Abstract

        Background and Objective: According to recent changes in diagnostic criteria for diabetes, the harmonization of results obtained from various methods and systems by considering their accuracy and precision is essential. This study aimed to evaluate the accuracy, precision and consensus of some routine laboratory glucose kits in comparison with Hexokinase reference method.

      

           Material and Methods: The participants were 38 diabetic patients with fasting blood sugar (FBS) ≥126 mg/dl, nine prediabetic patients with FBS of 100-125 mg/dl, 15 non-diabetic people with FBS of 60-100 mg/dl and 9 hypoglycemic patients with FBS of ≤60 mg/dl. Their FBS were measured by four routine laboratory glucose kits:    Glucose oxidase on BT3000 analyzer with an open system and Hexokinase reference method on a close system (COBAS INTEGRA^®400plus analyzer, Roche kit). Accuracy and precision were determined and compared with reference method.

         Results: Glucose oxidase methods showed a good agreement with the reference method, in Correlation Coefficient>0.99. based on  regression analysis, the  slope of 1.114 for Pars Azmoon, 1.105 for Bionik, 1.121 for Elitech and 1.087 for Human were reported (P<0.05). Error of the mean for ParsAzmoon was 12.79, for Bionik 10.86, for Elitech 12.58 and for Human were 8.46. Coefficient of Variation   showed more imprecision for Bionik and Human kits.

       Conclusion: given the same almost standard errors, standard devisions and regression analysis, the precision in four methods is the same but in comparison with Hexokinase, reference method has not the accuracy.

          Keywords: Blood Glucose, Glucose Oxidase, Hexokinase, Methods, Consensus

Abstract

      Background and Objectives: Diabetes induced oxidative stress plays an important role in pathological damage to the heart and liver by increased production of extracellular matrix. It is thought that the use of medicinal plants, particularly Portulaca oleracea. L and regular exercise are effective. The aim of this study was to evaluate the effects of Portulaca oleracea. L consumption along with resistance training on cathepsin S, cystatin C and C-reactive protein (CRP) levels on type 2 diabetes patients.

     Methods: In this semi-experimental study, 28 female type 2 diabetes patients with a mean age of 52 were randomly devided into 4 groups of control, exercise, supplement and supplement-exercise. Portulaca oleracea. L supplement was cosumed 7.5 g per day. Resistance training program was performed with a rubber band for 8 weeks, 3 days a week for 60 minutes with40-50% intensity, up to a maximum repetition. Blood samples were taken before and 48 hours after the last intervention.

     Results: After eight weeks, cathepsin S, cystatin C and CRP levels in the supplementation and supplementaion-exercise group were significantly reduced (P<0.05). There were also significant differences between the groups.

    Conclusion: Portulaca oleracea. L consumption and resistance training have each separate positive impacts on the cathepsin S, cystatin C and CRP levels, but the simultanous effect of Portulaca oleracea. L seed consumption and physical activity can lead to a better efficiency.

     Keywords: Portulaca oleracea, resistance training, cathepsin S,  cystatin C, C-reactive protein, type 2 diabetes mellitus.

Abstract

       Background and Objective: Various cellular factors affect the process of HIV activity. One of these cellular factors are structures known as microRN that are expected to be involved in controlling HIV replication and infectivity. The expression of one or a set of them may represent the patient's clinical conditions. In this study,  the expression of miR-29a and miR-29b involved in regulating viral genes’ expression was evaluated in three HIV-positive groups and a healthy control group. Later,  the expression level of these microRNAs  was compared between the cases and controls.

      Methods: Total RNA extraction was performed on the collected samples using RNx-plus kit and then the microRNA expression levels were evaluated using Relative Real-time PCR. The obtained data was entered into SPSS 22 and Graphpad softwares and analyzed using Kruskal-Wallis and Man-Whitney tests. P-value of less than 0.05 was considered as statistical significance level.

     Results: The expression level of miR-29a  was reduced in patients under treatment and drug-resistant patients ( P ≤ 0.05)  . All three HIV-positive groups including people without drug treatment, patients under treatment and drug-resistant patients showed reduced miR-29b expression level compared to  control group (P ≤ 0.05).

     Conclusion: the decreased expression of miR-29a and miR-29b in patients under treatment and drug-resistant patients indicates an  increased viral replication and reduced CD4 cell count. It may be possible to predict the progression of the disease by miRNA measurement or control viral replication using these mir-RNAs that requires further studies.

