四连接素在帕金森病脑脊液中的表达变化

目的探讨脑脊液中可能的帕金森病（PD）生物标记物。方法采用荧光差异凝胶电泳（DIGE）技术分离并筛选PD患者和正常对照者脑脊液中差异表达蛋白质，用基质辅助激光解吸电离飞行时间质谱（MALDl-TOFMS）技术进行鉴定并分析，应用Westernblotting进行验证。结果DIGE发现20个明显的差异蛋白点，共鉴定出8个蛋白质。DIGE分析和Westernblotting验证均显示蛋白四连接素在PD患者脑脊液中发生显著下调。结论四连接素可能参与了PD发生，有可能成为PD的生物标记物。     

儿童结核性脑膜炎脑脊液蛋白质组学初步研究

目的 结核性脑膜炎是致残、致死率较高的儿童时期常见中枢神经系统感染性疾病。本实验拟通过建立正常中国儿童及结核性脑膜炎患儿脑脊液蛋白质双向凝胶电泳图谱,筛选结核性脑膜炎特异性蛋白,为结核性脑膜炎的早期诊断提供线索,同时也为进一步探讨结核性脑膜炎的发病机制提供新思路。方法 收集结核性脑膜炎患儿和同年龄正常儿童脑脊液各4 ml,固相PH梯度二维聚丙烯酰胺凝胶电泳（2-DE）技术分离蛋白质,考马斯亮蓝染色,Imagescanner扫描仪以及LabScan扫描软件对凝胶进行扫描获取图像,利用图像分析软件PDQuest 7.0进行强度校正、点检测、背景消减、均一化和匹配等分析,以正常儿童脑脊液的2-DE凝胶为参考胶,将结核性脑膜炎患儿脑脊液的凝胶与之进行比较分析,寻找两组患儿脑脊液蛋白质谱的差异表达斑点。结果 正常儿童脑脊液分离获得蛋白质斑点546个,结核性脑膜炎患儿脑脊液分离获得蛋白质斑点533个,发现有64个蛋白质点存在质或量的差别,其中有14个蛋白质点仅在正常儿童CSF表达,有27个点仅在结脑患儿CSF中表达,与正常儿童CSF蛋白质点表达量比较发现,结脑患儿CSF有15个蛋白质点表达量有2倍以上上调,8个蛋白质点表达量有2倍以上下调。共选出20个差异蛋白质点进行MALDI-TOF-MS分析,获得了20张肽质量指纹图谱。进入数据库进行搜索后有15个差异蛋白质已经通过MALDI-TOF-MS鉴定。结论 本实验成功建立了正常儿童和结核性脑膜炎患儿脑脊液蛋白质2-DE图谱,通过凝胶图像分析和计算机信息处理,发现了一些与结核性脑膜炎相关的蛋白质差异位点。经质谱分析及生物信息学技术初步鉴定了部分差异蛋白质,发现载脂蛋白A-I、抗肿瘤坏死因子α抗体、HLA-II 类抗原DRB1-4、MRP14蛋白可能与结核性脑膜炎的发生发展密切相关,这些蛋白能否成为结核性脑膜炎诊断的标记物,值得进一步的研究。     

癫痫患者脑脊液差异蛋白质组学分析

目的应用蛋白质组学技术研究脑脊液的癫痫相关蛋白,以便从蛋白质水平揭示癫痫发生发展机制及可能的诊断标志物。方法双向凝胶电泳和质谱技术分离和鉴定癫痫组(n=8)和对照组(n=8)脑脊液的差异表达蛋白。结果约68个蛋白质点表达水平在两组间存在明显差异,选择18个差异蛋白质点质谱鉴定。除4个假想蛋白和未知功能蛋白及得到7个已知功能蛋白,涉及免疫反应、运输蛋白、能量代谢酶蛋白、细胞骨架蛋白和热休克蛋白。结论大部分被鉴定的蛋白为首次报道,这些蛋白与癫痫的发生发展相关,可能成为癫痫脑脊液分子标志物。     

