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Abbas Kazemi Ashtiani , Peyman Khoshnood , Mohammad Javad Fatemi , Seyed Jaber Mousavi , Seyed Aboozar Hoseini ,
Volume 74, Issue 10 (1-2017)
Volume 74, Issue 10 (1-2017)
Abstract
Background: The use of random flaps is one of the most common methods of reconstructive surgery because they are easy to use and quick to do. However, the absence of axial vessels especially in the distal areas can cause ischemia and loss of total or part of the flap. Different methods and systemic and topical medications have been recommended to prevent ischemia in random flaps. The aim of this study was to evaluate the effect of stem cells derived from umbilical cord blood in random flap survival in rats.
Methods: This experimental study was conducted in Animal Laboratory of Hazrat Fatemeh Hospital in 2012. In this study twenty Sprague-Dawley male rats weighing approximately 300 to 350 g were selected and divided randomly into two groups. In both groups after anesthesia, a flap was created in the posterior part of each rat with a size of 2 x 6 cm. In the intervention group we injected stem cells derived from umbilical cord blood into the flap, and after eight days the effects on the survival of flaps were examined by digital photography and then pathological examination was performed.
Results: The mean of viable flap in the stem cell group was 6.57 cm2 and in control group 4.71 cm2. The minimum and maximum flap survival in the intervention group were 4.71 and 8.75, and the minimum and maximum flap survived in control group were 1.86 and 7.77. This difference was significant and showed that the viable parts of flap were more in the intervention group (P=0.49). In pathologic examinations epidermal and muscle necrosis of the skin were reported in 3 cases in the intervention group and 5 cases in control group.
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Conclusion: This study showed that cord blood stem cells can be effective somehow in reducing ischemia and increasing random flap survival. However, similar studies are recommended in order to compare the results of this drug and placebo or other proven effective drugs. |
Elham Hoveizi , Tayebeh Mohammadi ,
Volume 74, Issue 11 (2-2017)
Volume 74, Issue 11 (2-2017)
Abstract
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Background: One of the major causes of death in the world is cancer and therefore any study in the field of cancer biology is of great importance. Head and neck cancers represent approximately 2-5% of neoplasms which is higher in some countries. The most appropriate therapy for various cancers is identifying effective and efficient ways that contribute to initiation of apoptosis. Cyclophosphamide is an alkylating agent that stops the replication of DNA and then, it stops the cell proliferation and viability. Therefore, cyclophosphamide is used to treat various types of cancer. In this study we evaluate the cytotoxic effects of cyclophosphamide on viability of (head and neck cancer cells) HN5 cell line and compare it with fibroblast cells as noncancerous cells. Methods: This experimental study was done in cell and developmental laboratory in faculty of science, Shahid Chamran University of Ahvaz in Spring of 2016. HN5 cell line and embryonic fibroblast cells were grown in DMEM supplemented with 10% fetal bovine serum (FBS), penicillin/streptomycin (100 U/ml, 100 µg/ml) at 37 °C, then the effects of different concentrations of cyclophosphamide on cell viability was evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. 4',6-diamidino-2-phenylindole (DAPI) staining was performed to determine the proportion of apoptotic cells by manually counting pyknotic nuclei. According to standard procedures from day 13 embryos of outbred strains naval medical research institute (NMRI), fibroblast cells were isolated. In this study HN5 cell line and fibroblasts were exposed to cytostatics for 72 hours. Results: Various concentrations of cyclophosphamide were effective in cytotoxicity of HN5 cancer and fibroblast cells. A significant cytotoxicity was observed with the examined concentration of 1 µg/ml of cyclophosphamide with 50% in 3th day and P< 0.001. Interestingly, at low concentrations, cyclophosphamide was more toxic than at higher concentrations. Conclusion: Totally cyclophosphamide had low toxicity effects on both of the cell lines but the toxicity effects of cyclophosphamide on HN5 were significantly greater than fibroblast cells. These results indicate that cyclophosphamide can be a potential anticancer agent. |
Mohsen Sheykhhasan , Mohsen Nikbakht , Mahdieh Ghiasi ,
Volume 74, Issue 11 (2-2017)
Volume 74, Issue 11 (2-2017)
Abstract
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Intervertebral disks (IVD) acts as shock absorber between each of the vertebrae in the spinal column by keeping the vertebrae separated when the shock caused by the action. They also serve to protect the nerves that run down the middle of the spine and intervertebral disks. The disks are made of fibrocartilaginous material. The outside of the disk is made of a strong material called the annulus fibrosus. Inside this protective covering is a jelly-like substance known as mucoprotein gel. This interior is known as the nucleus pulposus. The nucleus pulposus consists of large vacuolated notochord cells, small chondrocyte-like cells, collagen fibrils, and aggrecan, a proteoglycan that aggregates by binding to hyaluronan. Attached to each aggrecan molecule are glycosaminoglycan (GAG) chains of chondroitin sulfate and keratan sulfate. Intervertebral disks degeneration is frequently associated with low back and neck pain, which accounts as a disability. Despite the known outcomes of the Intervertebral disks degeneration cascade, the treatment of IVD degeneration is limited in that available conservative and surgical treatments do not reverse the pathology or restore the IVD tissue. Regenerative medicine for IVD degeneration, by injection of Intervertebral disks cells, chondrocytes or stem cells, has been extensively studied in the past decade in various animal models of induced IVD degeneration, and has progressed to clinical trials in the treatment of various spinal disease. Despite preliminary results showing positive effects of cell-injection strategies for IVD regeneration, detailed basic research on Intervertebral disks cells and their niche demonstrates that transplanted cells are unable to survive and adapt in the avascular niche of the IVD. For this therapeutic strategy to succeed, the indications for its use and the patients who would benefit need to be better defined. To surmount these obstacles, the solution will be identified only by focused research, both in the laboratory and in the clinic. In present paper, the potential utilization of different adult stem cells for intervertebral disc regeneration has been reported. Bone marrow mesenchymal stem cells, adipose tissue derived stem cells, synovial stem cells and committed IVD cells have been studied for this purpose either in vitro or in vivo. |
Mohammd Javad Fatemi , Shirin Chehroudi , Tooran Bagheri , Sahar Saleh , Amir Atashi , Mohsen Saberi , Seyed Aboozar Hoseini , Shirin Araghi ,
Volume 74, Issue 12 (3-2017)
Volume 74, Issue 12 (3-2017)
Abstract
Background: Acute and chronic wound healing has always been problematic. Stem cells with or without the scaffold carrying these cells have been proposed as new methods in the treatment of wounds. In this case study we have tried to examine the effect of scaffold made of polyether sulfone (PES) alone, with stem cells and along with stem cell and growth factor on wound healing in rats.
Methods: This experimental study was conducted in Animal Laboratory of Hazrat Fatemeh Hospital in 2012. In this study, 48 rats were randomly divided into four groups. A wound created on the back of each rat at the size of 3×3 cm. The surface of the wound in the first group is covered with PES seeded with adipose-derived stem cell (ASC) and growth factor (GF), in the second group with polyether Sulfone seeded with ASC, in the third group only with PEWS, and in the fourth group (control) with Vaseline gauze. On 20th and 35th days, the surface of the wound was assessed by photography in order to understand the process of healing. In addition, on days 20 and 45, the histopathology characteristics of the samples were studied with a biopsy of the wounds.
