shRNA-mediated downregulation of α-N-Acetylgalactosaminidase inhibits migration and invasion of cancer cell lines

Objective(s): Extracellular matrix (ECM) is composed of many kinds of glycoproteins containing glycosaminoglycans (GAGs) moiety. The research was conducted based on the N-Acetylgalactosamine (GalNAc) degradation of ECM components by α-N-acetylgalactosaminidase (Nagalase) which facilitates migration and invasion of cancer cells. This study aims to investigate the effects of Naga-shRNA downregulation on migration and invasion of cancer cell lines. Materials and Methods: In this study, MCF-7 cell line (human mammary carcinoma cell line) and A2780 (human ovarian carcinoma cell line) were used. The level of normalized Naga expression and Nagalase protein were evaluated by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay/western blotting, respectively. Migration and invasion were determined using transwell assays, and statistical analysis was carried out by ANOVA test. Results: Response to transduction by shRNA compared to the control group, migrative and invasive properties of the transfected cells were significantly inhibited. Conclusion: These results indicate that Nagalase may have an important role in migration and invasion of cancer cells and can be considered as a candidate for further studies

Repair of rat cranial bone defect by using amniotic fluid-derived mesenchymal stem cells in polycaprolactone fibrous scaffolds and platelet-rich plasma

Introduction: Tissue regenerative medicine strategies, as a promising alternative has become of major interest to the reconstruction of critical size bone defects. This study evaluated the effects of the simultaneous application of polycaprolactone (PCL), amniotic fluid mesenchymal stem cells (AF-MSCs) and platelet-rich plasma (PRP) on the repair of rat cranial bone defects. Methods: The AF-MSCs were isolated at the end of the second week of pregnancy in rats. PRP obtained from rat blood and the random PCL fibrous scaffolds were prepared using the electrospinning method. Circular full thickness (5 mm) bone defects were developed on both sides of the parietal bones (animal number=24) and the scaffolds containing AF-MSCs and PRP were implanted in the right lesions. Thereafter, after eight weeks the histological and immunohistochemistry studies were performed to evaluate the bone formation and collagen type I expression. Results: The spindle-shaped mesenchymal stem cells were isolated and the electron microscope images indicated the preparation of a random PCL scaffold. Immunohistochemical findings showed that collagen type I was expressed by AF-MSCs cultured on the scaffold. The results of hematoxylin and eosin (H & E) staining indicated the formation of blood vessels in the presence of PRP. Additionally, immunofluorescence findings suggested that PRP had a positive effect on collagen type I expression. Conclusion: The simultaneous application of fibrous scaffold + AF-MSCs + PRP has positive effects on bone regeneration. This study showed that PRP can affect the formation of new blood vessels in the scaffold transplanted in the bone defect

The Evaluation of Nerve Growth Factor Over Expression on Neural Lineage Specific Genes in Human Mesenchymal Stem Cells

Objective Treatment and repair of neurodegenerative diseases such as brain tumors, spinal cord injuries, and functional disorders, including Alzheimer’s disease, are challenging problems. A common treatment approach for such disorders involves the use of mesenchymal stem cells (MSCs) as an alternative cell source to replace injured cells. However, use of these cells in hosts may potentially cause adverse outcomes such as tumorigenesis and uncontrolled differentiation. In attempt to generate mesenchymal derived neural cells, we have infected MSCs with recombinant lentiviruses that expressed nerve growth factor (NGF) and assessed their neural lineage genes. Materials and Methods In this experimental study, we cloned the NGF gene sequence into a helper dependent lentiviral vector that contained the green fluorescent protein (GFP) gene. The recombinant vector was amplified in DH5 bacterial cells. Recombinant viruses were generated in the human embryonic kidney 293 (HEK-293) packaging cell line with the helper vectors and analyzed under fluorescent microscopy. Bone marrow mesenchymal cells were infected by recombinant viruses for three days followed by assessment of neural differentiation. We evaluated expression of NGF through measurement of the NGF protein in culture medium by ELISA; neural specific genes were quantified by real-time polymerase chain reaction (PCR). Results We observed neural morphological changes after three days. Quantitative PCR showed that expressions of NESTIN, glial derived neurotrophic factor (GDNF), glial fibrillary acidic protein (GFAP) and Microtubule-associated protein 2 (MAP2) genes increased following induction of NGF overexpression, whereas expressions of endogenous NGF and brain derived neural growth factor (BDNF) genes reduced. Conclusion Ectopic expression of NGF can induce neurogenesis in MSCs. Direct injection of MSCs may cause tumorigenesis and an undesirable outcome. Therefore an alternative choice to overcome this obstacle may be the utilization of differentiated neural stem cells