6-羟基多巴胺诱导SH-SY5Y细胞帕金森病模型中Peroxiredoxin6表达的研究

目的 应用蛋白质组学技术研究6-羟基多巴胺(6-OHDA)诱导SH-SY5Y细胞帕金森病(PD)模型中Peroxiredoxin6(Prx6)的表达变化.方法 在建立6-OHDA诱导SH-SY5Y细胞帕金森病模型的基础上,分别提取6-OHDA实验组和对照组孵育24h的细胞总蛋白,应用荧光差异双向凝胶电泳(DIGE)技术获得蛋白点的差异表达信息,运用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOF MS)鉴定出差异蛋白质.结果 DIGE分析发现实验组有一个表达明显上调的差异蛋白质点(P<0.01),经质谱分析鉴定确认为Prx6.结论 6-OHDA诱导SH-SY5Y细胞的帕金森病模型中Prx6表达上调,提示PD中有氧化应激存在,Prx6上调及氧化应激可能与PD的发病机制有关.     

帕金森病患者脑脊液蛋白质的双向凝胶电泳图谱初步分析

目的用双向凝胶电泳(2D-PAGE)技术分析帕金森病患者脑脊液中蛋白质的变化。方法用丙酮沉淀法提取脑脊液中的蛋白质,以固相pH梯度等电聚焦(IPG-IEF)为第一向,十二烷基磺酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)为第二向进行2D-PAGE,蛋白点经银染后用图像分析软件对图谱进行分析。结果比较帕金森病组和对照组的2D-PAGE图谱显示:9个蛋白点在帕金森病患者脑脊液中含量减少,8个蛋白点消失,另有8个蛋白点含量上调,初步推测其中部分蛋白点可能为血浆铜蓝蛋白及其前体,结合珠蛋白β链,以及载脂蛋白J片断和前体。结论2D-PAGE技术可用于脑脊液蛋白质分离,反映蛋白质表达的差异。帕金森病患者脑脊液蛋白质与对照相比有显著差异.     

骨髓源神经干细胞移植治疗中枢神经系统损伤的蛋白质组学研究

目的 利用同位素标记相对和绝对定量(iTRAQ)技术以及串联质谱技术分析中枢神经系统损伤患者干细胞移植前术后脑脊液蛋白质表达变化情况.方法 收集正常成人以及中枢神经系统损伤患者移植前后脑脊液,组内等量混合后利用高效液相色谱柱去除高丰度蛋白,样本经iTRAQ试剂标记后利用二维色谱分离,经串联质谱鉴定并相对定量.结果 质谱得到的多肽片段经谱库搜索鉴定的蛋白共59个,其中患者移植前脑脊液与正常对照组脑脊液存在差异的蛋白质有13个,5个呈高表达,8个呈低表达.患者移植后脑脊液有7种蛋白质与正常对照组表达无差异.结论 蛋白质组学研究方法可以用来寻找神经系统损伤标志物,为神经系统损伤以及修复的机制研究提供基础.     

蛋白质组学技术鉴定帕金森病模型中的3种新蛋白质

目的 利用蛋白质组学技术对MPP 诱导的PC12细胞帕金森病(PD)模型进行研究,以期发现PD发病机制中的新蛋白.方法 建立MPP 诱导的PC12细胞PD模型并提取细胞总蛋白,荧光差异凝胶电泳(D1GE)进行蛋白样品标记与分离,应用DeCyder软件分析蛋白质差异表达信息,运用MALDI-TOF质谱鉴定差异蛋白质.结果 质谱分析和数据库检索鉴定了3种可能同PD发病相关但在有关PD研究文献中尚未见报道的新蛋白质,他们分别足与线粒体功能相关的蛋白MPPs,具有分子伴侣活性的蛋白NAC和与免疫炎症相关的蛋白gc1qBP.结论 这些新发现蛋白可能与PD的发病机制密切相关.     

胶质瘤CD133阳性和阴性U87细胞蛋白质组学分析

目的 通过对人脑胶质瘤U87细胞中CD133+和CD133-细胞进行配对研究,寻找差异性蛋白.方法 通过磁珠选择分离U87中CD133+和CD133-细胞,利用双向凝胶电泳(2D - PAGE)检测上述细胞总体蛋白,分析差异性蛋白点.通过质谱( MALDI -TOF - MS)对蛋白点进行鉴定,建立两种细胞差异性蛋白文库.结果 CD133+和CD133-细胞配对比较,蛋白质组学检测结果中,差异有统计学意义(P＜0.05)的2D - PAGE蛋白点共有73个,＞1.5倍差异蛋白点有46个,＞2倍差异蛋白点有27个,＞3倍差异蛋白点有8个.在CD133+ U87细胞中,有10个蛋白表达增多(上调),63个蛋白表达下降(下调).共得到44张高质量质谱肽质量指纹图谱(PMF),鉴定了35种蛋白质.结论 CD133+和CD133-细胞是同一细胞株中两种不同的细胞,蛋白方面存在的差异,将成为进一步研究它们相互关系的重要靶点.     

