CircRNA3616 knockdown attenuates inflammation and apoptosis in spinal cord injury by inhibiting TLR4/NF-κB activity via sponging miR-137.

PURPOSE: The present work focused on exploring the role of circRNA3616 in neuronal inflammation and apoptosis in spinal cord injury (SCI). METHODS: The SCI mouse model and circRNA3616 knockdown SCI mouse model were established. This work focused on assessing the mouse locomotor function using Basso Mouse Scale (BMS) and BMS subscore. Hematoxylin-eosin (HE) staining and Tunel staining were conducted, while myeloperoxidase (MPO) activity was also detected on spinal cord tissues. We also knocked down circRNA3616 expression in NSC-34 cells. Meanwhile, the SCI cell model was established by oxygen glucose deprivation (OGD) in NSC-34 cells. Moreover, we conducted dual-luciferase reporter gene assay. Flow cytometry (FCM) was conducted to detect SCI cell apoptosis, whereas cell counting kit-8 (CCK-8) assay was performed to analyze cell viability. This study also implemented enzyme-linked immunosorbent assay to detect inflammatory factors in spinal cord tissues, serum, and cells. RESULTS: CircRNA3616 knockdown reduced the damage, inflammatory response, apoptosis, and MPO activity in SCI mouse serum and spinal cord tissues. CircRNA3616 knockdown increased BMS and BMS subscore of SCI mice. CircRNA3616 up-regulated TLR4 expression by sponging miR-137. CircRNA3616 knockdown inhibited the TLR4, p-IkBα, p-p65/p65 protein expression, while promoting IkBα protein expression within SCI mouse spinal cord. TLR4 reversed circRNA3616 knockdown-induced inhibition on NF-κB pathway activity in SCI cells. CircRNA3616 knockdown attenuated neuronal cell inflammation and apoptosis via TLR4/NF-κB pathway after SCI. CONCLUSION: CircRNA3616 silencing attenuates inflammation and apoptosis in SCI by inhibiting TLR4/NF-κB activity via sponging miR-137. CircRNA3616 is the possible anti-SCI therapeutic target.

Methylprednisolone Induces Neuro-Protective Effects via the Inhibition of A1 Astrocyte Activation in Traumatic Spinal Cord Injury Mouse Models.

Traumatic spinal cord injury (TSCI) leads to pathological changes such as inflammation, edema, and neuronal apoptosis. Methylprednisolone (MP) is a glucocorticoid that has a variety of beneficial effects, including decreasing inflammation and ischemic reaction, as well as inhibiting lipid peroxidation. However, the efficacy and mechanism of MP in TSCI therapy is yet to be deciphered. In the present study, MP significantly attenuated the apoptotic effects of H2O2 in neuronal cells. Western blot analysis demonstrated that the levels of apoptotic related proteins, Bax and cleaved caspase-3, were reduced while levels of anti-apoptotic Bcl-2 were increased. In vivo TUNEL assays further demonstrated that MP effectively protected neuronal cells from apoptosis after TSCI, and was consistent with in vitro studies. Furthermore, we demonstrated that MP could decrease expression levels of IBA1, Il-1α, TNFα, and C3 and suppress A1 neurotoxic reactive astrocyte activation in TSCI mouse models. Neurological function was evaluated using the Basso Mouse Scale (BMS) and Footprint Test. Results demonstrated that the neurological function of MP-treated injured mice was significantly increased. In conclusion, our study demonstrated that MP could attenuate astrocyte cell death, decrease microglia activation, suppress A1 astrocytes activation, and promote functional recovery after acute TSCI in mouse models.

Percutaneous Endoscopic Transforaminal Lumbar Discectomy via Eccentric Trepan foraminoplasty Technology for Unilateral Stenosed Serve Root Canals.

