nasrin rastegarvand
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Background
MicroRNAs (miRNAs) regulate the immune response to infectious diseases. They exhibit alterations during the early stages of disease development, even before the pathogen is detectable.
ObjectivesThe objective of this study was to analyze the expression levels of miR-1307-3p and miR-3613-5p at the onset of COVID-19, aiming to identify miRNAs that could function as early predictive biomarkers.
MethodsThis case-control study analyzed the expression levels of miR-1307-3p and miR-3613-5p at the onset of COVID-19 to identify early predictive biomarkers. Plasma miRNAs from 26 COVID-19 patients hospitalized in the ICU ward were sequenced and compared to a healthy control group of 26 individuals. Additionally, plasma biomarkers associated with the immune system were evaluated. Total RNA was extracted using an RNX solution and subjected to cDNA synthesis with a specific loop sequence for each miRNA. The samples were subsequently analyzed using relative real-time PCR.
ResultsRT-qPCR analysis revealed a significant reduction in the detection rate of miR-1307-3p (P < 0.0001) in the patient group compared to the control group. Conversely, no significant differences were observed in the levels of miR-3613-5p between the patient and control groups.
ConclusionsmiR-1307-3p shows potential as a biomarker for the diagnosis of COVID-19.
Keywords: COVID-19 Patients, Micrornas, Gene Expression -
مقدمه
انتقال ژن، یک ابزار قدرتمند برای تغییر عملکرد سلول ها است که از کاربردهای آن می توان به درمان انواع بیماری ها اشاره کرد. با این وجود، مطالعات مختلف نشان می دهد که علیرغم وجود تنوع در روش های انتقال ژن، محدودیت های قابل توجهی در کاربرد هر روش وجود دارد. هدف از این مطالعه، مقایسه دو روش انتقال ژن با استفاده از ترکیبات PEI و لیپوفکتامین در سلول های سرطانی است.
مواد و روش هادر این مطالعه، از Caco-2 و HCT-116 به عنوان سلول های هدف و از سلول HEK-293 نیز به عنوان سلول کنترل استفاده شد. بدین منظور، ابتدا انتقال ژن با استفاده از دو ماده PEI و لیپوفکتامین انجام شد و سپس نتایج آنها بر اساس بیان پروتیین GFP با یکدیگر مورد مقایسه قرار گرفت.
یافته هانتایج این مطالعه نشان داد که انتقال ژن در سلول های HEK-293، به میزان مطلوبی صورت گرفته است، اما این فرآیند در سلول های رده کلورکتال بسیار ناچیز بود. همچنین در این میان، تفاوت هایی نیز بین میزان انتقال ژن در سلول های کلورکتال مشاهده شد؛ به گونه ای که میزان ترنسفکت در سلول های Caco-2 از سلول های HCT-116 کمتر بود.
بحث و نتیجه گیریمیزان پذیرش ژن خارجی در سلول ها، به عوامل متعددی از جمله نوع و منشا سلول وابسته است. از آنجاییکه سلول های سرطانی دستخوش تغییرات متعدد ژنتیکی شده اند، در نتیجه دستکاری ژنتیکی آنها با محدودیت های بسیاری همراه است. به همین منظور توصیه می شود برای دستیابی به میزان بالای انتقال ژن در این سلول ها، از سایر روش ها مانند گزینش سلول به وسیله آنتی بیوتیک و تهیه رده سلولی پایدار استفاده شود.
کلید واژگان: ترنسفکشن، لیپوفکتامین، سرطان، کلورکتال، سیستم انتقال ژنYafteh, Volume:25 Issue: 3, 2023, PP 23 -35BackgroundGene transfection is a powerful tool for changing cell function, which is used for treating various diseases. Nevertheless, various studies show that despite the variety of gene transfer methods, there are significant limitations in the application of each method. The present study aimed to compare two methods of gene transfer using PEI and Lipofectamine compounds in cancer cell.
Materials and MethodsIn this study, Caco-2 and HCT-116 were used as target cells, and HEK-293 cells were used as control cells. For this purpose, initially, gene transfer was performed using PEI and Lipofectamine, and then their results were compared with each other based on the expression of GFP protein.
ResultsThe results of this study showed that although gene transfer was carried out in HEK-293 cells at a favorable rate, this process was very insignificant in colorectal cells. In addition, in the meantime, some differences were observed between the amounts of gene transfer in colorectal cells, so that the amount of transfection in Caco-2 cells was lower than that of the HCT-116 cells.
ConclusionThe degree of acceptance of foreign genes in cells depends on several factors, including the type and origin of the cell. Since cancer cells have undergone many genetic changes, their genetic manipulation is associated with many limitations. For this reason, it is recommended to use other methods, such as cell selection by antibiotics and preparation of stable cell lines to reach a high level of gene transfer in these cells.
