Real-time RT-PCR Technique Research Articles

A trend over time study of hepatic Farnesoid-X-activated receptor and its downstream targets modulation by valproic acid in mice

Valproic acid (VA) is a broad-spectrum anticonvulsant agent that acts through several molecular mechanisms to control different types of seizures. The main concern of the drug is its liver toxicity. Considering the regulatory roles of the Farnesoid nuclear receptors and the nuclear transcription factor Nrf2 in modifying and neutralizing the harmful effects of oxidative damage, the present study was designed to evaluate the role of FXR-Nrf2 and some downstream target gene alterations in hepatotoxicity induced by VA. Thirty-five eight-week-old male albino mice were randomly divided into five groups, including a control group, and four groups were assigned to receive VA (300 mg/kg/day; oral) for 3, 7, 10, and 14 days. Serum levels of ALT, AST, ALP, and total and direct bilirubin (TB, DB) were measured. Liver histology and the expression of FXR, Nrf2, α-GST, SOD, and TNF-α were assessed using H&E staining and real-time RT-PCR techniques. Maximum extent of biochemical and histopathological damage was observed on the 14th day, but changes in the expression of FXR, Nrf2, α-GST, and SOD were seen at three points: a significant upregulation on the 3rd day, a remarkable downregulation on the 10th day, and a second-time upregulation on the 14th day. In conclusion, considering the observed dysregulation in FXR-Nrf2 cascade expression during VA administration, it seems that downregulation in this pathway and consequently its downstream detoxification and antioxidant genes may play a role in liver toxicity.

Exportin 4 DNA promoter methylation in liver fibrosis.

A role for exportin 4 (XPO4) in the pathogenesis of liver fibrosis was recently identified. We sought to determine changes in hepatic XPO4 promoter methylation levels during liver fibrosis. The quantitative real-time RT-PCR technique was used to quantify the mRNA level of XPO4. Additionally, pyrosequencing was utilized to assess the promoter methylation status of XPO4. The methylation rate of the XPO4 promoter was significantly increased with fibrosis in human and mouse models, while XPO4 mRNA expression negatively correlated with methylation of its promoter. DNA methyltransferases (DNMTs) levels (enzymes that drive DNA methylation) were upregulated in patients with liver fibrosis compared to healthy controls and in hepatic stellate cells upon transforming growth factor beta (TGFβ) stimulation. The DNA methylation inhibitor 5-Aza or specific siRNAs for these DNMTs led to restoration of XPO4 expression. The process of DNA methylation plays a crucial role in the repression of XPO4 transcription in the context of liver fibrosis development.

Regulation of gene transcription in Escherichia coli O157:H7 in response to a natural derivate peptide of esculentin-1a used in combination with essential oils from plants of the Cymbopogon genus.

We analyzed the expression of several genes implicated in the pathogenicity of Escherichia coli O157:H7, treating bacteria with Esc(1-21), a derivative of peptide esculentin-1 in combination with three essential oils obtained from plants from the Cympopogon genus. We used the checkerboard assay to determine the antimicrobial activity of the combinations. We analyzed the expression of some genes implicated in the pathogenicity and quorum sensing system of E. coli O157:H7 by real-time RT-PCR technique. Treatment of the bacteria with the peptide combined with oils had an efficacious antimicrobial activity. The analysis of gene expression showed that all used combinations regulate positively the espAD and ler genes, located in the pathogenicity island, named the locus of enterocyte effacement. None of the combinations affects the quorum sensing genes: lsrABCFKR and qseBC. This study demonstrates that the use of essential oil/peptide combinations can be effective in fighting microbial infections.

