Ameliorate effects of resveratrol and l-carnitine on the testicular tissue and sex hormones level in busulfan induced azoospermia rats.

Busulfan (Bus), is an alkylating agent widely used in chemotherapy which has been proven to possess toxic side effects on testicles. This study was carried out to compare the probable treatment effects of resveratrol (Res) or/and l-carnitine (Lca), as strong antioxidants, on the testicular tissue as well as on the level of sex hormones in busulfan-induced azoospermic rat models. A total of 78 adult male rats, were divided into six different experimental groups including: 1) Control; 2) Lca + Res; 3) BUS; 4) Bus + Lca; 5) BUS + Res and 6) Bus + Lca + Res. Busulfan was intraperitoneally administered in a single dose (10 mg/kg b.w), while resveratrol (20 mg/kg b.w/day) and l-carnitine (200 mg/kg b.w/day) were orally administered by gavage during 48 consecutive days to the rats. At the end of the experiment in all groups the level of LH, FSH, and testosterone were biochemically analyzed by ELISA and the testicular tissue evaluated histologically using stereological technique. Results showed that Lca or/and Res, increased the body and testis weight, the volume of the testis, interstitial tissue, germinal epithelium, and seminiferous tubule, the number of the different cells of germinal epithelium and the level of testosterone. On the other hand, Lca, Res and their combination decreased the concentration of LH and FSH compared to the group treated with Bus. In conclusion, these results suggested that l-carnitine or/and resveratrol treatment significantly attenuated busulfan -induced changes of the rat reproductive system led to the recovery of both testis and sperm parameters. However, co-administration of L-ca and Res was more effective than their individual treatment. This combination may alleviate the side effects of alkylating drugs, such as busulfan and may be beneficial for spermatogenesis.

Methamphetamine Downregulates the Sperm-Specific Calcium Channels Involved in Sperm Motility in Rats.

Calcium channels play essential roles in sperm motility. A family of sperm-specific cation channels including CatSper1-4 has been identified as voltage-dependent ion channels that act as sperm motility regulators. Methamphetamine is known to cause apoptosis in seminiferous tubules and affect sperm quality. This research was conducted to investigate the effects of methamphetamine on expression of the CatSper family and Mvh genes. Thirty-six adult Wistar rats were divided into four groups of nine rats each: the control and experimental groups 1, 2, and 3. The control group received no solvents or drugs, but experimental groups 1, 2, and 3 were daily given 0.2 mL of a solution by gavage that contained 0.5, 1, and 2 mg of methamphetamine, respectively, for 45 days. The rats were then anesthetized, and one testis removed from each rat was used in a reverse transcription-polymerase chain reaction (RT-PCR). Analysis of variance (ANOVA) and Tukey's posthoc test were used to analyze the data at the P < 0.05 significance level. Treatment with methamphetamine resulted in decreased testis and epididymis weights compared to the control rats. The results showed that the mRNA fold expression level of the CatSper family and Mvh genes decreased significantly in experimental groups compared to that in the control (P < 0.05). Methamphetamine decreased the expression levels of the CatSper and Mvh genes, and thus, it seemed that it can increase the probability of infertility through sperm motility reduction by lowering the expression levels of these genes.

Ovulation Induction Changes Epigenetic Marks of Imprinting Genes in Mice Fetus Organs.

