ABSTRACT
OBJECTIVE@#To explore the appropriate procedures for preparing extracellular microvesicles (MV) derived from human mesenchymal stem cells (MSC).@*METHODS@#Human MSCs from umbilical cords were cultured in a serum-free medium and maintained in a basal medium for 72 hours after the cell confluence reached to 80%. The supernatants of cultured cells were collected and MVs were enriched. MVs were identified by flow cytometry and electron microscopy. The total protein amount in MVs was used as a parameter for the content of MVs. The supernatants were adjusted to different pH values, and the output of MVs was detected. The supernatants were also collected for enriching the MV and detecting the protein content of MV after the cells were maintained in the basic medium for different time.@*RESULTS@#Flow cytometric analysis showed that the MVs expressed CD9, CD63 and CD81, morphologically presented round under an electron microscope and the diameter of MV was around 100 nm. After enrichment of MV, the protein content of MVs in the supernatants was 416.8±128.1, 255.4±77.9 and 142.8±46.4 μg per 10 MSC，respectively at pH of supernatant 3, 7 and 9 (P＜0.05). The protein content of the supernatants per 10 MSC was 173.6±44.5, 262.4±49.6 and 364.2±37.8 μg respectively after starvation culture for 48, 72 and 96 hrs (P＜0.05).@*CONCLUSION@#MVs can be readily collected after MSCs were starved for 96 hours, and the pH of the supernatants is adjusted at 3.0.
Subject(s)
Humans , Cell-Derived Microparticles , Cells, Cultured , Flow Cytometry , Mesenchymal Stem Cells , Umbilical CordABSTRACT
BACKGROUND: Senile degenerative lumbar spinal stenosis typically manifests clinically as lower back and leg pain caused by compression of the nerve root. Conventional total laminectomy for degenerative lumbar spinal stenosis can quickly alleviate a patient's symptoms, but produces an unsatisfactory therapeutic effect because of spinal instability caused by degenerative spondylolisthesis, and also has many adverse reactions. OBJECTIVE: The purpose of this study is to investigate whether total laminectomy combined with lumbar pedicle screw fixation for treatment of senile degenerative lumbar spinal stenosis can effectively reduce lower back and leg pain caused by compression of the nerve root, increase lumbar spine stability, and reduce adverse reactions. METHODS: A prospective, single-center, self-control, interventional trial. One hundred and sixty older adult patients with degenerative lumbar spinal stenosis who will receive treatment at the Department of Orthopedics, Qinghai Provincial People's Hospital, China will be included in this study. All patients will undergo total laminectomy combined with lumbar pedicle screw fixation, with follow-up at 3, 6, 9, and 12 months post-surgery. RESULTS AND CONCLUSION: The primary outcome measure of this study is recovery rate in Japanese Orthopedic Society (JOA) score at 12 months post-surgery, which is used to evaluate improvements in patients lower back and leg pain. Secondary outcome measures of this study include changes in JOA score, spinal canal diameter, lumbar spine morphology displayed on computed tomography images, and incidence of adverse events post-surgery. Results of a preliminary study involving 71 older adult patients with degenerative lumbar spinal stenosis who received the same treatment showed that at 3 months post-surgery, JOA score and spinal canal diameter were significantly increased compared with before surgery (P < 0.05). Findings from this study may provide clinical evidence supporting that total laminectomy combined with lumbar pedicle screw fixation is a safe and reliable method for treatment of senile degenerative lumbar spinal stenosis because it rapidly alleviates lower back and leg pain and provides spine stability. This study was approved by Medical Ethics Committee of Qinghai Provincial People's Hospital of China(approval No.QHY201602G).This study will be performed in strict accordance with the Declaration of Helsinki formulated by the World Medical Association.Participants provided signed informed consent prior to participation in the study.This study was designed in December 2017. Patient recruitment and data collection will begin in August 2018. Data analysis will be performed in October 2019. The study will be completed in December 2019. Results will be disseminated through presentations at scientific meetings and/or by publication in a peer-reviewed journal. The version of this study protocol is (1.0). This trial was registered with the Chinese Clinical Trial Registry (registration number: ChiCTR1800014726).
ABSTRACT
@#ObjectiveTo observe the effect of Xiaoyao powder combined with prozac and psychological therapy on post-stroke depression (PSD).Methods85 PSD patients were randomly divided into the treatment group (n=43) and control group (n=42). All patients were treated with routine therapy, including prozac and psychological therapy. Patients in the treatment group were also given Xiaoyao powder. Scores of Hamilton Depression Scale (HAMD), modified Barthel index (MBI) and Scandinavian Stroke Scale (SSS) of all patients were evaluated before and after therapy.ResultsAfter treatment, the effective rate and MBI scores were significantly higher and scores of HAMD and SSS were significantly lower for patients of the treatment group compared with those of the control group (P<0.01).ConclusionXiaoyao powder combined with prozac and psychological therapy can improve depression and neural function of PSD patients significantly.