A importância da Biologia Molecular no diagnóstico do SARS-CoV-2

The pandemic caused by the etiologic agent SARS-CoV- 2, started at the beginning of 2020. And create a big challenge in the scientific community from all around the world. The symptoms can variate of levels manifestation to severs cases of pneumonia. Studies are seeking ways of prevention, diagnosis and treatment of the disease. In view of the pandemic scenario and the worsening number of cases and deaths due to the disease, the search for an accurate diagnosis is essential. Therefore, the aim of this study is demonstrating the importance of using molecular biology through the technique of Real Time Transcriptase Reverse Polymerase Chain Reaction (Real Time RT-PCR), for the detection of the genetic material of SARS-CoV-2 from data found in the literature. After a critical analysis of the literature, it was possible to confirm the importance and efficiency of using the Real Time RT-PCR technique in the diagnosis of infection, since this technique allows a good performance in the detection of viral genetic material, guaranteeing a high success rate when differentiating an infected patient from the non-infected. Concluding that until now this technique is considered as the main diagnostic method for the disease, remaining as a “gold standard”.

Application of multiplex realtime PCR detection for hemorrhagic fever syndrome viruses

BackgroundMultiplex real-time PCR is a quick and cost effective method for detection of various gene simultaneously. HFSV (Hemorrhagic Fever Syndrome Virus) is a newly emerging infectious disease because of globalization and climate change. We tried to develop a molecular diagnostic technique for various causative viruses and evaluate its usefulness for improving public health. MethodsMolecular diagnostic test method that qualitatively detects viruses causing viral hemorrhagic fevers hired Taq-Man Real-time RT-PCR technique. The Ct value was experimentally observed three or more times at the RNA concentration before and after the detection limit. After designing a multiplex real-time RT-PCR test for target gene of selected 17 viruses, the detection limit for each target and the presence or absence of cross-reaction and interference reaction were evaluated to determine its availability. ResultsSix kinds of viruses, including Crimean-Congo hemorrhagic fever virus, Omsk hemorrhagic fever virus, Sabia virus, Chapare virus, Yellow fever virus, and Variola virus (A4L gene, B12R gene), were able to confirm the detection limit of 0.5 copies/μl, and other Ebola virus, Marburg virus, Rift Valley fever virus, Kyasanur Forest disease virus, Junin virus, Guanarito virus, Machupo virus, Chikungunya virus, Hantavirus, Dengue virus types 1–4, and Lassa virus (L gene, GPC gene), and 11 kinds of viruses, the detection limit was confirmed at 5 copies/μl. No cross-reaction or interference between detected genes was observed. ConclusionThe virus test method developed through this study using multiplex is expected to be used for public health and quarantine as a test method that can be used when a hemorrhagic fever virus of unknown cause is introduced.

Comparative Study of Quantitative Real-Time Monitoring QRT-PCR for BCR-ABL Gene in Chronic Myelogenous Leukemia (CML) Between Blood and Bone Marrow Samples

Objectives: This study aimed to assess the quantitative real-time RT-PCR (QRT-PCR) technique as a diagnostic tool for molecular surveillance of the BCR-ABL duplicate in Chronic Myelogenous Leukemia (CML) using both peripheral blood (PB) and bone marrow (BM) samples. Methods: Prospective analysis has been conducted a by quantitative real-time RT-PCR (QRT-PCR) for both PB and BM specimens, from 25 patients with untreated CML. QRT-PCR investigation was carried out previous and during treatment with Imatinib for untreated CML. Statistical examinations showed useful agreement of PB and BM pre- treatment specimens. Nevertheless, using the SPSS statistical method that estimates the agreement between PB and BM data, Results: This study showed low correspond of BCR-ABL measurements in PB and BM for specimens acquired through treatment. PB values tended to be lower than the conformable BM values [average difference = -0.37 (p<0.001) in 36 coupled samples] and the 95% limits of agreement ranged from -1.23 to 0.48. Nevertheless, the present study showed that BM and PB QRT-PCR values followed a similar direction during treatment (Spearman correlation coefficient, 0.83; 95% CI, 0.70, 0.96). Conclusion: Our findings imply that PB and BM measures of BCR-ABL are frequently quantitatively different. The most accurate way to determine whether there is minimal residual disease is through BM sampling because BM results tend to be greater than PB values (MRD). Based on these findings, we advise avoiding switching BM and PB sampling for MRD monitoring during CML treatment because doing so could result in incorrect interpretation of treatment outcomes.