OBJECTIVE: Genomic imprinting is an epigenetic phenomenon that plays a critical role in normal development of embryo. Using exogenous hormones during assisted reproductive technology (ART) can change an organism hormonal profile and subsequently affect epigenetic events. Ovarian stimulation changes gene expression and epigenetic pattern of imprinted genes in the organs of mouse fetus. MATERIALS AND METHODS: For this experimental study, expression of three imprinted genes H19, Igf2 (Insulin-like growth factor 2) and Cdkn1c (Cyclin-dependent kinase inhibitor 1C), which have important roles in development of placenta and embryo, and the epigenetic profile of their regulatory region in some tissues of 19-days-old female fetuses, from female mice subjected to ovarian stimulation, were evaluated by quantitative reverse-transcription PCR (qRT-PCR) and Chromatin immunoprecipitation (ChIP) methods. RESULTS: H19 gene was significantly lower in heart (P<0.05), liver (P<0.05), lung (P<0.01), placenta (P<0.01) and ovary (P<0.01). It was significantly higher in kidney of ovarian stimulation group compared to control fetuses (P<0.05). Igf2 expression was significantly higher in brain (P<0.05) and kidney (P<0.05), while it was significantly lower in lung of experimental group fetuses in comparison with control fetuses (P<0.05). Cdkn1c expression was significantly higher in lung (P<0.05). It was significantly decreased in placenta of experimental group fetuses rather than the control fetuses (P<0.05). Histone modification data and DNA methylation data were in accordance to the gene expression profiles. CONCLUSION: Results showed altered gene expressions in line with changes in epigenetic pattern of their promoters in the ovarian stimulation group, compared to normal cycle.

Effect of ß-asarone in normal and ß-amyloid-induced Alzheimeric rats.

INTRODUCTION: ß-Asarone is a major component of Acorus tatarinowii Schott. It has pharmacological effects that include antihyperlipidemic, anti-inflammatory, and antioxidant activity. In the present study, the effect of ß-asarone on neurodegeneration induced by intrahippocampal administration of ß-amyloid was investigated in adult male Wistar rats. MATERIAL AND METHODS: The rats were randomly divided into 9 groups: normal control, sham-operated control, ß-asarone (12.5, 25, and 50 mg/kg intragastrically, daily) alone, Alzheimeric control rats (ß-amyloid, intrahippocampal), ß-asarone (12.5, 25, and 50 mg/kg intragastrically, daily) together with ß-amyloid, and treatment was performed accordingly. Animals were injected with ß-amyloid bilaterally. Animals received ß-asarone daily using an intragastric tube for 50 days, starting from 30 days before administration of the ß-amyloid. The rats were sacrificed and parameters of oxidative stress, superoxide dismutase (SOD) and glutathione peroxidase (GPX) activity were measured in hippocampus homogenate. Histopathological changes were examined by Bielschowsky staining. RESULTS: Our results showed that administration of ß-asarone (25 and 50 mg/kg) significantly increased the levels of antioxidant enzymes, including SOD (1.09 ±0.02, 1.21 ±0.02, p < 0.001, respectively) and GPX (58.94 ±0.78, 68.92 ±3.64, p < 0.001, respectively) in comparison with Alzheimeric control rats (SOD and GPX level for Alzheimeric control group: 0.44 ±0.01, 35.09 ±1.15, respectively). Histopathological examination showed that ß-asarone decreased cell loss in the cerebral cortex and hippocampus in Alzheimeric rats. CONCLUSIONS: These results indicate that ß-asarone is effective in providing protection against oxidative stress and neuronal damage induced by ß-amyloid.

Cytotoxic Effects of Juglone and Pterocarya fraxinifolia on Prostate Cancer Cells.

AIM: Juglone with naphthoquinone structure has medicinal properties and its anticarcinogenic and antioxidant effects have been proven. In this research, the cytotoxic and apoptosis effects of juglone and Pterocarya fraxinifolia (PF) methanolic extract on human prostate cancer cells were studied. MATERIALS AND METHODS: The PC3 and DU145 human cancer cells and normal cells of primary prostate epithelial cells (ATCC PCS-440-010) were treated with juglone and PF extract at the concentrations of 10, 50, 100, 500, and 1000 µg/mL for 24, 48, 72, and 96h. The morphological changes were examined by reversed microscope. The survival percentage of cell lines was evaluated by MTT (3,4,5-dimethylthiazole-2-yl-2,5-diphenyltetrazolium bromide) test. The rate of apoptosis and expression of AR and CLU genes were examined by flow cytometry and real-time polymerase chain reaction. RESULTS: All concentrations after 24h caused morphological changes in PC3 and DU145 cells, and these changes were intensified after 48, 72, and 96h. Also, concentrations of 100 and 500 µg/mL caused morphological changes in normal cells. The results of MTT test showed a significant decrease in PC3 and DU145 cell survival rate at 50, 100, and 500 µg/mL concentrations (P < 0.05). Juglone at 10 µg/mL concentration induced apoptosis in cancer cell lines. CONCLUSION: Juglone and PF could decrease the growth of cancer cell lines through the mitochondrial pathway. So PF could be considered as a potential candidate for therapeutic herbal medicine in treating cancers.