帕金森病患者脑脊液诊断标志物：基于双向电泳-质谱检测的蛋白质组学病例-对照

摘要 背景：国内外学者利用蛋白质组学技术发现了阿尔茨海默病的生物标志物可能为total tau proteins,amyloid-b(Ab42）,帕金森病尚无明确的诊断标志物。 目的：探讨采用比较蛋白质组学方法寻找帕金森病病人脑脊液中差异蛋白质作为诊断标志物。 设计、时间及地点：人脑脊液双向电泳图谱对照实验,于2008-9/2009-6在重庆神经病学重点实验室完成。 材料：病人脑脊液均采自重医医科大学附属第一医院神经内科病人,采用腰椎穿刺的方式取得。 方法：分帕金森病组共27人,采用改良webster,Hoehn和Yahr评分量表将此组病人为轻度,14人;中度8人、重度,5人这三个亚组;疾病对照组16人;正常对照组9人。应用PH4-7胶条进行双向电泳分离各组脑脊液蛋白质,用PDquest8.0分析蛋白质电泳图谱,运用MALD I-TOF-MS质谱鉴定差异蛋白,在Mascot中搜索蛋白质数据。 主要观察指标：帕金森病各亚组蛋白质电泳图谱上与实验对照组,空白对照组的差异蛋白点。 结果：本实验发现了4种蛋白质在帕金森各亚组与实验对照组﹑空白对照组比较中差异显著,表达下调的是：依赖DNA的RNA聚合酶III的亚基RPC(p<0.05);Dual serine/threonine and tyrosine protein kinase;活性调控细胞骨架关联蛋白;表达上调的是G2/mitotic-specific cyclin-B1（p<0.05）。 结论：此四种差异蛋白质点存在于帕金森病全程,可考虑作为帕金森病的诊断标志物,协助诊断帕金森病。     

颅内动脉瘤患者血清差异表达蛋白质的研究

目的 采用蛋白质组学方法研究散发性颅内动脉瘤和健康成人外周血清蛋白表达差异.方法 对散发性颅内动脉瘤和健康成人的血清样本进行双向凝胶电泳分离,通过对两种蛋白图谱的比较,确定差异表达的蛋白点;而后对差异点进行基质辅助激光解析电离飞行时间质谱分析和蛋白数据库信息检索;随机选取2种蛋白进行Western blot法验证蛋白质组学实验结果.结果 在颅内动脉瘤和健康成人中鉴定出14种差异表达蛋白,得到了11个肽质量指纹图谱(peptide mass fingerprinting,PMF),最终7种蛋白质得到确认.结合珠蛋白、C反应蛋白、B因子、淀粉样血清蛋白A和载脂蛋白E前体在动脉瘤组表达上调;载脂蛋白A-Ⅳ前体和间α胰蛋白酶抑制物H4重链前体表达下调.蛋白印迹实验结果与双向电泳结果一致.结论 成功鉴定出7个与颅内动脉瘤相关的蛋白,为进一步阐明颅内动脉瘤形成的病理生理学机制提供了新的思路.     

Tetranectin and apolipoprotein A‐I in cerebrospinal fluid as potential biomarkers for Parkinson’s disease