LAMP-Based Point-of-Care Nucleic Acid-Based Detection Method Can Be Useful for Quick Decision-Making for Diagnosis of Acute COVID-19 Emergency Cases in Hospital Settings

Real-time RT-PCR is used as a gold standard method for the diagnosis of COVID-19. Since real-time RT PCR is nucleic acid-based, it is a highly sensitive and specific test. However, this test takes 4–8 h to generate results and, in emergency settings, this delay may prove fatal for certain patients. The frequent surge in COVID cases increases patient load in emergency settings. Thus, a nucleic acid-based rapid POC test is required that can generate results quickly as well as being comparable to real-time RT-PCR. In this study, comparison of real-time RT-PCR was carried out using the rapid nucleic acid-based LAMP method. Nasopharyngeal swabs were taken in duplicate from patients visiting the kiosk and were analyzed for the presence of the SARS-CoV-2 virus by both real-time RT-PCR and LAMP techniques ID NOW(bbott). Out of 14 positive and 31 negative samples tested by real-time RT-PCR, 13 samples were identified as positive and 31 were observed as negative with the LAMP-based test. Hence, the sensitivity and specificity of this method were found to be 92.9% and 93.5%, respectively. Therefore, LAMP-based point-of-care testing has the potential to be used in hospital emergency settings for quick diagnosis of critically ill patients, and the information generated here will further draw the attention of policymakers toward such nucleic acid-based rapid tests.

Gender-based differences of inflammatory, coagulation, and cardiac markers in COVID-19 patients in Erbil city

In December 2019, a new coronavirus disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) appeared in Wuhan city and quickly became a global health issue. COVID-19 causes various symptoms ranging from no symptoms to potentially deadly pneumonia. The study aimed to understand the effects of SARS-CoV-2 infection on immune response and the differences in inflammatory, coagulation, and cardiac biomarkers between male and female patients. Between June 1st and November 1st, 2020, 95 cases of SARS-CoV-2 infected individuals were studied at Zanko Hospital. SARS-CoV-2 infection was confirmed using the real-time RT-PCR technique. All cases were analyzed for clinical, epidemiological, and laboratory data. On average, the patients were 50.64 (SEM= 2.359) years old, with 61 males and 34 females. The patients had elevated C-reactive protein (CRP), which was 43.96 (SEM= 6.154), while the erythrocyte sedimentation rate (ESR) was 50.50 (SEM= 5.498). The mean of D-Dimer, ferritin and lactate dehydrogenase (LDH) were 1.204 (SEM= 0.164), 534.7 (SEM= 61.48), and 366.6 (SEM= 36.81), respectively. There were no significant differences in the study's data mentioned above between male and female patients. In conclusion, inflammation is the most prominent symptom in COVID-19 patients, and males and females are nearly equally affected.

Effect of heat inactivation on CT value of realtime RT-PCR technique for SARS-CoV-2 detection

SARS-CoV-2 was the cause of the global COVID-19 pandemic in 2019. The cycle threshold (CT) value of the real-time RT-PCR technique is the gold standard in infections diagnosis, infections risk assessment, discharged criteria, and patient isolation for SARS-CoV-2. Heat inactivation is a simple, economical solution for safety in testing for SARS-CoV-2. The change of CT values after heat inactivation has not been thoroughly evaluated. This study evaluated the effect of heat inactivation on CT value in the realtime RT-PCR technique of SARS-CoV-2 detection. Four groups of samples consisted of one non-inactivated group, three groups that were inactivated at 65oC/30min, 80oC/10min, and 95oC/10min; each group included 16 tubes, from 16 nasopharyngeal swabs samples that were confirmed to be infected with SARS-CoV-2. Groups were inactivated and then analysed by realtime PCR technique. The results show that virus inactivation by heat increased the CT value of realtime PCR in SARS-CoV-2 positive samples. The higher temperature was in the inactivation, and the higher CT value was increased.

