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14-3-3 gamma and zeta protein expression in active microglia Immune response mechanisms of Parkinson’s disease

Neural Regeneration Research 2008年第3期3卷 233-236页

作者： Jing He shenggang sun Xiaowu Chen Department of Neurology the Third Hospital of Wuhan Wuhan 430060 Hubei Province China Department of Neurolog-y the Affiliated Union Hospital of Tongji Medical College Huazhong University of Science and Technology Wuhan410128 Hubei Province China

BACKGROUND： The progressive degeneration of dopaminergic neurons in Parkinson＇s disease is associated with an activated glial reaction, combined with an inflammatory process. These responses lead to the production of cytokines, such as interferon- γ, tumor necrosis factor- α （TNF- α ）, and interleukin-1 β. In addition, 14-3-3 protein is a component of Lewy bodies in Parkinson＇s disease. OBJECTIVE： To observe the expression of 14-3-3 γ and ζ protein, as well as TNF-α, in mouse microglia, as well as changes after lipopolysaccharide （LPS） activation. To investigate possible mechanisms of dopaminergic neuronal injury due to activated microglia. To and clarify the immune response mechanisms of Parkinson＇s disease. DESIGN： Randomized controlled observation, cell study.SETTING： Laboratory of Department of Neurology, the Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： The BV-2 immortalized murine microglia cell line was purchased from China Unit cell center. LPS was provided by Sigma Company. Cell cultures were purchased from Gibco. Phospho-（Ser） 14-3-3 binding motif antibody was purchased from Santa Cruz Biotechnologies. FITC was provided by Linfei Biotechnology, Wuhan, China. TNF- α ELISA was provided by Jingmei Biotech Co, Wuhan, China. The flow cytometer was provided by Becton Dickinson, Canada. METHODS： The present experiment was performed at the Laboratory of Department of Neurology, the Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from April to December 2006. The microglial cell line, BV-2, was cultured in vitro and stimulated with LPS for 2, 6, 12, and 24 hours. BV-2 cultures without LPS were used as controls. MAIN OUTCOME MEASURES： Expression of 14-3-3 γ protein was detected by flow cytometry. 14-3-3 ζ percentage expression and the mean fluorescence intensity was detected by immunofluorescence. TNF- α expression was detected by ELISA. RES

关键词： 14-3-3 protein microglia lipopolysaccharide TNF-α

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Proliferation changes in hippocampal neurons following pilocarpine-induced intractable epilepsy in adult rats

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Neural Regeneration Research 2010年第24期5卷 1858-1861页

作者： Lancai Xiong Xuejun Deng Fang Wang Chunling Du shenggang sun Department of Neurology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Hubei Province China

Epilepsy can lead to the changes in neurons residing in the dentate gyrus. The present study aimed to observe the cell dividing features following epilepsy in adult rat hippocampi, and to study difference in cell proliferation between adult rats with common epilepsy and intractable ***, male, Sprague Dawley rats ware randomly divided into control （n = 8, treatment with normal saline） and three expenmental groups： common epilepsy （n = 33）, intractable epilepsy （n = 11）, and drug-responsive （n = 25）. Pilocarpine （15 mg/kg） was intrapentoneally administered to establish epilepsy in the three experimental groups. Rats that developed epilepsy were treated with chloral hydrate. Rats that did not exhibit spontaneous seizures were enrolled in the common epilepsy group, and rats with spontaneous seizure were included in the spontaneous seizure group. At 6 hours after epileptic attack termination, rats ware intraperitoneally injected with bromodeoxyuridine （BrdU; 50 mg/kg）, an optimal marker forlabeling cell proliferation in vivo, four *** results at 48 hours after BrdU injection indicated that the number of BrdU-positive cells was the highest in the common epilepsy group, followed by the control group,and lastly the intractable group （P 〈 0.01）. In addition, the number of BrdU-positive cells in the common epilepsy group was similar to the drug-responsive group. The present findings demonstrated that intractable epilepsy led to decreased hippocampal neurons in adult rats when compared to common epilepsy.

