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AIM:To study the effect of centromere protein W ( CENP-W) down-regulation on human glioma U87 cells.METHODS:Small interfering RNA ( siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot .The proliferation of the cells was analyzed by MTT assay , BrdU staining and colony formation experiment .Transwell chamber assay was used to detect the invasion a-bility of the cells .The cell migration ability was measured by a scratch test .The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry .RESULTS:The results of MTT assay , colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group .The results of Transwell and scratch experiments showed that the migra-tion and invasion abilities in experimental group were weaker than those in blank control group and negative control group . The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G 0/G1 phase .The percentage of apoptotic cells in experimental group was higher than that in control group ( P<0.05 ) .CON-CLUSION:Down-regulation of CENP-W expression inhibits the proliferation , migration and invasion of human glioma cells and promotes the apoptosis of the cells , suggesting that CENP-W may be a potential target of gene therapy for human glioma.
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Objective AIM:To study whether the RWD containing sumoylation enhancer (RSUME) enhanced small ubiquitin related modifiers (SUMO) to competitively inhibit Ubiquitin B (UBB)-mediated degradation of hypoxia hypoxia-inducible factor 1α.(HIF-1α) and the invasive pituitary adenomas.Methods The expression of protein and mRNA levels of RSUME,SUMO-1,UBB and HIF-1α were detected by using immunohistochemistry,western blot and qPCR in 38 cases of non-invasive pituitary adenoma,38 cases of invasive pituitary adenomas and 10 cases of normal pituitary capsule.The expression of SUMO-1 was analyzed in different types of pituitary adenomas.Results The protein,and mRNA levels of RSUME,SUMO-HIF-1αwere significantly higher in the invasive pituitary adenomas than in the non-invasive pituitary adenomas and normal pituitary capsule (P＜0.01).The protein and mRNA levels of RSUME,SUMO-HIF-1αwas higher in non-invasive pituitary adenomas than in normal pituitary capsule(P＜0.01).However,the protein levels of UBB-HIF-1α were significantly lower in the invasive pituitary adenomas than in the non-invasive pituitary adenomas and normal pituitary capsule (P＜0.01).The protein of UBB-HIF-1α were lower in non-invasive pituitary adenomas than in normal pituitary capsule (P＜0.01).The expression levels of SUMO were not significantly different among different types of pituitary adenomas (P＞0.05).Conclusion RSUM may increase pituitary adenomas angiogenesis and promote tumor invasion through enhancement of SUMO of HIF-1α which competitively inhibits ubiquitination of HIF-1 α.
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Objective To evaluate the short-term effect of Pipeline embolization device (PED)for the treatment of unruptured wide-necked intracranial aneurysms. Methods From October 2015 to September 2016,15 consecutive patients with unruptured wide-necked intracranial aneurysm (aneurysm neck and aneurysm body ratio ≥0. 5)treated with PED at the Department of Neurosurgery,the Second Affiliated Hospital of Nanchang University were enrolled retrospectively. Their clinical and imaging data were analyzed. Kamran scale was used to evaluate the embolization rate of aneurysms and the changes of the parent arteries. DSA examination was performed again at 6 -12 months after operation. Results Fifteen PED were implanted in 15 patients with unruptured wide-necked intracranial aneurysms,including 13ophthalmic artery aneurysms,1 posterior communicating artery aneurysm,and 1 cavernous sinus aneurysm. The technical success rate was 100% . Immediately after PED implantation,Karman rating of 15 cases were aneurysm grade 2 embolization,parent artery grade A (grade 2a). DSA examination was performed again at 6 - 12 months after operation showed that 14 patients were aneurysm grade 4,parent artery was grade A (grade 4a). One patient (ophthalmic artery aneurysm)underwent the second DSA examinations at 6 and 12 months after operation showed that the residual development of aneurysms. The aneurysm embolization was grade 3, and the parent artery was grade A (grade 3a). No branch artery occlusion was observed. Non of them had neurological deficit. The modified Rankin scale score was 0 in all 15 patients. Conclusion The use of PED in the treatment of unruptured wide-necked intracranial aneurysms has a higher occlusion rate. Its long-term effect still needs further follow-up.