        Keywords: HIV, expression, mir-29a, mir-29b.

Abstract

        Background and Objective: Normal hemoglobin (Hb) is formed of a heme group and a protein group known as globin. Globin is made of four polypeptide chains and in hemoglobinopathies, the structure of one of these four polypeptide chain becomes abnormal. Cellulose acetate method is a common way to differentiate haemoglobinopathies. Inability to identify the components of Hb low concentrations and incapability to isolate all Hb types are among the disadvantages of this method. The aim of this study was to report the prevalence of hemoglobinopathies in the North of Iran by capillary electrophoresis method.

      Methods: All patients with suspected hemoglobinopathies, referred by physicians for electrophoresis, have been studied in a private center in the city of Gorgan, Iran. The level of HbA2, HbA, HbF and other Hb was recorded.

       Results: Overall, 725 blood samples were analyzed using the capillary method. HbE was reported in 2 patients, HbH was observed in 2 patients and Hb Barts was reported in 3 patients. Using the capillary method, among patients with the SDG area, only 4 of 38 (10.52%) had HbS and the majority of them (89.48%) had HbD.

      Conclusion: HbD is the most common hemoglobinopathy in the North of Iran.

        Keywords: Hemoglobinopathy; hemoglobin D; Capillary Electrophoresis; Iran

Abstract

      Background and objective: Hepatitis B virus (HBV) is a DNA virus with high tendency toward hepatic tissue. There are currently about 3 million HBV-infected people and 350 to 400 million chronic carriers of this virus in the world. X protein plays a role in the over-expression of oncogenes, carcinogenicity of liver cells and overlaps with the basal core promoter of the virus. Mutations at specific nucleotides of this region increase viral replication and liver disease progression. The aim of this study was to investigate the frequency of mutations at nucleotides 1762, 1764 and 1766 of HBV X gene in patients with chronic hepatitis B and hepatitis B-related cirrhosis.

      Methods: In this study, 102 patients including 68 chronic hepatitis patients and 34 patients with hepatitis B-related cirrhosis were enrolled. After DNA extraction, HBV X gene was amplified and sequenced using Semi Nested-PCR. Obtained gene sequences were compared with the standard sequence of HBV virus X gene available in the gene bank (Okamoto AB033559). Then, the mutations in the gene X of HBV were identified.

      Results: Comparison of the standard sequence with sequences obtained from patients showed the presence of A1762T / G1764A mutation in 12 chronic (17.64%) and 13 cirrhotic (38.23%) patients. Also, C1766G / G1764T mutations were found in 8.23% of chronic patients and 17.64% of cirrhotic patients.

      Conclusion: A1762T / G1764A mutations in the overlapping region of the basal core promoter with gene X C-terminal may lead to liver disease progression from chronic hepatitis to cirrhosis, by changing the amino acid sequence of the X protein.

    

ABSTRACT

          Background and Objective: Genetic variations in the gene encoding endothelial nitric oxide synthase (eNOS) enzyme affect the susceptibility to cardiovascular disease. Identification of the way these changes affect eNOS structure and function in laboratory conditions is difficult and time-consuming. Thus, it seems essential to perform bioinformatics studies prior to laboratory studies to find  the variants that are more important. This study aimed to predict the damaging effect of changes in the coding region of eNOS using homology- and structure-based algorithms (SIFT and PolyPhen).

           Methods: First, the single nucleotide polymorphisms in the coding region (cSNPs) of the human eNOS gene were extracted from dbSNP. Resulting amino acid changes were reported as primary data required for the study. Then, position and type of amino acid changes along with the complete amino acid sequence were separately entered into the SIFT and PolyPhen tools for analysis.

         Results: Of 144 single nucleotide changes, 38 changes by the SIFT, 47 changes by the PolyPhen and 18 amino acid substitutions by both tools were predicted as damaging.

          Conclusion: It is predicted that 18 amino acid changes may have damaging phenotypic effects on the structure of the eNOS enzyme that may affect its performance by potentially affecting the enzyme’s various functional regions. Therefore, computational prediction of potentially damaging nsSNPs and prioritizing amino acid changes may be useful for investigating protein performance using targeted re-sequencing and gene mutagenesis experiments.

