Lentiviral Mediated Overexpression of NGF in Adipose-derived Stem Cells

Introduction: Human adipose-derived stem cells (ADSCs) are multipotent stem cells that can self-renew and differentiate into various types of cells such as adipocytes, osteocytes, and neural cells. These stem cells can be isolated by minimally invasive technique in large amounts. ADSCs are a useful resource for cell therapy and regenerative medicine. Nerve growth factor (NGF) is the first neurotrophin factor discovered and characterized for its anti-apoptotic role in neural development. NGF can promote neuronal survival and neurite outgrowth and it also promotes neuron differentiation and migration. Moreover, research showed that NGF could protect axons from inflammatory damage, improve cognitive function in damaged brain models, and function in the prevention and treatment of neurological diseases like Alzheimer’s disease. In this study we use Lentiviral vector-mediated gene transfer technique to deliver NGF gene to ADSCs and overexpress this factor in ADSCs. Method and Materials: ADSCs extracted from human adipose tissue after lipoaspiration by digestion method. ADSCs characterized with Flowcytometry and differentiation assay in adipogenic and osteogenic differential media. The NGF gene was cloned in pCDH-513B-1 (System Bioscience, Mountain View, CA, United States) under a cytomegalovirus (CMV) promoter. Recombinant lentiviruses were produced according to the Prof. Trono lab protocol with some modifications in HEK 293T cells. The spinfection method was used to transduce ADSCs. NGF expression was assayed using fluorescent microscope to trace green fluorescent protein (GFP) marker, RT-PCR and western blotting. Results: Extracted ADSCs had mesenchymal morphology and differentiated into adipocytes and osteocytes in differentiating media. HEK293T easily transfected with pCDH-513B-1 and over 99% of them expressed GFP so we gathered pseudoviruses from the supernatant. ADSCs transduced with these pseudoviruses transferred NGF and after transduction expressed GFP, as seen under fluorescent microscope. RT-PCR and western blotting verified NGF overexpression in them

A Comparative Analysis of Clinical Characteristics and Laboratory Findings of COVID-19 between Intensive Care Unit and Non-Intensive Care Unit Pediatric Patients: A Multicenter, Retrospective, Observational Study from Iranian Network for Research in Viral

Introduction: To date, little is known about the clinical features of pediatric COVID-19 patients admitted to intensive care units (ICUs). Objective: Herein, we aimed to describe the differences in demographic characteristics, laboratory findings, clinical presentations, and outcomes of Iranian pediatric COVID-19 patients admitted to ICU versus those in non-ICU settings. Methods: This multicenter investigation involved 15 general and pediatrics hospitals and included cases with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection based on positive real-time reverse transcription polymerase chain reaction (RT-PCR) admitted to these centers between March and May 2020, during the initial peak of the COVID-19 pandemic in Iran. Results: Overall, 166 patients were included, 61 (36.7%) of whom required ICU admission. The highest number of admitted cases to ICU were in the age group of 1–5 years old. Malignancy and heart diseases were the most frequent underlying conditions. Dyspnea was the major symptom for ICU-admitted patients. There were significant decreases in PH, HCO3 and base excess, as well as increases in creatinine, creatine phosphokinase (CPK), lactate dehydrogenase (LDH), and potassium levels between ICU-admitted and non-ICU patients. Acute respiratory distress syndrome (ARDS), shock, and acute cardiac injury were the most common features among ICU-admitted patients. The mortality rate in the ICU-admitted patients was substantially higher than non-ICU cases (45.9% vs. 1.9%, respectively; p<0.001). Conclusions: Underlying diseases were the major risk factors for the increased ICU admissions and mortality rates in pediatric COVID-19 patients. There were few paraclinical parameters that could differentiate between pediatrics in terms of prognosis and serious outcomes of COVID-19. Healthcare providers should consider children as a high-risk group, especially those with underlying medical conditions

Mesenchymal Stem Cells: Rising Concerns over Their Application in Treatment of Type One Diabetes Mellitus