Wang E‐S, Sun Y, Guo J‐G, Gao X, Hu J‐W, Zhou L, Hu J, Jiang C‐C. Tetranectin and apolipoprotein A‐I in cerebrospinal fluid as potential biomarkers for Parkinson’s disease.
Acta Neurol Scand: 2010: 122: 350–359.
© 2010 The Authors Journal compilation © 2010 Blackwell Munksgaard. Objective – The application of biomarkers may potentially improve the efficiency of the diagnosis for Parkinson’s disease (PD). However, no reliable biomarker has been identified to date. This study is aimed to identify proteins that might serve as potential biomarkers for PD diagnosis or pathogenesis. Materials and methods – Two‐dimensional difference gel electrophoresis (2D DIGE) technique, in combination with matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry (MALDI‐TOF MS), was used to determine the differentially expressed cerebrospinal fluid (CSF) proteins in PD patients (n = 3) compared with normal controls (n = 3). Selected proteins were further confirmed by Western blotting analysis in the CSF of PD patients (n = 8), Alzheimer’s disease (AD) patients (n = 6) and normal control subjects (n = 7). Results – Eight proteins were identified after MS and protein database interrogation. In the CSF of PD patients, the expression levels of one isoform of apolipoprotein A‐I (apoA‐I), tetranectin, myosin phosphatase target subunit 1 (MYPT1), and two unknown proteins were down‐regulated, whereas the expression levels of another apoA‐I isoform, proapolipoprotein, and lipoprotein were up‐regulated. Western blotting indicates that the expression of tetranectin was reduced in the CSF from PD patients and elevated in AD, while the expression of apoA‐I was changed only in the CSF from PD patients. Conclusion – Our preliminary results suggest that tetranectin and apoA‐I may serve as potential biomarkers for PD, though further validation is needed.     

Diagnosis of disease-specific proteins in cerebrospinal fluid of children infected with tuberculous meningitis

BACKGROUND: Isolated mycobacterium tuberculosis from cerebrospinal fluid (CSF) is regarded as the "gold standard" for diagnosis of tuberculous meningitis (TBM). However, culture of CSF specimens is time-consuming and lacks sensitivity. There is a strong need to determine complementary disease-specific markers, which are essential for increasing early diagnosis and improving prognosis in patients with TBM. OBJECTIVE: To establish proteomic profiles of CSF in TBM and normal children using two-dimensional polyacrylamide gel electrophoresis, and to screen for disease-specific proteins. DESIGN, TIME AND SETTING: The case-control study was conducted at the Department of Pediatrics, Xiangya Hospital of Central South University and the Key Laboratory of Cancer Proteomics of Ministry of Public Health of China between January 2008 and January 2009. PARTICIPANTS: The TBM group included three patients with a strongly positive tuberculin skin test, as well as positive CSF mycobacterial staining and culture, who were admitted to the Department of Pediatrics, Xiangya Hospital from January 2008 to January 2009. Three healthy, age- and gender-matched children served as the control group. METHODS: CSF proteins were separated using two-dimensional polyacrylamide gel electrophoresis in both groups. Gels were scanned using Image scanner and LabScan software. Differentially expressed proteins were analyzed using PDQuest 7.0 software. The clearly discernible spots, which were expressed only in the TBM group, were chosen to perform matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis. MAIN OUTCOME MEASURES: Differentially expressed spots on CSF profiles of TBM and normal children were measured.RESULTS: Following comparison of two-dimensional polyacrylamide gel electrophoresis maps between TBM and control groups, 546 and 533 spots were detected, respectively. A total of 64 differentially expressed proteins were observed between the groups, including 15 upregulated spots, eight downregulated spots, 27 spots that were exclusively expressed in the TBM group, and 14 spots that were exclusively expressed in the control group. At total of 20 spots that were exclusively expressed in the TBM group were chosen for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis, and 20 peptide mass fingerprints were obtained. After searching the data base, 16 proteins were matched. CONCLUSION: Two-dimensional polyacrylamide gel electrophoresis profiles of the CSF proteome were successfully established in the TBM and normal children. Parts of these differentially expressed proteins were identified through mass spectrometry and bioinformatics. Results indicated that apolipoprotein A-I, anti-tumor necrosis factor-alpha antibody, crystal structure of MRP14 and HLA class II histocompatibility antigen DRB1-4 could be closely correlated with TBM pathogenesis.     

Cerebrospinal fluid proteome profile in multiple sclerosis

Cerebrospinal fluid (CSF) proteins may provide important information about the pathomechanisms present in multiple sclerosis (MS). Although diagnostic criteria for early MS are available, there is still a need for biomarkers, predicting disease subtype and progression to improve individually tailored treatment. Using the two-dimensional difference gel electrophoresis (2-D-DIGE) technology for comparative analysis, we compared CSF samples from patients with MS of the relapse-remitting type (RRMS, n = 12) and from patients with clinically isolated syndrome (CIS, n = 12) suggestive of a first demyelinating attack with neurologically normal controls. Protein spots that showed more than two-fold difference between patients and controls were selected for further analysis with MALDI-TOF mass spectrometry. Immunoblot analysis was performed to confirm the validity of individual candidate proteins. In RRMS, we identified 1 up-regulated and 10 down-regulated proteins. In CIS, 2 up-regulated and 11 down-regulated proteins were identified. One of these proteins (Apolipoprotein A1) was confirmed by immunoblot. Though the pathophysiological role of these proteins still remains to be elucidated in detail and further validation is needed, these findings may have a relevant impact on the identification of disease-specific markers.     