        

ABSTRACT

       Background and Objective: Herpes simplex encephalitis is a life-threatening consequence of the central nervous system (CNS) infection with Herpes simplex virus (HSV). Although it is a rare disease, mortality rates reach 70% in the absence of therapy and only a minority of individuals can return to normal function. The aim of this study was to determine possible correlation between HSV infection and the incidence of encephalitis in patients with neurological signs.

        Methods: Overall, 152 CSF samples were tested from patients with neurological signs referred to Mahdieh Clinical Laboratory in Isfahan from 2010 to 2013. After cerebrospinal fluid (CSF) collection, DNA was extracted and real-time polymerase chain reaction (PCR) was performed for HSV detection.

          Results: Of 152 patients tested, 50 were diagnosed with encephalitis. HSV DNA was present in the CSF of 13 patients with encephalitis. HSV was significantly higher (p< 0.05) in patients with encephalitis, which shows the significance of infection as an etiological factor of this disease. About 60% of the encephalitis cases were in age range of 1-24 months.

         Conclusion: According to the findings of the present study, Cesarean section is recommended for HSV-positive mothers. A routine real-time PCR test is suggested for HSV detection in patients with encephalitis to avoid unnecessary antiviral treatments.

       

ABSTRACT

          Background and Objective: Adipose tissue secrets various hormones including adiponectin, which is closely related to weight control and energy, balance. This study investigated the effects of resistance training on adiponectin, testosterone and cortisol levels in untrained men.  

           Methods: Forty untrained men (mean age of 23.8±2.66 years, mean weight of 67.43 ± 4.96 kg) were randomly and equally assigned into groups of upper extremity resistance training, lower extremity resistance training, combined resistance training, and control. The subjects performed eight weeks of weight training, three sessions per week (five sets of 60-85% one repetition maximum). Blood sampling was done prior to the start of the program, after the fourth week, and after the eight week. Alpha level was set to 0.05 for all statically analyses.

         Results: Repeated measures ANOVA showed that eight weeks of upper extremities training significantly decreased body fat percentage (p=0.002, 7.39%), and significantly increased adiponectin (p=0.000; 90.42%) and testosterone (p=0.002; 24.19%) levels. In the lower extremities training group, body fat percentage (p=0.006, 7.39%) decreased significantly, while adiponectin (p=0.012; 87.82%) and testosterone (p=0.000; 23.54%) levels increased significantly compared to the pretest. Eight weeks of combined training significantly increased BMI (p=0.006, 1.88%), muscle mass (p=0.007, 2.24%), and adiponectin (p=0.000, 91.56%) level. However, cortisol level decreased (p=0.017, 19.17%) after four weeks of training.  

         Conclusion: Upper and lower extremities resistance trainings significantly change testosterone levels. Different types of resistance training significantly increases serum adiponectin level and changes body composition, which are effective in prevention of cardiovascular diseases.

         Keywords: Resistance Training, Adiponectin, Testosterone, Cortisol.

Background and Objective: The cell division cycle 25 (CDC25)is a familyof highly conserved dual-specificity phosphatases that activate cyclin-dependent kinase complexes. These complexes are the main cell cycle regulators. Mammalian cells ,exposure to DNA damaging radiations such as ionizing radiation and ultraviolet light, prevent cell cycle progression by activation of checkpoint pathways and lead to cell death.

      Methods: In this study, mice were exposed to different doses of ionizing radiation. Their total cellular protein was extracted from the bone marrow. After determining and matching the protein concentrations, CDC25A phosphatase levels were measured by western blotting.

        Results: The results showed that exposure to different doses of ionizing radiation in vivo significantly increased the expression of CDC25A compared to control group (P <0.05).

        Conclusion: Exposure to ionizing radiation increases the expression of CDC25A phosphatase, which increases the possibility of tumorigenesis in that area by increasing bone marrow cell proliferation.

        Keywords: Cell Cycle, CDC25A, Ionizing Radiation, Cyclin-Dependent Kinase.