Changes in the Hypothalamic-Pituitary-Gonadal Axis in Adult Male Rats Poisoned with Proteus and Biscaya Insecticides.

BACKGROUND: Insecticides may have negative effects on reproductive organs. Given the interaction between leptin and the hypothalamic-pituitary-gonadal (HPG) axis, we sought to investigate the changes in leptin and the HPG axis in adult male rats poisoned with Proteus and Biscaya insecticides. METHODS: Our experimental subjects were 110 adult male Wistar rats (80-90 days of age; average weight=200-210 g). They were randomly split into 11 groups of 10 rats: control, sham, and 9 experimental groups namely treatment with 2.75, 5.5, and 11 mg/kg/BW of Proteus, treatment with 1.5, 3, and 6 mg/kg/BW of Biscaya, treatment with 2.75 mg/kg/BW of Proteus+1.5 mg/kg/BW of Biscaya, treatment with 5.5 mg/kg/BW of Proteus+3 mg/kg/BW of Biscaya, and treatment with 11 mg/kg/BW of Proteus+6 mg/kg/BW of Biscaya. Intraperitoneal injections were performed over a 14-day period. For bloodletting at the end of the experiment, blood samples were withdrawn from the rats in order to investigate the serum concentration of luteinizing hormone (LH), follicle-stimulating hormone (FSH), gonadotropin- releasing hormone (GnRH), testosterone, and leptin. The data were analyzed using SPSS, version 16, via ANOVA and the Duncan test. A P value equal to or less than 0.05 was considered statistically significant. RESULTS: Our comparisons between the experimental groups (average and maximum compound concentrations of Proteus and Biscaya) and the control group showed a significant decrease in the mean serum levels of FSH (P=0.001), LH (P=0.001), GnRH (P=0.001), testosterone (P=0.005), and leptin (P=0.001) in all the experimental groups in a dose-dependent manner. CONCLUSION: Proteus and Biscaya decreased GnRH, LH, FSH, and testosterone by reducing the serum level of leptin in the hypothalamus in a dose-dependent manner.

Spermatogenesis after Transplantation of Adipose Tissue-Derived Stem Cells in Azoospermic Guinea Pigs: A Histological and Histomorphometric Study.

BACKGROUND: The purpose of this research was to determine histomorphometric changes in busulfan-induced azoospermia after transplantation of Adipose Tissue-Derived Stem Cells (AdSCs) in guinea pig. AdSCs were isolated from adipose tissue around the testes of guinea pigs and characterized for mesenchymal properties. MATERIALS AND METHODS: Guinea pigs were allocated into three groups, including the control group without any intervention. To induce azoospermia, groups 2 and 3 received a dose of 40 mg/kg of busulfan with 21 days interval. Group 3 received 1×106 AdSCs in their seminiferous tubules of left testes, 35 days following last busulfan injection, while right testis in the group was considered for comparison as controls. Sixty days following transplantation of cell, histomorphometric and histopathologic changes of the experiments were assessed. RESULTS: After AdSCs' transplantation, normal spermatogenesis appearance was noticed compared to busulfan-induced azoospermia and AdSCs recovered spermatogenesis, and our findings can be added to the literature in treating azoospermic infertilities. CONCLUSION: The transplanted AdSCs could induce production of germinal cells using testicular seminiferous tubules and were an effective source in treating azoospermia.

Early oxygen therapy does not protect the brain from vasogenic edema following acute ischemic stroke in adult male rats.

