fatemeh roudbari

KRAS and BRAF genes are the biomarkers in Colorectal Cancer (CRC) which play prognostic and predictive roles in CRC treatment. Nowadays, the selection of rapid and available methods for studying KRAS and BRAF mutations in anti-EGFR therapy of patients suffering from CRC plays a significant role. In this study, the mutations of these two oncogenes were evaluated by different methods.

This study was performed on 50 Formalin-Fixed Paraffin-Embedded (FFPE) tissue blocks of patients diagnosed with colorectal cancer. After DNA extraction, KRAS and BRAF gene mutations were evaluated using reverse dot blot, and results were compared with PCR-RFLP and allele-specific PCR for KRAS and BRAF mutations, respectively.

KRAS gene mutations were detected in 42% of patients, of which 30% were in codon 12 region, and 12% in codon 13. The most frequent mutations of KRAS were related to G12D and 10% of patients had BRAF mutated genes. The type of KRAS gene mutations could be evaluated by reverse dot blot method. In general, the results of PCR-RFLP and allele-specific PCR were similar to the findings by reverse dot blot method. 

These findings suggest that PCR-RFLP and allele-specific PCR methods are suitable for screening the presence of the mutations in KRAS and BRAF oncogenes. In fact, another method with more sensitivity is needed for a more accurate assessment to determine the type of mutations. Due to higher speed of detection, reduced Turnaround Time (TAT), and possible role of some KRAS point mutations in overall survival, reverse dot blot analysis seems to be an optimal method.

The role of immune system in natural course of viral hepatitis has been drawn some attention. One of the main diagnostic markers of the immune system function in different diseases might be the ratio of CD4+ to CD8+ T lymphocytes (CD4+/CD8+ ratio).

This research aimed to measure and compare CD4+/CD8+ ratio in the patients with chronic hepatitis B (CHB) and control group.

In this cross-sectional study, thirty-three CHB patients and thirty age and sex-matched healthy controls were included. Immunophenotyping of isolated T cells was performed using specific anti-CD4 and anti-CD8 antibodies by flow cytometry. Consequently, CD4+, CD8+ and the ratio of CD4+/CD8+were counted and compared between the two groups.

CD4+ counts (%) were considerably reduced in patients with CHB compared to the healthy controls (51.22 ± 10.5 vs. 63.14 ± 9.9, P = 0.00), whereas CD8+ counts (%) were higher in the patients with CHB than healthy controls (48.8 ± 10.5 vs. 36.85 ± 9.86, P = 0.00). Moreover, CD4+/CD8+ ratio remarkably decreased in the patients with CHB (1.15 ± 0.5) than healthy controls (1.93 ± 0.9) (P = 0.00). Area under curve (AUC) of 0.79 (SE = 0.06, CI = 0.68 - 0.90, P value = 0.05) was reported for CD4+/CD8+ ratio with a sensitivity of 72.73% and specificity of 73.33% in 1.35 cut-off (likelihood ratio = 2.72).

The research indicated an impaired balance between T cell subsets associated with a higher proportion of CD8+ T cells and a lower proportion of CD4+ T cells and CD4+/CD8+ ratio in patients with CHB.

Kirsten rat sarcoma (KRAS) gene is a target of genetic alterations which are diagnostic and prognostic biomarkers in patients with metastatic colorectal cancer who are treated with monoclonal anti-EGFR antibodies such as cetuximab and panitumumab. KRAS mutations are seen in 35-42% of patients with colorectal cancer. The high frequency of these mutations in colorectal cancer represents their high potential as a biomarker in early diagnosis of cancer. This study was done to evaluate the frequency of KRAS gene mutations in a small population of Iranian patients suffering from colorectal cancer.
50 formalin-fixed paraffin-embedded tissue blocks with colorectal cancer (CRC), already confirmed by histopathology and immunohistochemistry testing, were received to Payvand Clinical and Specialty Laboratory, Tehran, from across the country in 2015. DNA was extracted from the tissue blocks and its quality was then evaluated. The reverse dot blotting method was used to evaluate KRAS gene mutations.
KRAS mutations were found in 42% of the study patients. 30% and 12% of the mutations were found in codon 12 and codon 13, respectively. Moreover, no mutation was found in codon 61. Results also showed that the most frequency of samples examined belonged to male with 68% (average age of 56 years old) and then to female with 32% (median age of 54.8 years old).
This study was performed to evaluate the frequency of KRAS gene mutations in Iranian colorectal cancer patients. According to the study results, the frequency of KRAS mutations was consistent with that of other countries, reported in previous studies. The high prevalence of these mutations in patients with colorectal cancer indicates the important role of these genes in this group of patients. Thus, the presence of these mutations can be used as a suitable biomarker for evaluation of response to targeted therapies in patients suffering from colorectal cancer.

Omentin-1 is an adipokine that is highly secreted in visceral adipose tissue compared to subcutaneous adipose tissue and increases insulin sensitivity. The aim of the present study was to investigate the effect of 8-week resistance training on omentin-1 plasma levels in insulin resistant male rats. Twenty-four Wistar male rats with average weight 161±23 gr were randomly divided into three groups: health control، insulin resistance control، insulin resistance training. After fructose-inducing insulin resistance to the two groups of insulin resistance control and insulin resistance training bleeding in all subjects was done then training group was exercised for 8 weeks (3d/wk). In the training protocol، a ladder was used on which rats carriedpen loads suspended from their tails. After the training session omentin-1، insulin، glucose، lipids profile plasma concentration and (HOMA-IR) index were measured. The results of this study indicate that 8 weeks of resistance training can cause significant increase of omentin-1 and HDL-C plasma concentration in insulin resistance training group (P≤0. 05) and insulin، glucose، cholesterol، LDL-C، TG plasma concentration and (HOMA-IR) index were decreased. This study indicates that resistance training increases omentin-1 plasma concentration in insulin resistance rats and improves lipids and metabolic profile.

