Publications by authors named "Shahram Darabi"

31 Publications

Role of cerebrospinal fluid in differentiation of human dental pulp stem cells into neuron-like cells.

Anat Cell Biol 2020 Sep;53(3):292-300

Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.

Human dental pulp stem cells (hDPSCs) could be differentiated into neuron like-cells under particular microenvironments. It has been reported that a wide range of factors, presented in cerebrospinal fluid (CSF), playing part in neuronal differentiation during embryonic stages, we herein introduce a novel culture media complex to differentiate hDPSCs into neuron-like cells. The hDPSCs were initially isolated and characterized. The CSF was prepared from the Cisterna magna of 19-day-old Wistar rat embryos, embryonic cerebrospinal fluid (E-CSF). The hDPSCs were treated by 5% E-CSF for 2 days, then neurospheres were cultured in DMEM/F12 supplemented with 10 μm retinoic acid (RA), glial-derived neurotrophic factor and brain-derived neurotrophic factor for 6 days. The cells which were cultured in basic culture medium were considered as control group. Morphology of differentiated cells as well as process elongation were examined by an inverted microscope. In addition, the neural differentiation markers (Nestin and MAP2) were studied employing immunocytochemistry. Neuronal-like processes appeared 8 days after treatment. Neural progenitor marker (Nestin) and a mature neural marker (MAP2) were expressed in treated group. Moreover Nissl bodies were found in the cytoplasm of treated group. Taking these together, we have designed a simple protocol for generating neuron-like cells using CSF from the hDPSCs, applicable for cell therapy in several neurodegenerative disorders including Alzheimer's disease.
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http://dx.doi.org/10.5115/acb.19.241DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7527124PMC
September 2020

The Effect of Low-Level Laser Therapy and Curcumin on the Expression of LC3, ATG10 and BAX/BCL2 Ratio in PC12 Cells Induced by 6-Hydroxide Dopamine.

J Lasers Med Sci 2020 21;11(3):299-304. Epub 2020 Jun 21.

Hearing Disorders Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Parkinson's disease (PD) is one of the most common neurodegenerative disorders. The neuroinflammation in the brain of PD patients is one of the critical processes in the immune pathogenesis of PD leading to the neural loss in the substantia nigra. Due to the anti-inflammatory effects of curcumin (CU) and low-level laser therapy (LLLT), we examined the protective effect of CU and LLLT on PC12 cells treated with 6-hydroxydopamine (6-OHDA) as a Parkinson model. PC12 cells were pretreated using various concentrations of 6-OHDA for 24 hours to induce oxidative and cellular damages. PC12-6-OHDA cells were co-treated with CU and LLLT. The effects of CU and LLLT on Bax/Bcl2 and LC3/ATG10 expression were analyzed by real-time PCR and cell viability was assessed by MTT assay. Cell A Software was used to calculate the length of the Neurite and cell body areas. The results of this study show that the combination of CU dose-dependently and LLLT has a significant neuroprotective effect on cells and cellular death significantly decreases by increasing CU concentration. CU+LLLT decreases Bax/Bcl2 ratio which is an indicator of apoptosis and it also rescued a decrease in LC3 and ATG10 expression in comparison with 6-OHDA group. This study shows that the combination of 5 μM CU and LLLT has the best neuroprotective effect on PC12 cells against 6-OHDA by decreasing the BAX/BCL2 ratio.
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http://dx.doi.org/10.34172/jlms.2020.50DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7369553PMC
June 2020

Association between ABCC8 Ala1369Ser Polymorphism (rs757110 T/G) and Type 2 Diabetes risk in an Iranian population: A Case-Control Study.

Endocr Metab Immune Disord Drug Targets 2020 Jul 12. Epub 2020 Jul 12.

School of Pharmacy, Faculty of Sciences, University of Rome Tor Vergata, Rome. Italy.

Objective: Glucose metabolism increases ATP/ADP ratio within the β-cells and causes ATP-sensitive K+ (KATP) channel closure and consequently insulin secretion. The enhanced activity of the channel may be a mechanism contributing to the reduced first-phase of insulin secretion observed in T2DM. There is no study to date in the Kurdish ethnic group regarding the relationship between SNP Ala1369Ser (rs757110 T/G) of SUR1 gene and T2DM, and additionally, the results of this association in other populations are inconsistent. Therefore, our aim in this study was to explore the possible association between SNP Ala1369Ser and type 2 diabetes in an Iranian Kurdish ethnic group.

Methods: In this study, we checked out the frequency of alleles and genotypes of SNP Ala1369Ser in T2DM individuals (207 patients; men/women: 106/101) and non-T2DM subjects (201 controls; men/women: 97/104), and their effects on anthropometric, clinical, and biochemical parameters. Genomic DNA was extracted from the leukocytes of blood specimens using a standard method. We amplified the ABCC8 rs757110 polymorphic site (T/G) using a polymerase chain reaction (PCR) method and a designed primer pair. To perform the PCR-RFLP method, the amplicons were subjected to restriction enzymes and the resulting fragments separated by gel electrophoresis.

Results: The frequency of the G-allele of Ala1369Ser polymorphism was significantly (0.01) higher in the case group than the control group (19% vs. 9%, respectively). In the dominant model (TT vs. TG+GG), there was a significant relationship between this SNP and an increased risk of T2DM (P = 0.00). T2DM patients with TG+GG genotypes had significantly higher fasting plasma insulin and HOMA-IR than those who had the TT genotype (P = 0.02 and 0.01, respectively).

Conclusions: Our study is the first study to investigate the association between Ala1369Ser ABCC8 genetic variation and T2DM in the Kurdish population of western Iran. The obtained results clearly show that Ala1369Ser polymorphism of ABCC8 is associated with an increased risk of T2DM in this population.
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http://dx.doi.org/10.2174/1871530320666200713091827DOI Listing
July 2020

Trehalose Neuroprotective Effects on the Substantia Nigra Dopaminergic Cells by Activating Autophagy and Non-canonical Nrf2 Pathways.

Iran J Pharm Res 2019 ;18(3):1419-1428

Department of Clinical Biochemistry, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.