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Results: The Results of wound healing in the control group was better than the other groups and its statistical difference between others was meaningful. (P=0.008, P=0.013, P=0.001) On day 20, by examining histopathological characteristics including epithelialization, the number of inflammatory cells, the amount of angiogenesis and collagen synthesis in control group, we gained better results. (P=0.000), But on day 45, the results in different parameters were not equal. Conclusion: polyether sulfone scaffold alone or with adipose-derived stem cells couldn’t improve the process of wound healing. Also adding vascular endothelial growth factor (VEGF) did not change the results significantly. |
Homa Mohseni Kouchesfahani , Somayeh Ebrahimi-Barough , Jafar Ai , Azam Rahimi ,
Volume 74, Issue 12 (3-2017)
Volume 74, Issue 12 (3-2017)
Abstract
Background: Small molecule Purmorphamin (PMA) is the agonist of smoothened protein in Sonic hedgehog (Shh) signaling pathway. Effect of purmorphamin small molecule on differentiation of mesenchymal cells into bone tissue has been studied previously. Use of Shh causes progression of neural differentiation, and the differentiated cells express specific neural markers. Neurofilament (NF) and acetylcholine esterase (Chat) are specific markers of motor neurons and their expression in differentiated cells indicates their conversion into motor neurons. The aim of this study was to evaluate the ability of PMA to differentiate the human endometrial stem cells (hEnSCs) into motor neurons.
Methods: This analytical study was done in Tehran University of Medical Sciences laboratory on September of 2015. In this study hEnSCs were enzymatically extracted from endometrial tissue. After third passages, the flow cytometry was done for mesenchymal stem cells markers. The mesenchymal stem cells were divided into control and differentiated groups. FBS 10%+DMEM/F12 was added to the culture medium of control group and the differentiating group was treated with differentiating medium containing N2, PMA, DMEM/F12, FBS, B27, IBMX, 2ME, FGF2, RA, BDNF. After 21 days immunocytochemistry (ICC) test was done for the expression of NF and Chat proteins and Real-time PCR analysis for expression of neural markers such as NF, Chat, Nestin and GFAP (as glial marker) at mRNA level.
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Results: The flow cytometry analysis showed that hEnSCs were positive for mesenchymal markers CD90, CD105 and CD146 and negative for endothelial marker CD31, and hematopoietic marker CD34. The immunocytochemistry and Real time-PCR results showed that the cells treated with PMA expressed motor neuron markers of NF and Chat. Conclusion: According to the results of this study, it can be concluded that small molecule PMA has the potency to induce the differentiation of hEnSCs into neural cells, specifically motor neurons by activating Shh signaling pathway. |
Maryam Ataie , Atefeh Solouk , Fatemeh Bagheri , Ehsan Seyed Jafari,
Volume 75, Issue 4 (7-2017)
Volume 75, Issue 4 (7-2017)
Abstract
An increase in the average age of the population and physical activities where the musculoskeletal system is involved as well as large number of people suffering from skeletal injuries which impose high costs on the society. Bone grafting is currently a standard clinical approach to treat or replace lost tissues. Autografts are the most common grafts, but they can lead to complications such as pain, infection, scarring and donor site morbidity. The alternative is allografts, but they also carry the risk of carrying infectious agents or immune rejection. Therefore, surgeons and researchers are looking for new therapeutic methods to improve bone tissue repair. The field of tissue engineering and the use of stem cells as an ideal cell source have emerged as a promising approach in recent years. Three main components in the field of tissue engineering include proper scaffolds, cells and growth factors that their combination leads to formation of tissue-engineered constructs, resulting in tissue repair and regeneration. The use of scaffolds with suitable properties could effectively improve the tissue function or even regenerate the damaged tissue. The main idea of tissue engineering is to design and fabricate an appropriate scaffold which can support cell attachment, proliferation, migration and differentiation to relevant tissue. Scaffold gives the tissue its structural and mechanical properties, for instance flexibility and stiffness that is related with the tissue functions. Biomaterials used to fabricate scaffolds can be categorized into natural or synthetic biodegradable or non-biodegradable materials. Polymers are the most widely used materials in tissue engineering. Growth factors are a group of proteins that cause cell proliferation and differentiation. Two main cell sources are specialized cells of desired tissue and stem cells. However, according to the low proliferation and limited accessibility to the cells of desired tissue, stem cells are better suggestion. Combination of mesenchymal stem cells harvested from bone marrow, adipose tissue and cord blood with proper scaffolds and growth factors could be a useful method in treatment of skeletal injuries. In this review paper, we focus on the application of mesenchymal stem cells in the repair of damaged bone, cartilage, meniscus, ligaments, tendons and spine tissue.
Mohammad Molavi , Rasoul Yousefi Mashouf , Mohammad Yousef Alikhani , Hassan Mahmoudi , Alireza Zamani , Fariba Keramat , Sara Soleimani Asl ,
Volume 75, Issue 6 (9-2017)
Volume 75, Issue 6 (9-2017)
Abstract
Background: Brucellosis is a systemic infection caused by gram-negative coccobacilli and facultative intracellular bacteria of the genus brucella. Brucellosis is a bacterial disease common to humans and livestock. Infection with Brucella spp. continues to pose a human health risk globally despite strides in eradicating the disease from domestic animals. The humoral and cellular immunity plays an important role in brucellosis. The effect of phagocytosis and cellular immunity has been demonstrated in brucellosis. The effect of cupping as a therapeutic method on bacterial diseases has been demonstrated. By considering this fact that cupping is an effective method on host immune system functions and has potential effect to regulate the inflammatory reactions.
Methods: This experimental study was carried out on 48 rats in 6 groups (48 rats were divided into 6 groups) during the first 6 months from January 2015 to July 2016 in the laboratory of microbiology and animal of Hamedan University of Medical Sciences. The rats were infected through the injection of Brucella melitensis with plenty 5×105 (cfu/ml) of bacteria. After a week, in order to approve the accuracy of the infection in the studied rats inoculated with coombos Brucella melitensis, Wright test, 2-Mercaptoethanol test and Coombs' Wright test were used. The rats were then treated with cupping method in their sacral area. IFN-γ was measured using enzyme-linked immunosorbent assay. And the study of tissues using hematoxylin and eosin (H&E) Staining.
Results: The results of this study showed that cupping leads to an increase in the mean serum level of interferon-gamma. The histopathological study of liver tissue showed that the radial arrangement was not observed in the infected group with brucellosis and the cells were acidophilus. While, the radial arrangement was observed in treated rats with cupping, but it was not complete. The number of enlarged sinusoids were reduced. But, cell infiltration was observed.
Conclusion: This study showed that cupping can increase serum level of IFN-γ, and thus help to the clearance of disease and improvement of injury in the brucellosis animals lab.
