Of course, more complementary studies considering other LRP4 gene subtypes with more individuals for better findings are needed.

Inflammatory bowel disease (IBD) includes two basic categories ulcerative colitis (UC) and Crohn's disease (CD) that the etiology of which remains unclear. Tumor necrosis factor alpha (TNFα) promoter polymorphisms are a good candidate for susceptibility to IBD as there is a significant relationship between them. The main aim of this study was to assess TNFα gene polymorphisms with IBD susceptibility at positions -1031in Iranian patients.
In this case-control study, were studied 101 patients with IBD (86 ulcerative colitis, 15 Crohn's disease) and 100 healthy controls were studied. PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) was used for determining of genotyping. In following, allele frequency and genotype distribution of polymorphism T> C in TNFα gene between the case and control groups were typed.
The frequency of genotype TT, TC and CC among patients was 64.4%, 28.7% and 6.9% and in control group was 63%, 29% and 8%, respectively. Also, allele frequency T-1031 of TNFα gene in IBD patients was high, while there is no statistical significant(p>0.05).
There was no significant correlation between TNFα gene polymorphisms and susceptibility to IBD at position -1031. Our results showed that TNFα gene polymorphisms cannot be considered as a potential prognostic marker cause of IBD in Iranian population.

There are different types of herbs in Iran as they can be applied for curing and controlling bacterial infection. In this context, Urtica dioica L. has shown great effects against gram negative and positive bacteria and antioxidant activity as well. This study showed that nettles leaf essence has high antibacterial and antioxidant activities. In this experimental study, the leaves of nettle were collected from Sehezar region, Tonekabon, and then were extracted by Clevenger. Moreover, the chemical compounds of nettles leaves essence were identified by gas chromatography–mass/mass spectrometry (GC/MS/MS). Also, MIC and MBC tests were carried out against some bacteria, including Staphylococcus aureus, methicillin resistance Staphyloccus aureus, Escherchia coli, Salmonella typhimurium, Bacillus subtilis, Pseudomonas aeruginosa, and Shigella dysenteria. The DPPH producer was used for evaluating antioxidant capacity as well. The essence of nettles leaves has acceptable antibacterial results against either gram negative and positive bacteria except Bacillus subtilis. Also, the antioxidant activity and the antioxidant activity index (AAI) of the essence were 21.53 and 2.116 &mug/ml, respectively. Moreover, the main components included phytol (35.0%), nonanal (15.3%), &beta-ionone (9.0%), phtaleic acid (7.4 %) and carvacrol (3.5%). For the first time, it has been demonstrated that nettle leaves essence can be used as a new broad spectrum antibacterial agent and efficient antioxidant for controlling and remedy of infectious diseases.

Hepatitis B virus is the cause of acute and chronic hepatitis، cirrhosis and hepatocellular carcinoma. In the present، the only secure way to prevent hepatitis B is vaccination، but the vaccination is not 100% affective. Considering the importance of this، the efficacy of vaccination in high risk groups and health-care workers are at risk being involved with all types of hepatitis viruses. For reduction of hepatitis B infection among hospital staff these groups should be vaccinated against HBV besides using other standards for HBV prevention. The purpose of this study is to evaluate the anti-hepatitis B antibody among students of nurse’s aid school in Be''sat Hospital. Anti-hepatitis B antibody levels were measured using ELISA in 64 nurse’s aid students. the vaccination history of study population was unclear. 37 (57. 8%) cases were non-immune and 27 (42. 2%) cases were immune against HBV. Our results showed similarity with other studies. Several factors affected HBV immunization such as age، genetics، obesity، and smoking. This study showed that more than half of the people are non-immune. It should be noted that these people are in continuous contact with their classmates and patients so they should be evaluated for HBs antibody titer in different intervals.

Beta-thalassemia is a genetic disorder manifested by the presence of anemia in adult patients. One approach to treatment of beta-thalassemia is induction of the fetal gamma-globin gene. One of the gamma-globin repressors is a complex called DRED (Direct repeat erythroid-definitive). DRED is composed of TR2 and TR4 DNA binding subunits. The aim of this study was to set up the RNAi system to increase the expression of the gamma-globin gene. In this study, lipofectaminTM2000 was used to transfect siRNA molecules and pSV-β-Galactosidase vector was used as a reporter to monitor transfection efficiency. Real-time PCR method was used to measure TR4 knockdown and gamma-globin expression levels. Our findings showed that K562 cells were transfected by 40% using LipofectamineTM 2000. Also, the level of TR4 expression decreased by 44% after using TR4siRNA, even though the expression of gamma-globin gene was induced by 1.18 times. Despite a 44% knockdown of TR4, no increase in gamma-globin mRNA was observed. Two factors may count for this observation; first, TR4 knockdown may have been limited by our transfection efficiency. Second, simultaneous knockdown of TR2 and TR4 may lead to increased gamma-globin levels. In conclusion, although knocking down of TR4 expression occurred by using RNAi system, this can not be a solitary efficient way to induce the gamma-globin expression.