Brain edema aggravates primary brain injury and increases its mortality rate after ischemic stroke. It is believed that normobaric oxygen therapy (NBO) may produce neuroprotective effects against ischemic stroke; however, reports have been controversial, and its effects on vasogenic brain edema as a major complication of brain ischemia have not been clarified. The present study investigates the effects of NBO on cerebral edema and blood - brain barrier integrity using rat model of ischemic stroke. Transient focal cerebral ischemia was induced in adult male Sprague-Dawley rats by left middle cerebral artery occlusion (MCAO) for 90 min followed by 24 h reperfusion. Early NBO supplementation was started 15 min after MCAO and continued for 90 min. The results of the present study show that early oxygen therapy following acute ischemic stroke does not reduce vasogenic brain edema, nor does it protect against oxidative stress-induced BBB destruction. Additionally, cerebral edema formation occurs in conjunction with an increased mortality rate, serious brain injury, and impairment of brain antioxidant power. These findings suggest that further experimental studies should be carried out to clarify the beneficial effects and potential side effects of early oxygen therapy in acute ischemic stroke before its clinical use.

The Regenerative Effect of Bone Marrow-Derived Stem Cells in Spermatogenesis of Infertile Hamster.

BACKGROUND: Infertility is a serious social problem in advanced nations, with male factor in half of all cases of infertility. This study was conducted to determine the regenerative effect of bone marrow-derived stem cells in spermatogenesis of infertile hamster. METHODS: Twelve adult male hamsters were equally divided into azoospermic and control groups. Busulfan was intraperitoneally used for induction of azoospermia, while the right testis was treated with bone marrow-derived stem cells (106 BM-SCs), labeled with sterile trypan blue, 35 days after busulfan injection. The left testis served as positive control for azoospermia. Sixty days after cell transplantation, the animals were euthanized and both testes were removed and evaluated histologically. RESULTS: BM-SCs were spindle-shaped, adherent to the culture flasks and had positive expression of CD29 and CD73 and negative expression of CD45. Alcian blue staining confirmed differentiation of BM-SCs into chondrocytes. Karyotyping denoted to stability of chromosomes. Treatment with busulfan in seminiferous tubules resulted into distruption of spermatogenesis. After two months in busulfan treatment group, seminiferous tubular atrophy and germinal epitheliums degenerations were noticed with no spermatozoa in epididymis. After treatment of busulfan group with BM-SCs, spermatogonia, primary spermatocytes, spermatids and sperms were present in seminiferous tubules. CONCLUSION: As cell transplantation in seminiferous tubules resulted into a rapid repair of pathological changes, BM-SCs can be recommended an effective treatment measure in azoospermia. It seems that more studies are necessary to confirm the use of this technique in treatment of azoospermia and infertility in human.

Isolation, Characterization and Growth Kinetic Comparison of Bone Marrow and Adipose Tissue Mesenchymal Stem Cells of Guinea Pig.

BACKGROUND: Mesenchymal stem cells (MSCs) from different sources have different characteristics. Moreover, MSCs are not isolated and characterized in Guinea pig for animal model of cell therapy. AIM OF THE WORK: was the isolating of bone marrow MSCs (BM-MSCs) and adipose tissue MSCs (AT-MSCs) from Guinea pig and assessing their characteristics. MATERIAL AND METHODS: In this study, bone marrow and adipose tissue were collected from three Guinea pigs and cultured and expanded through eight passages. BM-MSCs and AT-MSCs at passages 2, 5 and 8 were seeded in 24-well plates in triplicate. Cells were counted from each well 1~7 days after seeding to determine population doubling time (PDT) and cell growth curves. Cells of passage 3 were cultured in osteogenic and adipogenic differentiation media. RESULTS: BM-MSCs and AT-MSCs attached to the culture flask and displayed spindle-shaped morphology. Proliferation rate of AT-MSCs in the analyzed passages was more than BM-MSCs. The increase in the PDT of MSCs occurs with the increase in the number of passages. Moreover, after culture of BM-MSCs and AT-MSCs in differentiation media, the cells differentiated toward osteoblasts and adipocytes as verified by Alizarin Red staining and Oil Red O staining, respectively. CONCLUSION: BM-MSCs and AT-MSCs of Guinea pig could be valuable source of multipotent stem cells for use in experimental and preclinical studies in animal models.