Methods: This experimental study was carried out on 48 rats in 6 groups (48 rats were divided into 6 groups) during the first 6 months from January 2015 to July 2016 in the laboratory of microbiology and animal of Hamedan University of Medical Sciences. The rats were infected through the injection of Brucella melitensis with plenty 5×105 (cfu/ml) of bacteria. After a week, in order to approve the accuracy of the infection in the studied rats inoculated with coombos Brucella melitensis, Wright test, 2-Mercaptoethanol test and Coombs' Wright test were used. The rats were then treated with cupping method in their sacral area. IFN-γ was measured using enzyme-linked immunosorbent assay. And the study of tissues using hematoxylin and eosin (H&E) Staining.
Results: The results of this study showed that cupping leads to an increase in the mean serum level of interferon-gamma. The histopathological study of liver tissue showed that the radial arrangement was not observed in the infected group with brucellosis and the cells were acidophilus. While, the radial arrangement was observed in treated rats with cupping, but it was not complete. The number of enlarged sinusoids were reduced. But, cell infiltration was observed.
Conclusion: This study showed that cupping can increase serum level of IFN-γ, and thus help to the clearance of disease and improvement of injury in the brucellosis animals lab.
Razieh Zarifian Yeganeh , Abbas Shakoori Garakani , Saman Mehrabi , Nader Ebadi, Maziar Motiee Langroudi , Mohammad Reza Noori Daloii,
Volume 75, Issue 7 (10-2017)
Volume 75, Issue 7 (10-2017)
Abstract
Background: Head and neck squamous cell carcinoma (HNSCC) is the malignancy of squamous cells (the epidermal layer of skin) in cavities in head and neck includes: larynx, pharynx, paranasal sinuses and oral cavity. The main goal of this research was to understand the effect of mutations in two important genes (KRAS and BRAF) in RAS/MAP kinase (EGFR) signaling pathway in tumor cells with head and neck squamous cell carcinoma in Iran.
Methods: The present cross-sectional study performed from October 2015 to September 2016 on 40 patients suffering from head and neck squamous cell carcinoma, all confirmed by pathology department of Imam Khomeini hospital. Tumor samples were achieved from the surgical cancer department of Imam Khomeini hospital and stored in liquid nitrogen until starting tests. The tests done in genetic laboratory of Tehran University of Medical Sciences. Techniques we used in this research, were DNA extraction based on phenol-chloroform approach, Multiplex PCR (M-PCR) to amplify mentioned exons and KRAS/BRAF strip assays to detect mutations in mutated hotspots in exon 2 of KRAS and codon V600E in BRAF gene.
Results: In this study, we observed 7 mutations in codons 12 and 13 exon 2 in KRAS gene (about 17.5%) and 4 mutations in codon V600E in BRAF gene (about 10%) of obtained tumor samples. The hotspot mutation in codon 12 were Asp (10%) and Ser (5%) respectively. In BRAF, the most common mutation, as we expected according to other researches, was observed in codon V600E. We also observed that 29 people of these patients were male (about 72.5%) and 11 patients were female (about 27.5%). Moreover, 28 patients were over 50 years, while 7 patients were below the age of 50.
Conclusion: The results of this study showed that mutations in genes KRAS and BRAF especially in studied hotspots, and the effects on their molecules in EGFR signaling pathway are important in involving head and neck squamous cell carcinoma, as other cancers. These findings may be considered in choosing drugs for targeted chemotherapy.
Methods: The present cross-sectional study performed from October 2015 to September 2016 on 40 patients suffering from head and neck squamous cell carcinoma, all confirmed by pathology department of Imam Khomeini hospital. Tumor samples were achieved from the surgical cancer department of Imam Khomeini hospital and stored in liquid nitrogen until starting tests. The tests done in genetic laboratory of Tehran University of Medical Sciences. Techniques we used in this research, were DNA extraction based on phenol-chloroform approach, Multiplex PCR (M-PCR) to amplify mentioned exons and KRAS/BRAF strip assays to detect mutations in mutated hotspots in exon 2 of KRAS and codon V600E in BRAF gene.
Results: In this study, we observed 7 mutations in codons 12 and 13 exon 2 in KRAS gene (about 17.5%) and 4 mutations in codon V600E in BRAF gene (about 10%) of obtained tumor samples. The hotspot mutation in codon 12 were Asp (10%) and Ser (5%) respectively. In BRAF, the most common mutation, as we expected according to other researches, was observed in codon V600E. We also observed that 29 people of these patients were male (about 72.5%) and 11 patients were female (about 27.5%). Moreover, 28 patients were over 50 years, while 7 patients were below the age of 50.
Conclusion: The results of this study showed that mutations in genes KRAS and BRAF especially in studied hotspots, and the effects on their molecules in EGFR signaling pathway are important in involving head and neck squamous cell carcinoma, as other cancers. These findings may be considered in choosing drugs for targeted chemotherapy.
Mohsen Sheykhhasan , Mahdieh Ghiasi ,
Volume 75, Issue 9 (12-2017)
Volume 75, Issue 9 (12-2017)
Abstract
Stem cells are undifferentiated biological cells that can differentiate into more specialized cells and divide (through mitosis) to produce more stem cells (self-renew). In mammals, there are two broad types of stem cells: embryonic stem cells, which are isolated from the inner cell mass of blastocysts, and adult stem cells, which are found in various tissues. Mesenchymal stem cells (MSCs) are multipotent cells that are called as one of the most adult stem cells. Due to their highly proliferative potential and their suitable self-renewal capacity, these cells have provided a powerful and promising source for use in the field of regenerative medicine. Also, mesenchymal stem cells are known for their important properties involving multilineage differentiation potential, trophic factor secretion and localization along various organs and tissues. So that MSCs can differentiate into a variety of cell lineages, including: Osteoblasts (bone cells), chondrocytes (cartilage cells), adipocytes (fat cells), myocytes (muscle cells), hepatocytes (liver cells) and endothelial cells. Efficacy of differentiated MSCs to regenerate cells in the injured tissues requires the ability to maintain the differentiation toward the desired cell fate. Since MSCs represent an attractive source for autologous transplantation, cellular and molecular signaling pathways and micro-environmental changes have been studied in order to understand the role of cytokines, chemokines, and transcription factors on the differentiation of MSCs. The differentiation of MSC into a mesenchymal lineage is genetically manipulated and promoted by specific transcription factors associated with a particular cell lineage. Recent studies have explored the integration of transcription factors, including Runx2, Sox9, PPARγ, MyoD, GATA4, and GATA6 in the differentiation of MSCs. Therefore, the overexpression of a single transcription factor in MSCs may promote trans-differentiation into specific cell lineage, which can be used for treatment of some diseases. In this review, we critically discussed and evaluated the role of transcription factors and related signaling pathways that affect the differentiation of MSCs toward adipocytes, chondrocytes, osteocytes, skeletal muscle cells, cardiomyocytes, and smooth muscle cells.