ABSTRACT
            Background and Objectives: Diagnosis of hepatitis E virus (HEV) infection could be missed in some cases if serological tests are used solely. Molecular characterization of HEV is essential for diagnosis of acute and chronic HEV infections, and evaluating the chronic HEV infection status in immunocompromised patients. The aim of this study was to prepare a suitable HEV positive control, determine the limit of detection (LOD) of HEV RNA for a specific molecular test, and evaluate the efficiency and precision of the test.
           Methods: Genomic region of HEV NCBI reference sequence was constructed. LOD, intra-assay precision, and inter-assay precision were calculated to evaluate the efficiency and precision of the test. Then, tenfold serial dilutions of the HEV positive control were prepared. Real time PCR was performed three times for each dilution. Mean, standard deviation, and coefficient of variation of cycle thresholds obtained in three independent and simultaneous tests were calculated, and the results were analyzed.
          Results: The LOD of this test was determined as 1.4×104 copy/ml or 42 copy/reaction or 14 copy/µl. Intra-assay precision and inter-assay precision for all assays were lower than 2.5% and 10%, respectively.
          Conclusion: We propose that the real time PCR assay targeting the ORF2/3 overlapping conserved region is suitable for detection of a wide range of different HEV genotypes found in acute and chronic HEV infections. However, the precision of the test should be improved for detecting HEV RNA lower than 103 copy/ml.
          Keywords: Hepatitis E virus, Limit of Detection, Real Time PCR.

ABSTRACT
       Background and Objective: Escherichia coli is one of the most common causes of hospital-acquired infections. Extended-spectrum β-lactamase (ESBL)-producing E. coli strains are resistant to third-generation cephalosporins. The three main genes involved in ESBL production are TEM, SHV and CTX-M. Detection of ESBL-producing E. coli is of importance for infection control, reduction of excessive antibiotic use and epidemiological surveillance. This study aimed to detect ESBL-producing E. coli strains isolated from wound infections using phenotypic and molecular methods.
       Methods: During 2013- early 2015, 86 strains were collected from three hospitals in Isfahan, Iran. Antibiotic susceptibility testing was done using ceftazidime and ceftazidime + clavulanic acid discs. Polymerase chain reaction was used for the detection of the three resistance genes.
      Results: The resistance genes SHV, CTX-M and TEM were detected in 49 isolates (56.9%). In addition, 39 isolates (45%) were ESBL-producing strains. According to the results, 5 (5.8%), 14 (16.2%), 19 (22%) and 11 (12.7%) isolates contained the SHV, CTX-M, TEM and CTX-M + TEM genes, respectively. The frequency of CTX and TEM were significantly higher than that of SHV gene (P <0.05). Most of the isolated bacteria were resistant to cefazolin and sensitive to nitrofurantoin.
       Conclusions: There is a difference between the frequency of ESBL-positive isolates reported in the phenotypic and genotypic methods, which could be due to the lower sensitivity of the phenotypic method and impact of environmental factors on the emergence of antibiotic resistance.
       Keywords: Antibiotic resistance genes, ESBL, TEM, SHV, CTX-M, Escherichia coli.

ABSTRACT
       Background and Objective: Toxoplasmosis is a common parasitic infection worldwide. The infection can be caused via consumption of contaminated meat and mother-to-child (congenital) transmission, causing changes in central nervous system tissue, eye irritation and sometimes death. The human form of the disease is often asymptomatic and may be accompanied with general discomfort and swelling of the lymph nodes when associated with chorioretinitis. Acute infection in immunocompromised individuals could lead to mortality. The purpose of this study was to determine the prevalence of anti-Toxoplasma gondii antibodies in serum of patients with multiple sclerosis (MS) referred to the MS Center in city of Ahvaz, southeastern Iran.
        Methods: Blood samples were taken from 100 patients with MS and 100 healthy control participants. After separating the serum, presence of anti-Toxoplasma antibodies (IgG, IgM) was evaluated by enzyme-linked immunosorbent assay.
        Results: Frequency of anti-Toxoplasma IgG was 38% and 21% in the patients and controls, respectively. Toxoplasma IgM antibodies were not detected in any of the study groups. Pearson correlation coefficient showed a significant association between Toxoplasma antibodies and MS.
       Conclusion: Due to high prevalence of toxoplasmosis in MS patients, it is recommended to measure serum titers of the patients regularly, and placed them under antiparasitic therapy when necessary.
      Keywords: Toxoplasma, Multiple sclerosis, MS, Ahvaz.