The effect of Aloe vera gel on viability of dental pulp stem cells.

BACKGROUND: Dental pulp stem cells (DPSCs) can play a prominent role in tissue regeneration. Aloe vera L. (Liliaceae) contains the polysaccharide of acemannan that was shown to be a trigger factor for cell proliferation, differentiation, mineralization, and dentin formation. AIM: This study sought to determine the viability of DPSCs in Aloe vera in comparison with Hank's balanced salt solution (HBSS). MATERIALS AND METHOD: Twelve rabbits underwent anesthesia, and their incisor teeth were extracted; the pulp tissue was removed, chopped, treated with collagenase and plated in culture flasks. DPSCs from passage 3 were cultured in 24-well plates, and after 3 days, the culture media changed to 10, 25, 50, and 100% concentrations of Aloe vera at intervals of 45 and 90 min and 3 and 6 h. Distilled water was used as negative and HBSS as positive control for comparison. The cell morphology, viability, population doubling time (PDT), and growth kinetics were evaluated. RT-PCR was carried out for characterization and karyotyping for chromosomal stability. RESULTS: Aloe vera showed a significant higher viability than HBSS (74.74%). The 50% Aloe vera showed higher viability (97.73%) than other concentrations. PDT in 50% concentration was 35.1 h and for HBSS was 49.5 h. DPSCs were spindle shaped and were positive for CD73 and negative for CD34 and CD45. Karyotyping was normal. CONCLUSIONS: Aloe vera as an inexpensive and available herb can improve survival of avulsed or broken teeth in emergency cases as a transfer media.

Growth Kinetics, Characterization, and Plasticity of Human Menstrual Blood Stem Cells.

One of the readily available sources of mesenchymal stem cells (MSCs) is menstrual blood-derived stem cells (Men-SCs), which exhibit characteristics similar to other types of MSCs. This study was performed to determine the growth kinetics, plasticity, and characterization of Men-SCs in women. During spring 2014 in the southern Iranian city of Shiraz, menstrual blood (5 mL) was obtained from 10 women on their third day of menstruation in 2 age groups of 30 to 40 and 40 to 50 years old. Ficoll was used to separate the mononuclear cell fraction. After the Men-SCs were cultured, they were subcultured up to passage 4. Growth behavior and population doubling time were evaluated by seeding 5×10(4) cells into 12- and 24-well culture plates, and the colonies were enumerated. The expression of CD44, CD90, and CD34 was evaluated. The osteogenic potential was assessed by alizarin red staining. The Men-SCs were shown to be plastic adherent and spindle-shaped. Regarding the growth curves in the 12- and 24-well culture plates, it was demonstrated that in the women aged between 30 and 40 years, population doubling time was 55.5 and 62 hours, respectively, while these values in the women aged between 40 and 50 years were 70.4 and 72.4 hours, correspondingly. Positive expression of CD44 and CD90 and negative expression of CD34 were noted. In the osteogenic differentiation medium, the cells differentiated toward osteoblasts. As human Men-SCs are easily collectable without any invasive procedure and are a safe and rapid source of MSCs, they can be a good candidate for stem cell banking and cell transplantation in women.

Additive effect of BLA GABAA receptor mechanism and (+)-MK-801 on memory retention deficit, an isobologram analysis.

There is a near correlation between N-methyl-d-aspartate (NMDA) and γ-aminobutyric acid (GABA) receptors in the modulation of learning and memory in the basolateral amygdala (BLA). In this study, we investigated the involvement of GABAA receptors in the BLA in amnesia induced by (+)-MK-801, a noncompetitive antagonist of NMDA receptors, in male Wistar rats. After guide cannulae were bilaterally placed in the BLA, animals were trained in a step-through type passive avoidance task and then tested 24h after training to measure memory retrieval and locomotor activity. Post-training intra-BLA microinjection of (+)-MK-801 (0.5 µg/rat) and GABAA receptor agonists (muscimol at doses 0.05 and 0.1 µg/rat) or antagonist (bicuculline at doses 0.05 and 0.1 µg/rat) decreased step-through latency during retrieval but did not alter locomotor activity. Results also showed that a subthreshold dose of muscimol (0.025 µg/rat) potentiated impairment induced by (+)-MK-801, whereas bicuculline (0.025 µg/rat) restored it. Furthermore, the highest dose of muscimol (0.5 µg/rat) increased locomotor activity induced by (+)-MK-801. Isobologram analysis showed that there was an additive but not synergistic effect between muscimol and (+)-MK-801 on memory retention deficits in the BLA. In conclusion, muscimol and bicuculline decreased retention of memory formation in the BLA, and GABAA receptors in the BLA may be involved in the additive effect on (+)-MK-801-induced memory retention deficits.