Proteomics analysis of MPP+-induced apoptosis in SH-SY5Y cells

Accumulating evidence suggests that oxidative stress plays a pivotal role in dopaminergic neurodegeneration. However, the kinds of proteins involved in the response to oxidative stress remain unclear. In the present study, SH-SY5Y cells were treated with neurotoxin 1-methyl-4-phenyl-pyridinium ion (MPP+) to induce apoptotic neuronal injury. 2D-DIGE followed by MALDI-TOF-MS was used to determine the changing protein levels. Proteomics analysis revealed that 22 proteins were differentially altered in MPP⁺-treated SH-SY5Y cells, of which 7 were up-regulated proteins and 15 were down-regulated proteins, respectively. Three protein spots were unambiguously identified as sorcin, annexin V, and ribosomal protein P0. The three proteins showed a significant increase in level, suggesting a role in MPP⁺-induced apoptosis. The functional roles of these three proteins collectively indicate that multiple mechanisms are pertinent in the underlying pathogenesis of Parkinson’s disease (PD), such as apoptosis, calcium homeostasis, and DNA insults.     

Alterations in cerebral metabolomics and proteomic expression during sepsis

The cause of brain dysfunction during sepsis and septic encephalopathy is still under ongoing research. Sepsis induced changes in cerebral protein expression may play a significant role in the understanding of septic encephalopathy. The aim of the present study was to explore cerebral proteome alterations in septic rats. Fifty-six male Wistar rats were randomly assigned to a sepsis group (coecal ligature and puncture, CLP) or a control group (sham). Surviving rats were killed 24 or 48 hours after surgery and whole-brain lysates were used for two-dimensional gel electrophoresis and subsequent protein identification. Differentially expressed proteins were identified by mass spectrometry. Using the Ingenuity Pathways Analysis (IPA) tool, the relationship and interaction between the identified proteins was analyzed. Mortality was 53 % in septic rats. No rat of the control group was lost. More than 1,100 spots per gel were discriminated of which 29 different proteins were significantly (2-fold, P<0.01) changed: 24 proteins down-regulated after 24 hours; two proteins up-regulated and three down-regulated after 48 hours. IPA identified 11 of 35 differentially regulated proteins allocating them to an existing inflammatory pathway. In the analysis of septic rat brains, multiple differentially expressed proteins associated with metabolism, signaling, and cell stress can be identified via proteome analysis, that may help to understand the development of septic encephalopathy.     

Different apolipoprotein E,apolipoprotein A1 and prostaglandin-H2 D-isomerase levels in cerebrospinal fluid of schizophrenia patients and healthy controls

Abstract

Objectives. To identify proteins differentially expressed in schizophrenia patients, we collected 50 µl cerebrospinal fluid from 17 first-episode schizophrenia patients and 10 healthy controls. Methods. Their proteins were separated by two-dimensional gel electrophoresis without using any depletion method and identified by mass spectrometry. Results. Approximately 550 spots were detected, six of which had significantly different intensities in schizophrenia compared to control specimens. We were able to validate in individual samples the upregulation of apolipoprotein E, apolipoprotein A1 and prostaglandin-H2 D-isomerase by Western blot analyses and detect the downregulation of transthyretin, TGF-β receptor type-1 and coiled-coil domain-containing protein 3 precursor. Conclusions. These findings may help to elucidate the disease mechanisms and confirm the hypothesis of disturbed cholesterol and phospholipid metabolism in schizophrenia, and thus reveal the final role players. Moreover, a grouped protein expression analysis of apolipoprotein E, apolipoprotein A-I, and prostaglandin-H2 D-isomerase in cerebrospinal fluid from patients might be a potential diagnostic tool for schizophrenia.     