The purpose of the current study was to investigate the effect of a high- intensity treadmill running (8 weeks) with or without aqueous extraction of black Crataegus-Pentaegyna (Siyah -Valik) on liver ABCA1 mRNA expression and plasma HDL-C, total, and direct bilirubin concentration in male rats. Twenty Wistar male rats (4-6 weeks old, 140-170 g weight) were used. Animals were randomly assigned into training (n =10) and control (n =10) groups and further divided into saline-control (SC, n=5), saline-training (ST, n=5), black Crataegus-Pentaegyna (Siyah-Valik) Control (SVC, n=5), and black Crataegus-Pentaegyna (Siyah-Valik)-training (SVT, n=5) groups. Training groups have performed a high-intensity running program (34 m/min on 0% grade, 60 min/day and 5 days/week) on a motor-driven treadmill for 8 weeks. Animals were orally fed with black Crataegus-Pentaegyna (Siyah –Valik) extraction (500 mg/kg body weight) and saline solution for last six weeks. A significant differences have found in liver ABCA1 gene expression between SVC and SVT (P<0.003) and between SVC with SC groups (P< 0.038). HDL-C levels were significantly (P<0.036) between groups. A higher HDL level was found in SV treated groups and between SVC and SC groups. The levels of bilirubin total and bilirubin direct remained unchanged. The current results show that high-intensity treadmill running affected liver ABCA1 mRNA expression in different directions in saline (increase) and SV (decrease) treated animals. Findings also indicate an opposite pattern of change in saline and SV treated animals at rest. It seems the existence of opposite effect of exercise with supplementation of SV might be attributed to the suppression of lignds which is provided by SV supplementation at rest. This in turn might be also taking in account in lower liver ABCA1 mRNA expression and its related nuclear receptors such as LXR in SVT not ST groups.

Heart as a high metabolic and aerobic tissue is consuming lipid as a fuel for its energy provision at rest during light and moderate exercise, except when lactate level is higher in blood circulation. It has been shown that any type of regular exercise and crataegus species would improve cardiovascular function and minimizes several risk factors via stimulating lipid metabolism by acting on enzymes and genes expression such as ABCA1 and PPAR α which are involving in this process.. Twenty Wistar male rats (4-6 weeks old, 140-173 g weight) were used. Animals were randomly classified into training (n = 10) and control (n = 10) groups and then divided into saline-control (SC), saline-training (ST), Crataegus-Pentaegyna -control (CPC), and Crataegus-Pentaegyna -training (CPT) groups. Training groups have performed a high-intensity running program (at 34 m/min (0% grade), 60 min/day, 5 days/week) on a motor-driven treadmill for eight weeks. Animals were orally fed with Crataegus-Pentaegyna extraction (500mg/kg) and saline solution for six weeks. Seventy- two hours after the last training session, rats were sacrificed, hearts were excised, cleaned and immediately frozen in liquid nitrogen and stored at -80 °C until RNA extraction. Plasma also was collected for plasma variable measurements. Statistical analysis was performed using a two way analysis of variance, and significance was accepted at P < 0.05.. A non-significant (P < 0.4, P < 0.79, respectively) increase in ABCA1 and PPAR α genes expression was accompanied by a significant (P < 0.01, P < 0.04, P < 0.04, respectively) reduction in TC, TG, and VLDL-C levels in Crataegus-Pentaegyna groups.. Our findings show that a high intensity treadmill running was able to express ABCA1 and PPAR α in rat heart. Data also possibly indicate that the Crataeguse-Pentaegyna supplementation solely could mimic training effect on the mentioned genes and lipid profiles via different mechanism(s).

The global HIV epidemic continues to expand and exceeding previous predictions. An effective vaccine represents the best hope to curtail the HIV epidemic. DNA vaccines induce humoral and cellular responses and mimic live vaccines without their pathogenic potential. The importance of CD8+ CTL responses in controlling HIV and SIV viremia has led to production of a series of vaccine candidates that effectively induce these responses. It is now widely believed that an HIV vaccine strategy must stimulate both a strong humoral (antibody) as well as cell-mediated (CTL) immune response.The p24 and gp41 play many important roles in host-virus interaction and pathogenesis. These proteins are considered as attractive vaccine candidate in which their immunogenecity and immunomodulatory effects have been confirmed. In this study, a construct, pcDNA3.1Hygro- (p24-gp41), was evaluated as a DNA vaccine candidate in Balb/C mice for generation of effective cellular immune responses. For immunizing, we used dendrosome, a novel family of vehicles for transfection and therapy. IFN-γ cytokine production and total antibody were detected by ELISA. Lymphoprolifration assay was performed by MTT test. ELISA and MTT assays confirmed that the cited p24-gp41 fusion gene is able to enhance immune responses in mice. The construct that was used in this research can be a good candidate for DNA vaccine against HIV-1, if the future complementary tests demonstrate the same trends of immunogenic responses shown in this study.