Keywords: Cancer, Colorectal, Gene delivery system, Lipofectamine, Transfection -
ObjectivesThe aim of this study was to evaluate the etiologic agents of a hand, foot, and mouth disease (HFMD) outbreak in Ahvaz, Southwest Iran and their evolutionary analysis by phylogenetic construction and Simplot analysis.MethodsWe collected 16 serum samples in an outbreak of HFMD in Ahvaz in October 2013. RNA was extracted from the samples and subjected to reverse transcription polymerase chain reaction for detection of Enterovirus group A and B. Positive cases were sequenced and subjected to phylogenetic and Simplot analysis for detecting the signs of recombination.ResultsOf the 16 specimens, nine (56.25%) were PCR-positive with the universal primers for the Enterovirus 5’UTR region. Coxsackievirus A6 was detected as a predominant agent of HFMD with two cases of Echovirus 6 and Echovirus 30. In the case of Echovirus 6, the signs of recombination in the 5’UTR region were observed based on phylogenetic and Simplot analysis.ConclusionsCoxsackievirus A6 is the main agent of HFMD in Ahvaz. The evidence of recombination in this isolate of Echovirus 6 in this study emphasizes common hygiene practices and sanitation to prevent the circulation of this isolate in community and the advent of new strains.Keywords: Hand, Foot, and Mouth Disease, Polymerase Chain Reaction, Echovirus 6, Echovirus 30, Coxsackievirus A6, Co-circulation
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BackgroundHepatitis B virus (HBV) is a major problem for healthcare workers worldwide, and among them, dentists are at risk of acquiring HBV infection. The prevalence of HBV infection has been reported among the dentists in different regions of the world. Since none of the available drugs can clear HBV infection, the presence of effective immunity against HBV infection is important to prevent HBV infection.ObjectivesThis study aimed at determining HBs antibody and specific HBV gamma interferon among the dentists, who received hepatitis B vaccine.MethodsThe blood samples were collected from 40 dentists, including 7 endodontics, 2 oral and maxillofacial radiologist, 4 periodontics, 11 oral and maxillofacial surgeons, 6 implantologists, 3 orthodontics, 1 oral and maxillofacial pathologist, 2 esthetic and restorative dentists, and 4 doctors of dental surgery (DDS) at from dental college of Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran during December, 2013. Overall, 31 (77.5%) dentists had already received 3 doses of recombinant hepatitis B vaccine, and 9 (22.5%) had received only two doses of the vaccine. Their sera were tested for HBsAb and anti-HBc-IgG by the Enzyme Linked Immunosorbent Assay (ELISA) test. The lymphocyte of individuals was separated from their blood sample by Ficoll-Hypaque, cells were washed with phosphate buffered saline (PBS) by centrifugation, and finally the pellet cells was resuspended in RPMI-1640 media. Separated cells were exposed to 2.5 µg of purified recombinant HBs antigen, and supernatants were collected after 72 hours and tested for detection of specific interferon γ level by ELISA test.ResultsOverall, 97.5% of dentists showed positive HBs antibody test results while 36 showed (90%) positive test results for specific interferon γ against hepatitis B virus infection.ConclusionsHigh coverage of 97.5% immune response against hepatitis B infection was found, indicating high efficacy of recombinant HBV vaccine among the dentists.Keywords: Hepatitis B Virus_Hbsab_Interferon Gamma_Health Care Workers
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Background
Health care workers are at high risk of acquiring hepatitis B virus (HBV) infection through occupational exposure to blood or body fluids. Thus, the assessment of anti-HBs status after immunization is very important.
ObjectivesThis study aimed to evaluate the measurement of HBsAb titer and specific gamma interferon response among the vaccinated health care workers in Golestan Hospital, Ahvaz, Iran.Patients and
MethodsThe blood samples of 39 health care workers, including 13 general surgeons, 10 anesthesiologists, 5 neurosurgeons, 3 general physicians, 1 orthopedist, 2 urologist and 5 nurses were collected during June 2013. All the participants had received HBV vaccine. They had received last vaccine dose from 2 months to 14 years ago. Their sera were tested for anti-hepatitis B antibody and HBc-IgG by the ELISA. Also, the evaluation of specific interferon γ response against HBsAg was carried out using ELISA test. The age of health care workers were between 24 and 58 years with the mean age of 34.3 ± 7.4 y.
ResultsOut of 39 sera, 22 (56.41%) had HBsAb titer above 100 IU/mL, 17 (43.6%) had titer below 100 IU/mL, 27 (69.2%) had positive specific HBsAg interferon γ, 8 (20.5%) cases had positive antibody response above 100IU, but negative for specific interferon γ and 3 (7.6%) cases were positive for HBc-IgG.
ConclusionsOverall, 87.2% of the health care workers had immunity against HBV infection, which showed remarkable immunity response following HBV vaccination. Booster dose of HBV vaccine is recommended for those whose immunity are below 100 IU/mL.
Keywords: Hepatitis B Virus_Interferon Gamma_Workers
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