Mohsen Sheykhhasan , Mahdieh Sadat Ghiasi ,
Volume 76, Issue 5 (8-2018)
Volume 76, Issue 5 (8-2018)
Abstract
The cartilage is a connective tissue that, due to the strength of its extracellular matrix, allows the tissue to tolerate mechanical stress without undergoing permanent deformation. It is responsible for the support of soft tissues and due to its smooth surface and elasticity, gives the joints the ability to slip and bend. excessive weight, excessive activity, or trauma can all cause cartilage to injury. The injury can lead to swelling, pain and varying degrees of mobility loss. The process of repairing musculoskeletal (orthopedic) injuries has led to problems in the medical field, which can be attributed to the inherent weakness of adult cartilage tissue. Therefore, this necessitates research focused on the development of a new restructuring strategy by combining chondrocytes or stem cells with scaffolds and growth factors to address these problems. Correspondingly, the recent tissue engineering strategies strongly support the simultaneous use of stem cells, scaffolds and growth factors. It has also been observed that due to the relatively low proliferation of transplanted chondrocytes, new cartilage models construction have examined the use of adipose-derived stem cells. Mature adipose tissue is produced as an important source of multi-functional stem cells that can be easily separated from the stromal vascular fraction (SVF) by adipose liposuction digestion. The adipose-derived stem cells are easily accessible without any serious complications and have the power to differentiate into several cell lines, including chondrocytes as well as, they evidence self-renewal when trapped in gel scaffolds such as collagen. Also, recent studies demonstrate some of the mechanisms involved in the process of making cartilage of adipose-derived stem cells in vitro and their restorative ability in bio-engineered scaffolds in the presence of growth factors. In addition, the important role of non-encoding mRNA molecules (miRNAs) has been identified in the process of chondrogenic differentiation of adipose-derived stem cells. Furthermore, in several studies, the effect of several miRNAs has been confirmed on the regulation of the cartilage differentiation of the adipose-derived stem cells and has also been associated with effective results. In this article, we will present an overview of the advance in adipose-derived stem cells application in cartilage regeneration.
Davoud Farajzadeh , Parisa Jalali ,
Volume 76, Issue 5 (8-2018)
Volume 76, Issue 5 (8-2018)
Abstract
The natural killer group 2D (NKG2D) is a transmembrane protein and a member of the CD94/NKG2 family of C-type lectin-like receptors. NKG2D is encoded by the KLRK1 gene, which is located in the NK-gene complex (NKC) placed on chromosomes 6 and 12 in mice and humans, respectively. NKG2D forms a homodimer structure and binds through ectodomains with its related ligands. Each of its monomers consists of two β-sheets, two α-helices, and four disulfide bands and also contains a β-strand that distinguishes it from other C-type lectin-like receptors. NKG2D ligands are homologs of major histocompatibility complex (MHC) class I molecules in mice and humans. MHC class I chain-related protein A (MICA) and B (MICB) and human cytomegalovirus UL16-binding proteins (ULBP1-6) are recognized by the human NKG2D. In Natural Killer (NK) cells, NKG2D-mediated cytotoxicity can be elicited via two different systems by signaling from immunoreceptor tyrosine-based activation motifs in DAP12 or via a Syk-independent pathway activated by DAP10. Therefore, NKG2D is an activating immunoreceptor which was first recognized on NK cells but subsequently found on γδT cells, CD8+ αβT cells, and macrophages. NKG2D-ligand diversity may facilitate the detection of the presence of a broad range of viruses and may provide protection against rapidly evolving cancers. NKG2D ligand recognition induces and/or improves immune responses to cancer cells. NK cells recognize a wide range of stressed cells. The activation of NKG2D receptor can lead to the lysis of the target cell and the production of various cytokines and chemokines depending on the nature of the stimulation as a result of NK and myeloid-mediated innate immunity and as well as T γδ and CD8+ mediated-adaptive immune system. However, inappropriate expression of NKG2D ligands could cause autoimmune diseases in healthy cells, including rheumatoid arthritis, colitis, celiac disease, multiple sclerosis, alopecia areata, type 1 diabetes, and chronic obstructive pulmonary disease. Therefore, a precise understanding of the structure and function of NKG2D receptor and its interaction with various ligands may lead to the development of strategies to treat autoimmune diseases. Hence, the purpose of this review is to examine the detailed studies on the function of NKG2D receptor and their related ligands.
Fahimeh Kalbkhani , Mohammad Reza Sam ,
Volume 76, Issue 6 (9-2018)
Volume 76, Issue 6 (9-2018)
Abstract
Background: Using natural compounds with low toxicity on normal cells and high efficacy on malignant cells is highly appreciated for treatment of colorectal cancer (CRC). In the present study, the effect of fish-oil derived eicosapentaenoic acid (EPA) on the cell number, cell proliferation rate and caspase-3 enzyme activity in LS174T human colorectal cancer cell line was investigated.
Methods: This experimental study was performed in cell culture lab, Institute of Biotechnology, affiliated to the Urmia University, Urmia, Iran from April to September 2017. LS174T colorectal cancer cells at a density of 5×105 cells per well were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and kept at 37 °C in a humidified incubator with 5% CO2 for 24 hours. Thereafter, the cells were treated with 50, 100, 150 and 200 μmol EPA for 48 hours and cell numbers were counted using neobauer chamber and caspase-3 activities were measured by performing the caspase-3 colorimetric assay (Abcam, Cambridge, MA, USA). Furthermore, 5×103 LS174T colorectal cancer cells were cultured and treated with the above-mentioned EPA concentrations for 24, 48 and 72 hours, after which cell proliferation rate was evaluated by WST-1 proliferation assay (Roche Diagnostics, Mannheim, Germany).
Results: Treatment of LS174T colorectal cancer cells with 50, 100, 150 and 200 μmol EPA decreased the number of cells in a dose-dependent manner. We also found that treatment of malignant cells with increasing EPA concentrations (50 to 200 μmol) significantly decreased cell proliferation in a dose and time dependent manner. After a 72 hours treatment of LS174T cells with 200 μmol EPA, cell proliferation was calculated to be 30.3% compared to untreated control cells. Following 48 hours treatment, caspase-3 activity increased with increasing EPA concentrations in which at 200 μmol EPA, caspase-3 activity increased by 3.4 fold compared to untreated control cells.
Conclusion: Fish-oil derived eicosapentaenoic acid as a safe compound decreases the number of colorectal cancer cells and their proliferation rate and activates caspase-3 enzyme, as an executor protein in apoptosis.
Methods: This experimental study was performed in cell culture lab, Institute of Biotechnology, affiliated to the Urmia University, Urmia, Iran from April to September 2017. LS174T colorectal cancer cells at a density of 5×105 cells per well were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and kept at 37 °C in a humidified incubator with 5% CO2 for 24 hours. Thereafter, the cells were treated with 50, 100, 150 and 200 μmol EPA for 48 hours and cell numbers were counted using neobauer chamber and caspase-3 activities were measured by performing the caspase-3 colorimetric assay (Abcam, Cambridge, MA, USA). Furthermore, 5×103 LS174T colorectal cancer cells were cultured and treated with the above-mentioned EPA concentrations for 24, 48 and 72 hours, after which cell proliferation rate was evaluated by WST-1 proliferation assay (Roche Diagnostics, Mannheim, Germany).