关键词： intractableepiiepsy hippocampal neurons cell proliferation pilocarpine

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Influence of endogenous ciliary neurotrophic factor on neural differentiation of adult rat hippocampal progenitors

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Neural Regeneration Research 2013年第4期8卷 301-312页

作者： Jun Ding Zhili He Juan Ruan Ying Liu Chengxin Gong shenggang sun Honghui Chen Department of Neurology Union HospitalTongji Medical CollegeHuazhong University of Science and Technology Mental Health Center Tongji Medical CollegeHuazhong University of Science and Technology New York State Institute for Basic Research in Developmental Disabilities Staten IslandNYUSA

Ciliary neurotrophic factor is the only known neurotrophic factor that can promote differentiation of hippocampal neural progenitor cells to glial cells and neurons in adult rats. This process is similar to spontaneous differentiation. Therefore, ciliary neurotrophic factor may be involved in spontaneous differentiation of neural stem cells. To verify this hypothesis, the present study isolated neural progenitor cells from adult male rats and cultured them in vitro. Results showed that when neural progenitor cells were cultured in the absence of mitogen fibroblast growth factor-2 or epidermal growth factor, they underwent spontaneous differentiation into neurons and glial cells. Western blot and immunocytochemical staining showed that exogenous ciliary neurotrophic factor strongly induced adult hippocampal progenitor cells to differentiate into neurons and glial cells. Moreover, passage 4 adult hippocampal progenitor cells expressed high levels of endogenous ciliary neurotrophic factor, and a neutralizing antibody against ciliary neurotrophic factor prevented the spontaneous neuronal and glial differentiation of adult hippocampal progenitor cells. These results suggest that the spontaneous differentiation of adult hippocampal progenitor cells is mediated partially by endogenous ciliary neurotrophic factor.

关键词： neural regeneration stem cells spontaneous differentiation neural progenitor cells endogenous neurotrophic factors ciliary neurotrophic factor regeneration grants-supported paper photographs-containing paper neuroregeneration

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Changes of dynamical balance of free radicals induced by levodopa in rat glia-containing mesencephalic culture fluid

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Neural Regeneration Research 2006年第5期1卷 401-404页

作者： Wenxia Li shenggang sun Jingru Wang Baoliang sun Yanbo Zhang Deparlment of Neurology Affiliated Hospital of Taishan Medical University Taian 271000Shandong Province China Department of Neurology Union Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430022 Hubei Province. China

BACKGROUND ： Parkinson disease is neurodegenerative disorders, characterized by a progressive and selective degeneration of nigrostdatal dopaminergic pathway. Its main clinical symptoms include bradykinesia, dgidity, rest tremor and disturbances in balance. Levodopa （L-DOPA） is the ＂gold standard＂ for the symptomatic treatment of Parkinson disease, but L-DOPA is toxic to dopaminergic neurons and the chronic administration of L-DOPA often causes the side effects of motor such as ＂on-off ＂, etc., and its mechanism still has not been completely clarified. OBJECTIVE ： To observe the changes of the content of glutathione （GSH）, activities of glutathione peroxidase （GSH-Px） and superoxide dismutase （SOD） and content of malondialdehyde （MDA） in gila-containing mesencephalic culture fluid after L-DOPA of different concentrations were added. DESIGN： A comparative observation. SETTINGS ： Department of Neurology, Affiliated Hospital of Taishan Medical University; Department of Neurology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS ： The experiment was carried out in the Basic Research Institute, Taishan Medical University from December 2003 to September 2004. Ten postnatal newborn SD rats （within 2 days） were selected, either male or female. Fetal bovine serum （DMEM/F12） was from Gibco Company; L-DOPA and glial fibrillary acidic protein （GFAP） from Sigma Company. Kits for determination of GSH, GSH-Px, SOD and MDA were purchased from Nanjing Jiancheng Bioengineering Research Institute. METHODS ： Gila-containing mesencephalic culture fluid were placed in 24-well culture plate, and L-DOPA of 50, 100 and 500 μmot/L was added to each group, the blank control group was also set. The glia-containing mesencephalic culture fluid selected at 4, 24, 48 and 72 hours respectively to determine the GSH content with colorimetdc quantitative technique, GSH-Px activity with colorimetdc method, SOD activity with xanthine oxidase meth

关键词： free radicals Parkinson disease neurodegenerative disorders Science and Technology glutathione peroxidase superoxide dismutase dopaminergic neurons colorimetric method dynamical balance xanthine oxidase determination of Basic Research acidic protein side effects time point

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Increased Neuronal Hypoxic Tolerance Induced by Repetitive Chemical Hypoxia

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2002年第2期22卷 132-134页

作者： LI Hongge(李红戈) LIU Chngqin(刘昌勤) sun shenggang(孙圣刚) Department of Neurology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Department of Neurology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Department of Neurology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022