Type 1 diabetes mellitus (T1DM) is an autoimmune disorder that leads to beta cell destruction and lowered insulin production. In recent years, stem cell therapies have opened up new horizons to treatment of diabetes mellitus. Among all kinds of stem cells, mesenchymal stem cells (MSCs) have been shown to be an interesting therapeutic option based on their immunomodulatory properties and differentiation potentials confirmed in various experimental and clinical trial studies. In this review, we discuss MSCs differential potentials in differentiation into insulin-producing cells (IPCs) from various sources and also have an overview on currently understood mechanisms through which MSCs exhibit their immunomodulatory effects. Other important issues that are provided in this review, due to their importance in the field of cell therapy, are genetic manipulations (as a new biotechnological method), routes of transplantation, combination of MSCs with other cell types, frequency of transplantation, and special considerations regarding diabetic patients’ autologous MSCs transplantation. At the end, utilization of biomaterials either as encapsulation tools or as scaffolds to prevent immune rejection, preparation of tridimensional vascularized microenvironment, and completed or ongoing clinical trials using MSCs are discussed. Despite all unresolved concerns about clinical applications of MSCs, this group of stem cells still remains a promising therapeutic modality for treatment of diabetes

Wavefront-guided laser-assisted subepithelial keratectomy in low myopia, myopic astigmatism and high myopia

AIM: To compare the safety, efficacy, predictability, stability and complications of wavefront-guided laser-assisted subepithelial keratectomy(LASEK)in low myopia, myopic astigmatism and high myopia correction.<p>METHODS: A retrospective analysis of 416 eyes were assigned to 3 groups: 159 eyes with low myopia(LM)and mean refractive spherical equivalent(MRSE)of -3.68±1.33 dioptre(D); 161 eyes with myopic astigmatism(MA)and MRSE of -5.99±2.24D and mean cylinder of 2.41±1.07D; and 96 eyes with high myopia(HM)and MRSE of -7.41±0.80D. After an epithelial flap creation, a wavefront-based excimer laser ablation was performed. Safety, efficacy, predictability and stability were evaluated at day 10, 2, 6 and 12mo postoperatively.<p>RESULTS:At 12mo, the MRSE was -0.36±0.31D in LM group, 0.15±0.41D in MA group and 0.58±0.68D in HM group. The uncorrected visual acuity(UCVA)was 20/20 in 90.60% of patients in LM group, 78.90% in MA group and 67% in HM group. Efficacy indices were 0.98, 1.04 and 0.92 in LM, MA and HM groups, respectively. Safety indices were 1.00, 1.07 and 1.05 in LM, MA and HM respectively. Five eyes(3.1%)in the LM group gained 1 line. Forty-four eyes(27.3%)in MA gained 1-3 lines and eighteen eyes(19.2%)of HM group gained 1-2 lines of BSCVA. Only 2 eyes in LM group developed corneal haze. There were not statistically significant differences in efficacy and safety indices amongst three groups. <p>CONCLUSION: Wavefront-guided LASEK is an effective and safe procedure for the treatment of LM, MA, and HM.although in myopic astigmatism the predictability, efficacy and safety indices had been better

Intravenous Injection of Human Umbilical Cord Matrix Stem Cell (Wharton Jelly Stem Cell) Provides Functional Recovery in a Rat Model of Traumatic Brain Injury

Objective: This study was designed to examine the effects of human umbilical cord matrixstem cell (hUCMSC) administration in rats for 6 weeks after traumatic brain injury (TBI).Materials and Methods: Adult male Wistar rats (n = 30) were injured with controlled corticalimpact device and divided into three groups. The treatment group (n = 10) was injectedwith 2 × 106 hUCMSC intravenously, the vehicle group (n=10) received phosphate bufferedsaline (PBS) whereas the control group (n = 10) receive nothing. All injections wereperformed one day after injury into the tail veins of the rats. All cells were labelled withBrdu before injection. Evaluation of the neurological function of the rats was performedbefore and after injury using Neurological Severity Scores (NSS). The rats were sacrificed6 weeks after TBI and brain sections were stained using Brdu immunohistochemistry.Results: Statistically significant improvement in functional outcome was observed in thetreatment group compared with the control group (p < 0.01). This benefit was visible 1 weekafter TBI and persisted for six weeks (end of trial). Histological analysis showed that hUCMSCwere present in the lesion boundary zone at 6 weeks in all cell injected animals.Conclusion: Rats injected with hUCMSC after TBI survive for at least six weeks and showfunctional improvemnt