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Monoamine alterations and rotational asymmetry in a rat model of Parkinson's disease following lateral ventricle transplantation of human amniotic epithelial cells

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Neural Regeneration Research 2009年第12期4卷 1007-1012页

作者： Shouru Xue Xinxin Yang Wanli Dong guozhen hui Lihe GUO Department of Neurology First Affiliated Hospital of Soochow University Suzhou 215006 Jiangsu Province China Department of Neurosurgery First Affiliated Hospital of Soochow University Suzhou 215006 Jiangsu Province China Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Shanghai China Shanghai Celstar Institute of Biotechnology Shanghai 201203 China

BACKGROUND： Human amniotic epithelial cells （HAECs） can differentiate into neurons, astrocytes and oligodendrocytes. They biologically secrete many active neurotrophins and have the capacity to metabolize dopamine enzymes. These features underlie a theoretical basis for the treatment of Parkinson＇s disease （PD）. OBJECTIVE： To investigate the survival and differentiation of transplanted HAECs in the lateral ventricle of PD model rats, and to explore its effect on circling behavior, as well as levels of dopamine （DA）, the metabolite homovanillic acid, dihydroxyphenyl acetic acid, 5-hydroxyindoleacetic acid, and 5-hydroxytryptamine in the striatum. DESIGN, TIME AND SETTING： A randomized, controlled, animal study was performed at the Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, and Shanghai Celstar Institute of Biotechnology from May 2007 to December 2008. MATERIALS： HAECs were derived from the placental chorion following caesarean delivery at the Shanghai International Matemal and Child Health Hospital. 6-hydroxydopamine （6-OHDA）, and mouse anti-human Vimentin monoclonal antibody were purchased from Sigma, USA; mouse anti-human nestin and tyrosine hydroxylase （TH） monoclonal antibodies were purchased from Chemicon, USA. METHODS： A total of 114 healthy, adult, Sprague Dawley rats were randomly assigned to two groups： PD model [n = 90, stereotactic microinjection of 2 μL 6-OHDA （3.5 μg/uL） into the striatum] and control （n = 24, no treatment）. The 51 successful PD model rats were randomly divided into 3 subgroups （n = 17）： HAEC, PBS, and model. The HAEC and PBS groups were respectively injected with 10 μL PBS solution containing 1 × 10^5/mL HAECs or 10 pL PBS into the lateral ventricle. The model group was not treated. MAIN OUTCOME MEASURES： TH protein expression in the striatum was evaluated by immunohistochemistry 5 weeks after HAEC transplantation. At 10 weeks, HAEC survival in the lateral ventric

关键词： human amniotic epithelial cells Parkinson's disease model lateral ventricle cell transplantation dopamine

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Upregulation of stromal cell-derived factor-1 alpha/CXCR4 axis-induced migration of human neural progenitors by tumor necrosis factor-alpha and interleukin-8

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Neural Regeneration Research 2009年第11期4卷 832-837页

作者： Jing Qu Hongtao Zhang guozhen hui Xueguang Zhang Huanxiang Zhang Department of Cell Biology Medical College of Soochow University Suzhou 215123 Jiangsu Province China Jiangsu Province Key Laboratory of Stem Cell Research Suzhou 215016 Jiangsu Province China Department of Orthopaedics First Affiliated Hospital of Soochow University Suzhou 215007 Jiangsu Province China Department of Neurosurgery First Affiliated Hospital of Soochow University Suzhou 215007 Jiangsu Province China

BACKGROUND： Studies of several animal models of central nervous system diseases have shown that neural progenitor cells （NPCs） can migrate to injured tissues. Stromal cell-derived factor 1 alpha （SDF-la）, and its primary physiological receptor CXCR4, have been shown to contribute to this process. OBJECTIVE： To investigate migration efficacy of human NPCs toward a SDF-1α gradient, and the regulatory roles of tumor necrosis factor-α （TNF-α） and interleukin-8 （IL-8） in SDF-1α/CXCR4 axis-induced migration of NPCs. DESIGN, TIME AND SETTING： An in vitro, randomized, controlled, cellular and molecular biology study was performed at the Laboratory of Department of Cell Biology, Medical College of Soochow University between October 2005 and November 2007. MATERIALS： SDF-1α and mouse anti-human CXCR4 fusion antibody were purchased from R＆D Systems, USA. TNF-αwas purchased from Biomyx Technology, USA and IL-8 was kindly provided by the Biotechnology Research Institute of Soochow University. METHODS： NPCs isolated from forebrain tissue of 9 to 10-week-old human fetuses were cultured in vitro. The cells were incubated with 0, 20, and 40 ng/mL TNF-α, or 0, 20, and 40 ng/mL IL-8, for 48 hours prior to migration assay. For antibody-blocking experiments, cells were further pretreated with 0, 20, and 40 μg/mL mouse anti-human CXCR4 fusion antibody for 2 hours. Subsequently, the transwell assay and CXCR4 blockade experiments were performed to evaluate migration of human NPCs toward a SDF-1α gradient. Serum-free culture medium without SDF-1α served as the negative control. MAIN OUTCOME MEASURES： The transwell assay was performed to evaluate migration of human NPCs toward a SDF-1α gradient, which was blocked by fusion antibody against CXCR4. In addition, CXCR4 expression in human NPCs stimulated by TNF-α and IL-8 was measured by flow cytometry. RESULTS： Results from the transwell assay demonstrated that SDF-1α was a strong chemoattractant for human NPCs （P 〈 0.01）, and 20 ng/mL produced

