Expression of AEBP1 and its prognostic role in glioma based on the data-mining of TCGA and GTEx databases / 西安交通大学学报(医学版)

【Objective】 To explore the expressions of adipocyte enhancer binding protein 1 （AEBP1） gene and its isoforms in different types of gliomas， and the influence of AEBP1 gene on the prognosis of patients with different types of gliomas. 【Methods】 We used the GEPIA2 visual network analysis tool to analyze AEBP1 gene expression levels in the tumor tissues of glioblastomas （GBM， including classical， mesenchymal， neural， and proneural ones） and low-grade gliomas （LGG， including astrocytoma， oligoastrocytoma， oligodendroglioma） in the TCGA database and normal human tissue samples in the TCGA and GTEx databases by one-way ANOVA. The distribution trend of isoforms of AEBP1 gene in gliomas was analyzed using the violin plot. The Kaplan-Meier survival curve was drawn and the Logrank test was used to analyze the influence of AEBP1 gene expression in GBM and LGG tumor tissues on the prognosis of glioma patients. 【Results】 The expression of AEBP1 in the tumor tissues of overall GBM and the four types of GBM was higher than that in the normal control tissues （P<0.05）. The expression of AEBP1 in astrocytoma and oligodendrocyte astrocytoma tumor tissues was higher than that in normal control tissues （P<0.05）. There were nine isoforms of AEBP1 gene in GBM and LGG， and the expression level in GBM was higher. The overall survival （OS） of the AEBP1 low expression group of GBM patients and the proneuronal GBM patients was better than that of the high expression group （P<0.05）. The OS and progression-free survival of LGG patients and the AEBP1 low-expression group of astroglioma were better than those of the high-expression group （P<0.05）. 【Conclusion】 AEBP1 has an important clinical value in the pathogenesis and development of GBM and LGG， and thus can be used as a diagnostic marker and a candidate gene for targeted therapy.

Regulatory effect of annexin A5 on glioma cell invasion andmigration and its mechanisms / 西安交通大学学报(医学版)

【Objective】 To investigate the regulatory effect of annexin A5 on glioma cell invasion and migration and its mechanism. 【Methods】 The expression of annexin A5 in 100 cases of glioma tissues and 20 cases of normal brain tissues was detected by immunohistochemistry. The expression of annexin A5 was downregulated by transfection with siRNA targeting annexin A5 (si-Annexin A5) in human glioma cell line (U251). The expression of annexin A5 was confirmed by RT-PCR and Western blot analysis. The proliferation ability of U251 cells was detected by MTT test and colony formation test, the apoptosis of U251 cells was detected by flow cytometry and Hoechst 33258 staining, and the migration and invasion ability of U251 cells was examined by wound healing test and Matrigel Transwell invasion test. The expressions of Raf, p-Raf, MEK1/2, p-MEK1/2, ERK1/2, p-ERK1/2, c-Myc and E-Cadherin in U251 cells were analyzed by Western blot. 【Results】 Compared with those of normal brain tissues, the mRNA and protein expression levels of Annexin A5 in glioma tissues increased by 2.45 times and 2.87 times, respectively (P<0.05). Pearson correlation analysis showed that with the increase of tumor grade, the positive rate of Annexin A5 gradually increased, and the tumor grade and positive rate were significantly positively correlated (r=1.000, P=0.000). The cell viability of U251 cells in the si-Annexin A5 group after 48 h and 72 h of culture was significantly reduced by 29.46% and 40.43%, respectively, compared with that in the control group (P<0.05). Compared with the control group, in the si-Annexin A5 group the colony formation rate was reduced by 68.58%, while the apoptosis rate was increased by 24.41 times (P<0.05); the cell migration rate and invasion rate were reduced by 65.35% and 68.80% (P<0.05). The protein expression of p-Raf, p-MEK1/2, p-ERK1/2 and c-Myc in the si-Annexin A5 group were significantly reduced by 54.67%, 70.37%, 60.26% and 54.95%, respectively, and that of E-Cadherin was increased by 3.58 times (P<0.05). 【Conclusion】 Downregulation of Annexin A5 inhibits the growth and motility of glioma cells and induces cell apoptosis by inhibiting the Raf/MEK/ERK signaling pathway.

miR-139-5p targeting DNMT1 inhibits the growth and metastasis of glioblastoma / 西安交通大学学报(医学版)