Results: Treatment of LS174T colorectal cancer cells with 50, 100, 150 and 200 μmol EPA decreased the number of cells in a dose-dependent manner. We also found that treatment of malignant cells with increasing EPA concentrations (50 to 200 μmol) significantly decreased cell proliferation in a dose and time dependent manner. After a 72 hours treatment of LS174T cells with 200 μmol EPA, cell proliferation was calculated to be 30.3% compared to untreated control cells. Following 48 hours treatment, caspase-3 activity increased with increasing EPA concentrations in which at 200 μmol EPA, caspase-3 activity increased by 3.4 fold compared to untreated control cells.
Conclusion: Fish-oil derived eicosapentaenoic acid as a safe compound decreases the number of colorectal cancer cells and their proliferation rate and activates caspase-3 enzyme, as an executor protein in apoptosis.
Mohammad Moradi , Kamran Atarodi , Mahshid Mohammadipour , Kamran Mousavi Hosseini ,
Volume 76, Issue 6 (9-2018)
Volume 76, Issue 6 (9-2018)
Abstract
Background: Thrombopoietin (TPO) is an important cytokine that has a critical role in regulating hematopoietic stem cells (HSCs) proliferation and megakaryocyte differentiation. Because of scares amount of this protein in human plasma, in many biotechnological centers around the world, recombinant production of this protein has been carried out. This study was aiming to gene cloning and expression of recombinant thrombopoietin.
Methods: This research is an experimental laboratory study carried out in Blood Transfusion Research Center, Tehran, Iran, from July 2016 to August 2017. At the beginning HepG2 cell line was cultured and RNA extraction was performed. Extracted RNA was used as template for cDNA synthesis and subsequently the synthesized cDNA was adopted to isolate TPO gene through polymerase chain reaction (PCR) reaction using designed primers. After isolating the TPO sequence from HepG2 cell line, the designated sequence was inserted into pET32 vectors. Recombinant plasmid was amplified by meriting from DH5α replicating system. The amplified plasmids were sequenced via chain termination method. Next step was transforming the recombinant plasmid into Rosetta-gami bacteria to express the recombinant protein. In order to induce protein expression, an appropriate amount of isopropyl β-D-1-thiogalactopyranoside (IPTG) was added to growth media, then bacterial lysate of expression host was prepared and assayed via polyacrylamide gel electrophoresis and western blotting test.
Results: After sequencing of recombinant plasmid, it was confirmed that TPO sequence has been successfully colonized in adopted vector. Subsequent to induction of recombinant protein, total cell protein analysis affirmed that recombinant protein has been expressed in its soluble form at cytoplasmic condition. Location of expected recombinant protein band on polyacrylamide gel and reaction of recombinant protein with His-tag monoclonal antibody at western blotting was asserting that expressed protein is the one of interest.
Conclusion: Rosetta-gami bacteria has capability of expressing recombinant thrombopoietin in its soluble form. By harnessing this method of recombinant protein expression, it would be possible to take advantage of high throughout bacterial expression system which would not produce inclusion body and its product doesn’t need further processing and refolding.
Methods: This research is an experimental laboratory study carried out in Blood Transfusion Research Center, Tehran, Iran, from July 2016 to August 2017. At the beginning HepG2 cell line was cultured and RNA extraction was performed. Extracted RNA was used as template for cDNA synthesis and subsequently the synthesized cDNA was adopted to isolate TPO gene through polymerase chain reaction (PCR) reaction using designed primers. After isolating the TPO sequence from HepG2 cell line, the designated sequence was inserted into pET32 vectors. Recombinant plasmid was amplified by meriting from DH5α replicating system. The amplified plasmids were sequenced via chain termination method. Next step was transforming the recombinant plasmid into Rosetta-gami bacteria to express the recombinant protein. In order to induce protein expression, an appropriate amount of isopropyl β-D-1-thiogalactopyranoside (IPTG) was added to growth media, then bacterial lysate of expression host was prepared and assayed via polyacrylamide gel electrophoresis and western blotting test.
Results: After sequencing of recombinant plasmid, it was confirmed that TPO sequence has been successfully colonized in adopted vector. Subsequent to induction of recombinant protein, total cell protein analysis affirmed that recombinant protein has been expressed in its soluble form at cytoplasmic condition. Location of expected recombinant protein band on polyacrylamide gel and reaction of recombinant protein with His-tag monoclonal antibody at western blotting was asserting that expressed protein is the one of interest.
Conclusion: Rosetta-gami bacteria has capability of expressing recombinant thrombopoietin in its soluble form. By harnessing this method of recombinant protein expression, it would be possible to take advantage of high throughout bacterial expression system which would not produce inclusion body and its product doesn’t need further processing and refolding.
Investigation the effect of copper nanoparticles on the toxicity and migration of keratinocyte cells
Sanaz Alizadeh , Nasser Aghdami , Bagher Seyedalipour , Parvaneh Mohammadi ,
Volume 76, Issue 8 (11-2018)
Volume 76, Issue 8 (11-2018)
Abstract
Background: Re-epithelialization has an important role in skin wound healing. Delays in re-epithelialization are more likely to create the chronic wound. Impaired wound healing leads to a large burden of morbidity and mortality. Current treatments based on the use of autografts, allografts and xenografts, suffer from limitations such as, quantity of donor skin available, donor-site infection, potential risk of disease transmission and rejection of the graft. Given this problems, nanomaterial such as copper nanoparticles has attracted considerable research interest because of their high surface area to volume ratio, high stability, clinical safety, and antibacterial effects. Epithelialization involves keratinocyte migration and proliferation to the wound site. Therefore, this study was conducted to investigate the effect of copper nanoparticles on keratinocyte cell migration and proliferation.
Methods: This experimental study was performed in Royan Institute, Tehran in 2016. In this study we investigated the effect of copper nanoparticles on viability, migration and proliferation of keratinocyte cells. Cultured human foreskin Keratinocyte cells were exposed to various concentration (1, 10 and 100 µmol) and sizes (40 and 80 nm) of copper nanoparticles for 24, 48 and 72 hours. The copper nanoparticles toxicity was examined by MTS assay. Cell migration has also been investigated with the Scratch assay.
Results: The results showed that the 1, 10 and 100 µmol concentrations of 40 and 80 nm copper nanoparticles were not toxic for cultured human foreskin keratinocyte cells after 24h. It was also found keratinocyte cell proliferation was increased by 1 µmol concentration of 80 nm copper nanoparticles after 72h. The results of the Scratch assay showed that the 1 µmol concentration of 80 nm copper nanoparticles significantly (P<0.05) increased keratinocyte cell migration compared to deionized water as of control group after 24h.
Conclusion: It seems the 1 µmol concentration of 80 nm copper nanoparticle could stimulate keratinocyte cell migration and proliferation. However, in vivo studies conducted on animal model wound healing subjects are needed for determining re-epithelialization.