Patients with Active Rheumatoid Arthritis Have Lower Frequency of nTregs in Peripheral Blood.

BACKGROUND: Patients with rheumatoid arthritis (RA) suffer from wide ranges of autoimmune reactions in joints. The mechanism of which is generally unknown and maybe associated with Treg deregulation. OBJECTIVE: To compare the frequency of nTregs in peripheral blood of patients with active rheumatoid disease with healthy individuals. METHODS: Twenty five newly diagnosed patients with active RA disease were selected based on the clinical and laboratory criteria before starting their therapies. Treg cells in peripheral blood samples were enumerated by immune staining and flowcytometry analysis. RESULTS: Clinical and laboratory results were in favor of active disease in all the studied patients although they showed variations in Disease Activity Score-28 (DAS-28). Compared to the healthy controls, RA patients had significantly lower frequency of CD4+CD25hi or CD4+CD25+FoxP3+ natural regulatory T cells. In spite of that, there were no significant differences between patients and healthy controls in respect to the CD4/CD8 ratio. Interestingly, more CD4+CD25-FoxP3+ cells were found in peripheral blood of patients. The frequencies of the Tregs did not show strong associations with the DAS-28. CONCLUSION: We showed lower abundance of nTregs in peripheral blood of RA patients which highlights the significance of these cells in RA.

Differentiation of human-induced pluripotent stem cells into insulin-producing clusters.

OBJECTIVES: In diabetes mellitus type 1, beta cells are mostly destroyed; while in diabetes mellitus type 2, beta cells are reduced by 40% to 60%. We hope that soon, stem cells can be used in diabetes therapy via pancreatic beta cell replacement. Induced pluripotent stem cells are a kind of stem cell taken from an adult somatic cell by "stimulating" certain genes. These induced pluripotent stem cells may be a promising source of cell therapy. This study sought to produce isletlike clusters of insulin-producing cells taken from induced pluripotent stem cells. MATERIALS AND METHODS: A human-induced pluripotent stem cell line was induced into isletlike clusters via a 4-step protocol, by adding insulin, transferrin, and selenium (ITS), N2, B27, fibroblast growth factor, and nicotinamide. During differentiation, expression of pancreatic ß-cell genes was evaluated by reverse transcriptase-polymerase chain reaction; the morphologic changes of induced pluripotent stem cells toward isletlike clusters were observed by a light microscope. Dithizone staining was used to stain these isletlike clusters. Insulin produced by these clusters was evaluated by radio immunosorbent assay, and the secretion capacity was analyzed with a glucose challenge test. RESULTS: Differentiation was evaluated by analyzing the morphology, dithizone staining, real-time quantitative polymerase chain reaction, and immunocytochemistry. Gene expression of insulin, glucagon, PDX1, NGN3, PAX4, PAX6, NKX6.1, KIR6.2, and GLUT2 were documented by analyzing real-time quantitative polymerase chain reaction. Dithizone-stained cellular clusters were observed after 23 days. The isletlike clusters significantly produced insulin. The isletlike clusters could increase insulin secretion after a glucose challenge test. CONCLUSIONS: This work provides a model for studying the differentiation of human-induced pluripotent stem cells to insulin-producing cells.

miR-375 induces human decidua basalis-derived stromal cells to become insulin-producing cells.