Treatment of new cases of Acute Promyelocytic Leukemia With Arsenic Trioxide

Introduction: Arsenic Trioxide is effective and approved for treatment of relapsed or refractory APL cases to ATRA but its effects in new cases of APL is not clear and needs long term follow up to dis¬close the role of this drug in treatment of APL in combination with chemotherapy/ATRA or alone. Material and methods: we studied 111cases of APL (94 new case and 17 relapsed) diagnosed by mor¬phological criteria and confirmed by cytogenetic and/or RT-PCR for the presence of PML/RARA fu¬sion gene."nArsenic Trioxide was infused as 0.15mg/kg/day doses, until complete remission by morphological cri¬teria or till 60 days. In case of complete remission, after 28 days do rest, 0.15mg/kg/days Arsenic Tri-oxide was infused for an additional 28 days as consolidation. Also, we studied minimal residual disease by semi-sensitive RT- PCR on peripheral blood samples up to a year after complete remission. Results: Complete remission was observed in 95 patients (85.6%) and median time to complete remis¬sion was 30 days. There was no significant difference between remission rate in new and relapsed cases."nDuring the induction phase, the most common cause of toxicity and mortality was APL differentiation syndrome (23 cases or 20.7%). Other toxicities were serosistis (7.2%) and hepatotoxicity (19.8%). With a median follow up of 16.5(1-57) months for patients in complete remission, one and two year disease free survival (DFS) was 88.3% and 63.7%, respectively. We observed 24 relapses and 19 of them achieved second complete remission, again by Arsenic Trioxide. Median time to relapse was 17 months (4-33) and median time of second DFS after re-treatment with Arsenic Trioxide was 18 months. We observed a third and fourth remission for some patients, who relapsed, again by Arsenic Trioxide."nFor patients in complete remission, one and three years survival was 95.5% and 87.6%, respectively. Minimal residual disease was positive in 4 (8.3%) out of 48 cases up to a year after remission induction and 3 of these patients clinically relapsed."nConclusion: Arsenic Trioxide is effective as a first line treatment of APL. Results of Arsenic Trioxide combination with chemotherapy/ATRA needs further study. Also it seems that Arsenic Trioxide is ap¬plicable for relapsed patients again and drug resistance is an unusual even

Increase of CXCR4 Expression on Expanded Non-enriched Cord Blood CD34+ Cells Using MSCs

Introduction: A number of potential cell adhesion molecules, which mediate essential cell-to-cell or cell-to-matrix interactions, are expressed on the surface of CD34+ hematopoietic progenitor cells (HPCs), including integrins, CD44, and CXCR4. These molecules are essential for homing process. In this study, we compared the changes of expression of CD44 and CXCR4 on the CD34+ hematopoietic progenitor cells expanded on MSCs in the presence of cytokines. Material and Methods: Cord blood CD34+ cells were expanded using human bone marrow mesenchymal stem cells and cytokines (TPO, SCF, FLt-3, IL-6, and IL-3), and then expression of CD44 and CXCR4 on CD34+ cells were evaluated by flow cytometric analysis. Results: After 2 weeks of serum free culture of CD34+ cells in the presence of cytokines, the expression of CXCR4 on CD34+ cells was decreased 3.4 fold (p<0.05). In contrast, the expression of CXCR4 on CD34+ cells expanded on hMSCs was increased (p<0.05). The expression of CD44 on expanded CD34+ cells in both methods did not differ significantly. Conclusions: Our results indicated that co-culture of cord blood stem cells on hMSCs significantly increased CXCR4 expression on cord blood CD34+ cells

Beneficial effects of angiotensin converting enzyme inhibition on scopolamine-induced learning and memory impairment in rats, the roles of brain-derived neurotrophic factor, nitric oxide and neuroinflammation

