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Objective:To investigate the effects of over-expression of E2F transcription factor 1 (E2F1) on proliferation, invasion, apoptosis and radiosensitivity of glioma cell U251.Methods:Real-time quantitative PCR (qRT-PCR) were used to detect the differential expression of E2F1 mRNA in glioma cells LN18, SW1088, U251 and normal brain glial cells. The stable over-expression of E2F1 plasmid was constructed and transfected into U251 cells. qRT-PCR and Western blot test were used to detect the expression of E2F1, pituitary tumor transforming gene 1(PTTG1), C-Myc, B-cell lymphoma-2 (Bcl-2), Bcl2-associated X (Bax) mRNA and protein expression in the control group and E2F1 over-expression group.U251 cells were divided into control group(no X-ray irradiation), irradiation group(6 Gy dose of X-ray), and irradiation + E2F1 over-expression group(transfected with E2F1 first, then irradiated by 6 Gy of X-ray). Cell proliferation ability was detected by cell counting Kit-8(CCK-8) cell viability detection reagent, and cell invasion and migration ability were detected by Transwell chamber. Apoptosis and cell cycle were detected by flow cytometry.GraphPad Prism 8.0 was used for data analysis.The statistical methods were one-way ANOVA and independent sample t-test. Results:qRT-PCR showed that there was statistical difference in the mRNA levels of E2F1( F=201.92, P<0.05) in different cell lines.The expression levels of E2F1 mRNA in LN18(4.04±0.29), SW1088(3.19±0.16)and U251(4.66±0.20) cells were higher than those in HEB(1.02±0.07)cells ( q=27.00, 19.40, 32.52, all P<0.05). After successfully constructing U251 cells with stable over-expression of E2F1 plasmid, qRT-PCR and Western blot detection results showed that: the mRNA and protein levels of E2F1, PTTG1, C-Myc and Bcl-2 in E2F1 over-expression group were higher than those in control group ( t=77.16, 57.88, 4.63, 51.13, 7.50, 70.85, 8.38, 48.81, all P<0.05). Bax mRNA(0.20±0.01) and protein(0.66±0.01) levels were lower than those in control group((1.00±0.02), (0.94±0.01)), and the differences were statistically significant ( t=1.74, 54.65, both P<0.05). After X-ray irradiation (6 Gy), CCK8 detection results showed: the proliferation ability of the three groups at 24, 48, 72 and 96 h were significantly different ( F=95.41, 187.53, 1 158.49, 7 883.78, all P<0.05). The proliferation capacity of the irradiation group were lower than those of the control group at 24, 48, 72 and 96 h ( q=19.51, 27.20, 66.60, 174.9, all P<0.05). The proliferation capacity of irradiation + E2F1 over-expression group at 24, 48, 72 and 96 h were higher than those of irradiation group ( q=10.63, 10.81, 21.11, 60.90, all P<0.05). Transwell assay results showed that there were significant differences in cell invasion and migration ability among the three groups ( F=315.38, 681.10, both P<0.05). The invasion and migration ability of cells in the irradiation group were lower than those in the control group ( q=35.09, 12.76, both P<0.05), and the invasion and migration ability of cells in the irradiation + E2F1 over-expression group were higher than those in the irradiation group ( q=52.06, 22.81, both P<0.05). Flow cytometry showed that there were significant differences in apoptosis rate and percentage of cells in each cycle among the three groups ( F=667.63, 3 213.30, 3 011.26, 861.98, all P<0.05). The percentage of the apoptosis rate, S phase and G2 phase cells in the irradiation group were higher than those in the control group ( q=51.10, 89.39, 51.82, all P<0.05), while the percentage of G1 phase cells in the irradiation group was lower than that in the control group ( q=141.2, P<0.05). The apoptosis rate and percentage of S phase and G2 phase cells in the irradiation + E2F1 over-expression group were lower than those in the irradiation group ( q=18.87, 41.42, 29.31, all P<0.05), while the number of G1 phase cells in the irradiation + E2F1 over-expression group was lower than that in the irradiation group ( q=70.73, P<0.05). Conclusion:Over-expression of E2F1 can reduce the radiosensitivity of glioma U251 cells by regulating the expression of mRNA and protein of genes related to cell cycle and apoptosis, and E2F1 may be involved in the radioresistance of glioma cells.
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Objective:To investigate the clinicopathological features, treatment programs and prognosis of patients with primary diffuse large B-cell lymphoma (DLBCL) in cavernous sinus.Methods:The clinical data of a patient with primary DLBCL in cavernous sinus who were admitted to Wuhan No.1 Hospital in December 2020 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was a 63-year-old female who underwent resection of the cavernous sinus lesion, and the pathological diagnosis was DLBCL. The patient received 6 courses of R-CHOP regimen chemotherapy, lumbar puncture + intrathecal injection of chemotherapy drugs, and twice additional rituximab immunochemotherapy, and no tumor cells were found in the results of liquid-based thin layer cytology for cerebrospinal fluid exfoliated cells; twice magnetic resonance imaging (MRI) re-examination after the operation showed no recurrence and adjacent metastasis of the tumor. The patient's symptoms were significantly improved without residual neurological sequelae.Conclusions:Primary DLBCL in cavernous sinus is rare in clinical practice, early diagnosis is crucial for the prognosis of patients, and different protein expression may indicate the prognosis. Biopsy, complete resection of the tumor under the premise of preserving important anatomical structures and functions, and standardized chemotherapy combined with intrathecal injection local chemotherapy can effectively prolong the survival time of patients and improve the quality of life.