Methods: This experimental study was performed in Royan Institute, Tehran in 2016. In this study we investigated the effect of copper nanoparticles on viability, migration and proliferation of keratinocyte cells. Cultured human foreskin Keratinocyte cells were exposed to various concentration (1, 10 and 100 µmol) and sizes (40 and 80 nm) of copper nanoparticles for 24, 48 and 72 hours. The copper nanoparticles toxicity was examined by MTS assay. Cell migration has also been investigated with the Scratch assay.
Results: The results showed that the 1, 10 and 100 µmol concentrations of 40 and 80 nm copper nanoparticles were not toxic for cultured human foreskin keratinocyte cells after 24h. It was also found keratinocyte cell proliferation was increased by 1 µmol concentration of 80 nm copper nanoparticles after 72h. The results of the Scratch assay showed that the 1 µmol concentration of 80 nm copper nanoparticles significantly (P<0.05) increased keratinocyte cell migration compared to deionized water as of control group after 24h.
Conclusion: It seems the 1 µmol concentration of 80 nm copper nanoparticle could stimulate keratinocyte cell migration and proliferation. However, in vivo studies conducted on animal model wound healing subjects are needed for determining re-epithelialization.
Saba Sorayyayi , Sogand Vahidi , Mohammad Mohammadzadeh , Sayyed Saied Hosseini-Asl ,
Volume 76, Issue 8 (11-2018)
Volume 76, Issue 8 (11-2018)
Abstract
Background: Gastric cancer is among the most common malignancies in certain parts of the world, such as northwest Iran. miRNAs are small and single-stranded noncoding RNAs with about 19-23 nucleotides. Several studies have shown that miRNAs play important roles in gastric tumorigenesis. The aim of this study was to determine the effect of miRNA-1266-5p repression on the cell survival and alterations of the cell cycle in gastric cancer cell line of AGS (NCBI Code: C131, Gastric epithelial cell line).
Methods: This experimental study was performed from April to December 2017 in Cellular-Molecular Research Center of Ardabil University of Medical Sciences, Iran. In this study, AGS cells were cultured in RPMI-1640 medium containing 10% serum and 1% antibiotic. The cells were transfected with miR-1266-5p mimic, miR-1266-5p inhibitor and HiPerFect reagent alone as negative control. The miR-1266-5p expression and transfection efficiency were analyzed by Stem-loop TaqMan qRT-PCR. The cell proliferation and cell cycle alterations were determined using MTT calorimetric assay and flow cytometry, respectively. The results were analyzed using SPSS 19.0 statistics software (SPSS Inc., Chicago, IL, USA) and presented as the means±standard deviation (SD).
Results: miR-1266-5p expression was increased in AGS cells transfected with miR-1266-5p mimic compared to control cells (P=0), while miR-1266-5p expression was decreased in transfected cells with the inhibitor compared to controls (P=0). Among different time points, the most effects of miR-1266-5p mimic and inhibitor were noticed after 48 hours of transfection. The upregulated miR-1266-5p significantly decreased cell growth, in contrast, inhibitor promoted cell proliferation (P=0). In addition, miR-1266-5p upregulation induced cell cycle arrest at the transition of G1 to S phase and led to G0/G1 entry (P=0), while of miR-1266-5p led to G2/M entry (P=0.001).
Conclusion: According to the results obtained from this study, miR-1266-5p can reduce cell survival and induce cell cycle arrest and act as a tumor suppressor in AGS cells. While its inhibition can increase cell survival and reduce apoptosis.
Methods: This experimental study was performed from April to December 2017 in Cellular-Molecular Research Center of Ardabil University of Medical Sciences, Iran. In this study, AGS cells were cultured in RPMI-1640 medium containing 10% serum and 1% antibiotic. The cells were transfected with miR-1266-5p mimic, miR-1266-5p inhibitor and HiPerFect reagent alone as negative control. The miR-1266-5p expression and transfection efficiency were analyzed by Stem-loop TaqMan qRT-PCR. The cell proliferation and cell cycle alterations were determined using MTT calorimetric assay and flow cytometry, respectively. The results were analyzed using SPSS 19.0 statistics software (SPSS Inc., Chicago, IL, USA) and presented as the means±standard deviation (SD).
Results: miR-1266-5p expression was increased in AGS cells transfected with miR-1266-5p mimic compared to control cells (P=0), while miR-1266-5p expression was decreased in transfected cells with the inhibitor compared to controls (P=0). Among different time points, the most effects of miR-1266-5p mimic and inhibitor were noticed after 48 hours of transfection. The upregulated miR-1266-5p significantly decreased cell growth, in contrast, inhibitor promoted cell proliferation (P=0). In addition, miR-1266-5p upregulation induced cell cycle arrest at the transition of G1 to S phase and led to G0/G1 entry (P=0), while of miR-1266-5p led to G2/M entry (P=0.001).
Conclusion: According to the results obtained from this study, miR-1266-5p can reduce cell survival and induce cell cycle arrest and act as a tumor suppressor in AGS cells. While its inhibition can increase cell survival and reduce apoptosis.
Jafar Mohammadshahi , Soheila Refahi , Bahareh Yousefipour , Mehran Sardari , Roghayeh Teimourpour ,
Volume 76, Issue 9 (12-2018)
Volume 76, Issue 9 (12-2018)
Abstract
Hepatitis B virus (HBV) is an etiological agent of hepatitis B infection. Hepatitis B is a life-threatening disease that affects the liver. The clinical outcomes of the disease are varied from asymptomatic disease to serious complication such as cirrhosis and hepatocellular carcinoma (HCC). Despite availability of the vaccine and appropriate treatment, hepatitis B infection still remains a major public health problem worldwide. Based on WHO reports, over 887.000 people die annually from hepatitis B complication including cirrhosis and hepatocellular carcinoma. Hepatitis B is very contagious and spreads through infected blood, body fluids, mother to baby during birth, contaminated needle and between sexual partners. HBV uses sodium taurocholate cotransporting polypeptide (NTCP) receptor to enter hepatocytes and by replicating in these cells interferes with liver functions. In fact liver damage is as result of virus multiplication and activation of immune responses especially virus-specific cytotoxic T lymphocytes (CTLs) against infected cells. CTLs and CD4Th1 cells by killing infected cells and releasing antiviral cytokines control virus replication in infected individuals. Also, the functions of these cells in patients who successfully clear the infection are potentially strong. In contrast to acute self-limited HBV infection in persistent HBV infection, these cells are exhausted. Several studies have showed that the great challenge in clearance of the HBV infection is related to stability of covalently closed circular DNA (cccDNA). cccDNA produce in viral life cycle and remains inside the infected cells for a long time and act as a template for generating new pre-genomic RNA and virus propagation. So far, no antiviral treatment has been effective in the complete elimination of this structure. Prevention of the disease can be achieved by using effective vaccine. Previous studies indicated that neutralizing antibodies against surface antigen of the virus known as S antigen have protective properties. Therefore, a subunit vaccine containing S antigen is available. Currently S antigen is produced in recombinant form and WHO recommended the first dose should be given within a day of birth. Pegylated IFN-γ and nucleotide-nucleoside analogues are effective drugs against HBV infection, but they may have severe side effects. Ineffectiveness of the vaccine on premature infants and immunocompromised people and also drug side effects has made HBV infection a great trouble.