Trehalose, as a natural disaccharide, is known as an autophagy inducer. The neuroprotective effects of trehalose in the rat model of Parkinson's disease were the aim of the present study. Parkinson's disease model was induced by injecting 6-hydroxydopamine (6-OHDA) in the striatum of male Wistar rats. Apomorphine-induced behavior and substantia nigra neuronal counts were applied to evaluate the neuroprotective effects of trehalose. The autophagy was studied using the expression of p62 and LC3II/LC3I ratio. In addition, the antioxidant effects of trehalose were assessed by analyzing the levels of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and also glutathione reductase (GR), glutathione peroxidase (GPx) and Catalase (CAT) enzymes. Moreover, the levels of 3, 4-dihydroxyphenylacetic acid (DOPAC) and dopamine (DA) were assessed.The behavioral test showed that trehalose in the treatment group reduced the damage to the substantial nigra dopaminergic neurons, which was characterized by improved motor and reduced rotations in the treatment group as compared with the lesion group. In the histological examinations of the treatment group, trehalose prevented the destruction of dopaminergic neurons. Trehalose treatments increased autophagy (high LC3II/LC3I ratio) and the expression of the p62 protein as well. Through p62-dependent manner, it led to increased nuclear translocation of Nrf2 transcription factor and elevated expression of downstream antioxidant enzymes, such as GR, GPx, and CAT, restoring DA and DOPAC contents of the cells. In the current study, trehalose simultaneously protects substantia nigra dopaminergic cells by activating both non-canonical p62/SQSTM1-Keap1-Nrf2 and autophagy pathways.
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http://dx.doi.org/10.22037/ijpr.2019.2387DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6934986PMC
January 2019

Antimicrobial peptides-loaded smart chitosan hydrogel: Release behavior and antibacterial potential against antibiotic resistant clinical isolates.

Int J Biol Macromol 2020 Dec 6;164:855-862. Epub 2020 Jul 6.

Cellular and Molecular Research Centre, Iran University of Medical Sciences, Tehran, Iran; Department of Tissue Engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran. Electronic address:

In this study, we synthesized thermo-responsive chitosan (TCTS) hydrogels, and loaded with different concentrations of antimicrobial peptide (AMP) (0, 4, 8 and 16 μg·ml) to fabricate an antibacterial wound dressing against resistant clinical isolates. Physico-chemical properties, release behavior, cytobiocompatibility and antibacterial activity of the AMP-TCTS hydrogels against standard strain and resistant Acinetobacter baumannii were fully determined in vitro. The TCTS-40% β-glycerolphosphate hydrogels showed a gelation time of 15 min at 37 °C. 80% weight loss at day 35 with no changes in pH value was observed. AMP-TCTS hydrogels showed a burst release of AMP (around 40%) at day 1, and a controlled release up to day 7. A dramatic water uptake was observed at first 4 h, and then continued for 10 h in a steady manner. All the AMP-TCTS hydrogels showed excellent cytobiocompatibility for human fibroblasts. The TCTS showed no antibacterial activity against both standard strain and clinical isolates. All the AMP-TCTS hydrogels had strong antibacterial activity against standard strains, but only 16 μg·ml showed antibacterial behavior against resistant A. baumannii. Our results strongly suggest the 16 μg·ml AMP-TCTS hydrogel as an excellent antibacterial wound dressing against resistant A. baumannii, and now promises to proceed with pre-clinical investigations.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.07.011DOI Listing
December 2020

Trans-Differentiation of Human Dental Pulp Stem Cells Into Cholinergic-Like Neurons Via Nerve Growth Factor.

Basic Clin Neurosci 2019 Nov-Dec;10(6):609-617. Epub 2019 Nov 1.

ENT and Head & Neck Research Center and Department, Hazrat Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran.

Introduction: Cell therapy has been widely considered as a therapeutic approach for neurodegenerative diseases and nervous system damage. Cholinergic neurons as one of the most important neurons that play a significant role in controlling emotions, mobility, and autonomic systems. In this study, Human Dental Pulp Stem Cells (hDPSCs) were differentiated into the cholinergic neurons by β-mercaptoethanol in the preinduction phase and also by the nerve growth factor (NGF) in the induction phase.

Methods: The hDPSCs were evaluated for CD73, CD31, CD34, and Oct-4. Concentration-time relationships for NGF were assessed by evaluating the viability rate of cells and the immune response to nestin, neurofilament 160, microtubule-associated protein-2, and choline acetyltransferase.

Results: The hDPSCs had a negative response to CD34 and CD31. The optimal dose for the NGF was 50 ng/mL seven days after the induction when the highest percentage of expressing markers for the Cholinergic neurons (ChAT) was detected.

Conclusion: The results of this study provided a method for producing cholinergic neurons by hDPSCs, which can be used in cytotherapy for degenerative diseases of the nervous system and also spinal cord injury.
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http://dx.doi.org/10.32598/bcn.10.6.609DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7253808PMC
November 2019

LC3 and ATG5 overexpression and neuronal cell death in the prefrontal cortex of postmortem chronic methamphetamine users.

J Chem Neuroanat 2020 09 19;107:101802. Epub 2020 May 19.

Hearing Disorders Research Center, Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

Methamphetamine (METH) abuse is accompanied by oxidative stress, METH-induced neurotoxicity, and apoptosis. Oxidative stress has devastating effects on the structure of proteins and cells. Autophagy is an evolutionarily conserved intracellular regulated mechanism for orderly degradation of dysfunctional proteins or removing damaged organelles. The precise role of autophagy in oxidative stress-induced apoptosis of dopaminergic neuronal cells caused by METH has not clarified completely. In this study, we sought to evaluate the effects of METH abuse on autophagy in the prefrontal cortex of postmortem users, mainly focusing on the ATG5 and LC3 during neuroinflammation. Postmortem molecular and histological examination was done for two groups containing 12 non-addicted and 14 METH addicted cases. ATG5 and LC3 expression were analyzed by real-time PCR and immunohistochemistry (IHC) methods. Histopathological analysis was performed by stereological cell counting of neuronal cells using Hematoxylin and Eosin (H & E) staining technique. In order to detect DNA damage in the prefrontal lobe, Tunnel staining was performed. Real-time PCR and IHC assay showed overexpression of ATG5 and LC3 protein in the prefrontal cortex of Meth users. The cell death and neuronal degeneration were increased significantly based on Tunel assay and the stereological analysis in the Prefrontal cortex. Chronic METH exposure probably induces ATG5 and LC3 overexpression and neuronal cell death in the Prefrontal cortex of the postmortem cases.
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http://dx.doi.org/10.1016/j.jchemneu.2020.101802DOI Listing
September 2020

The Effect of Low-Power Laser Therapy on the TGF/β Signaling Pathway in Chronic Kidney Disease: A Review.

J Lasers Med Sci 2020 15;11(2):220-225. Epub 2020 Mar 15.

Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

The purpose of this study is to investigate the effects of low-power lasers on kidney disease by investigating several studies. A number of articles from 1998 to 2019 were chosen from the sources of PubMed, Scopus, and only the articles studying the effect of low-power lasers on kidney disease were investigated. After reviewing the literature, 21 articles examining only the effects of low-power lasers on kidney disease were found. The results of these studies showed that the parameter of the lowpower laser would result in different outcomes. So, a low-power laser with various parameters can be effective in the treatment of kidney diseases such as acute kidney disease, diabetes, glomerulonephritis, nephrectomy, metabolic syndrome, and kidney fibrosis. Most studies have shown that low-power lasers can affect TGFβ1 signaling which is the most important signaling in the treatment of renal fibrosis. Lasers can be effective in reducing or enhancing inflammatory responses, reducing fibrosis factors, and decreasing reactive oxygen species (ROS) levels in kidney disease and glomerular cell proliferation.
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http://dx.doi.org/10.34172/jlms.2020.36DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7118498PMC
March 2020

Postmortem Study of Molecular and Histological Changes in the CA1 Hippocampal Region of Chronic Methamphetamine User.

Iran J Pharm Res 2019 ;18(4):2067-2082

Cellular and Molecular Research Center, Faculty of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.

Methamphetamine (Meth) is recognized as one of the most important new distributed abused drug that causes severe damage to the different parts of the brain, especially hippocampus. Previous studies have demonstrated that Meth can induce apoptosis and cell death in the brain. In this study, we evaluated the long-term effects of Meth abuse in the CA1 region of postmortem hippocampus. Postmortem molecular and histological analysis was performed for five non-addicted subjects and five Meth addicted ones. Iba-1 (microglia) and glial fibrillary acidic protein, GFAP (astrocytes) expression were assayed by western blotting and immunohistochemistry (IHC) methods. Histopathological assessment was done with stereological counts of hippocampal cells stained with hematoxylin and eosin (H and E). Tunel staining was used to detect DNA damage in human brains. In addition, protein-protein interaction analysis network was investigated. Western blotting and immunohistochemistry assay showed overexpression of GFAP and Iba-1 protein in the CA1 hippocampal region of Meth users' brain. Stereological analysis in the CA1 region revealed increased neuron degeneration. Furthermore, significant apoptosis and cell death were confirmed by Tunel assay in the hippocampus. The prominent role of TLR4, IL1B, CASP1, and NLRP3 in the molecular mechanism of Meth was highlighted via PPI network analysis. Chronic Meth use can induce GFAP and Iba-1 upregulation and neuronal apoptosis in the CA1 region of the postmortem hippocampus.
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http://dx.doi.org/10.22037/ijpr.2019.15483.13123DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7059073PMC
January 2019

The Effect of Photobiomodulation Therapy on the Differentiation, Proliferation, and Migration of the Mesenchymal Stem Cell: A Review.

J Lasers Med Sci 2019 1;10(Suppl 1):S96-S103. Epub 2019 Dec 1.

Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

The purpose of this study is to investigate the effect of a low-power laser on the proliferation, migration, differentiation of different types of mesenchymal stem cells (MSCs) in different studies. The relevant articles that were published from 2004 to 2019 were collected from the sources of PubMed, Scopus, and only the articles specifically examining the effect of a lowpower laser on the proliferation, differentiation, and migration of the MSCs were investigated. After reviewing the literature, only 42 articles were found relevant. Generally, most of the studies demonstrated that different laser parameters increased the proliferation, migration, and differentiation of the MSCs, except the results of two studies which were contradictory. In fact, changing the parameters of a low-power laser would affect the results. On the other hand, the source of the stem cells was reported as a key factor. In addition, the combination of lasers with other therapeutic approaches was found to be more effective. The different parameters of lasers has been found to be effective in the proliferation, differentiation, and migration of the MSCs and in general, a low-power laser has a positive effect on the MSCs, helping to improve different disease models.
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http://dx.doi.org/10.15171/jlms.2019.S17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6983866PMC
December 2019

Protective effect of Photobiomodulation Therapy and Bone Marrow Stromal Stem Cells Conditioned Media on Pheochromocytoma Cell Line 12 Against Oxidative Stress Induced by Hydrogen Peroxide.

J Lasers Med Sci 2019 6;10(3):163-170. Epub 2019 Jul 6.

Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Bone marrow stromal stem cells (BMSCs), a type of adult stem cells, secrete bioactive molecules such as trophic factors, growth factors, chemokine and cytokines that may be effective against oxidative stress in neurodegenerative diseases. In this study, we examined the protective effect of BMSCs conditioned media (CM) and photobiomodulation therapy (PBMT) on PC12 cells exposed to H2O2 as an oxidative injury model. BMSCs were cultured and confirmed by flow cytometry analysis and underwent osteogenic and adipogenic differentiation. Then, PC12-H2O2 cells were co-treated with BMSCs-CM and PBMT. The effect of BMSCs-CM and PBMT (He-Ne laser, 632.8nm, 3mW, 1.2J/ cm , 378s) on Bax/Bcl2 expression, cell viability, was assessed by real-time PCR and MTT assay. The length of the Neurite and cell body areas were assessed by Cell A software. Flowcytometry analysis, as well as osteogenic and adipogenic staining, confirmed the BMSCs. The length of the Neurite was the highest in the group which received CM+PBMT and cell body areas were significant in CM+PBMT compared to other groups. Based on our results, elevating H2O2 concentration increased cell death significantly and using concentrations of 250 µM resulted in a dramatic increase in the mortality compared to the other groups. Our result demonstrated that the combination of CM +PBMT has a protective effect on PC12 cells against oxidative stress.
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http://dx.doi.org/10.15171/jlms.2019.26DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6817795PMC
July 2019

Resistance of human primary mesenchymal stem cells to cytotoxic effects of nutlin-3 in vitro.

J Cell Biochem 2020 01 26;121(1):788-796. Epub 2019 Aug 26.

Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.

Background: The small-molecule nutlin-3 was found to be an effective therapeutic compound and p53 activator, and acts as a murine double minute 2 antagonist, although these findings need to be clinically confirmed. The essential components of the bone marrow include mesenchymal stem cells (MSCs), which play a key role in protecting, regenerating, and proliferating hematopoietic stem cells (HSCs). This feature is vital for HSC after exposure to myelotoxic anticancer agents; nevertheless, the effects of nutlin-3 on MSCs remain to be disclosed. The present research study was conducted to examine the antiproliferative and proapoptotic effectiveness of nutlin-3 in bone marrow MSCs (BMSCs).

Materials And Methods: Human-derived BMSCs were cultured for different durations, that is, 24, 48, and 72 hours, and treated using various concentrations of nutlin-3, including 5, 10, 25, 50, and 100 μΜ. To investigate the effect of nutlin-3 on the apoptosis, cell vitality and proliferation in BMSCs, the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), thiazolyl blue tetrazolium bromide, propidium iodide (PI) and annexin V assay, as well as real-time polymerase chain reaction, were used.

