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Objective:To investigate the clinical manifestations and pathogenic gene mutation sites of familial cavernous hemangioma by a pedigree study of this disease.Methods:A family of cerebral cavernous hemangioma who was admitted to the Department of Neurology of the First Affiliated Hospital of Henan University of Science and Technology in April 2019 was diagnosed as cerebral cavernous hemangioma type 1 based on clinical manifestations and head magnetic resonance imaging (MRI), diffusion weighted imaging and susceptibility weighted imaging screening. According to Zabramski classification criteria, the family′s clinical data were collected and genes were sequenced.Results:A 58-year-old female proband had dizziness and headache as the main symptoms, her daughter and son had no clinical symptoms, and her granddaughter had clinical manifestations of cerebral hemorrhage and seizures. The proband and her family members showed multiple cavernous hemangioma on cranial MRI,and the p.L436fs mutation in the KRIT1 gene of familial cerebral cavernous malformation type 1 was confirmed through genetic examination, which was consistent with the Zabramski typing results based on head MRI. The mutation site of the familial spongiform malformation type 1 pathogenic gene was found to be p.L436fs in KRIT1 gene, which has not been reported in familial cerebral cavernous hemangioma type 1 until now.Conclusion:A new p.L436fs mutation of KRIT1 gene was found in familial cerebral cavernous malformation type 1, which expands understanding of the clinical manifestations and pathogenic gene mutation sites of familial cavernous hemangioma.
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Objective:To evaluate the role of silent information regulator 1 (SIRT1)/nuclear factor E2 related factor 2 (Nrf2) signaling pathway in sleep deprivation-induced cognitive dysfunction in rats.Methods:Thirty-six adult male Sprague-Dawley rats, aged 54-56 weeks, weighing 600-750 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), sleep deprivation group (group SD) and sleep deprivation+ SIRT1 agonist Srt1720 group (group SD+ Srt). Sleep deprivation model was established by modified multi-platform water environment method.Srt1720 10 mg/kg was injected intraperitoneally every 12 h starting from 24 h before establishing the model in group SD+ Srt, while the equal volume of 0.9% normal saline was injected intraperitoneally in C and SD groups.After the end of sleep deprivation, Morris water maze test was performed to evaluate the cognitive function.Animals were then sacrificed, and their hippocampi were removed for determination of neuronal degeneration rate in hippocampal CA1 region (using HE staining), the apoptosis rate in hippocampal CA1 (using TUNEL assay ), the expression of SIRT1 and Nrf2 in hippocampal CA1 (by immunohistochemistry) and the contents of reactive oxygen species (ROS) and superoxide dismutase (SOD) (by microplate method). Results:Compared with group C, the escape latency was significantly prolonged, the time of staying at the platform quadrant was shortened, and the frequency of crossing the platform was decreased on 2-4 days, the apoptotic rate and neuronal degeneration rate were increased, the expression of SIRT1 and Nrf2 was down-regulated, ROC content was increased, and SOD content was decreased in SD and SD+ Srt groups ( P<0.05). Compared with group SD, the escape latency was significantly shortened, the time of staying at the platform quadrant was prolonged, and the frequency of crossing the platform was increased on 3 and 4 days, the apoptotic rate and neuronal degeneration rate were decreased, the expression of SIRT1 and Nrf2 was up-regulated, ROC content was decreased, and SOD content was increased in group SD+ Srt ( P<0.05). Conclusions:Sleep deprivation can induce oxidative stress response in hippocampus by inhibiting the activation of SIRT1/Nrf2 signaling pathway, leading to cognitive dysfunction in rats.