This paper focuses on the development of renewable sources of isletreplacement tissue for the treatment of type I diabetes mellitus. Placental tissue-derived mesenchymal stem cells (MSCs) are a promising source for regenerative medicine due to their plasticity and easy availability. They have the potential to differentiate into insulin-producing cells. miR-375 is a micro RNA that is expressed in the pancreas and involved in islet development. Human placental decidua basalis MSCs (PDB-MSCs) were cultured from full-term human placenta. The immunophenotype of the isolated cells was checked for CD90, CD105, CD44, CD133 and CD34 markers. The MSCs (P3) were chemically transfected with hsa-miR-375. Total RNA was extracted 4 and 6 days after transfection. The expressions of insulin, NGN3, GLUT2, PAX4, PAX6, KIR6.2, NKX6.1, PDX1, and glucagon genes were evaluated using real-time qPCR. On day 6, we tested the potency of the clusters in response to the high glucose challenge and assessed the presence of insulin and NGN3 proteins via immunocytochemistry. Flow cytometry analysis confirmed that more than 90% of the cells were positive for CD90, CD105 and CD44 and negative for CD133 and CD34. Morphological changes were followed from day 2. Cell clusters formed during day 6. Insulin-producing clusters showed a deep red color with DTZ. The expression of pancreatic-specific transcription factors increased remarkably during the four days after transfection and significantly increased on day 7. The clusters were positive for insulin and NGN3 proteins, and C-peptide and insulin secretion increased in response to changes in the glucose concentration (2.8 mM and 16.7 mM). In conclusion, the MSCs could be programmed into functional insulin-producing cells by transfection of miR-375.

Effect of Platelet-Rich Plasma on CCl4-Induced Chronic Liver Injury in Male Rats.

Platelet-rich plasma (PRP) has been of great concern to the scientists and doctors who are involved in wound healing and regenerative medicine which focuses on repairing and replacing damaged cells and tissues. Growth factors of platelet-rich plasma are cost-effective, available, and is more stable than recombinant human growth factors. Given these valuable properties, we decided to assess the effect of PRP on CCl4-induced hepatotoxicity on rats. The rats received CCl4 (1 mL/kg, i.p. 1 : 1 in olive oil) twice per week for 8 weeks. Five weeks after CCl4 injection, the rats also received PRP (0.5 mL/kg, s.c.) two days a week for three weeks. Twenty-four hours after last CCl4 injection, the animals bled and their livers dissected for biochemical and histopathological studies. Blood analysis was performed to evaluate enzyme activity. The results showed that PRP itself was not toxic for liver and could protect the liver from CCl4-induced histological damages and attenuated oxidative stress by increase in glutathione content and decrease in lipid peroxidative marker of liver tissue. The results of the present study lend support to our beliefs in hepatoprotective effects of PRP.

Role of music in morphine rewarding effects in mice using conditioned place preference method.

OBJECTIVES: This research aims at studying the neuroendocrine effects of music on creating morphine dependence in mice using conditioned place preference (CPP). METHODS: The mice treated with 10 mg/kg morphine subcutaneously, fast music and slow music. Morphine was used to create dependence. In order to recognize the morphine rewarding effects, CPP technique was used. In the conditioning stage that lasted for 8 days, different groups of mice, after receiving the treatment were randomly placed in compartment for 30 minutes. The post-conditioning stage included the fourth day, the ninth day, the 12th day and the 16th day. RESULTS: Comparing place preference between morphine group and the control group, a significant increase (p<0.05) was observed in the place preference of morphine group, while a significant decrease (p<0.05) was demonstrated in the place preferences of morphine + taxi girl music group compared with morphine group alone. In addition morphine + alone in the rain music group demonstrated a significantly increased conditioned place preference (p<0.05) compared with the morphine group. CONCLUSIONS: Alone in the rain music acts as a positive pleasant emotion increasing the dopaminergic activity in the Nucleus Accumbens (NAc) and Ventral Tegmental Area (VTA) and through associated learning mechanisms of reward-related behavior increases morphine addiction. However, taxi girl music may act as unpleasant experiences producing negative emotions and reducing morphine addiction.