Results: BMSCs viability significantly decreased (P < .05) in the cells treated at concentrations of 50 and 100 μM for 24 hours and concentrations of 25, 50, and 100 μM for 48 hours and at all concentrations for 72 hours. The apoptosis of BMSCs (TUNEL positive) was significantly more visible at concentrations of 25 and 50 μM compared with that in the controls (P < .05), while this increased through dose-dependent processes. Annexin V/PI staining revealed negligible dose-dependent increases in all the apoptotic cells after 72 hours of incubation, and this apoptosis elevation was significant at 25 and 50 μM (P < .05).

Conclusion: Resistance to nutlin-3 was observed in human bone marrow-derived MSCs; nevertheless, further clinical data are required to be obtained with long-duration exposure to confirm the present findings.
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http://dx.doi.org/10.1002/jcb.29324DOI Listing
January 2020

Evidence for hyperprolactinemia in migraineurs: a systematic review and meta-analysis.

Neurol Sci 2020 Jan 23;41(1):91-99. Epub 2019 Aug 23.

Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran.

Background: One of the hypothalamus-pituitary axis hormones which may play a crucial role in pathophysiology of migraine is prolactin which is secreted from anterior pituitary gland and synthesized by various immune system cells as well. Whether prolactin blood levels can affect the migraine pathogenesis is an open question. Therefore, investigating prolactin circulatory levels in migraineurs may pave the way to underpin the mechanisms of migraine pathophysiology at biochemical levels. In the current investigation, the prolactin blood levels in the migraine subjects were investigated using systematic review and meta-analysis.

Methods: Using online and specialized biomedical databases including Google Scholar, Medline, Pubmed, Pubmed Central, Embase, and Scopus, without the beginning date restriction until Feb 2019, the systematic review retrieved 11 publications in this systematic review after fulfilling for the inclusion and exclusion criteria. For heterogeneity, extent calculation statistical testing was applied. In the present study, the levels of circulatory prolactin in migraineurs assessed using standardized mean difference (SMD) as the effect size.

Results: Q quantity and I% statistic index showed a high heterogeneity in the 13 selected publications (188.370 and 92.568, respectively) and random-effects model was chosen for further analyses. The meta-analysis on a total number of 460 migraineurs and 429 healthy controls found that the weighted pooled SMD for the effects of prolactin blood concentrations on migraine pathogenesis was as follows: SMD = 1.435 (95% confidence interval, 0.854-2.015).

Conclusion: The current investigation presents evidence that prolactin blood levels are higher in migraineurs than healthy subjects.
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http://dx.doi.org/10.1007/s10072-019-04035-7DOI Listing
January 2020

The Combined Effects of Mesenchymal Stem Cell Conditioned Media and Low-Level Laser on Stereological and Biomechanical Parameter in Hypothyroidism Rat Model.

J Lasers Med Sci 2018 17;9(4):243-248. Epub 2018 Sep 17.

Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Many studies have shown the positive effect of laser radiation and application of the mesenchymal stem cells (MSCs) and their secretion in stimulating bone regeneration. The aim of this study was determining effects of MSC conditioned media (CM) and low-level laser (LLL) on healing bone defects in the hypothyroid male rat. We assigned 30 male Wistar rats randomly to 3 groups: control, hypothyroidism, CM+LLL. Four weeks after surgery, the right tibia was removed. Biomechanical examination and histological examinations were performed immediately. Our results showed significant increase in bending stiffness (116.09±18.49), maximum force (65.41±8.16), stress high load (23.30±7.14), energy absorption (34.57±4.10), trabecular bone volume (1.34±0.38) and the number of osteocyte, osteoblast, and osteoclast (12.77±0.54, 6.19±0.80, 1.12±0.16 respectively) in osteotomy site in the LLL+CM group compared to the hypothyroidism group (<0.05). The results indicated that using the LLL + CM may improve fracture regeneration and it may hasten bone healing in the hypothyroid rat.
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http://dx.doi.org/10.15171/jlms.2018.44DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6499561PMC
September 2018

Effects of Sertoli Cell Transplantation on Spermatogenesis in Azoospermic Mice.

Cell Physiol Biochem 2019 8;52(3):421-434. Epub 2019 Mar 8.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran,

Background/aims: The aim of this study was to evaluate the potential and significant applications of Sertoli cells (SCs) transplantation, and to explore the effect of transplantation on spermatogenesis process, in azospermic mice.

Methods: In this study, we utilized 18 adult mice (28‒30 g), divided into four experimental groups: (1) control, (2) vehicle (DMSO 2%) (10 µl) (3) busulfan and (4) busulfan+ SCs (1×104 cells/μL). SCs were isolated from the testis of 4-week-old mouse and after using anesthetics, 10 μl of SCs suspension (1×104 cells/μL) was injected over 3-5 min, into each testis and subsequently, sperm samples were collected from the tail of the epididymis. Afterward, the animals were euthanized and testis samples were taken for histopathology experiments, and RNA extraction, in order to examine the expression of c-kit, STRA8 and PCNA genes.

Results: Our data showed that SCs transplantation could notably increase the total sperm count and the number of testicular cells, such as spermatogonia, primary spermatocyte, round spermatid, SCs and Leydig cells, compared to the control, DMSO and busulfan groups. Furthermore, the result showed that the expression of c-kit and STRA8 were significantly decreased in busulfan and busulfan/SCs groups, at 8 weeks after the last injection (p<0.001), but no significant decrease was found for PCNA, compared to the control and DMSO groups (P<0.05).

Conclusion: These findings suggest that SCs transplantation may be beneficial as a practical approach for therapeutic strategies in reproductive and regenerative medicine. We further highlighted the essential applications that might provide a mechanism for correcting fertility in males, suffering from cell deformity.
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http://dx.doi.org/10.33594/000000030DOI Listing
March 2019

Intra-articular ozone therapy efficiently attenuates pain in knee osteoarthritic subjects: A systematic review and meta-analysis.

Complement Ther Med 2019 Feb 28;42:240-247. Epub 2018 Nov 28.

Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran.

Objectives: Ozone (O) gas is being used for chronic pain relief in knee osteoarthritis (KOA). However, there are controversies whether this gas can be medically useful in KOA pain treatments. The aim of this study was to evaluate the effectiveness of intra-articular ozone therapy for pain relief in KOA subjects using a systematic review and meta-analysis and standardized mean difference (SMD) as the effect size.

