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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 therapeutic efficacy of enteral nutrition through percutaneous endoscopic gastrostomy/jejunostomy (PEG/PEJ) in patients with severe cerebral hemorrhage.Methods:Eighty-five patients with severe cerebral hemorrhage admitted to our hospital from January 2015 to December 2018 were enrolled into this retrospective study. According to ways of enteral nutrition, all 85 patients were divided into nasogastric tube group ( n=44) and PEG/PEJ tube group ( n=41). The clinical data were analyzed retrospectively, and the enteral nutrition treatment efficacy, incidence of complications, and length of hospital stays between the two groups were compared. Results:The incidences of diarrhea (14.6%, 6/41), gastric retention (34.1%, 14/41), and hypoproteinemia (26.8%, 11/41) in PEG/PEJ tube group were significantly lower than those in nasogastric tube group (38.6% [17/44], 59.1% [26/44], and 47.7% [21/44], P<0.05). However, the rate of obstruction ducts in PEG/PEJ tube group (34.1%, 14/41) was significantly higher than that in nasogastric tube group (11.4%, 5/44, P<0.05). As compared with the patients in nasogastric tube group, patients in the PEG/PEJ tube group had significantly shorter average length of hospital stays ([35.2±4.7] d vs. [37.6±5.4] d, P<0.05). The NRS2002 scores of patients in the nasogastric tube group and PEG/PEJ tube group after enteral nutrition treatment were 1.73±0.52 and 1.87±0.64, respectively, without significant difference ( P<0.05). Conclusion:The enteral nutrition treatment through PEG/PEJ could significantly reduce the incidences of diarrhea, gastric retention and hypoproteinemia, and shorten the average length of hospital stays in patients with severe cerebral hemorrhage; rate of obstruction of percutaneous endoscopic jejunostomy ducts should be reduced.
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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 explore the predictive value of neuron-specific enolase (NSE) in patients with mild cognitive impairment (MCI) secondary to mild,moderate craniocerebral injury.Methods Seventy-six patients with mild,moderate craniocerebral injury,admitted to our hospital from March 19,2014 to September 1,2015,were chosen in our study;16 of them had secondary MCI during follow up (experimental group) and 60 did not show cognitive dysfunction (control group).Their clinical data between the two groups were compared.The predictive value of different NSE levels for secondary MCI was analyzed.Logistic regression analysis was performed to analyze the risk of secondary MCI in patients with different NSE levels.Multiple linear regression analysis was used to analyze the influence of mini-mental state examination (MMSE) scores in cognitive level of secondary MCI patients.Results (1) There were significant differences in age,salvage time,proportion of hypertension,ratio of skull fracture,injury severity scale (ISS) scores and total cholesterol (TC) between the experimental group and control group (P＜0.05).(2) A lowest quartile group,second quartile group,third quartile group and highest quartile group were divided using NSE levels as independent variables (9.31 ng/mL-12.08 ng/mL,12.09 ng/mL-15.68 ng/mL,15.69 ng/mL-19.65 ng/mL and 19.66 ng/mL-23.47 ng/mL);following the increase of NSE levels,the age,salvage time,proportion of hypertension,and ISS scores were significantly increased (P＜0.05);Single-factor and multivariate Logistic regression analyses showed that the risk of secondary MCI in the highest quartile group was 1.42 and 1.21 folds,respectively,as compared with that in the lowest quartile group.(3) Multiple linear regression analysis showed that baseline NSE level,age,salvage time,and ISS scores were the nfluence factors of MMSE scores in patients with secondary MCI;when the NSE content increased 1 ng/mL,MMSE decreased 0.369 points.Conclusion NSE level in patients with traumatic brain injury is an independent risk factor for secondary MCI,and its level is significantly associated with cognitive impairment.
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Objective To study the Homer1a protein expression and its relationship with neurological deficit and neuronal apoptosis in craniocerebral trauma patients. Methods Forty-two craniocerebral trauma patients, admitted to our hospital from May 2012 to March 2016, were selected as craniocerebral trauma group; 50 healthy subjects accepted physical examination at the same period in our hospital were selected as normal control group (n=50). Immediately after admission, serum contents of Homer1a protein and nerve function damage indices (neurospecific estrogenase [NSE]), fatty acid binding protein [FABP], insulin-like growth factor [IGF-1], and S100B protein) were measured by enzyme linked immunosorbent assay (ELISA). Serum apoptotic indices (soluble apoptotic factor [(sFas)], sFas ligand [sFasL], and cell lymphoma-2 [Bcl-2]) were detected by radioimmunoassay. Results Immediately after admission, serum content of Homer1a protein content in craniocerebral trauma group ([113.27±12.19] pg/mL) was significantly higher than that in normal control group ([53.93±4.06] pg/mL, P<0.05); the median serum Homer1a protein level was 115.302 pg/mL, and according to this level, the patients from the craniocerebral trauma group were further divided into high Homer1a group and low Homer1a group. Serum NSE, FABP, S100B, sFas and sFasL levels in the high Homer1a group, low Homer1a group and normal control group were decreased in sequence, and IGF-1 and Bcl-2 levels increased in sequence, with significant differences (P<0.05). Conclusion Expression of Homer1a protein is increased in patients with traumatic brain injury, and its content is directly related to nerve injury and neuron apoptosis.
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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.