OBJECTIVE: To evaluate the clinical outcomes of percutaneous lumbar foraminoplasty for unilateral stenosed nerve root canals. METHODS: The article is a retrospective analysis. From May 2016 to April 2017, 32 patients with lumbar spinal stenosis syndrome (unilateral stenosed nerve root canals) were treated with percutaneous endoscopic transforaminal lumbar discectomy (PETLD). The study included 15 men and 17 women, with an average age of 53.8 ± 15.4 years, ranging from 24 to 78 years. The indexes used for preoperative and postoperative 1 day, 3 months, and final follow up were the visual analogue scale (VAS) for lumbar and leg, the Oswestry disability index (ODI), and the modified Macnab criteria. All patients were followed up for an average 6 months after the operation. RESULTS: The average operative time was 75.82 ± 10.58 min, the average blood loss was 15.83 ± 3.75 mL, and the average hospital stay after surgery was 6.2 ± 4.6 days. The VAS score (leg) decreased from 6.94 ± 0.50 preoperatively to 1.16 ± 0.45 at the final follow up (P < 0.05) and ODI were obviously improved, from preoperative evaluation of 80.19 ± 5.55 to 9.44 ± 1.16 at the final follow up (P < 0.05). However, the postoperative VAS score (lumbar) did not show an improvement, reducing from 1.78 ± 0.49 preoperatively to 1.62 ± 0.55 at the final follow-up (P > 0.05). According to the modified Macnab criteria, the outcome showed that the excellent and good rate was 90.6%. There were three patients with hip soreness, and nerve root symptoms were relieved. CONCLUSION: Percutaneous endoscopic transforaminal lumbar discectomy has a satisfactory clinical effect in the treatment of lumbar spinal stenosis syndrome, especially for unilateral stenosed nerve root canals, and in decompressing the lateral recess and relieving the nerve root symptoms.

PTEN-GSK3ß-MOB1 axis controls neurite outgrowth in vitro and in vivo.

Mps One binder 1 (MOB1) is a core component of NDR/LATS kinase and a positive regulator of the Hippo signaling pathway. However, its role in neurite outgrowth still remains to be clarified. Here, we confirmed, for the first time, that MOB1 promoted neurite outgrowth and was involved in functional recovery after spinal cord injury (SCI) in mice. Mechanistically, we found that MOB1 stability was regulated by the PTEN-GSK3ß axis. The MOB1 protein was significantly up-regulated in PTEN-knockdown neuronal cells. This effect was dependent on the lipid phosphatase activity of PTEN. Moreover, MOB1 was found to be a novel substrate for GSK3ß that is phosphorylated on serine 146 and degraded via the ubiquitin-proteasome system (UPS). Finally, in vivo lentiviral-mediated silencing of PTEN promoted neurite outgrowth and functional recovery after SCI and this effect was reversed by down-regulation of MOB1. Taken together, this study provided mechanistic insight into how MOB1 acts as a novel and a necessary regulator in PTEN-GSK3ß axis that controls neurite outgrowth after SCI.

PTEN silencing enhances neuronal proliferation and differentiation by activating PI3K/Akt/GSK3ß pathway in vitro.

The failure of neuronal proliferation and differentiation is a major obstacle for neural repair and regeneration after traumatic central nervous system (CNS) injury. PTEN acts as an intrinsic brake on the neuronal cells, but its roles and mechanism still remain to be clarified. Herein, for the first time we confirmed that PTEN had a dual effect on the neuronal cells in vitro. Firstly, we found that PTEN knockdown significantly promoted cell proliferation and differentiation. Then, PTEN knockdown activated PI3K/Akt and Wnt/ß-catenin pathways in vitro. Further evidence revealed that GSK3ß as a key node involved in PTEN controlling cell proliferation and differentiation in PC12 cells. In addition, we identified that PTEN-GSK3ß pathway modulated neuronal proliferation via ß-catenin. Taken together, these results suggest that PTEN silencing enhances neuronal proliferation and differentiation by activating PI3K/Akt/GSK3ß pathway that it may be a promising therapeutic approach for CNS injury.

Unilateral vertebral artery injury in a patient with displaced upper cervical spine fractures: the treatment for one case of vertebral artery embolism.