The Effect of Pantoprazole on Tumor Growth and Apoptosis in Gastric Cancer Stem-Like Cells

Background: Gastric cancer is a daunting global problem with unsatisfactory treatment. Due to the key role of Gastric Cancer Stem-like Cells (GCSCs) in all stages of gastric cancer and the failure of contemporary anticancer therapies, many research efforts are focusing on these treatmentresistant cells. Pantoprazole, as recently considered antitumor agent with well-documented effects on tumorigenesis inhibition, has seldom been investigated in GCSCs in previous studies. We aimed to study the influence of pantoprazole on cell proliferation, apoptosis, and the transcription of genes involved in the cell proliferation and apoptosis pathways. Materials and Methods: Herein, we isolated GCSCs from MKN-45 cell line, on a non-adherent surface and then evaluated the effect of pantoprazole on cell growth and apoptosis of GCSCs by means of MTT, DNA laddering and quantitative real-time RT-PCR techniques. Results: Our findings showed that treated cells with pantoprazole decreased cell proliferation and induced apoptosis. PCNA (Proliferating Cell Nuclear Antigen) and antiapoptotic Bcl2 genes were downregulated and Bax and CASP3 proapoptotic genes, as well as tumor suppressor p53 gene, were overexpressed. Conclusion: Our results revealed that pantoprazole induced apoptosis and declined tumor growth and support the idea that pantoprazole played as a promising breakthrough in gastric cancer therapy.

Association study of Bif-1 gene expression with histopathological characteristics and hormone receptors in breast cancer

BackgroundBreast cancer is a heterogeneous disease that has various clinical outcomes. Bax-interacting factor-1 (Bif-1) is a member of the endophilin B family that generates the pro-apoptotic BCL2-Associated X (BAX) protein in response to apoptotic signals. Lack of Bif-1 inhibits the intrinsic pathway of apoptosis and enhancements the risk of tumor genesis. The present study aimed to investigate the relationship between hormone receptors (ER, PR, and HER2) status and different levels of Bif-1 gene expression in breast cancer patients.MethodsBif-1 gene expression was evaluated in 50 breast cancer tumors and 50 normal breast mammary tissues using the SYBR Green real-time RT-PCR technique. Multivariate and univariate analyses were used to appraise the relationship between the prognostic significance of the Bif-1 gene using SPSS software. In this study, the Bif-1 was selected as a candidate for a molecular biomarker and its expression status in breast cancer patients with hormone receptors (ER, RR, and HER2) compared to patients without these hormone receptors.ResultsThe study showed that the relative expression of the Bif-1 gene in tissues of patients with hormone receptors in breast cancer compared to those without hormone receptors was not statistically significant. The expression levels of the Bif-1 gene in different groups were evaluated for hormone receptor status. No significant relationship was found between the Bif-1 gene expression and hormone receptors (ER, PR, and HER2) (p > 0.05).ConclusionBif-1 gene expression may be a useful prognostic marker in breast cancer.

Inhibition of dengue virus infection by trifluoperazine.

Dengue virus (DENV), a member of the genus Flavivirus, family Flaviviridae, is the most widespread viral pathogen transmitted to humans by mosquitoes. Despite the increased incidence of DENV infection, there are no antiviral drugs available for treatment or prevention. Phenothiazines are heterocyclic compounds with various pharmacological properties that are very adaptable for drug repurposing. In the present report, we analyzed the antiviral activity against DENV and the related Zika virus (ZIKV) of trifluoperazine (TFP), a phenothiazine derivative in clinical use as an antipsychotic and antiemetic agent. TFP exhibited dose-dependent inhibitory activity against the four DENV serotypes and ZIKV in monkey Vero cells at non-cytotoxic concentrations with 50% effective concentration values in the range 1.6-6.4 µM. A similar level of antiviral efficacy was exhibited by TFP against flavivirus infection in the human cell lines A549 and HepG2. Mechanistic studies, performed using time-dependent infectivity assays, real-time RT-PCR, Western blot, and immunofluorescence techniques, indicated that uncoating of the virus during penetration into the cell was the main target for TFP in infected cells, but the compound also exerted a minor effect on a late stage of the virus multiplication cycle. This study demonstrates that TFP, a pharmacologically active phenothiazine, is a selective inhibitor of DENV multiplication in cell culture. Our findings open perspectives for the repositioning of phenothiazines like TFP with a wide spectrum of antiviral efficacy as potential agents for the control of pathogenic flaviviruses.