Proteomic studies of potential cerebrospinal fluid protein markers for Alzheimer's disease

By comparing the cerebrospinal fluid (CSF) proteome between Alzheimer's disease (AD) patients and controls, it may be possible to identify proteins that play a role in the disease process and thus to study the pathogenesis of AD. Two-dimensional gel electrophoresis (2-DE), SYPRO Ruby staining and mass spectrometry were used for clinical screening of disease-influenced CSF proteins in AD patients compared to controls. In order to increase the detection of CSF proteins and to improve the separation of protein isoforms in a 2-D gel, micro-narrow range IPG strips were used. The levels of eight proteins and their isoforms, including apolipoprotein A1, apolipoprotein E, apolipoprotein J, beta-trace, retinol-binding protein, kininogen, alpha-1 antitrypsin, cell cycle progression 8 protein, and alpha-1beta glycoprotein were significantly altered in CSF of AD patients compared to controls. Furthermore, we also used liquid-phase IEF, as a prefractionation step, prior to 2-DE for comparison of CSF proteins between individual AD patients and controls. The levels of 37 proteins spots were altered in AD patients. One of the identified proteins, alpha-2-HS glycoprotein, has not previously been linked to AD. Our study shows that several glycoproteins are altered in AD.     

Cerebrospinal fluid diagnostic markers for two-dimensional electrophoresis-mass spectrometry in Parkinson's disease patients

BACKGROUND: Previous studies have confirmed the existence of specific proteins in body fluid of Parkinson's disease (PD) patients. However, the existing research has contained several interference factors with poor reproducibility and has not focused on patients grouped according to disease duration.OBJECTIVE: To verify differential expression of proteins in cerebrospinal fluid of PD patients grouped in order of disease severity through the use of two-dimensional electrophoresis-mass spectrometry methods. DESIGN, TIME AND SETTING: The proteomic-based, case-control study was performed between September 2008 and June 2009 at the Key Laboratory of Neurology in the First Affiliated Hospital of Chongqing Medical University. PARTICIPANTS: A total of 52 outpatients and/or inpatients, who were admitted to the Department of Neurology in the First Affiliated Hospital of Chongqing Medical University between 2008 and 2009, were randomized into the present study. Among them, 27 PD patients served as the PD group and were assigned to three subgroups according to modified Webster, Hoehn, and Yahr rating scales: 14 = mild, 8 = moderate, and 5 = severe; non-PD group of 16 patients included 5 cases of viral meningitis, 3 cases of acute myelitis, 1 case of Guillain-Barre syndrome, 2 cases of tuberculous meningitis, 2 cases of restless legs syndrome, and 3 cases of essential tremor; control group (n = 9) consisted of muscular tension headache in 6 cases, as well as syncope, trigeminal neuralgia, idiopathic orthostatic hypotension in 1 case. METHODS: Cerebrospinal fluid was collected from the involved patients using the lumbar puncture method. Proteins in the cerebrospinal fluid were separated by two-dimensional electrophoresis. MAIN OUTCOME MEASURES: Characteristics of protein electrophoresis patterns were analyzed, differentially expressed proteins were detected using matrix-assisted laser desorption ionization time of flight mass spectrometry, and protein data were analyzed in the Mascot database. RESULTS: Five protein electropherograms were analyzed by PDQuest 8.0, and (789 ± 32) protein spots were observed. There were significant differences in four protein spots in each of the PD sub-groups compared with the non-disease and control groups. Expression was down-regulated in three protein spots and up-regulated in one protein spot; 100% repetition rate was observed in four protein spots. According to the Mascot database, protein spots with down-regulated expression were as follows: DNA-guided RNA polymerase Ⅲ subunit RPC5 (score: 50 points); double serine, threonine, and tyrosine protein kinase (score: 64 points, P < 0.05); activity-regulated cytoskeleton-associated protein (score: 58 points, P < 0.05). However, G2 mitotic-specific cyclin was up-regulated (score: 84 points, P < 0.05). CONCLUSION: Differential protein expression in the cerebrospinal fluid of PD patients was detected by two-dimensional electrophoresis-mass spectrometry, revealing changes in DNA-guided RNA polymerase III subunit RPC5, double serine, threonine, and tyrosine protein kinase, activity-regulated cytoskeleton-associated protein, and G2 mitotic cell cyclin, with good reproducibility.