The effect of the brain-derived neurotrophic factor (BDNF), cytokines, and renin angiotensin system (RAS) on memory function have been demonstrated. In this study, the effects of RAS inhibitor captopril (Capto) on hippocampal BDNF, interleukin −6 (IL-6), oxidative stress indicators, and nitric oxide (NO) in scopolamine (Sco)-induced memory impairment in rats were examined. The groups were (1) control, (2) Sco in which Sco was applied 30 min prior to the behavioral tests, and (3–5) Sco-Capto 10, 50, and 100 groups, where Capto (10, 50, or 100 mg/kg), were applied 2 weeks prior to the experiment, as well as 30 min prior to each Sco injection. The Morris Water Maze (MWM) test was conducted, and BDNF, IL-6, NO metabolites, malondialdehyde (MDA), thiol, superoxide dismutase (SOD), and catalase (CAT) were measured. Sco increased the delay and distance to the platform in the MWM test (P < .01 to P < .001), while shortening the time and distance in the target area (P < .01 to P < .001). Additionally, Sco increased IL-6, NO metabolites, and MDA, while decreasing BDNF, thiol, SOD, and CAT (P < .01 to P < .001). Although the Capto reduced the latency and distance traveled to the platform (P < .05 to P < .001), it elevated the time and distance traveled in the target area (P < .05 to P < .01). Furthermore, Capto improved BDNF, thiol, SOD, and CAT levels, and decreased IL-6, NO metabolites, and MDA (P < .05 to P < .001). RAS has a role in learning and memory impairment due to cholinergic system dysfunction. The possible mechanism(s) are including its effects on BDNF, neuro-inflammation and oxidative stress

The combination effect of dexamethasone with either metoclopramide or propofol on nusea and vomiting afther general anesthesia

Background and Objective: A common complication after general anesthesia is nausea and vomiting followed by different problems such as spasm, hypoxia and pulmonary aspiration. This complication is more common in patients with full stomach, Eye injury, head trauma, cesarean and laparoscopy. Propofol and metoclopramide are two common drugs to prevent nausea and vomiting after operation. On the other hand adding dexamethasone to the above drug, has an important effect on decreasing nausea and vomiting. In this study, the effect propofol and metoclopramide associated with dexamethasone on nausea and vomiting after operation was investigated. Materials and Methods: In this clinical trial study, 100 patients with ASA I, II classes, aged 16-60 years with selective orthopedic surgery randomly have divided into two groups. In group one, 48 patients received metoclopramide (10mg) with dexamethasone (8mg) and in group two, 52 patients received propofol (20mg) with dexamethasone (8mg), five minutes before the end of operation. Prevalence of nausea and vomiting in both groups was considered after 4 hours and results were analyzed by Chi-Square, t-student and Fisher exact tests. Results: The rate of nausea in group 1 and 2 was 35.4% and 11.5% respectivly (P<0.05). The rate of vomiting was 27.7% and 7.7% in group 1 and 2 respectivly (P<0.05). Conclusion: This study showed that the antiemetic effect of propofol with dexamethasone is more effective to prevent nausea and vomiting than metoclopromide with dexamethasone

The study of psychiatric comorbidity in patient with pemphigus

Background: The relationship between skin, neuron system and mental status is complicated. Pemphigus is severe and painful autoimmune skin disorder that occurs in 0.5-1 person in 100000. Regarding the critical role of mental status in autoimmune diseases such as pemphigus, the aim of this research was to investigating psychiatric comorbidity in patient with pemphigus. Methods: This research was a descriptive survey study. The society of this research included the patients with pemphigus referred to Razi Dermatology University Hospital in Tehran. Participants were 200 persons who were selected through available sampling. Finally, 198 persons filled out the demographic questionnaire and symptom checklist-90 (SCL-90). Results: This research showed that the prevalence of psychiatric disorders in this sample was 62.62%. 28.78% (57 persons) of patients were male and 33.84% (67 persons) were female. In both genders the symptoms of paranoia were the most prevalent disorders (45% in female and 60% in male) and phobia was the least prevalent in our sample (8.2% in female and 24% in male). The upper grade in education, the less prevalent disorders. 81.45% of participants with mental disorders were married and 18.55% of them were single. Of patients only 26.32% had visited the psychiatrist or psychologist and 95.15% of them had used medications. This research also showed that there was a significant difference between two genders. Men showed higher rate of prevalence in psychiatric symptoms than women. Conclusion: The results showed that psychiatric disorders in patients with pemphigus are high compared to general population. This research congruent with other researches showed that higher age, lower grade of education and marriage related to worse prognosis in psychological status in patient with pemphigus. Thus dermatologists must recognize and manage these psychiatric comorbidities to treat patients effectively and to improve the quality of life in patients with autoimmune diseases such as pemphigus.&nbsp