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[Objective] To detect the expression of NAD(P)H oxidase (Nox) and the content of ROS in different malignant gliomas,and explore the influence of Nox and intracellular ROS levels in the survival,proliferation,and malignant phenotype of gliomas.[Methods]Thirty human glioma specimens (10 with grade Ⅰ and I1,10 with grade II and 10 with grade IV),performed resection in our hospital from August 2007 to August 2010,were collected in our study;another 10 normal brain tissues were collected as controls.Real time PCR was used to detect the mRNA expressions of Nox(1-5);ROS level was detected by flow cytometry and Nox4 protein level was detected by immunofluorescence and Western blotting.[Results] The Noxl-5 mRNA and protein levels and ROS content were significantly different between each 2 groups (P＜0.05);Nox4 mRNA expression and ROS level significantly increased following the increment of malignancy degrees (controls＜low-grade gliomas＜grade III gliomas＜ grade IV gliomas,P＜0.05);the Nox protein expression was low in controls while that in gliomas was high,and the higher malignancy of the tumors,the higher expression levels of the tumors.[Conclusion] Nox expression and ROS content have significantly positive relations with the malignancy of the tumors,which indicates that Nox expression and ROS content might paly important roles in the occurrence of glioma and malignant proliferation.
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Objective To describe a simple and efficient method to obtain large quantities of higllly purified Schwann cells(SCs).Methods SCs were isolated from the sciatic and brachial nerves of 3-to 5-day-old newborn SD rats by collagenase digestion.The isolated SCs were plated at the density of3x105/mL for primary culture,several rounds oftrypsin digestion were performed within 72 h to purify SCs. Results Compared with the purification using 1.25 g/L trypsin digestion in serial differential detachment procedures, Our protocol allowed easier and more efficient separation of the SCs from the fibroblasts.Immunocytochemical staining showed that the purity of the SCs exceeded 95%.Conclusion The purification protocol of the SCs we established can be easily carried out and yields well reproducible results to obtain large quantities ofhighly purified SCs for transplantation studies.
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Objective To compare the differentiation potentials of bone marrow-derived stromal cells (BMSCs) and adipose-derived stromal cells (ADSCs) into neuron-like cells in vitro. Methods The fifth-passage of cultured adult SD rat BMSCs and ADSCs were induced with 10 ng/mL epidermal growth factor (EGF) and 20 ng/mL basic fibroblast growth factor (bFGF). After induction for 6, 12, 24, 72 h and 1 and 2 weeks, the cells were harvested to examine the expressions of the neural markers nestin and β-tubulin Ⅲ using immunohistoChEnistry and Western blot. Results Both the BMSCs and ADSCs underwent morphological changes into neuron-like cells and expressed the neuron-specific markers after the induction. The two cells exhibited significantly different positivity rates for nestin and β-tubulin Ⅲ after the induction (P<0.05). The ADSCs exhibited stronger ability than BMSCs to differentiate into neuron-like cells shown by greater nestin and β-tubulin Ⅲ positivity rates after the induction. Conclusion ADSCs possess stronger capacity of induced differentiation into neuron-like ceils than BMSCs in in vitro culture.
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Objective To explore the reliability of acquiring Schwann cells from bone marrow stromal cells (BMSCs) by inducing their differentiation in vitro. Methods BMSCs isolated from the tibial and femural bone marrow of SD rats by adherent culture were passaged and induced with beta-mercaptoethanol followed by retinoic acid, forskolin, basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) and heregulin. Immunocytochemistry was performed to identify the expression of the Schwann cell markers P75, S100 and glial fibrillary acidic portein (GFAP), and fluorescent real-time quantitative RT-PCR was used to detect the mRNA expressions of P75, S100 and CD104 in the induced BMSCs. Results The induced BMSCs exhibited morphological changes into cells resembling primary cultured Schwann ceils, and expressed P75, S100 and GFAP proteins, as shown by immunofluorescent staining, at the levels similar to those in Schwann cells. The induced cells, however, remained negative for P75 mRNA expression as demonstrated by RT-PCR. Conclusion The BMSCs can be induced to partially acquire the phenotypic features of Schwann cells, suggesting the strong plasticity of the BMSCs. The protocol of pre-induction followed by induction with an array of agents still needs further modification to induce the in vitro differentiation of BMSCs into Schwann-like cells.
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Objective To analyze the effect of NexusTM coils for endovascular occlusion of intracranial aneurysms. Methods In 41 patients with intracranial aneurysms, endovascular occlusion of 43 aneurysms was performed using NexusTM coils. The follow-up data of the patients for 6 to 12 months were reviewed, and the imaging data from digital subtraction angiography (DSA), CT angiography (CTA) or magnetic resonance arthrography (MRA) alter the treatment were analyzed. Results In the 41 patients, 1 died, 1 had aneurysm recurrence, 3 had cerebral infarction, 1 showed ocular paralysis, and 2 developed hydrocephalus after the surgery. Evaluation with modified Rankin Scale showed grade 0 in 8 cases, grade 1 in 19 cases, grade 2 in 7 cases, grade 3 in 3 cases, grade 4 in 1 case, grade 5 in 1 ease and grade 6 in 1 case. Conclusion Endovascular embolization with NexusTM coils is effective for treatment of intracranial aneurysms especially in cases of small aneurysms and parent artery occlusion. Caution should be taken with the coil for endovascular occlusion of the neck of anterior and middle cerebral artery aneurysms with thin parent arteries, as the fibers in the coil may cause thrombosis and potential cerebral infarction.