Marjan Ghorbani-Anarkooli , Sara Dabirian, Hasan Moladoust, Adib Zendedel, Mohammad Hadi Bahadori,
Volume 77, Issue 1 (4-2019)
Volume 77, Issue 1 (4-2019)
Abstract
Background: Evaluation of cell viability is momentous in pharmacologic and oncological research. Cell viability evaluation determines cell sensitivity and consequently treatment outcome. Various methods are available to determine cell survival. Each of these methods evaluates different endpoints. Accordingly, determining the correlation between these methods is important. In this study, in order to determine the viability of human anaplastic thyroid cancer cell line, the sensitivity of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, trypan blue test and clonogenic assay were compared.
Methods: This experimental study was performed in the Cellular and Molecular Research Center at Guilan University of Medical Sciences, Rasht, Iran from October 2016 to March 2017. The human anaplastic thyroid cancer cell line was cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). The cultured cells were treated with melatonin, for 24 hours. Then, the viability of the cells was evaluated by MTT assay, trypan blue test and clonogenic assay. Furthermore, plating efficiency and surviving fraction were used in order to draw survival curve in the clonogenic assay.
Results: The concentration of melatonin at IC50 point was 4.794±0.117 millimolar (mM) in MTT assay, 4.375±0.894 mM in trypan blue test and 2.246±0.326 mM in clonogenic assay. Comparing the IC50 values of these test revealed that C50 values obtained from MTT assay and trypan blue test had no significant difference (P=0.6446), while there was a significant difference between IC50 values obtained from MTT and clonogenic assays (P=0.0032). Moreover, the IC50 values obtained from trypan blue test and clonogenic assay were also significantly different (P=0.0078). The results of the regression analysis of cell viability were shown a linear, positive and significant correlation between these three methods and MTT assay and trypan blue test showed higher correlation (r=0.99, P<0.001).
Conclusion: Based on our results, all these methods were effective to identify cytotoxicity in human anaplastic thyroid cancer cell line, while MTT assay and trypan blue test were more sensitive than clonogenic assay.
Methods: This experimental study was performed in the Cellular and Molecular Research Center at Guilan University of Medical Sciences, Rasht, Iran from October 2016 to March 2017. The human anaplastic thyroid cancer cell line was cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). The cultured cells were treated with melatonin, for 24 hours. Then, the viability of the cells was evaluated by MTT assay, trypan blue test and clonogenic assay. Furthermore, plating efficiency and surviving fraction were used in order to draw survival curve in the clonogenic assay.
Results: The concentration of melatonin at IC50 point was 4.794±0.117 millimolar (mM) in MTT assay, 4.375±0.894 mM in trypan blue test and 2.246±0.326 mM in clonogenic assay. Comparing the IC50 values of these test revealed that C50 values obtained from MTT assay and trypan blue test had no significant difference (P=0.6446), while there was a significant difference between IC50 values obtained from MTT and clonogenic assays (P=0.0032). Moreover, the IC50 values obtained from trypan blue test and clonogenic assay were also significantly different (P=0.0078). The results of the regression analysis of cell viability were shown a linear, positive and significant correlation between these three methods and MTT assay and trypan blue test showed higher correlation (r=0.99, P<0.001).
Conclusion: Based on our results, all these methods were effective to identify cytotoxicity in human anaplastic thyroid cancer cell line, while MTT assay and trypan blue test were more sensitive than clonogenic assay.
Arash Abdolmaleki, Mohammad-Bagher Ghayour, Saber Zahri, Asadollah Asadi , Morteza Behnam-Rassouli ,
Volume 77, Issue 2 (5-2019)
Volume 77, Issue 2 (5-2019)
Abstract
Background: Tissue engineering is a developing multidisciplinary and interdisciplinary field involving the use of bioartificial implants for tissue remodeling with the target for repair and enhancing tissue or organ function. Acellular nerve has been used in experimental models as a peripheral nerve substitute. The purpose of the present study was to evaluate the mechanical and histological characteristics of acellular nerve scaffolds compared to the fresh nerve for application in environmental nerve repair.
Methods: This experimental study was conducted in Ferdowsi University of Mashhad Regeneration Research Laboratory, Mashhad, Iran, from May 2017 to October 2018. In this study for preparing the scaffold. The rats were sacrificed by intraperitoneal anesthesia with 10 % Chloral Hydrate solution. Then sciatic nerve fragments of the rats were removed above the nerve branching site and after cleansing of the tissues were decellularized by Sondell method, briefly nerves were treated with a series of detergent baths consisting of distilled water for 8 h, Triton X-100 for 12 h, and sodium deoxycholate for 24 hours according to the Sondell protocol. All acellularization steps were performed at room temperature. Then decellularized scaffolds were evaluated histologically and mechanically.
Results: The results of tissue evaluations showed that decellularization of scaffolds were done completely, this was demonstrated by hematoxylin and eosin staining and DAPI staining. Also the specialized tissue evaluations by picro-fuchsin staining and evaluation the scaffolds by scanning electron microscopy (SEM) micrographs showed that the collagen and elastin strands are relatively preserved in the extracellular matrix in comparison with control groups. As well as mechanical examination of scaffolds in tensile test showed that extracellular matrix of scaffolds was relatively preserved the main components of tissue compared to control group and scaffolds have good mechanical resistance quality for use in tissue engineering.
Conclusion: The results of the present study showed that decellularized scaffolds that prepared with Sondell decellularization method by preserving the main components of the tissue can be a good platform for investigating cellular behaviors.
Methods: This experimental study was conducted in Ferdowsi University of Mashhad Regeneration Research Laboratory, Mashhad, Iran, from May 2017 to October 2018. In this study for preparing the scaffold. The rats were sacrificed by intraperitoneal anesthesia with 10 % Chloral Hydrate solution. Then sciatic nerve fragments of the rats were removed above the nerve branching site and after cleansing of the tissues were decellularized by Sondell method, briefly nerves were treated with a series of detergent baths consisting of distilled water for 8 h, Triton X-100 for 12 h, and sodium deoxycholate for 24 hours according to the Sondell protocol. All acellularization steps were performed at room temperature. Then decellularized scaffolds were evaluated histologically and mechanically.
Results: The results of tissue evaluations showed that decellularization of scaffolds were done completely, this was demonstrated by hematoxylin and eosin staining and DAPI staining. Also the specialized tissue evaluations by picro-fuchsin staining and evaluation the scaffolds by scanning electron microscopy (SEM) micrographs showed that the collagen and elastin strands are relatively preserved in the extracellular matrix in comparison with control groups. As well as mechanical examination of scaffolds in tensile test showed that extracellular matrix of scaffolds was relatively preserved the main components of tissue compared to control group and scaffolds have good mechanical resistance quality for use in tissue engineering.