Summary: To investigate the effects of time interval and cumulative dosage of repetitive mild cellular hypoxia on shape of neurodegeneration and neuroprotection in mice, population spike amplitude (PSA) was measured during hypoxia and posthypoxic recovery in hippocampal slices from untreated control and mice pretreated in vivo with a single or repeatedly intraperitoneal injection of 3-nitropropionate (3-NP). Posthypoxic recovery of PSA was dose-dependent in single pretreated slices, with maximal recovery on pretreatment attained with 20 mg/kg 3-NP (82±32%, P< 0.01). Upon 5 and 9 treatments with 20 mg/kg 3-NP (dosage interval 3 days), PSA recovered to (38±9) % with the difference being not significant vs control group and (72±45) % with the difference being significant (P< 0.05 to control, P<0.05 to 5 treatments), respectively. In contrast, with 2 days time interval, recovery after 5 and 9 treatments was (30±25) % and (16±14) %, respectively (without significant difference from control). Continued neuroprotection was also observed upon increase of dosage interval to 4 and 5 days. It was suggested that repetitive chemical hypoxia is a model for neurodegenerative disease and continued neuroprotection depending on time interval between repetitive hypoxic episodes rather than cumulative dosage. At appropriate time intervals increased neuronal hypoxic tolerance could be induced with number of hypoxic episodes.

关键词： hypoxia electrophysiology Huntington's disease 3-nitropropionate

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Experimental Study on Heterograft of Glomus Cells of Carotid Body for Hemiparkinsonian Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2002年第2期22卷 129-131页

作者： CAO Xuebing(曹学兵) sun shenggang(孙圣刚) TONG E＇tang(童萼塘) Department of Neurology Xiehe Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430022 Department of Neurology Xiehe Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430022 Department of Neurology Xiehe Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430022

Summary: To observe the effects of heterograft of glomus cells of carotid body on hemiparkinsonian rat models, rats with unilateral 6-hydroxydopamine (6-OHDA)-induced lesions of the right dopaminergic neurons of substantia nigra received intrastriatal glomus cells heterograft. Apomorphine-induced rotation was monitored for 30 min at various time points after grafting. The striata were cut and examined for dopamine content by HPLC and for immunohistochemical staining of tyrosine hydroxylase positive neurons (TH +) at the end of the experiments. The results showed that apomorphine-induced rotational behavior was significantly reduced for 12 weeks and the dopamine contents were significantly elevated after grafting (P<0.01), and TH + cells survived better. The present study demonstrates that intrastriatal heterograft of glomus cells within carotid body in rats with 6-OHDA-elicited lesions could reduce apomorphine-induced rotational behavior and elevate the dopamine contents and numbers of TH + cell surviving within striatum, and can serve as a new and effective alternative for Parkinson disease.

关键词： carotid body glomus cell heterograft Parkinson disease

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一种基于PIO改进强化学习的航天器复杂多约束姿态机动规划新方法

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中国科学：技术科学 2023年第2期53卷 200-209页

作者：华冰孙胜刚吴云华陈志明南京航空航天大学航天学院南京210016

本文针对多个姿态约束条件下的航天器姿态机动规划问题进行了研究,提出了一种基于鸽群算法的改进的策略梯度强化学习算法(PIOPGRL).首先,针对强制指向约束和禁止指向约束,建立了基于角度的姿态约束模型,根据约束模型建立了强化学习的回... 详细信息

本文针对多个姿态约束条件下的航天器姿态机动规划问题进行了研究,提出了一种基于鸽群算法的改进的策略梯度强化学习算法(PIOPGRL).首先,针对强制指向约束和禁止指向约束,建立了基于角度的姿态约束模型,根据约束模型建立了强化学习的回报函数.然后,使用适应度函数替代策略评价函数,将鸽群算法与强化学习相融合.针对策略梯度强化学习算法计算量大、收敛速度慢的问题,使用鸽群算法求解策略梯度,极大减少了计算量.仿真结果表明,相比于策略梯度强化学习算法,基于自PIO改进强化学习的航天器姿态机动规划算法(PIOPGRL)在极大减少计算量的同时,有更优的规划结果,更小的机动代价,适用于微小航天器解决多个姿态约束条件下的姿态机动规划问题.