Relative Expression of mRNA BARF1 Epstein-Barr Virus from Tumor Tissue Biopsy in Formalin-Fixed Paraffin-Embedded in Nasopharyngeal Carcinoma

Nasopharyngeal carcinoma (NPC) is a malignant tumor originating from nasopharyngeal epithelial cells. NPC is endemic in some regions, especially in Southeast Asia countries. NPC is also a multifactorial disease involving environmental factors, genetic factors, and infection from Epstein-Barr virus (EBV). According to WHO classification, Undifferentiated NPC is histopathologically associated with EBV infection and categorized into WHO subtype 3. EBV has two phases in its infection cycle: the lytic and latent phases. The BARF1 gene is a mediator of the transition from the latent phase to the lytic phase. Previous studies suggest measurement of EBV mRNA activity at the primary tumor site in the nasopharyngeal reflects the pathogenesis of NPC rather than measuring circulating EBV DNA or serological diagnosis. This study aimed to determine the relative expression potential of BARF1 mRNA at different tumor stages in NPC patients as a predictor of NPC pathogenesis. This research design was a descriptive research method in the form of a cross-sectional study. The samples used were 22 patients diagnosed as NPC WHO class III at the Anatomical Pathology Section of Prof. Dr. Margono Soekarjo, Purwokerto, who met the inclusion criteria. The relative expression of BARF1 mRNA was carried out using the one-step real-time RT-PCR technique and then calculated using a formula of 2-ΔCt. The T-test was used to compare the relative expression of early and late-stage BARF1 mRNA. The relative expression of BARF1 mRNA in the late-stage advanced stage (n = 6; 0.708292-0.840177; med = 0.7164655) was increased compared to the early stage (n = 2; 0.708841-0.712423; med = 0. 710632).

A review of intelligent medical imaging diagnosis for the COVID-19 infection

Due to the unavailability of specific vaccines or drugs to treat COVID-19 infection, the world has witnessed a rise in the human mortality rate. Currently, real time RT-PCR technique is widely accepted to detect the presence of the virus, but it is time consuming and has a high rate of eliciting false positives/negatives results. This has opened research avenues to identify substitute strategies to diagnose the infection. Related works in this direction have shown promising results when RT-PCR diagnosis is complemented with Chest imaging results. Finally integrating intelligence and automating diagnostic systems can improve the speed and efficiency of the diagnosis process which is extremely essential in the present scenario. This paper reviews the use of CT scan, Chest X-ray, lung ultrasound images for COVID-19 diagnosis, discusses the automation of chest image analysis using machine learning and deep learning models, elucidates the achievements, challenges, and future directions in this domain.

A Significant Reduction in the Plasma Levels and Gene Expression of CCL2 in Patients with Osteoarthritis following Intervention with Krocina™.

inflammatory chemokines such as CCL2 and CCL5 are involved in the progress of osteoarthritis. Crocin with antioxidant and anti-inflammatory properties can reduce the symptoms of osteoarthritis (OA). This study was performed investigate the effect of Krocina™, on the gene expressions and plasma levels of CCL2 and CCL5 in OA patients. The study included 35 patients that were randomized in the Krocina™ and placebo groups. The intervention was Krocina™ 15mg daily for four months. Clinical and paraclinical parameters were measured. CCL2 and CCL5 genes expression and plasma levels were determined using the SYBR Green Real-Time RT-PCR and Enzyme-linked Immunosorbent Assay (ELISA) techniques. The C-reactive protein (CRP) value in the Krocina™ group and the visual analogue scale (VAS) value in the Krocina™ and placebo groups decreased significantly after the intervention. The gene expression of CCL2 in the Krocina™ and placebo groups decreased significantly. On the contrary, the gene expression of CCL5 in the Krocina™ and placebo groups increased significantly. Moreover, the plasma levels of CCL2 in the Krocina™ and placebo groups decreased meaningfully. There was no difference regarding the plasma levels of CCL5 within the Krocina™ and placebo groups before and after the intervention in either of the groups. Administration of Krocina™ reduced the clinical signs of inflammation and CRP and VAS value. Also, Krocina™ significantly decreased the plasma levels and gene expression of CCL2 in osteoarthritis patients.