【Objective】 To investigate the expression and biological role of miR-139-5p in glioblastoma and the regulatory effect of miR-139-5p on DNA methyltransferase 1 (DNMT1). 【Methods】 qRT-PCR was used to detect the expression of miR-139-5p in glioblastoma tumor tissue, paired paracancerous tissue, human normal glioma cell line HEB, and human glioma cell line U251. The expression of miR-139-5p in U251 cells was up-regulated by transfection of miR-139-5p mimetics, and the expression of DNMT1 was down-regulated by transfection of DNMT1-targeted siRNA (DNMT1-siRNA). The expression of DNMT1 and neurofibromatosis type 2 (NF2) in tissues and cells was detected by qRT-PCR, Western blotting, immunohistochemistry and immunofluorescence. The cell counting kit-8 (CCK-8), flow cytometry and Matrigel Transwell were used to evaluate the proliferation, apoptosis and invasion ability of U251 cells. 【Results】 Compared with paracancerous tissues or HEB cells, miR-139-5p expression in glioblastoma tissues and U251 cells was suppressed (P<0.05). Compared with control cells, transfection of miR-139-5p mimic significantly down-regulated the expression of DNMT1 and up-regulated the expression of NF2 (P<0.05). Compared with control cells, transfection of DNMT1-siRNA significantly promoted the expression of NF2 (P<0.05). Transfection of miR-139-5p mimetics or DNMT1-siRNA significantly induced U251 cell apoptosis and inhibited cell invasion (P<0.05). 【Conclusion】 miR-139-5p plays an anti-cancer role in glioblastoma, and it inhibits tumor proliferation and metastasis by targeting negative regulation of DNMT1.

Clinical effects of Zero-P vs.traditional titanium plate for single level cervical spondylosis / 西安交通大学学报(医学版)

ABSTRACT:Objective To compare the clinical effects of Zero-P and traditional titanium plate with cage in treating single level cervical spondylosis.Methods We retrospectively analyzed the clinical data of 28 patients with single level cervical spondylosis who underwent anterior cervical discectomy and fusion (ACDF)by traditional titanium plate with cage (group A,n=16)or Zero-P implant (group B,n=16)in our department between January 2012 and January 2014.The intraoperative blood loss,operation time,postoperative JOA scores,JOA recovery rate,NDI scores,and dysphagia incidence in both groups were measured and compared.The changes of the midpoint interbody height (IBH),the cervical Cobb angle of the surgical segment and the cervical Cobb angle of C2 and C7 vertebral body were measured on the standing lateral cervical spine X-ray.Results The operation time was significantly longer in group A than in group B (P 0 .0 5 ).The incidence of dysphagia was lower in group B than in group A.The two groups did not significantly differ in JOA score or JOA improvement rate during the same period after operation (P>0.05).The NDI score in group B was significantly lower than that in group A (P0 .0 5 ).The midpoint interbody height in group B was significantly greater than that in group A (P0.05).Conclusion Zero-p and traditional titanium plate with cage have similar effects in treating single level cervical spondylotic myelopathy,and Zero-P system has the advantages of lower incidence of dysphagia after operation,shorter operation time and better recovery of postoperative cervical physiological structure.

Microsurgery of intra-spinal tumor via the quadrant pathway / 西安交通大学学报(医学版)

Objective To investigate the indications,surgical principles and advantages and disadvantages of microsurgery of intra-spinal tumor via the quadrant pathway.Methods We made a retrospective analysis of the clinical data of patients with spinal canal tumors treated via the quadrant pathway from October 2014 to July 201 5. Results Totally 1 6 patients were included.Their tumors were fully removed by microscopic resection,in which there were three epidural cases,twelve subdural extra-medullar cases,and one intra-medullar case.In terms of stage,there were three cases of cervical segment (C3 - C7 ),four cases of thoracic (T1 - T10 ),nine cases of thoracic and lumbar ones (T1 1 - S2 ). In pathology, there were nine cases of neurilemmoma, two cases of neurofibroma,four cases of meningeal tumor and one case of neuroepithelial cyst.The postoperative symptoms and signs of the patients were significantly improved,and no recurrence or complications occurred in the follow-up 2-10 months.Conclusion Quadrant pathway can be used in removing spinal epidural and subdural tumor,which is shorter than two vertebral segments in length,and the cross-sectional area is less than 2/3 of the spinal cord. However,surgery with extensive adhesion and intra-medullary lesions should be performed carefully,and the surgical techniques are more demanding and a longer learning curve is needed.