关键词： human neural progenitor cells migration stromal cell-derived factor 1 alpha CXCR4 tumor necrosis factor-α interleukin-8

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Therapeutic effects of human amniotic epithelial cell transplantation on double-transgenic mice co-expressing APPswe and PS1ΔE9-deleted genes

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Science China(Life Sciences) 2012年第2期55卷 132-140页

作者： XUE ShouRu CHEN ChongFang DONG WanLi hui guozhen LIU TianJun GUO Lille Department of Neurology First Affiliated Hospital of Soochow UniversitySuzhou 215006China Department of Neurosurgery First Affiliated Hospital of Soochow UniversitySuzhou 215006China Institute of Biochemistry and Cell Biology Shanghai Institute for Biological SciencesChinese Academy of SciencesShanghai 200031China

Human amniotic epithelial cells (HAECs), which exhibit characteristics of embryonic and pluripotent stem cells, could be utilized for cell therapy without legal or ethical problems. Double-transgenic (TG) mice (n=20) and wild-type (WT) mice (n=20) were randomly assigned to two groups, respectively. The transplantation group was treated with HAECs and the control group with PBS. A six-radial arm water maze was used to assess spatial memory. Immunofluorescence was utilized to track HAEC survival. Immunohistochemistry was used to determine octamer-binding protein 4 (oct-4) and nanog expression in the HAECs. High-performance liquid chromatography (HPLC) was used to measure acetylcholine levels in the hippocampus. The density of cholinergic neurons in the basal forebrain and nerve fibers in the hippocampus was measured following acetylcholinesterase staining. Results showed that transplanted HAECs survived for at least eight weeks and migrated to the third ventricle without immune rejection. Graft HAECs also expressed the specific stem cell markers oct-4 and nanog. Compared with the control group, HAEC transplantation significantly ameliorated spatial memory deficits in TG mice, as well as increased acetylcholine levels and the number of hippocampal cholinergic neurites. Intracerebroventricular HAEC transplantation improved spatial memory in double-TG mice, and results suggested that increased acetylcholine levels in the hippocampus, released by surviving cholinergic neurites, were responsible for this improvement.

关键词： Alzheimer's disease human amniotic epithelial cells spatial memory deficit transgenic mice

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Transformation of human amniotic epithelial cells into neuron-like cells in the microenvironment of traumatic brain injury in vivo and in vitro

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Neural Regeneration Research 2011年第10期6卷 744-749页

作者： Yi Lu guozhen hui Zhiyuan Wu Tianjin Liu Xunhe Ji Lihe Guo Department of Neurosurgery First Affiliated Hospital of Soochow University Suzhou 215006 Jiangsu Province China Department of Neurosurgery First Hospital of Jiaxing City Jiaxing 314000 Zhejiang Province China Institute of Biochemistry and Cell Biology Shanghai Institute of Life Sciences Chinese Academy of Sciences Shanghai 200031 China

Survival and differentiation of transplanted cells is closely related to the local *** present study cultured human amniotic epithelial cells（HAECs） in a simulated microenvironment in vitro comprising RPMI 1640 culture medium and the solution extracted from injured brain *** HAECs were round,triangular in form or irregularly shaped,with extended neuron-like processes;some of the processes were interconnected,representing neuron-like morphology and some HAECs were microtubule-associated protein *** survived for at least 4 weeks following transplantation into the center and edges of the trauma focus with traumatic brain injury,and were microtubule-associated protein ***,the motor function of rat hind limbs was significantly improved.

关键词： traumatic brain injury microenvironment transplantation human amniotic epithelialcells neural cell differentiation neural regeneration

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Human amniotic epithelial cells express specific markers of nerve cells and migrate along the nerve fibers in the corpus callosum

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Neural Regeneration Research 2012年第1期7卷 41-45页

作者： Zhiyuan Wu guozhen hui Yi Lu Tianjin Liu Qin Huang Lihe Guo Department of Neurosurgery Changzhou Second People＇s Hospital Nanjing Medical University Nanjing 213000 Jiangsu Province China Department of Neurosurgery the First Affiliated Hospital of Soochow University Suzhou 215006 Jiangsu Province China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes （microtubule-associated protein-2, glial fibrillary acidic protein and nestin） in human amniotic epithelial cells. The conditioned medium of human amniotic epithelial cells promoted the growth and proliferation of rat glial cells cultured in vitro, and this effect was dose-dependent. Human amniotic epithelial cells were further transplanted into the corpus striatum of healthy adult rats and the grafted cells could integrate with the host and migrate 1 2 mm along the nerve fibers in corpus callosum. Our experimental findings indicate that human amniotic epithelial cells may be a new kind of seed cells for use in neurograft.