ABSTRACT
        Background and Objectives: Human cytomegalovirus (HCMV) is the most common viral cause of morbidity and mortality in immunocompromised patients. The aim of this study was to evaluate the frequency of active CMV infection in hemodialysis patients in Gorgan, Iran.
        Methods: Plasma samples were obtained from 149 hemodialysis patients at Hemodialysis Unit of Panje-Azar Medical Centre in Gorgan, Iran. Presence of CMV-DNA in plasma samples was evaluated by polymerase chain reaction (PCR) using specific primers for highly conserved regions of major capsid protein gene of HCMV. In addition, level of CMV-IgM antibody was measured by serological testing. Demographic information and past medical history of patients were also recorded. Data was analyzed by SPSS software (version 18).
       Results: Total prevalence of CMV infection was 6.7% (10/149) among the patients receiving hemodialysis. CMV-DNA and anti-CMV IgM antibody were detected in 2.68% and 4.69%, of the samples, respectively. One case was found positive for both CMV-DNA and anti-CMV IgM antibody. CMV infection did not have any correlation with gender, age, ethnicity, duration of hemodialysis, and history of blood transfusion.
        Conclusion: A notable proportion of hemodialysis patients in Gorgan have active CMV infection. Accurate detection of these individuals is important for preventing infection spread, especially in immunocompromised individuals. Simultaneous diagnosis of CMV infection using serological testing and PCR assay could help reduce the risk of infection spread.
          Keywords: HCMV, Hemodialysis, PCR, Iran.

ABSTRACT
          Background and Objectives: Resistance training is a key component of exercise recommendations for weight control, yet very little is known about the effects of resistance training on appetite and related peptides. Hence, the aim of the present study was to investigate the effect of 8 weeks of resistance training on appetite and circulating acyl ghrelin, neuropeptide Y (NPY), and orexin in sedentary men.
           Methods: This study included 20 sedentary men (mean age: 21.6±3.5 year, body mass index: 23.1±2.7 kg/m²) who were equally divided into a control group and a resistance training group. Participants in the training group performed the whole body exercises three sessions per week with 3-4 sets of 8-10 repetitions at 60-85% one-repetition maximum. Participants in the control group performed no resistance training. Fasting blood samples were taken before starting the study and 72 hours after the last session of resistance training for evaluation of serum acyl ghrelin, NPY, and orexin levels. In addition, perceived appetite was assessed by visual analogue scale while fasting.
           Results: Statistical analysis showed that fasting acyl ghrelin and NPY were not changed by resistance training, but serum orexin level elevated by 40% in response to training (P=0.01). Appetite was not significantly different between the two groups at baseline (P=0.9). However, appetite significantly increased after resistance training (P=0.001).
           Conclusion: Results of the present study show that the 8-week resistance training increases perceived appetite by orexin promotion in previously sedentary men.
          Keywords: Appetite, Ghrelin, NPY, Orexins, Resistance Training.

ABSTRACT
          Three major hepatitis B virus (HBV) antigens include HBcAg, HBeAg and HBsAg. HBeAg is the extracellular form of HBcAg, and is seen almost exclusively in people who have circulating serum HBV DNA. Presence of HBsAg in serum indicates that the individual has contracted HBV infection. Chronic hepatitis HBeAg-negative/anti-HBe–positive is known as an important form of chronic hepatitis B in the Mediterranean region. In this report, we used Real-Time PCR and ELISA for detection of HBV and HBeAg/HbsAg, respectively. In our investigation on 4743 HBV cases referred to the Mahdieh Clinical Laboratory between 2008 and 2016, we found a 53-year-old man with clinical symptoms of hepatitis and abnormal molecular and serological features. Despite the presence of clinical symptoms and high viral load (128 × 10⁵ iu/ml), the patient was HBsAg-positive and HBeAg-negative. Identifying this type of HBV could indicate spread of this type of hepatitis in Isfahan, Iran.
           Keywords: Hepatitis B, HbsAg, HBeAg.

ABSTRACT
         Background and Objectives: Elevated blood lipid levels are correlated with risk of cardiovascular disease, especially in overweight individuals. The beneficial effect of strength training on lipid profile has been well documented. However, the effect of strength training with flow blood restriction has not been established. The purpose of this study was to compare the effect of eight weeks of strength training with and without blood flow restriction on some of the cardiovascular risk factors in overweight females.
         Methods: In this study, 28 overweight females aged 18 to 24 years were randomly assigned into three groups of blood flow restriction strength training (BFRT), traditional strength training (TRT) and control. Participants in the BFRT group performed eight weeks of strength training three sessions per week at 30% of 1-repetetion maximum, while participants in the TRT group performed the training at 65-80% of 1-repetetion maximum. Cardiovascular variables, systolic blood pressure and diastolic blood pressure were measured before and after the exercise training. Data was analyzed using descriptive statistics, paired t-test and one-way analysis of variance.
          Results: Triglyceride level decreased significantly in the BFRT (p=0.022) and TRT (p=0.038) groups compared with the control group. Moreover, there were no significant differences in the body mass, waist-to-hip ratio, body mass index, high-density lipoprotein, low-density lipoprotein, and cholesterol levels between the study groups (P≥0.05).
           Conclusion: Our findings suggest that the BFRT and TRT may not be suitable for preventing or even reversing the physiological changes induced by obesity.
         Keywords: Resistance Training, Lipids, lipoprotein, overweight.