Amphibian skins possess various antibacterial compounds that are effective against some microbial pathogens and are mostly released in response to environmental stress. In fact, the skin of Rana ridibunda, a large green frog, is a rich source of antimicrobial compounds that can be developed for therapeutic use. In the present study, the skin extract of Iranian Rana ridibunda was evaluated for its antimicrobial, hemolytic and cytototoxic activities. The frog specimens were collected from Minoodasht located in Golesten province in Iran, during 2009. Subsequently, their skins were removed and the intended compounds were extracted. The crude extract was partially purified by gel filtration chromatography. The antimicrobial effects of skin extract were assessed against various microorganisms such as Escherchia coli, methicillin-resistant and -sensitive Staphyloccus aureus, vancomycin-resistant and -susceptible Enteroccus fecalis, Pseudomonas aeroginosa and Candida albicans. In addition, its minimum inhibition concentration, cytotoxic and hemolytic activities were determined. The crude extract of Rana ridibunda skin had valuable antimicrobial effects against methicillin-resistant and -susceptible S. aureus in comparison with E.coli and vancomycin-resistant and -susceptible E. fecalis. Besides, no antimicrobial activities were seen against P. aeroginosa or C. albicans. Moreover, the hemolytic and cytotoxic activities of the skin extract were minimal. The antimicrobial activity of Iranian Rana ridibunda was comparable to those isolated from other Rana species. In conclusion, the skin extract of Rana ridibunda had the potential for a new therapeutic agent against the emerging drug-resistant bacteria, particularly methicillin-resistant and -sensitive S. aureus.

Hydroxyurea (HU) is currently used for β thalassemia treatment through induction of fetal γ-globin. In this study, effects of different concentrations of hydroxyurea on induction of γ-globin gene in K562 cells, and inhibitory effect of siRNA against candidate gene which may be involved in HU mediated γ-globin induction were assessed. In this experimental study, K562 cells were treated by different concentrations of HU (0, 50, 100 and 200 μM). siRNA against candidate gene in HU mediated γ-globin gene induction was transfected to K562 cells using lipofectamine 2000. The level of γ-globin gene induction and inhibition were determined by quantitative real-time PCR. There were 1.75-, 2.45- and 2.55- fold inductions in γ- globin gene using 50, 100 and 200μM HU, respectively. There has been 79.5% down regulation on candidate gene in siRNA transfected K562 cells. This study showed that γ-globin induction in 100μM HU is similar to 200 μM HU treated K562 cells. There was also efficient inhibitory effect on candidate gene which may be involved in HU mediated γ-globin induction.

CREB1 is an important downstream protein for many signaling pathways. By designing efficient siRNAs against CREB1, it may be possible to assess the role of molecules involved in signaling pathways in different cell types. In this research the efficiency of CREB1 knockdown by two different siRNAs in K562 cells has been studied. siRNAs have been designed according to the criteria suggested by Reynolds et al. K562 cells were transfected by siRNA using Lipofectamine 2000. The efficiency of CREB knockdown has been assessed by quantitative relative Real-time PCR. Our results have shown that only one of the siRNAs has a high level of inhibitory effect on CREB1 gene expression. The expression of CREB1 by this siRNA was knocked-down by 87% in K562 cells. In this research, although two siRNAs were designed according to the Reynolds et al. criteria, only one showed an inhibitory effect. Reasons other than the aforementioned criteria may be involved in effectiveness of siRNAs.

Understanding of the mechanisms involved in γ- to β-globin switching may be important for development of treatment options for b-thalassemia. Such studies require the availability of relevant cellular model systems. One such cell type is Hu11, a mouse erythroleukemia (MEL) cell line containing the human b-locus. MEL and Hu11 cells differentiate in the presence of the chemical dimethyl sulfoxide (DMSO). Nevertheless, levels of γ-globin induction in Hu11 cells after DMSO treatment have not been determined. In the present study, we determined γ-globin levels in Hu11 cells after treatment with various DMSO concentrations. Hu11 cells were cultivated in various DMSO concentrations and the levels of γ-globin were determined by real-time PCR. Our study showed that hemoglobin in Hu11 cells treated with 1% and 2% DMSO was increased by approximately 5 and 10 folds. Moreover real-time PCR results showed that g-globin levels using the indicated DMSO levels were increased by 66 and 298 folds, respectively. Hu11 cells differentiate in the presence of DMSO, and in doing so, their γ-globin levels are increased. Therefore these cells can be used to study the mechanisms of γ-globin induction. Such studies may be beneficial for the treatment of β-thalassemia.

CREB is an important downstream protein for many signaling pathways. By having efficient siRNAs against CREB, it may be possible to assess the role of molecules involved in signaling pathways in different cell types. In this research the efficiency of CREB knockdown by 2 different siRNAs in HeLa cells has been studied. HeLa cells were transfected by siRNAs using Lipofectamine 2000. RNA extraction and cDNA synthesis have been done. The efficiency of CREB knockdown has been assessed by quantitative relative real time PCR. According to our results only one of the siRNAs has a high level of inhibitory effect on CREB gene expression in HeLa cells. The expression of CREB by this siRNA was knocked-down by 64.4% in HeLa cells. By using efficient siRNA specifically for knocking down of CREB gene, we can apply this efficient siRNA in different cell types due to study of signaling pathway which CREB may be involved.