关键词： human amniotic epithelial cells cell transplantation microtubule-associated protein-2 glial fibrillary acidic protein nestin neurotrophin central nervous system injury neural regeneration

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Cerebrospinal fluid used as culture medium prior to autologous olfactory ensheathing cell transplantation

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Neural Regeneration Research 2010年第1期5卷 21-27页

作者： Weijiang Wu Qing Lan Hua Lu Aihua Zhu Yunzhao Jiang Ge Chen guozhen hui Department of Neurosurgery Wuxi Third People's Hospital Wuxi 214041 Jiangsu Province China Second Affiliated HospitaI Soochow University Suzhou 215004 Jiangsu Province China First Affiliated Hospital Soochow University Suzhou 215006 Jiangsu Province China

BACKGROUND： Cerebrospinal fluid can be an inducer for neural stem cells in vitro, but few studies employ cerebrospinal fluid to culture olfactory ensheathing cells. OBJECTIVE： To investigate the growth of nasal mucosa olfactory ensheathing cells in normal cerebrospinal fluid, and to analyze the feasibility of cerebrospinal fluid for culturing olfactory ensheathing cells used for transplantation. DESIGN, TIME AND SETTING： A completely randomized, block design study was performed at the Cell Laboratory, Wuxi Third People＇s Hospital, and Jiangsu Institute of Parasitic Diseases, China, in August 2008. MATERIALS： Dulbecco＇s modified Eagle＇s medium/F12 （DMEM/F12） and fetal bovine serum （Gibco BRL, USA）, mouse anti-rat P75 monoclonal antibody and rabbit anti-glial fibrillary acidic protein polyclonal anti body （＇Santa Cruz Biotechnology, USA）, mouse anti-rat myelin basic protein monoclonal antibody （Cymbus, UK）, mouse anti-rat microtubule-associated protein-2 monoclonal antibody （Transduction Laboratories, USA）, FITC conjugated rabbit anti-mouse monoclonal antibody （Boster, China）, TRITC conjugated goat anti-rabbit monoclonal antibody （Sigma, USA） were used. METHODS： Nasal mucosa olfactory ensheathing cells were separately incubated in DMEM/F12, cerebrospinal fluid, and changing DMEM/F12 into cerebrospinal fluid. Adult female Sprague Dawley rat models of spinal hemisection were established. Nerve injury was repaired by transplantation of nasal mucosa olfactory ensheathing cells cultured in cerebrospinal fluid or DMEM/F12. MAIN OUTCOME MEASURES： The proliferative ability of olfactory ensheathing cells cultured in cerebrospinal fluid was determined by a 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide assay. The morphology and purity of olfactory ensheathing cells were detected using immunohistochemistry. Animal behavior was evaluated by the Basso, Beattie and Bresnahan locomotor rating scale. Morphological repair was assessed by a horseradish peroxidase-tetra

关键词： cerebrospinal fluid olfactory ensheathing cells in vitro culture cell transplantation

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Brain-derived neurotrophic factor induces neuron-like cellular differentiation of mesenchymal stem cells derived from human umbilical cord blood cells in vitro

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Neural Regeneration Research 2011年第13期6卷 972-977页

作者： Lei Chen Zhongguo Zhang Bing Chen Xiaozhi Liu Zhenlin Liu Hongliang Liu Gang Li Zhiguo Su Junfei Wang guozhen hui Department of Neurosurgery Tianjin Fifth Central Hospital Tianjin 300450 China Department of Colorectal Cancer Surgery Liaoning Provincial CancerHospitail Shenyang 110042 Liaoning Province China Department of Gynaecology and Obstetrics Shanghai Pudong New District People's Hospitall Shanghai 201200 China Department of Neurosurgery First Affiliated Hospital of Soochow University Suzhou 215006 Jiangsu Province 'China

Human umbilical cord blood was collected from full-term deliveries scheduled for cesarean section. Mononuclear cells were isolated, amplified and induced as mesenchymal stem cells. Isolated mesenchymal stem cells tested positive for the marker CD29, CD44 and CD105 and negative for typical hematopoietic and endothelial markers. Following treatment with neural induction medium containing brain-derived neurotrophic factor for 7 days, the adherent cells exhibited neuron-like cellular morphology. Immunohistochemical staining and reverse transcription-PCR revealed that the induced mesenchymal stem cells expressed the markers for neuron-specific enolase and neurofilament. The results demonstrated that human umbilical cord blood-derived mesenchymal stem cells can differentiate into neuron-like cells induced by brain-derived neurotrophic factor in vitro.