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Objective:To investigate whether idebenone can improve behavioral disorders in mice with Parkinson disease (PD) by increasing PHB2 mediated mitophagy.Methods:In the first small experiment, thirty mice were randomly divided into normal control group, model group and treatment group according to the random number table method, with 10 animals in each group.The aim of this study was to observe the effect of idebenone on the behavior of Parkinson disease model mice. In the second experiment, 20 mice were randomly divided into blank control group, MPTP group, shRNA-PHB2 group and shRNA-PHB2+ MPTP group, with 5 mice in each group. The changes of tyrosine dehydrogenase (TH) in brain tissue were detected by immunofluorescence assay. In the third experiment, 30 mice were randomly divided into blank control group, shRNA-PHB2 group, MPTP+ idebenone group, shRNA-PHB2+ MPTP group, shRNA-PHB2+ idebenone group and shRNA-PHB2+ MPTP+ idebenone group, with 5 animals in each group. The aim of this study was to investigate the effect of idebenone on mitochondrial autophagy in mouse brain.C57BL-6 mice were intraperitoneally injected with MPTP to establish the animal model of chronic PD. Then 200 mg / kg idebenone was given by gavage for 21 days. And the expression of PHB2 in brain was inhibited by microinjection of adeno-associated virus 9 (AAV9) shRNA inhibin 2(PHB2) into lateral ventricle. The behavioral changes of the PD mice were detected by Morris water maze, and the changes of tyrosine dehydrogenase (TH) induced by inhibiting PHB2 were detected by immunohistochemistry. The protein expression of LC3 and PHB2 in substantia nigra of midbrain was detected by Western blot.The data were analyzed by GraphPad 7.0 and SPSS 22.0.Results:(1) In the water maze test data of the first small experiment, the repeated measurement ANOVA showed that the group-time interaction effects of latency of mice from 1 to 7 days were significant ( Ftime×group=20.51, P<0.05). Simple effect analysis showed that on the 5th, 6th and 7th day, the incubation period of the treatment group was significantly shortened (all P<0.05). Univariate analysis of variance showed that on the 7th day of the test, the differences between the control group and the model group, the model group and the treatment group, the control group and the treatment group were all statistically significant( t=-49.95, -21.81, 28.14; all P<0.01). In the third small experiment, repeated measurement analysis of variance showed that the interaction between time and group was significant ( Ftime×group=42.11, P<0.01). Simple effect analysis showed that compared with MPTP+ idebenone group, the latency of shRNA-PHB2+ MPTP+ idebenone group was significantly prolonged (all P<0.05). There were no significant difference between shRNA-PHB2+ MPTP+ idebenone group and shRNA-PHB2+ MPTP group except the 4th day ( P<0.05). On the 7th day, compared with MPTP+ idebenone group, the residence time of shRNA-PHB2+ MPTP+ idebenone group was significantly increased ( t=-34.36, P<0.001), but there was no significant difference between shRNA-PHB2+ MPTP group and shRNA-PHB2+ MPTP+ idebenone group ( t=2.94, P>0.05). (2)The results of immunofluorescence experiment showed that the relative expression of TH in the control group, model group, shRNA-PHB2 group and shRNA-PHB2+ MPTP group were (41.03±3.01), (24.20±4.18), (38.39±3.31) and (13.12±2.65), respectively. Compared with the control group, the expression of TH in the midbrain of the MPTP group was significantly down-regulated, the difference was statistically significant( t=7.98, P<0.01). Compared with the MPTP group, the expression of TH in shRNA-PHB2 group was down regulated ( t=-6.73, P<0.05). (3) Western blot results showed that the relative expression of LC3 in midbrain tissue of control group, shRNA-PHB2 group, MPTP+ idebenone group, shRNA-PHB2+ MPTP group, shRNA-PHB2+ idebenone group and shRNA-PHB2+ MPTP+ idebenone group were (0.86±0.07), (0.77±0.08), (0.42±0.05), (0.21±0.05), (0.66±0.09) and (0.27±0.07). The relative expression of PHB2 were (1.13±0.14), (0.56±0.11), (1.08±0.14), (0.27±0.07), (0.68±0.14) and (0.24±0.10). Compared with MPTP+ idebenone group, the relative expression of LC3 and PHB2 in shRNA-PHB2+ MPTP+ idebenone group was significantly decreased ( F=1.96, P<0.01). Conclusion:Idebenone can increase the level of mitophagy in PD mice through PHB2, thus improving the behavioral disorder.
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Objective:To observe the effect of ginsenoside Rd pretreatment on the expressions of N-methyl-D-aspartate(NMDA)receptor subunit NR2 B protein and endonuclease G(EndoG)in basal ganglia region after cerebral focal ischemia-reperfusion in rats and to investigate possible mechanism of ginsenoside Rd in the treatment of ischemic stroke.Methods:A rat model of middle cerebral artery occlusion(MCAO)was induced by intraluminal filament method.The expressions of NR2B and EndoG in basal ganglia region for focal cerebral iSChemia 1 hour,and 1,6,24 and 72 hours reperfusion were detected by immunohistochemical staining and image analysis method.The effects of ginsenoside Rd on the expressions of FaxioG and NR2B and the volume of cerebral infarction were evaluated.Results:The positive expression of NR2B in basal ganglia region on the ischemic side in ischemia-reperfusion group was increased significantly.The expression of EndoG in the nucleus was notable;the positive expressions of NR2B and EndoG at different reperfusion time points in ginsenoside Rd pretreatment group were decreased significantly(P<0.05 or P<0.01),and the volume of cerebral infarction was reduced significantly(P<0.01).Conclusions:The expressions of NMDA receptor subunit NR2B protein and apoptosis-inducing factor EndoG were increased significantly after cerebral focal ischemia reperfusion;ginsenoside Rd pretreatment may significantly reduce the expressions of NR2B and EndoG.It reduces the volume of cerebral infarction by inhibiting excitatory neurotoxicity and blocking neuronal apoptosis,and thus plays a role in neuroprotection.