关键词：姿态机动姿态约束路径规划强化学习鸽群算法航天器

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Minocycline protects the apoptosis of PC12 cells induced by 1-methyl-4-phenylpyridinium

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Journal of Nanjing Medical University 2005年第5期19卷 247-250页

作者： Wei SHEN shenggang sun Xuebing CAO Department of Neurology the affiliated Union Hospital of Tongji Medical Collage Huazhong University of Science and Technology Wuhan 430022. China

Objective： To explore the protective effect of minocycline on the apoptosis of cellular parkinsonism models induced by MPP^＋ . Methods： Using PC12 cells as the apoptotic model of dopaminergic neurons, MC and MPP^＋ were added into the culture medium of PC12 cells, and using MTr to assay the cell viability and metabolic state; The cells apoptosis was assayed by electrophoresis method and using flow cytometry FACS to assay the apoptosis ratio. Results： Added the MPP^＋ to get the concentration of 10μmol/L, the cellular parkinsonism model of apoptosis had been prepared. The pre-treatment of MC （ 100/μmol/L） could significantly increase the PC12 cell viability. The apoptosis ratio of MC＋MPP^＋ group was significantly lower than that of MPP^＋ group, but was still significantly higher than that of control group. Conclusion： MC may protect the cell apoptosis induced by MPP^＋ to some extent.

关键词： minocycline PC12 cell apoptosis parkinson disease 1-methyl-4-mhenylpyridinium

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Study on the effects of thrombin on AQP4 mRNA and AQP4 protein expression in rat primary astrocytes

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Journal of Nanjing Medical University 2006年第4期20卷 220-224页

作者： Jinghua Zhou Xuebing Cao shenggang sun Department of Neurology Union Hospital Tongii Medical College Huazhong University of Science and Technology Wuhan 430022 China

Objective： To study the biologic effects of various concentrations of thrombin on aquaporin 4 （AQP4） expression in rat primary cultured astrocytes, and to explore the regulation mechanism of transmembrane water transportation in astrocytes after intracerebral hemorrhage （ICH）. Methods： Primary cultured astrocytes were incubated in culture mediums containing various concentrations of thrombin for 24 h and harvested. AQP4 mRNA and AQP4 protein expression were determined by reverse transcription polymerase chain reaction （RT-PCR） and immunohistochemical technique. Cell apoptosis was detected by TdT- mediated dUTP nick end labeling （TUNEL） technique. Cell morphology was observed by phase contrast microscope, and cell viability was assayed by MTT. Results： AQP4 mRNA and AQP4 protein showed a low expression in normal astrocytes. The expression of AQP4 mRNA and AQP4 protein significantly increased in the astrocytes treated with 100 U/ml or 200 U/ml thrombin （P 〈 0.01 ）,and these astrocytes swelled. The number of TUNEL positive cells significantly increased. On the other hand, AQP4 mRNA and AQP4 protein expression were downegulated in the astrocytes treated with 0.5 U/ml or 1 U/ml thrombin （P 〈 0.05）, and the cell morphology did not change. Few TUNEL positive cells were observed. Conclusion： AQP4 overxpression induced by high concentrations of thrombin causes an increased permeability of water in astrocytic membrane. On the contrary, the decreased AQP4 expression prevents the astrocytes from swelling and apoptosis.

关键词： thrombin astrocytes aquaporin-4 apoptosis

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Study of apoptosis pattern of dopaminergic neurons and neuroprotective effect of nicotine in MPTP mouse model

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Journal of Nanjing Medical University 2007年第4期21卷 248-252页

作者： Dan Hua Wei Cao shenggang sun Department of Neurology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Hukei Province China Department of Nuclear Medicine Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Hukei Province China

Objective：To investigate the apoptosis of dopaminergic neurons and the protective effect of nicotine in 1-methyl-4-phenyl- 1, 2,3,6-tetrahydropyridine （MPTP）-induced mouse model of Parkinson＇s disease. Methods：The mouse model of Parkinson＇s disease were formed by MPTP （30 mg/kg/d×7, i.p. ）; and the loss and apoptosis of dopaminergic neurons was observed by Tyrosine Hydroxylase（TH） and TUNEL *** ＂Nicotime plus MPTP＂ group, mice were pretreated with nicotine before MPTP injection. The putative protective effect of nicotine was analyzed. Results：The number of TH-positive cells decreased during MPTP treatment. Apoptotic neurons began to appear after three injections of MPTP and peaked on the 8th day. In the MPTP-intoxicated mice treated with nicotine, the loss of TH-positive cells was significantly less than that of MPTP- treated group （30 mg/kg/d×7）（P 〈 0.05）. Conclusion：The chronic treatment of MPTP can induce the apoptosis of dopaminergic neurons in substantia nigra, and nicotine might have a neuroprotecitve effect on dopaminergic neurons against MPTP toxicity.

关键词： Parkinson's Disease apoptosis 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyridine Nicotine

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