Upregulation of Neurotrophic Factors and Myelin Basic Protein in Schwann-like Cells by T3 Hormone Following Transdifferentiation of Human Adipose-derived Stem Cells.

Peripheral nerve regeneration is a complicated phenomenon. Thyroid hormones are known as critical regulators in the nervous system development. The Schwann cells have the regenerative potency in the peripheral nervous system. In this study, the human adipose-derived stem cells were assessed in vitro, for transdifferentiation potency into Shwann-like cells (SLCs) as a candidate source for clinical cell therapy, under the treatment of triiodothyronine (T3) hormone, and compared with the untreated cells. The cell viability rate, myelination and neurotrophic factors expression of SLCs were evaluated two weeks post- induction by MTT assay, immunocytochemistry and real-time RT-PCR techniques, respectively. The obtained results revealed a significant decrease in SLCs viability, compared to the adipose-derived stem cells (P < 0.001). Immunocytochemistry technique was applied to detect SLCs markers, such as S100β, GFAP and myelin basic proteins (MBP) in the presence and absence of T3 treatment. The results indicated that administering T3 can significantly increase the differentiation and myelination potency of SLCs (P < 0.01). The findings of real-time RT-PCR technique indicated that the expression of Schwann cells markers, MBP, brain-derived neurotrophic factor and glial cell-derived neurotrophic factor were upregulated significantly with T3 hormone administration in comparison with the untreated cells (P < 0.05). The SLCs were able to express the neurotrophic factors and myelination related genes in the presence of T3 hormone. Furthermore, T3 administration improved myelination potency of adipose-derived stem cells, in vitro. Further in vivo experiments are necessary to confirm the advantages of using a combination of autologous SLCs and T3 hormone for peripheral nerve injury recovery.

Serum microRNA-486-5p expression in obese Egyptian children and its possible association with fatty liver

AimsSeveral microRNAs (miRNAs) are involved in regulating the process of adipogenesis. White adipose tissue is a major source for these miRNAs. We aimed to evaluate the expression of miR-486-5p in children with obesity and its possible association with nonalcoholic fatty liver disease (NAFLD). MethodThis case-control study included 100 obese and overweight children and 100 normal-weight children of matched age and sex. All children were subjected to anthropometric measurements and evaluation of miR-486-5p expression levels using the SYBR green-based real-time RT-PCR technique. ResultsObese children showed significantly up-regulated miR-486-5p gene expression (p value < 0.001) when compared to control group. MiR-486-5p gene expression showed significant positive correlation with weight (r = 0.924), BMI (r = 0.497), waist circumference (r = 0.387), fat mass (r = 0.361), LDL(r = 0.351), TG (r = 0.867), TC (r = 0.875) and presence of fatty liver (r = 0.760). The best cutoff value of miR-486-5p gene expression in the prediction of obesity was 0.44 with AUC 0.736 that has a sensitivity 60% and specificity 90%, ConclusionThe serum level of the miR-486-5p gene is up-regulated in obese and overweight children and might be an independent predictor for obesity and fatty liver susceptibility.

MiRNAs through β-ARR2/p-ERK1/2 pathway regulate the VSMC proliferation and migration

BackgroundmiRNAs are involved in plaque formation of atherosclerosis and vessel restenosis. In this study, we investigated the effects of miR-599, miR-204, and miR-181b on VSMC proliferation, and migration through TGFβ receptor 2 (TGFβR2), β-arrestin 2 (β-ARR2), SMAD2/p-SMAD2, and ERK1/2/p-ERK1/2. Materials & methodsGenes and miRNAs were predicted by bioinformatics tools and were transfected by PEI-miRNAs (miR-599, miR-204, and miR-181b) complexes into VSMCs. The gene and protein expression levels were evaluated by real-time RT-PCR and western blotting techniques, respectively. The VSMC proliferation and migration were studied by MTT and scratch assay, respectively. ResultsThe miR-181b and miR-204 downregulated significantly β-ARR2 gene and protein expression levels and p-ERK1/2 values. Moreover, TGFβR2 gene and protein expression levels and p-SMAD2 values were not significantly affected by miR-181b and miR-204. The VSMC proliferation (p = 0.0019, p = 0.0054, respectively) and migration (p < 0.0001, p < 0.0001, respectively) were inhibited by the miR-181b and miR-204. The miR-599 inhibited VSMC proliferation (p = 0.044) and migration (p = 0.0055) but it did not affect significantly the β-ARR2 and TGFβR2 gene and protein expression levels. ConclusionThe results suggested that the inhibitory effects of miR-181b and miR-204 on VSMC proliferation and migration are mediated by the β-ARR2/p-ERK1/2 pathway. Since VSMC proliferation and migration are involved in plaque growth, therefore this pathway can be a therapeutic target for atherosclerosis.