PURPOSE: To report a novel treatment method for vertebral artery injury. Vertebral artery injuries may be caused during trauma by fracture and excessive motion with subluxation from C2 to C6 in spite of vertebral artery deeply seated and normally well protected inside the transverse foramen. Optimal medical management of the occluded vertebral artery has yet to be determined. METHODS: We report on a severely displaced C2-C3 fracture that was found to have a vertebral artery injury. Medical records and imaging were reviewed. RESULTS: A 50-year-old lady was hit by steel tube without loss of consciousness, but complaining of severe cervical and bilateral periscapular pain. Physical examination identified a neurologically intact patient with frontotemporal ecchymosis and posterior cervical tenderness. MRA and DSA showed an occluded left vertebral artery. After 3 days of observation, the patient showed no symptoms of brain ischemia or abnormal sensation and motor at four limbs. To ensure safety, we took the left vertebral artery embolism at the C2 and C5 levels before operation. CONCLUSIONS: To our knowledge, this is the first report of a displaced C2-C3 fracture in which transcatheter unilateral VAI embolization was used to prevent VAI bleeding during operation.

Development and validation of nomogram based on miR-203 and clinicopathological characteristics predicting survival after neoadjuvant chemotherapy and surgery for patients with non-metastatic osteosarcoma.

BACKGROUND: Recently, nomograms have been used as models for risk prediction in malignant tumor because they can predict the outcome of interest for a certain individual based on many variables. This study aimed to establish an effective prognostic nomogram for osteosarcoma based on the clinicopathological factors and microRNA-203. RESULTS: The results showed that miR-203 expression was significantly lower in osteosarcoma tissues compared with the corresponding adjacent tissues (P < 0.001). Patients with low miR-203 expression had poor overall survival (OS) in osteosarcoma. The histological type, tumor size, AJCC stage and miR-203 expression were integrated in the nomogram. The nomogram showed significantly better prediction of OS than for patients with non-metastatic osteosarcoma. The ROC curve also showed higher specificity and sensitivity for predicting 3- and 5-year osteosarcoma patients' survival compared with AJCC stage. The decision curve analysis also indicated more potential of clinical application of the nomogram compared with AJCC staging system. Moreover, our findings were supported by the validation cohort. MATERIALS AND METHODS: We retrospectively investigated 301 patients with non-metastatic osteosarcoma. Data from primary cohort (n = 198) were used to develop multivariate nomograms. This nomogram was internally validated for discrimination and calibration with bootstrap samples and was externally validated with an independent patient cohort (n = 103). CONCLUSIONS: Our proposed nomogram showed more accurate prognostic prediction for patients with non-metastatic osteosarcoma.

MicroRNAs with prognostic significance in osteosarcoma: a systemic review and meta-analysis.

INTRODUCTION: This study aimed to elucidate the prognostic value of microRNAs (miRNAs) in patients with osteosarcoma. MATERIALS AND METHODS: Studies were recruited by searching PubMed, Embase, the Cochrane Library, China National Knowledge Infrastructure, and Wanfang data-bases (final search update conducted January 2017). Eligible studies were identified and the quality was assessed using multiple search strategies. RESULTS: A total of 55 articles that investigated the correlation between miRNA expression and either patient survival or disease recurrence in osteosarcoma was initially identified. Among these, 30 studies were included in the meta-analysis. The results of our meta-analysis revealed that elevated levels of miR-21, miR-214, miR-29, miR-9 and miR-148a were associated with poor prognosis in osteosarcoma. Additionally, downregulated miR-382, miR26a, miR-126, miR-195 and miR-124 expression indicated poor prognosis in osteosarcoma. CONCLUSIONS: miRNAs may act as independent prognostic factors in patients with osteosarcoma and are useful in stratifying risk.

Expression of MicroRNA-29a Regulated by Yes-Associated Protein Modulates the Neurite Outgrowth in N2a Cells.

Yes-associated protein (YAP) is proved to increase miR-29a in the present study, but the relevant molecular mechanism is not clear. Also, growing evidence indicates that the high-level miR-29a promotes the neurite outgrowth by decreasing PTEN (phosphatase and tensin homologue deleted on chromosome 10). Results show that the expression of miR-29a increases but the PTEN decreases during transfecting the N2a cells with the YAP plasmid. Meanwhile, the advancement of neurite outgrowth is presented via using multiple methods to detect the expression of GAP-43 and NF-200, which have a strong association with neurite outgrowth. The expression of miR-29a, GAP-43, and NF-200 shows an opposite tendency compared to the PTEN when YAP is downregulated. By treating N2a cells with miR-29a mimic and inhibitor, we also find the same conclusion. For in silico analysis of miR-29a, its promoter may have a binding site for YAP. Based on a luciferase reporter assay and a chromatin immunoprecipitation (ChIP) experiment, we demonstrate that YAP could increase the expression of miR-29a by targeting the promoter of miR-29a. In conclusion, the results identify that YAP promotes the neurite outgrowth via targeting the promoter of miR-29a, and it may be an effective therapeutic medicine for the neural disease.