Method: Using specialized biomedical online databases of Pubmed Central, Pubmed, Medline, Google scholar, Scopus and Embase databases without the beginning date restriction until July 2018, the systematic review retrieved 10 studies for meta-analysis after fulfilling the inclusion and exclusion criteria.

Results: Analysis of Q and I% indices showed a high heterogeneity in the selected studies (2600.330 and 99.654, respectively), thus, the random-effects model was chosen for SMD calculation. The primary analysis for the main hypothesis found that the weighted pooled effect size for the impact of intra-articular ozone therapy for pain reduction was as follows: SMD= -28.551 (95% confidence interval, -32.553 to -24.549). The P-value for the significance of the combined SMD examined by the z-test was 0.000 and thus, it was clearly considered statistically significant.

Conclusion: This meta-analysis presents evidence that intra-articular ozone therapy is an effective way for chronic pain management in KOA.
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http://dx.doi.org/10.1016/j.ctim.2018.11.023DOI Listing
February 2019

Elevated blood apelin levels in type 2 diabetes mellitus: A systematic review and meta-analysis.

Diabetes Res Clin Pract 2019 Feb 21;148:43-53. Epub 2018 Dec 21.

Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran.

Aims: Apelin is a circulatory blood peptide acting as a ligand for the orphan G protein-coupled receptor known as APJ. Whether apelin blood levels can affect the pathogenesis of type 2 diabetes mellitus is an open question. In the present study, we aimed to assess the levels of circulatory apelin peptide in the type 2 diabetic subjects using systematic review and meta-analysis under random-effects model and standardized mean difference (SMD) as the effect size. For heterogeneity testing, Q and I% statistic indices as well as meta-regression were applied.

Methods: Using specialized biomedical online databases of Pubmed, Pubmed Central, Medline, Google scholar, Scopus and Embase databases without the beginning date restriction until July 2018, the systematic review retrieved nine studies for meta-analysis after fulfilling the inclusion and exclusion criteria.

Results: Analysis of Q and I% statistic indices as well as meta-regression showed a high heterogeneity in the 16 selected studies (737.578 and 96.475, respectively), thus, the random-effects model was chosen. The primary analysis for the main hypothesis on a total number of 1102 cases and 1078 healthy control subjects found that the weighted pooled SMD for the impact of apelin blood concentration in type 2 diabetes mellitus was as follows: SMD = 2.136 (95% confidence interval, 1.580-2.693). The P-value for the significance of the combined SMD examined by the z-test was 0.000 and thus, it was clearly significant.

Conclusions: This meta-analysis presents evidence that apelin circulatory levels are higher in type 2 diabetic subjects than normal controls.
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http://dx.doi.org/10.1016/j.diabres.2018.12.012DOI Listing
February 2019

17 β-Estradiol Oxidative Stress Attenuation and Autophagy-Induced Dopaminergic Neuroprotection.

Cell J 2019 Apr 18;21(1):1-6. Epub 2018 Nov 18.

Department of Clinical Biochemistry, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.

Objective: Degeneration of dopaminergic neurons in the substantia nigra of the brain stem is the main pathological aspect of Parkinson's disease (PD). 17 β-estradiol (E2) has neuroprotective effects on substantia nigra, however, the underlined mechanism is not well-known. In this study, we evaluated the neuroprotective effects of E2 in the ovariectomized 6-hydroxydopamine- (6-OHDA) rat model of PD.

Materials And Methods: In this experimental study, all animals were ovariectomized to avoid any further bias in E2 levels and then these ovariectomized rats were randomly assigned into three experimental groups (10 rats in each group): ovariectomized control group (OCG), ovariectomized degeneration group receiving 25 μg of 6-OHDA into the left corpus striatum (ODG), and ovariectomized E2 pretreatment group pretreated with 0.1 mgkg-1 of 17 β-estradiol for three days prior to the destruction of corpus striatum with 6-OHDA (OE2PTG). The apomorphine behavioral test and Nissl staining were performed in all experimental groups. The expressions of Sequestosome-1 (P62), Unc- 51 like autophagy activating kinase (Ulk1), and microtubule-associated proteins 1A/1B light chain 3B (Lc3) genes were evaluated using reverse transcriptionpolymerase chain reaction (RT-PCR).

Results: E2 administration reduced the damages to the dopaminergic neurons of the substantia nigra. The motor behavior, the number of rotations, and histological tests in the treatment group showed the cell survival improvement in comparison with the control groups indicating that E2 can inhibit the neurodegeneration. P62 and Lc3 were expressed in all experimental groups while Ulk1 was not expressed in ODG group. Moreover, Ulk1 was expressed after the treatment with E2 in OE2PTG group.

Conclusion: E2 prevents neurodegeneration in dopaminergic neurons of the midbrain by over-expression of Ulk1 gene and augmenting the induction of autophagy.
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http://dx.doi.org/10.22074/cellj.2019.5799DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275422PMC
April 2019

The effects of Nutlin-3 on morphology, cellular proliferation, and apoptosis in rat primary mesenchymal stem cells.

J Cell Physiol 2019 07 27;234(7):11424-11430. Epub 2018 Nov 27.

Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.

Introduction: Nutlin-3 is a powerful antagonist of murine double minute 2/p53 interaction demonstrating promising therapeutic anticancer activity, which has not been clinically approved yet. Mesenchymal stem cells (MSCs) are an important part of the bone marrow niche and support regeneration and proliferation of hematopoietic stem cells after exposure to myelotoxic anticancer agents; however, the effect of Nutlin-3 compounds on MSCs themselves remains to be elucidated.

Materials And Methods: Rat-derived bone marrow-derived MSCs (BMSCs) were cultured and treated with different concentrations (5, 10, 25, 50, and 100 μM) and times (24, 48, and 72 hr) of Nutlin-3. The microculture tetrazolium test, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and propidium iodide and annexin-V assays, and quantitative real-time reverse-transcription polymerase chain reaction were performed to assess the effects of Nutlin-3 on the cell viability, proliferation, and apoptosis in BMSCs.

Results: The viability of BMSCs in the treated cells with concentrations of 100 μM at 24 hr, 50 and 100 μM at 48 hr, and in all concentrations at 72 hr was significantly (p < 0.05) low. The apoptotic index showed that the TUNEL-positive BMSCs were significantly higher in concentrations of 25 and 50 μM in comparison to control group ( p < 0.05) and augmented in a dose-dependent manner. Annexin-V-PI staining showed after 72 hr of incubation, there was a slight dose-dependent increase in total apoptotic cells at 10 and 25 μM of Nutlin-3, but a massive significant increase at 50 μM.