Changes in the Expression of SGO1 and SGO1-AS1 Genes in Colorectal Tumor Tissues, Compared to Healthy Tissues

Background and Aim: The protein encoded by the SGO1 gene is a member of the shugoshin family of proteins and protects the centromere during mitosis. lncRNAs are non-coding RNA with 200 nucleotides lengths, i.e., involved in regulating gene expression. The current study aimed to evaluate the expression of SGO1 and SGO1-AS1 in different stages of disease progression; we also compared their expression pattern in tumor tissues with healthy tissues in colorectal cancer patients. Methods &amp; Materials: In total, 40 tissue samples of patients with colorectal cancer were reported according to the examination and criteria with the approval of a pathologist. Besides, 40 normal tissues were sampled from a completely healthy part of the intestine of the same patients. After RNA extraction and cDNA synthesis, the Real-time RT-PCR technique was used to evaluate the expression of the desired genes in the study groups. ROC curve analysis was also used to determine the ability of each selected gene to diagnose the disease. Ethical Considerations: This study was approved by the Ethics Committee of Shahrekord Azad University (Code: IR.IAU.SHKREC.1398.020). Results The obtained data suggested that SGO1 significantly decreased in the colorectal cancer tumor samples (P&lt;0.001) and SGO1-AS1 LncRNA significantly increased expression, compared to adjacent healthy tissues. Additionally, in the age group of below 60 years, compared to the age group of over 60 years, SGO1 expression increased and SGO1-AS1 expression decreased. Based on the AUC obtained from the ROC diagram, it was found that the SGO1 gene with AUC=0.8041 and SGO1-AS1 with AUC=0.6364 could significantly distinguish a healthy population from patients with colorectal cancer. Conclusion: According to the collected results, SGO1 -AS1 and SGO1 were significantly reduced and increased in tumor tissue, respectively; however, only the SGO1 gene was introduced as a good marker for diagnosing colorectal cancer.

Differential inflammatory responses associated with Leishmania major and Ltropica in culture.

Anthroponotic cutaneous leishmaniasis (ACL) due to Leishmania tropica and zoonotic CL (ZCL) due to Lmajor have different clinical and epidemiological features. To determine whether pro-inflammatory cytokines are involved in diverse pathogenicity of Leishmania species causing CL. The capacity of Lmajor/Ltropica to modulate expression of IL-1β, IL-8 (CXCL8), IFN-γ, TNF-α and MCP-1 (CCL2) in peripheral blood mononuclear cells (PBMCs) and monocyte-derived macrophages (MDMs) was evaluated by real-time RT-PCR technique. PBMCs from both ZCL and ACL cases expressed significantly higher IFN-γ (P<.001) and TNF-α (P<.05) compared with healthy controls (HC). PBMCs from ACL patients expressed significantly higher IL-1β and IL-8 compared with ZCL patients and HC when stimulated with live Lmajor or Ltropica promastigotes (P<.001). After 4 and 10hours, Lmajor-infected MDMs expressed significantly higher IFN-γ (P<.05), and after 10hours, Ltropica-infected MDMs expressed significantly higher IL-1β, IFN-γ and IL-8 compared with noninfected cells (P<.05). This study shows differential parasite-mediated stimulations of the inflammatory response with Lmajor vs Ltropica ex vivo. Pro-inflammatory cytokines particularly IL-8 (CXCL8) and IL-1β might contribute in diverse clinical features of CL such as longer duration of lesion persistence in ACL patients.