Rat Nasal Respiratory Mucosa-Derived Ectomesenchymal Stem Cells Differentiate into Schwann-Like Cells Promoting the Differentiation of PC12 Cells and Forming Myelin In Vitro.

Schwann cell (SC) transplantation as a cell-based therapy can enhance peripheral and central nerve repair experimentally, but it is limited by the donor site morbidity for clinical application. We investigated weather respiratory mucosa stem cells (REMSCs), a kind of ectomesenchymal stem cells (EMSCs), isolated from rat nasal septum can differentiate into functional Schwann-like cells (SC-like cells). REMSCs proliferated quickly in vitro and expressed the neural crest markers (nestin, vimentin, SOX10, and CD44). Treated with a mixture of glial growth factors for 7 days, REMSCs differentiated into SC-like cells. The differentiated REMSCs (dREMSCs) exhibited a spindle-like morphology similar to SC cells. Immunocytochemical staining and Western blotting indicated that SC-like cells expressed the glial markers (GFAP, S100ß, Galc, and P75) and CNPase. When cocultured with dREMSCs for 5 days, PC12 cells differentiated into mature neuron-like cells with long neurites. More importantly, dREMSCs could form myelin structures with the neurites of PC12 cells at 21 days in vitro. Our data indicated that REMSCs, a kind of EMSCs, could differentiate into SC-like cells and have the ability to promote the differentiation of PC12 cells and form myelin in vitro.

MicroRNA-29c/PTEN pathway is involved in mice brain development and modulates neurite outgrowth in PC12 cells.

Growing evidence indicates that microRNAs (miRNAs) are important mediators of brain development and neurite growth. However, the affected signaling mechanisms are not clearly clarified. In the present study, we confirm that miR-29c is expressed during mice brain development and increases neurite outgrowth via decreasing PTEN expression. We first screen the picked-out miR-29c up-regulated in PC12 cells induced by nerve growth factor (NGF). In silico analysis of possible miR-29c targets, VEGFA, MAPK3, PDGFB, and PTEN mRNA are proposed as relatively likely putative binding sites for miR-29c. Subsequently, we detect that miR-29c is involved in brain development and has a negative relationship with the expression of PTEN. Then, using luciferase reporter assay,we demonstrate that miR-29c could directly target to the 3'-UTR of PTEN mRNA and result in down-expression of PTEN. By infecting PC12 cells with lentiviral pLKO-miR-29c or control, we also find that increasing levels of miR-29c markedly increase Akt phosphorylation level, and thus, promote neurite outgrowth of PC12 cells. Together, our results identify that miR-29c is required for mice brain development and modulates neurite outgrowth in PC12 cells via targeting PTEN and has a promising therapeutic target for neural disease.

Identification of rat respiratory mucosa stem cells and comparison of the early neural differentiation potential with the bone marrow mesenchymal stem cells in vitro.

The aim of this study is to identify rat nasal septum respiratory mucosa-derived mesenchyme stem cells (RM-MSCs) and to compare its neural lineage differentiation capacity with bone marrow-derived mesenchyme stem cells (BM-MSCs) after a short period of neural induction culture in vitro. The cell morphology was observed with light microscopy; cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The characteristics of the cells were evaluated with flow cytometry, immunofluorescence, real-time quantitative PCR (RT-PCR), and Western blotting. The results showed that rat nasal respiratory mucosa contains RM-MSCs that exhibited similar proliferation rate as BM-MSCs in vitro. Both RT-PCR and Western blotting analyses demonstrated that RM-MSCs showed higher expression of neural lineage markers than BM-MSCs after a short period of neural induction culture, and secreted higher level of brain-derived neurotrophic factor. RM-MSCs were more amenable to differentiate into neural or glial cell after a short period of neural induction culture than BM-MSCs in vitro; and it could be considered as another optimal source of stem cells for cell-based therapy to neurological diseases.