ABSTRACT
        Background and Objective: Lophomonas blattarum is a multi-flagellate protozoan that is commensal in hindgut of cockroaches and other insects. The protozoan can cause respiratory infection in humans. Most cases of the infections with this protozoan have been reported in China. Here, we present a case with chronic respiratory allergy caused by L. blattarum in Golestan province, Iran.
         Case Description: The case was a 37-year-old male with history of respiratory conditions and he was immunocompetent. An athlete. L. blattarum was detected in direct smear examination of sputum,
        Conclusion: Since we found respiratory infection in an immunocompetent individual who was also an athlete, it is necessary to study this parasite and its life cycle and transmission methods. It is also suggested to consider L. blattarum infection and treatment with metronidazole in cases of chronic allergies, especially those that do not respond to treatment.
        Keywords: Lophomonas blattarum, Respiratory Allergies, Iran.

ABSTRACT
          Background and Objectives: C-C chemokine receptor type 5 (CCR5) is a chemokine receptor expressed at high levels on the surface of T-cells. A 32-bp deletion in the coding region of the CCR5 (CCR5Δ32) leads to production of an incomplete protein that is not expressed on the cell surface. CCR5Δ32 may be involved in development of autoimmune disease, such as systemic lupus erythematosus. We investigated frequency of the CCR5Δ32 polymorphism in SLE patients and healthy controls, and evaluated the relationship between the CCR5Δ32 polymorphism and susceptibility to SLE in Golestan Province, Iran.
          Methods: Whole blood samples were taken from 80 SLE patients admitted to Shahid Sayyad Shirazi hospital and 80 healthy controls (from a blood bank) in the Golestan Province, in 2016. Baseline clinical and laboratorial characteristics were evaluated regarding the CCR5Δ32 genotypes. The CCR5Δ32 polymorphism was determined from genomic DNA by polymerase chain reaction.
          Result: Genotype frequencies of both groups were in the Hardy-Weinberg equilibrium. The frequencies of the CCR5 and the CCR5Δ32 alleles were 98.13% and 1.88% among the patients, and 98.75% and 1.25% among the controls, respectively. Homozygote CCR5Δ32 was not observed in the subjects. The frequency of heterozygous Δ32 was 3.8% and 2.5% among the SLE patients and controls, respectively (P-value>0.05). There was no significant association between the CCR5 status and clinical signs of SLE (P>0.05).
          Conclusion: Our data suggest that the CCR5Δ32 polymorphism has no correlation with SLE in our study population. In addition, the frequency of the Δ32 polymorphism in SLE patients and controls does not follow the Hardy-Weinberg equilibrium
          Keywords: CCR5, Homozygote CCR5Δ32, Heterozygote CCR5Δ32, CCR5Δ32 allele, SLE.

ABSTRACT
          Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transgenes expression depends on some experimental variables including cell type and density, Lipofectamine and DNA concentrations and Lipofectamine-DNA complexing time. The main objective of this study was to optimize transfection of SW480 colon cancer cells with Lipofectamine 2000.
          Methods: In this study, SW480 cells were transfected with plasmid containing green fluorescent protein reporter gene using Lipofectamine 2000. Green fluorescent protein expression was studied under a reverse fluorescence microscope and the results were analyzed with the ImageJ software. Effect of different quantities of plasmid DNA and different Lipofectamine 2000 volumes on cell transfection efficiency was evaluated.
          Results: The optimal volume of Lipofectamine and quantity of plasmid was 2 µl and 1µg, respectively, which showed 59% efficiency for the transfection of SW480 cells at 24 hours post-transfection.
          Conclusion: This study shows that Lipofectamine 2000 is an efficient reagent for the delivery of genes into SW480 cells. According to the results, the quantity of DNA per transfection and reagent concentrations are essential factors for a successful transfection.
          Keywords: Optimization; pEGFP-NI; Lipofectamine; SW480.