Conclusion: Here, we show that rat BMSCs are relatively resistant to Nutlin-3; however, further in vivo data with long-term exposure may help to corroborate our findings.
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http://dx.doi.org/10.1002/jcp.27798DOI Listing
July 2019

Stem cell transplantation and functional recovery after spinal cord injury: a systematic review and meta-analysis.

Anat Cell Biol 2018 Sep 28;51(3):180-188. Epub 2018 Sep 28.

Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Spinal cord injury is a significant cause of motor dysfunctions. There is no definite cure for it, and most of the therapeutic modalities are only symptomatic treatment. In this systematic review and meta-analysis, the effectiveness of stem cell therapy in the treatment of the spinal cord injuries in animal models was studied and evaluated. A systematic search through medical databases by using appropriate keywords was conducted. The relevant reports were reviewed in order to find out cases in which inclusion and exclusion criteria had been fulfilled. Finally, 89 articles have been considered, from which 28 had sufficient data for performing statistical analyses. The findings showed a significant improvement in motor functions after cell therapy. The outcome was strongly related to the number of transplanted cells, site of injury, chronicity of the injury, type of the damage, and the induction of immune-suppression. According to our data, improvements in functional recovery after stem cell therapy in the treatment of spinal cord injury in animal models was noticeable, but its outcome is strongly related to the site of injury, number of transplanted cells, and type of transplanted cells.
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http://dx.doi.org/10.5115/acb.2018.51.3.180DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6172584PMC
September 2018

SMER28 Attenuates Dopaminergic Toxicity Mediated by 6-Hydroxydopamine in the Rats via Modulating Oxidative Burdens and Autophagy-Related Parameters.

Neurochem Res 2018 Dec 4;43(12):2313-2323. Epub 2018 Oct 4.

Raymond Chang School, Ryerson University, Toronto, Canada.

Parkinson's disease is the second most common neurodegenerative disease that occurs due to cellular autophagy deficiency and the accumulation of alpha-synuclein in the dopaminergic neurons of the substantia nigra pars compacta (SNc) of the brainstem. The SMER28 (also known as 6-Bromo-N-prop-2-enylquinazolin-4-amine) is an autophagy inducer. In this study, the neuroprotective effects of SMER28 were evaluated on autophagy induction, antioxidant system activation, and microgliosis attenuation. The Parkinson's disease model was developed in the male Wistar rats by injection of 6-OHDA into the left striatum. Apomorphine-induced behavior assessment test and SNc cell counting were performed to investigate the neuroprotective effects of SMER28. This study examined the pharmacological roles of SMER28, especially by focusing on the autophagy (p62/ SQSTM1 and LC3II/LC3I ratio where LC3 is microtubule-associated protein 1A/1B-light chain 3), inhibiting free radicals, and activating the antioxidant system. The levels of malondialdehyde (MDA), reactive oxygen species (ROS), glutathione (GSH), GSH/glutathione peroxidase (GP), superoxide dismutase (SOD) activity and nuclear factor-erythroid 2-related factor-2 (Nrf2) were measured to evaluate the antioxidant activity of SMER28. Moreover, Iba-1 (ionized calcium binding adaptor molecule, indicating microgliosis) and tyrosine hydroxylase immunoreactivities were evaluated in the SNc. In the behavioral assessment, SMER28 (50 µg/kg) attenuated damages to the SNc dopaminergic neurons, characterized by improved motor function. The tissue observations revealed that SMER28 prevented the destruction of SNc neurons and attenuated microgliosis as well. It also reduced MDA and ROS production and increased GSH, GP, SOD, and Nrf2 activities by inducing autophagy (decreasing p62 and increasing LC3II/LC3I ratio). Consequently, possibly with further studies, it can be considered as a drug for neurodegenerative diseases with proteinopathy etiology.
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http://dx.doi.org/10.1007/s11064-018-2652-2DOI Listing
December 2018

Trehalose Activates Autophagy and Prevents Hydrogen Peroxide-Induced Apoptosis in the Bone Marrow Stromal Cells.

Iran J Pharm Res 2018 ;17(3):1141-1149

Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran.

Bone marrow stromal stem cells (BMSCs) play a significant role in cell therapy. These cells quickly die after transplantation to the affected area due to oxidative stress. The natural disaccharide, trehalose which can be known as autophagy inducer. The present study aimed to investigate the role of trehalose in preventing BMSCs from oxidative stress caused by HO. BMSCs were isolated from the adult rats. The cells were divided into three groups: (a) control; (b) 100 µM HO; (c) 100 µM HO and trehalose 3%. The morality rate was analyzed by viability test. Immunocytochemistry and Western blot was used in order to evaluate p62 protein and LC3II/LC3I ratio, respectively. In order to evaluate apoptosis, cleaved caspase-3 protein was used. In viability test, the survival rate for BMSCs after 8 h were 82%, 72%, 49%, and 39% (for groups who received 50, 100, 200, and 400 µM HO, respectively) compared to the control group. Pre-treatment with the use of trehalose 3% increased cell survivals. The levels of p62 protein, were increased in the cells under HO treatment, while the levels of p62 protein in the cytoplasm, as autophagy inclusions, reduced for the group with trehalose pre-treatment. In addition, trehalose caused to increase LC3II/LC3I ratio and decreased the expression of cleaved caspase-3. Trehalose decreased apoptosis and increased the autophagy and survival levels of the cells against HO. Due to the unique properties of trehalose and its low toxicity, it can be used as a pharmaceutical agent in cellular transplantation to reduce oxidative stress.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6094425PMC
January 2018

Therapeutic Effects of Laser on Partial Osteotomy in the Rat Model of Hypothyroidism.

J Lasers Med Sci 2018 20;9(2):121-127. Epub 2018 Mar 20.

Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Several experimental studies have displayed positive result for laser radiation on stimulating bone regeneration in recent years. The purpose of this experimental study was to determine low-level laser (LLL) effects on partial bone defects in hypothyroidism male rat. Forty male Wistar rats were randomly distributed as below groups: hypothyroidism + laser (Hypo + laser), hypothyroidism (Hypo), and control. Four weeks after surgery, the tibia bone was removed. Biomechanical and histological examinations were performed immediately. Our results showed significant reduction in the absorption of energy, resistance in bending deformation (bending stiffness), maximum force, high stress load, trabecular bone volume, and number of osteocytes, osteoblasts and osteoclasts in the osteotomy site in hypothyroidism rats compared to hypothyroidism + laser group (P<0.05). The results indicated that using laser may improve fracture regeneration and it may accelerate bone healing in hypothyroidism rat.
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http://dx.doi.org/10.15171/jlms.2018.23DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6046390PMC
March 2018

Low Level of Autophagy-Related Gene 10 (ATG10) Expression in the 6-Hydroxydopamine Rat Model of Parkinson's Disease

Iran Biomed J 2018 01 23;22(1):15-21. Epub 2017 Jul 23.