Conclusion: The results of the present study showed that decellularized scaffolds that prepared with Sondell decellularization method by preserving the main components of the tissue can be a good platform for investigating cellular behaviors.
Mahsa Nazari, Farid Zayeri , Seyed Saeed Hashemi Nazari , Sara Jambarsang, Ali Nikfarjam , Alireza Akbarzadeh Baghban ,
Volume 77, Issue 2 (5-2019)
Volume 77, Issue 2 (5-2019)
Abstract
Background: The Multi state Markov models have extensively application with categorization of laboratory marker of CD4 cells for evaluation of HIV disease progression. These models with different states result in different effects of covariates and prediction of HIV disease trend. The main purpose of this study was comparison of four and five states models with the three- state in order to select the model with better prediction ability of occurrence of HIV and finally death in HIV positive people.
Methods: A total of 305 HIV positive people were included in this cohort study in the Iran AIDS center in Imam Khomeini Hospital in Tehran that entered during March 1995 to January 2005 and then fallowed up to October 2014. The three continuous- time Markov models of three-, four- and five- state models were fitted to data to describe the evolution of a HIV disease Trend over different states.
For comparison of models, two criteria of modification of Akaike’s criterion (DRAIC) and likelihood cross-validation criterion (DRLCV) along with their 95% tracking interval was used. For fitting of these models and estimation of transition matrix and the hazard ratio of gender and treatment independent variables, the msm package of R project for statistical computing, version R 3.2.4 (www.r-project.org) was used.
Results: The results showed that the four- state model has more prediction ability than five-state model for evaluation of HIV disease Trend. In the four-state model, the progression hazard ratio to death for people who received highly active antiretroviral therapy (HAART) was 0.64 lower than who didn’t get this therapy. Moreover, the progression hazard ratio for men was 2.33 fold in comparison to women. The disease progression hazard ratio to death was 4.9 fold for men in comparison to women.
Conclusion: The (DRAIC) and (DRLCV) criterions showed that the four-state model has more predictive ability of the progression trend of HIV disease in comparison of five-state model.
Methods: A total of 305 HIV positive people were included in this cohort study in the Iran AIDS center in Imam Khomeini Hospital in Tehran that entered during March 1995 to January 2005 and then fallowed up to October 2014. The three continuous- time Markov models of three-, four- and five- state models were fitted to data to describe the evolution of a HIV disease Trend over different states.
For comparison of models, two criteria of modification of Akaike’s criterion (DRAIC) and likelihood cross-validation criterion (DRLCV) along with their 95% tracking interval was used. For fitting of these models and estimation of transition matrix and the hazard ratio of gender and treatment independent variables, the msm package of R project for statistical computing, version R 3.2.4 (www.r-project.org) was used.
Results: The results showed that the four- state model has more prediction ability than five-state model for evaluation of HIV disease Trend. In the four-state model, the progression hazard ratio to death for people who received highly active antiretroviral therapy (HAART) was 0.64 lower than who didn’t get this therapy. Moreover, the progression hazard ratio for men was 2.33 fold in comparison to women. The disease progression hazard ratio to death was 4.9 fold for men in comparison to women.
Conclusion: The (DRAIC) and (DRLCV) criterions showed that the four-state model has more predictive ability of the progression trend of HIV disease in comparison of five-state model.
Adeleh Poursaleh, Mohammad Najafi , Farahnaz Sadegh Beigee ,
Volume 77, Issue 3 (6-2019)
Volume 77, Issue 3 (6-2019)
Abstract
Background: The immune-mediated responses in vascular cells may include the increased expression of endothelial adhesion molecules, leukocyte rolling and infiltration, cellular lipid dysregulation and vascular smooth muscle cells (VSMCs) differentiation. Investigating the cellular and molecular events involved in the rolling process is useful for treatment or prevention of the vessel stenosis especially in coronary arteries. MiRNAs are small and single-stranded noncoding RNAs with about 19-23 nucleotides. In this study, the role of microRNA-125 was predictably selected and experimentally investigated on the changes of expression level of adhesion molecule in endothelial cells isolated from human aorta and on the monocyte cells isolated from whole blood human with endothelial cells adhesion. The aim of this study was to determine the effect of miRNA-125 repression on cell adhesion in leukocyte rolling process to reduce or suppress artery stenosis in susceptible individuals.
Methods: This experimental study was performed in Cellular-Molecular Research Center of Iran University of Medical Sciences, Tehran, Iran from July to December 2017. Normal aortic samples were prepared from subjects with brain death in Masih Daneshvari Hospital and under strictly sterile conditions, it was transferred as soon as possible. The endothelial cells were isolated from aorta of subjects with brain death using collagenase. The monocytes were isolated from whole blood. The microRNA-125 was transfected into ECs with use of polyethyleneimine (PEI). The expression level of adhesion molecule and monocyte recruitment were identified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique and CytoSelect™ leukocyte-endothelium adhesion assay kit (Cell Biolabs, San Diego, CA, USA), respectively.
Results: The results showed the microRNA-125 suppresses significantly integrin beta 2 (ITGB2) expression level (P=0.008). In addition, the monocyte-EC adhesion was shown in the aortic miRNA-treated endothelial cells. The adhesive rate between cells reduced significantly with microRNA-125 as compared with miR-synthetic (P=0.02). Thereby, there were the associations between the ITGB2 and miR-125a. Downregulation of ITGB2 may be reduced the adhesion of endothelial cells and moderating the process rolling.
Conclusion: This study suggested that the suppression of leukocyte rolling process might be more due to the function of ITGB2. However, the functional effects of this miRNA should be directly investigated on the studied gene.
Methods: This experimental study was performed in Cellular-Molecular Research Center of Iran University of Medical Sciences, Tehran, Iran from July to December 2017. Normal aortic samples were prepared from subjects with brain death in Masih Daneshvari Hospital and under strictly sterile conditions, it was transferred as soon as possible. The endothelial cells were isolated from aorta of subjects with brain death using collagenase. The monocytes were isolated from whole blood. The microRNA-125 was transfected into ECs with use of polyethyleneimine (PEI). The expression level of adhesion molecule and monocyte recruitment were identified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique and CytoSelect™ leukocyte-endothelium adhesion assay kit (Cell Biolabs, San Diego, CA, USA), respectively.
Results: The results showed the microRNA-125 suppresses significantly integrin beta 2 (ITGB2) expression level (P=0.008). In addition, the monocyte-EC adhesion was shown in the aortic miRNA-treated endothelial cells. The adhesive rate between cells reduced significantly with microRNA-125 as compared with miR-synthetic (P=0.02). Thereby, there were the associations between the ITGB2 and miR-125a. Downregulation of ITGB2 may be reduced the adhesion of endothelial cells and moderating the process rolling.
Conclusion: This study suggested that the suppression of leukocyte rolling process might be more due to the function of ITGB2. However, the functional effects of this miRNA should be directly investigated on the studied gene.
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