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Objective To investigate the expression of CENP-W in gliomas and its relationship with clinicopathological parameters and prognosis and to explore the effects of centromere protein W (CENP-W)on the invasion of gliomas cells.Methods The expressions of CENP-W in high-grade glioma tissues,low-grade glioma tissues,and adjacent brain tissues were detected by real-time fluorescence quantitative PCR and Western blotting. The correlation of the expression of CENP-W with clinicopathological parameters and prognosis was analyzed statistically.Human gliomas U2 5 1 cells in vitro were transfected with small interfering RNA to downregulate the expression of CENP-W.The invasion and migration capabilities of gliomas cancer cells were assessed by Transwell assays.Results The expression level of CENP-W was significantly higher in glioma tissues than in normal tissues. There was a positive correlation between the three protein expression levels and the pathological grade of gliomas. CENP-W siRNA was successfully transfected into U2 5 1 cells.Compared with those of the cells transfected with the scramble siRNA and control cells,the invasive and migration activities were inhibited in the U2 5 1 cells transfected with CENP-W siRNA.The Kaplan-Meier analysis and Log-Rank test showed significant differences in progress free survival (PFS)between the CENP-W high-expression and low-expression groups.Conclusion The expression level of CENP-W was positively correlated with the pathological grade of gliomas and CENP-W can promote glioma cell invasion.It implicates that CENP-W can be a novel target in gliomas treatment.
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Objective To compare the efficacies between interventional embolization and surgical clipping in treatment of ruptured intracranial aneurysms to provide an evidence‐based basis for selecting the clinical treatment scheme .Methods The related randomized controlled trail(RCT) literatures on the effects of interventional embolization and surgical clipping were retrieved from the databases of Pubmed ,Cochrane ,Medline and Embase .The screening was independently performed by two researchers according to the including and excluding criterion .The occurrence rate of adverse reactions ,postoperative 1‐year mortality rate ,re‐bleeding rate ,occurrence rate of vasospasm and ischemic cerebral infarction served as the measurement indicators .The data were extracted and performed the meta analysis by the RevMan5 .3 software .Results Sixteen RCT literatures were included for conducting analy‐sis ,involving 7 373 patients ,in which 3 092 cases adopted interventional embolization and 4 281 cases adopted surgical clipping .The occurrence rate of adverse events(OR=1 .25 ,95% CI ,1 .12-1 .40 ;P<0 .000 1) and re‐bleeding rate(OR=0 .43 ,95% CI ,0 .28 -0 .66 ;P=0 .000 1) in the interventional embolization group were lower than those in the surgical clipping group ;however ,there were no statistical differences between the interventional embolization group and surgical clipping group in the postoperative 1‐year mortality rate(OR=1 .13 ,95% CI ,0 .92-1 .39 ;P=0 .23) ,incidence rate of vasospasm (OR=1 .41 ,95% CI ,0 .99-2 .02 ;P=0 .06) and incidence rate of ischemic cerebral infarction(OR=0 .66 ,95% CI ,0 .42 -1 .05 ;P=0 .08) .Conclusion The current clinical re‐search evidences indicate that using the interventional embolization in treating ruptured intracranial aneurysms can obviously reduce the occurrence rate of adverse events than the surgical clipping ,but increases the re‐bleeding rate .The postoperative 1‐year mortali‐ty rate ,incidence rate of vasospasm and incidence rate of ischemic cerebral infarction have no obvious difference between these two kinds of operation .
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AIM:To investigate the effect of RWDD3 gene silencing on the biological characteristics of human glioma U251 cells.METHODS: A lentiviral vector expressing RWDD3 shRNA was constructed and transfeeted into the U251 cells.The expression of RWDD3 at mRNA and protein levels was detected by real-time PCR and Western blot , re-spectively .The cell activity was determined by MTT assay .The colony formation ability was detected by the colony forma-tion assay .The cell proliferation ability was detected by BrdU incorporation assay .The cell invasion and migration were evaluated by Transwell assay .Flow cytometry was used to monitor the changes of cell cycle distribution and apoptosis .RE-SULTS:Recombinant lentivirus was successfully transfected into U 251 cells.Compared with the cells transfected with the scrambled shRNA and control cells, the cell activity, colony formation ability, and the invasive and migratory activities were inhibited, the cell cycle was arrested in G 0/G1 phase, and the apoptosis was increased in the U 251 cells transfected with RWDD3 shRNA ( P <0.05 ) .CONCLUSION: RWDD3 plays a vital role in proliferation and invasion of glioma cells.It may serve as a potential target of gene therapy for glioma .