Hearing Disorder Research center, Shahid Beheshti University of Medical Science,Tehran, Iran

Background: Autophagy is a mechanism disassembling the damaged organelles from the cell. This study attempted to examine the expression of several autophagy-related genes in Parkinson’s disease (PD) rat model.

Methods: The male Wistar rats were divided into three groups as control, sham, and lesion. In the latter group, the PD rat model was induced by the injection of 6-hydroxydopamine in the striatum. The behavioral test was conducted one (baseline) and four weeks after the surgery through apomorphine hydrochloride. Then the RT-PCR technique was employed to evaluate the expressions of p62/SQSTM, autophagy-related genes (ATG)5, ATG12, ATG16L1, ATG10, as well as GAPDH and LC3.

Results: By injecting apomorphine, the striatal lesion group showed a significant contralateral rotation at fourth week as compared to the baseline. The examination of p62, ATG5, ATG12, ATG16L1, and LC3 expressions using RT-PCR revealed that p62, ATG5, ATG12, LC3, and ATG16L1 were expressed in the substantia nigra of PD rat model, while ATG10 was not expressed.

Conclusion: ATG10 expression is necessary for the initiation of autophagy. Thus, these results show that autophagy deregulation occurs in the initiation stages of the process in the rat model of PD.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5712380PMC
http://dx.doi.org/10.22034/ibj.22.1.15DOI Listing
January 2018

Condition medium of cerebrospinal fluid and retinoic acid induces the transdifferentiation of human dental pulp stem cells into neuroglia and neural like cells.

Anat Cell Biol 2017 Jun 27;50(2):107-114. Epub 2017 Jun 27.

Department of Medical Genetics, Faculty of Medicine & Dentistry, University of Alberta, Edmonton, Alberta, Canada.

Cerebrospinal fluid (CSF) contains several molecules which are essential for neurogenesis. Human dental pulp stem cells (hDPSCs) are putatively neural crest cell-derived that can differentiate into neurons and glial cells under appropriate neurotrophic factors. The aim of this study was to induce differentiation of hDPSCs into neuroglial phenotypes using retinoic acid (RA) and CSF. The hDPSCs from an impacted third molar were isolated by mechanical and digestion and cultured. The cells have treated by 10 µM RA (RA group) for 8 days, 10% CSF (CSF group) for 8 days and RA with CSF for 8 days (RA/CSF group). Nestin, microtubule-associated protein 2 (MAP2), and glial fibrillary acidic protein immunostaining were used to examine the differentiated cells. Axonal outgrowth was detected using Bielschowsky's silver impregnation method and Nissl bodies were stained in differentiated cells by Cresyl violet. The morphology of differentiated cells in treated groups was significantly changed after 3-5 days. The results of immunocytochemistry showed the presence of neuroprogenitor marker nestin was seen in all groups. However, the high percentage of nestin positive cells and MAP2, as mature neural markers, were observed at the pre-induction and induction stage, respectively. Nissl bodies were detected as dark-blue particles in the cytoplasm of treated cells. Our findings showed the RA as pre-inducer and CSF as inducer for using in vitro differentiation of neuron-like cells and neuroglial cells from hDPSCs.
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http://dx.doi.org/10.5115/acb.2017.50.2.107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5509894PMC
June 2017

In vitro non-viral murine pro-neurotrophin 3 gene transfer into rat bone marrow stromal cells.

J Neurol Sci 2017 Apr 21;375:137-145. Epub 2017 Jan 21.

Shefa Neurosciences Research Center, Khatam Al-Anbia Hospital, Tehran, Iran.

Neurotrophin 3 (NT-3) is an important factor for promoting prenatal neural development, as well as regeneration, axogenesis and plasticity in postnatal life. Therapy with NT-3 was reported to improve the condition of patients suffering from degenerative diseases and traumatic injuries, however, the disadvantage of NT-3 protein delivery is its short half-life, thus our alternative approach is the use of NT-3 gene therapy. In this study, the bone marrow stromal cells (BMSCs) were isolated from adult rats, cultured for 4 passages and transfected with either pEGFP-N1 or a constructed vector containing murine proNT-3 (pSecTag2/HygroB-murine proNT-3) using Lipofectamine 2000 followed by Hygromycin B (200mg/kg). The transfection efficiency of the transiently transfected BMSCs was evaluated using the green fluorescence protein containing vector (pEGFP-N1). A quantitative evaluation of the NT-3 expression of mRNA using real time qRT-PCR shows that there was double fold increase in NT-3 gene expression compared with non-transfected BMSCs, also, the culture supernatant yielded double fold increase in NT-3 using ELISA technique, the data were supported by immunoblotting technique. This suggests that the use of this transfection technique can be useful for gene therapy in different neurological disorders with neurodegenerative or traumatic origins.
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http://dx.doi.org/10.1016/j.jns.2017.01.058DOI Listing
April 2017

Efficacy of Compound Therapy by Ginseng and Ciprofloxacin on Bacterial Prostatitis.

Cell J 2016 4;18(1):103-11. Epub 2016 Apr 4.

Department of Anatomical Sciences, Iran University of Medical Sciences, Tehran, Iran; Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran.

Objective: Genitourinary tract infections play a significant role in male infertility. Infections of reproductive sex glands, such as the prostate, impair function and indirectly affect male fertility. The general aim of this study is to investigate the protective effect of Korean red ginseng (KRG) on prostatitis in male rats treated with ciprofloxacin (CIPX).

Materials And Methods: In this experimental study, we randomly divided 72 two male Wistar rats into 9 groups. The groups were treated as follows for 10 days: i. Control (no medication), ii. Sham [(normal saline injection into the vas deferens and oral administration of phosphate-buffered saline (PBS)], iii. Ginseng, iv. CPIX, v. CIPX+ginseng, vi. Uropathogenic Escherichia coli (E. coli) (UPEC), vii. UPEC+ginseng, viii. UPEC+CIPX, and ix. UPEC+ginseng+CIPX. The rats were killed 14 days after the last injection and the prostate glands were removed. After sample preparation, routine histology was performed using hematoxylin and eosin staining. The terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) method was used to determine the presence of apoptotic cells.

Results: The severity score for acinar changes and inflammatory cell infiltration in the UPEC+CIPX group did not significantly different from the UPEC group. However this score significantly decreased in the UPEC+CIPX+ginseng group compared to the UPEC group. Apoptotic index of all ginseng treated groups significantly decreased compared to the UPEC and CPIX groups.