关键词： human umbilical cord blood purification and culture brain-derived neurotrophic factor neuron-like cells neural regeneration

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早期胃肠外营养对重症脑外伤预后及氮代谢的影响

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中华神经外科杂志 1995年第6期11卷 344-347页

作者：王之敏惠国桢苏州医学院附一院神经外科

４０例严重脑外伤（ＧＣＳ４～７分）随机分成两组：早期胃肠外营养组（ＰＮ组）和传统延迟性胃肠内营养组（ＥＮ组），比较两组蛋白质、氨基酸代谢和免疫功能的改变，并进行临床预后分析。结果ＰＮ组获得较高的氮平衡，其血清氨基酸、... 详细信息

４０例严重脑外伤（ＧＣＳ４～７分）随机分成两组：早期胃肠外营养组（ＰＮ组）和传统延迟性胃肠内营养组（ＥＮ组），比较两组蛋白质、氨基酸代谢和免疫功能的改变，并进行临床预后分析。结果ＰＮ组获得较高的氮平衡，其血清氨基酸、转铁蛋白、前清蛋白和纤维连接蛋白及ＣＤ４细胞均高于ＥＮ组，ＰＮ组的死亡率明显低于ＥＮ组，且其伤后三个月时的生存质量优于ＥＮ组。

关键词：脑外伤胃肠外营养预后氮代谢

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小脑动脉的临床解剖探讨

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中华神经外科杂志 1997年第6期13卷 338-341页

作者：惠国桢夏春林苏州医学院附一院脑神经外科

目的：为神经外科临床提供解剖学资料。方法：手术显微镜下观察５０例成人脑标本小脑动脉的起始、行径、主要分支、穿动脉及大致分布，检查各小脑动脉与出入脑干的颅神经的接触关系。结果：５０例人脑有小脑下后动脉（ＰＩＣＡ）９４支... 详细信息

目的：为神经外科临床提供解剖学资料。方法：手术显微镜下观察５０例成人脑标本小脑动脉的起始、行径、主要分支、穿动脉及大致分布，检查各小脑动脉与出入脑干的颅神经的接触关系。结果：５０例人脑有小脑下后动脉（ＰＩＣＡ）９４支，小脑下前动脉（ＡＩＣＡ）９７支和小脑上动脉（ＳＣＡ）１１２支。２侧ＰＩＣＡ和１２侧ＳＣＡ接触三叉神经根，２侧ＡＩＣＡ接触面神经根，动脉与神经根接触多形成压迹。结论：小脑动脉的局部解剖有助于神经外科医生在颅后窝手术时，对这些动脉尤其是行程可能有变异的动脉及穿支要特别谨慎，避免损伤，并保护与小脑动脉关系密切的脑神经根；对某些脑干血管综合征患者及某些三叉神经痛、面肌痉挛患者采取相应的治疗。

关键词：小脑动脉颅后窝手术解剖神经外科

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Treatment of pituitary prolactinomas with bromocriptine

Treatment of pituitary prolactinomas with bromocriptine

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中国抗癌协会神经肿瘤专业委员会第八届学术会议

作者： Xiangdong Li, guozhen hui Department of Neurosurgery, the First Hospital Affiliated to Suzhou University, Su Zhou 215006, China

Objective: Clinical effects and mechanisms of bromocriptine treatment on pituitary prolactinomas were explored in this clinical and pathological study. Method: Twenty-six patients with prolactinomas were selected. Bromocriptine was administrated in 16 patients before operation, while 7 patients were not treated with bromocriptine before operation. Tumor specimens were resected during operation and examined under a light microscope and an electron microscope. Three cases of giant adenoma were only treated with bromocriptine, and changes in tumor volume were observed. Results: In 16 cases with bromocriptine treatment, the cell size decreased while karyoplasmic ratio increased. Observation under an electron microscope revealed that secretory granules, mitochondria, and rough endoplasmic reticulum were markedly reduced. When treated with bromocriptine for a long period, margination of the nuclear chromatin occurred. Some chromatin detached from the nuclear membrane and spread into the cytoplasm in a budding manner forming an apoptotic body. Some tumor cells exhibited vacuolar degeneration, necrosis, or autolysis. In patients who received only bromocriptine treatment, the tumor volume decreased by 98.52%. Conclusion: Bromocriptine could cure prolactinomas. The main underlying mechanism was that bromocriptine could induce apoptosis and secondary necrosis of tumor cells.

关键词： bromocriptine pituitary prolactinomas apoptosis

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