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Objective To explore the expressions and correlation of RWD containing sumoylation enhancer (RSUME),small ubiquitin-like modifier (SUMO-1),hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor (VEGF)in gliomas of different pathologic grade.Methods We investigated the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA with reverse transcription-polymerase chain reaction (RT-PCR),and investigated the immunohistochemical staining to determine the expressions of SUMO-1,HIF-1α and VEGF in 63 cases of human gliomas of different pathologic grade and 9 cases of normal brain tissues.We studied its correlation with the pathologic grade and the relationship between the expression of RSUME promoter sumoylation and HIF-1α/VEGF pathway in gliomas.Results There were significant differences (P <0.01)in the expressions of RSUME mRNA,HIF-1αmRNA and VEGF mRNA in glioma tissues.With the increasing degree of pathologic grade in tumor specimens,the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA increased markedly (P <0.01 ).There was a positive correlation of the expression levels of RSUME mRNA with HIF-1αmRNA and VEGF mRNA.There were significant differences (P <0.01 )in the expressions of SUMO-1,HIF-1αand VEGF in glioma tissues by immunohistochemical staining.With the ascending of pathologic grade of tumor specimens,the expression levels of SUMO-1,HIF-1α and VEGF increased markedly (P < 0.01 ).There was a positive correlation between the expression level of SUMO-1 and HIF-1α(r =0.857,P <0.01).The Kaplan-Meier analysis and Log-rank test showed significant differences in progress free survival (PFS)between the RSUME high-expression and low-expression groups (χ2 =36.032,P <0.01).Conclusion RSUME may enhance HIF-1α/VEGF pathway through sumoylation in gliomas.It implicates that RSUME is related to angiogenesis in gliomas and can promote tumor invasion and progression,indicating that RSUME can be a novel target in gliomas treatment.
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Objective To investigate the clinical treatment of severe hypertensive intracerebral hemorrhage in basal ganglia and broken into ventricles .Methods 16 patients with severe hypertensive intracerebral hemorrhage in basal ganglia and broken into ventricles received microsurgical treatment using posterior central gyrus posterior -lateral fissure upper approach .The hematoma cavity and ventricle were opened ,and brain surgical department drainage was placed in the hematoma cavity in the operation .The curative effect was analyzed retrospectively 6 months after operation.Results According to Glasgow Outcome Scale ( GOS):6 cases had good recovery;4 cases had moderate disability;2 cases had severe disability;1 case was in persistent vegetative state and 3 cases were dead .Conclusion Craniotomy microsurgical operation using posterior central gyrus posterior -lateral fissure upper approach is an effec-tive treatment of severe hypertensive intracerebral hemorrhage in basal ganglia and broken into ventricles .The fatality rate was reduced .
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Objective:To detect the expression of miR-106b~25 cluster in glioma cell line and tissues. Methods:Real-time PCR was performed to determine the expression of miR-106b~25 cluster members (miR-106b, miR-93, and miR-25) in different human glio-blastoma cell lines. Different pathological grade glioma specimens were surgically removed. In-situ hybridization was performed to de-tect the expression of miR-106b~25 cluster members in different pathological levels of glioma tissues. Results:In the expression of the benchmark on normal brain tissues, three kinds of miRNAs in all test cell lines have a tendency to increase. Based on the expression of the pathological level I average rate in 43 cases of glioma specimens collected after neurosurgical operations, the real-time PCR results showed that the average expression quantity of the three kinds of miRNAs in each group gradually increase. The increase in tumor path-ological levels results in statistically significant expression differences of miR-106b and miR-93 between the groups (F=4.479, P=0.018 and F=3.493, P=0.040, respectively). However, miR-25 expression differences between the groups have no statistically signifi-cant differences (F=2.766, P=0.075). In situ hybridization results show that the expressions of three miRNAs in high grade gliomas are significantly higher than that in the low-level tumor. Spearman rank correlation analysis results indicate that the expression of these miRNAs signal-intensity distribution is positively correlated with glioma, in accordance with WHO pathology classification. The corre-lation coefficient for miR-106b, miR-93, and miR-25 are 0.617, 0.438, and 0.463, respectively (P<0.001). Conclusion:The expression of miR-106b~25 cluster members is up-regulated in the glioma and is positively correlated with tumor grade.
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To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.