Conclusion: These results suggested that ginseng might be an effective adjunct in CIPX treatment of prostatitis. The combined use ginseng and CIPX was more effective than ginseng or CIPX alone.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4819379PMC
http://dx.doi.org/10.22074/cellj.2016.3993DOI Listing
April 2016

Creatine Enhances Transdifferentiation of Bone Marrow Stromal Cell-Derived Neural Stem Cell Into GABAergic Neuron-Like Cells Characterized With Differential Gene Expression.

Mol Neurobiol 2017 04 24;54(3):1978-1991. Epub 2016 Feb 24.

Middle Euphrates Neuroscience Center-Kufa College of Medicine, Annajaf, Iraq.

Creatine was reported to induce bone marrow stromal cells (BMSC) into GABAergic neuron-like cells (GNLC). In a previous study, creatine was used as a single inducer for BMSC into GNLC with low yield. In this study, BMSC-derived neurospheres (NS) have been used in generating GABAergic phenotype. The BMSC were isolated from adult rats and used in generating neurospheres and used for producing neural stem cells (NSC). A combination of all-trans-retinoic acid (RA), the ciliary neurotrophic factor (CNTF), and creatine was used in order to improve the yield of GNLC. We also used other protocols for the transdifferentiation including RA alone; RA and creatine; RA and CNTF; and RA, CNTF, and creatine. The BMSC, NSC, and GNLC were characterized by specific markers. The activity of the GNLC was evaluated using FM1-43. The isolated BMSC expressed Oct4, fibronectin, and CD44. The NS were immunoreactive to nestin and SOX2, the NSC were immunoreactive to nestin, NF68 and NF160, while the GNLC were immunoreactive to GAD1/2, VGAT, GABA, and synaptophysin. Oct4 and c-MYC, pluripotency genes, were expressed in the BMSC, while SOX2 and c-MYC were expressed in the NSC. The activity of GNLC indicates that the synaptic vesicles were released upon stimulation. The conclusion is that the combination of RA, CNTF, and creatine induced differentiation of neurosphere-derived NSC into GNLC within 1 week. This protocol gives higher yield than the other protocols used in this study. The mechanism of induction was clearly associated with several differential pluripotent genes.
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http://dx.doi.org/10.1007/s12035-016-9782-9DOI Listing
April 2017

Regulatory T cell number in multiple sclerosis patients: A meta-analysis.

Mult Scler Relat Disord 2016 Jan 10;5:73-6. Epub 2015 Nov 10.

Department of Anatomy, School of Medicine, Zanjan University of Medical Sciences (ZUMS), Zanjan, Iran.

Regulatory T cells (Treg cells), defined as CD4(+) CD25(+) FoxP3(+) T cells by expression of CD4, high-affinity IL-2 receptor and the transcription factor, forkhead box P3 (FoxP3). They play a pivotal role in protecting individuals from autoimmunity and a growing body of evidence suggests their role in the prevention of multiple sclerosis development. However, there are discrepancies about the type of defect in the Treg cells of multiple sclerosis patients and especially whether the Treg number alteration could be contributed to multiple sclerosis pathogenesis. Indeed, whether low number of Treg cells can be a risk factor contributing to multiple sclerosis pathogenesis is the matter of debate and there is not any comprehensive agreement on it. Thus, the objective of this systematic review and meta-analysis was to precisely quantify the nature and magnitude of the association between Treg cell number and the risk ratio/odds ratio (OR) of multiple sclerosis in the case-control studies. Hence, medical databases of Embase, PubMed/Medline, PubMed, PubMed Central and SCOPUS were searched for empirical papers using "Regulatory T cell frequency", "Treg frequency" in combination with "multiple sclerosis". In the case-control studies, papers were reviewed for inclusion/exclusion criteria and 8 publications were included. Under random-effect model meta-analysis the data showed that the frequency of Treg cells was not a risk factor in multiple sclerosis using current laboratory methods.
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http://dx.doi.org/10.1016/j.msard.2015.11.004DOI Listing
January 2016

Polarized neural stem cells derived from adult bone marrow stromal cells develop a rosette-like structure.

In Vitro Cell Dev Biol Anim 2013 Sep 15;49(8):638-52. Epub 2013 Jun 15.

Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran,

Bone marrow stromal cells (BMSCs) were reported to form floating aggregation of cells with expression of nestin, a marker for neural stem cells (NSCs). The purpose of this investigation is to evaluate the morphology and the molecular markers expressed by NSCs derived from these neurospheres. The BMSCs were isolated from Sprague Dawley rats and evaluated for osteogenesis, lipogenesis, and expression of fibronectin, CD90, CD106, CD31, and Oct4. The BMSCs were cultured with Dulbecco's modified Eagle's medium (DMEM)/F12 containing 15% fetal bovine serum, then with DMEM/F12 containing 2% B27, basic fibroblast growth factor, and epidermal growth factor. The cell aggregates or spheres were stained with acridine orange, which showed that the neurospheres comprised aggregated cells at either premitotic/postsynthetic (PS), postmitotic/presynthetic (PM) phases of cell cycle, or a mixture of both. The NSCs harvested from the neurospheres were polar with eccentric nucleus, and at either a PS or a PM cell cycle phases, some cells at the latter phase tended to form rosette-like structures. The cells were immunostained for molecular markers such as nestin, neurofilament 68 (NF68), NF160, and NF200 and glial fibrillary acidic protein (GFAP). Myelin basic protein (MBP), the pluripotency (Oct4, Nanog, and SOX2), and the differentiation genes (NeuroD1, Tubb4, and Musashi I) were also evaluated using reverse transcription polymerase chain reaction (RT-PCR). Nestin, NF68, NF160, NF200, GFAP, O4, and N-cadherin were expressed in the NSCs. The percentage of immunoreactive cells to nestin was significantly higher than that of the other neuronal markers. MBP was not expressed in BMSCs, neurospheres, and NSCs. The neurospheres were immunoreactive to GFAP. RT-PCR showed the expression of NeuroD1 and Musashi I. The pluripotency gene (SOX2) was expressed in NSCs. Oct4 and Nanog were expressed in BMSCs, while Oct4 and SOX2 were expressed in the neurosphere. This indicates that a pluripotency regularity network existed during the transdifferentiation of BMSCs into NSCs. Image processing of the neurospheres showed that the cells tended to form radial patterns. The conclusion of this study is that the NSCs generated from the BMSC-derived neurospheres have the morphology and the characteristics of neuroepithelial cells with tendency to forming rosette-like structures.
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http://dx.doi.org/10.1007/s11626-013-9628-yDOI Listing
September 2013