Comparison of the modified expanding suspended laminoplasty and posterior pedicle screw fixation for lumbar intraspinal tumors / 西安交通大学学报(医学版)

Objective To compare the modified expanding suspended laminoplasty and posterior pedicle screw fixation applied in lumbar intraspinal tumors and evaluate operation,complications,postoperative neurofunctional improvement and lumbar lumbar stability in the two groups.Methods We made a retrospective analysis of the clinical data of 1 14 cases of lumbar intraspinal tumors, including 66 cases of modified expanding suspended laminoplasty and 48 cases of posterior pedicle screw fixation.We compared the operation time,operation bleeding volume,and incidence of postoperative cerebrospinal fluid leakage in the two groups.We also analyzed the lumbar instability and spinal cord injury score standard by the Japanese Orthopedic Association (JOA)between the two groups at 3,6, 12,and 24 months after operation.Results The results of modified expanding suspended laminoplasty group were significantly better than those of posterior pedicle screw fixation group in operation time, operation bleeding volume and incidence of postoperative cerebrospinal fluid leakage (P 0.05).At 3,6 or 12 months after the operation, changes in the increase of JOA score of the two groups had no significant difference (P > 0.05 ).However,24 mouths after the operation,the group of posterior pedicle screw fixation had significantly improved JOA score compared with that in the modified expanding suspended laminoplasty group.Conclusion Both the modified expanding suspended laminoplasty and posterior pedicle screw fixation have a favorable outcome of postoperative lumbar instability and neurofunction.And posterior pedicle screw fixation is superior to modified expanding suspended laminoplasty in improving neurofunction.

Application of intraoperative nerve electrophysiological monitoring inlumbosacral spinal cord tumor resection / 西安交通大学学报(医学版)

Objective To study intraoperative neural electrophysiological monitoring applied in lumbosacral spinal cord tumor resection.Methods We retrospectively reviewed the clinical data of 212 patients undergoing lumbosacral spinal cord tumor resection with or without intraoperative neural electrophysiological monitoring in our hospital.The patients were divided into two groups:124 patients in the monitored group received intraoperative neural electrophysiological monitoring while 88 ones in the control group did not.The monitoring was performed by recording the cortical somatosensory evoked potential (CSEP),dermatomal somatosensory evoked potential (DSEP) and electromyography (EMG).The patients were followed up for 3-6 months and their postoperative outcome was analyzed.Results There were significant differences in the outcome (P <0.05),but no difference was found in the incidence of complications between the monitored group and the control group.The sensitivity of CSEP +DSEP+EMG was 100%,and the specificity was 55.9% in the former group.Conclusion Combined monitoring with CSEP,DSEP and EMG during lumbosacral spinal cord tumor resection is valuable in protecting the spinal nerve roots and ensuring better operation safety.

The value of pedicled fat and capsule-packed nerve root in operation of sacral canal cysts / 西安交通大学学报(医学版)

Objective To evaluate the value of pedicled fat and capsule-packed nerve root in operation of sacral canal cysts.Methods We collected the information of 14 cases of sacral canal cysts for operation in our department and analyzed the operation indications and skills as well as the prognosis.Results All the operations were performed with the help of the microscope and the electrophysiological monitor.The capsules were removed in 9 cases and wrapped in 6 cases,with the total resection rate of 64.3%.The clinical symptoms were improved markedly after the operation.Conclusion The operation with pedicled fat and the capsule-packed nerve root is valuable in treating sacral canal cysts because it can protect the nerve root and get lower recurrence rate.

Characterization of glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and its association with cell proliferation and apoptosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and the association between cell proliferation and apoptosis and the metabolic status.</p><p><b>METHODS</b>An in vitro hypoxic cell model was established in SHG44 cells using CoCl2. Real-time PCR and Western blotting were used to assess the expressions of hypoxia-inducible factor-1α (HIF-1α) and the enzymes involved in glycolysis including PDK1, PKM2, and LDHA. Intracellular ATP levels were measured by bioluminescence assay to assess the energy metabolic status of SHG44 cells. The viability and apoptosis of the cells were examined using MTT assay and flow cytometry, respectively.</p><p><b>RESULTS</b>The cells in hypoxic culture showed obviously increased expressions of HIF-1α, LDHA, PDK1, and PKM2 at both the mRNA and protein levels as compared to those in normal cell culture. Hypoxia of the cells also resulted in a lowered cell proliferative activity and an increased apoptosis rate with lowered intracellular ATP concentrations and elevated mitochondrial membrane potential.</p><p><b>CONCLUSION</b>Hypoxia can induce a glycolytic phenotype of tumor cells. The sensitivity of tumor cells to hypoxia-induced cell death is directly correlated with their metabolic status.</p>