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Objective To investigate the application value of CT perfusion (CTP) imaging for the revascularization treatment in adult patients with Moyamoya disease.Methods Adult patients with Moyamoya disease underwent revascularization in the Department of Neurosurgery,Wuhan No.1 Hospital from July 2009 to December 2016 were analyzed retrospectively.CTP and clinical evaluation were performed before and after 3-6 months of procedure.The modified Rankin Scale (mRS) was used to assess the functional outcomes.Results A total of 20 patients were enrolled in the study,including 9 females and 11 males,aged 29 to 73 years,with an average of 53.5 years.The initial symptom was ischemic stroke in 10 patients,transient isehemic attack in 7 patients,and hemorrhagic stroke in 3 patients.All patients underwent superficial temporal artery-middle cerebral artery bypass grafting plus encephalomyo-synangiosis under general anesthesia.All patients have different degrees of improvement in cerebral blood flow after procedure,and the CTP parameters were significantly improved compared with those before procedure (all P ＜0.05).The clinical symptoms were significantly improved in 3 cases (15％) and recovered in 13 cases (65％) at 6 months after procedure.The proportion of the mRS score 0-2 was significantly higher than that before procedure (90.0％ [18/20] 对 50.0％ [10/20];x2 =7.619,P =0.006).Conclusion CTP can evaluate the cerebral perfusion status in various vascular areas through hemodynamic parameters in early stage,which can effectively guide the operation mode of Moyamoya disease,and evaluate the changes of cerebral perfusion status after procedure as a means of follow-up of the disease.
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Objective To observe the expression of indoleamine 2,3-dioxy-genase(IDO) in hippocampus of rats with posttraumatic stress disorder (PTSD) and the protective effect of IDO inhibitor on neurons,and to explore the role of IDO in the pathogenesis of PTSD.Methods Adult male Wistar rats were randomly divided into the normal control group,PTSD model group and IDO inhibitor treatment group.The expression of IDO was detected by immunohistochemistry,RT-PCR and Western-blot.The apoptosis of rat hippocampal neurons was assayed by Tunel staining.Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by ELISA.Moreover behavioral evaluation was performed,including central residence time,percentage of open arm residence time and stage latency.Results Comparing with the control group,PTSD rats showed decreased central residence time ((22.65± 1.54)s),decreased percentage of open arm residence time((10.55± 1.96) ％),prolonged stage latency ((56.38±4.21) s) (P＜0.05),increased TNF-α ((8.58±0.6) pg/ml),IL-6 ((15.72±1.42) pg/ml) and IDO mRNA (0.8278±0.0796),increased IDO protein (1.2329±0.1148) expression and apoptosis rate ((81.47± 6.86) ％) in hippocampus (P＜ 0.05) (P＜ 0.05).However,rats treated with IDO inhibitor showed increased central residence time((30.78±3.20) s),increased percentage of open arm residence time ((10.55± 1.96)％),shortened stage latency ((56.38 4.21) s),meanwhile reduced expression of TNF-α((3.69±0.41) pg/ml),IL-6((7.45±0.58) pg/ml),IDO mRNA(0.2236 ±0.0387) and IDO protein(0.4235±0.0411) was detected in hippocampus(P＜0.05).Apoptosis rate ((42.54± 3.98)％) was also decreased in hippocampus(P＜0.05).Conclusion The content of TNF-α,IL-6 and IDO are increased significantly in the hippocampus of PTSD rats.IDO may participate in the pathogenesis of PTSD,and the IDO inhibitor may play a neuroprotective role in hippocampus of PTSD.
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Objective To explore the dynamic changes and the meaning of T lymphocyte subpopulation in peripheral blood and its clinical significance in patients with acute cerebral infarction.Methods The numbers of CD3 +,CD4 + and CD8 + T lymphocyte in peripheral blood were examined with flow cytometry in 20 patients with acute cerebral infarction(CI) at the 3rd,5th,7th and 14th day after the onset.Results The numbers of CD3 + and CD4 + T lymphocytes and the ratio of CD4 +/CD8 + in the patients with acute CI were lower than those in normal group at the 3rd and 5th day after the onset (P0.05).Conclusion The immunological function of the cells in the patients with acute CI decreased obviously at the 3th and 5th day, and then restored partly at the 7th day, at last restored completely at the 14th day after the onset. It suggests that the focus time for the prevention of infective complications is in 10 days after the acute CI.