轻度认知障碍及阿尔茨海默病患者外周血淋巴细胞中蛋白磷酸酯酶-2A的改变

目的 探讨轻度认知功能障碍(MCI)及阿尔茨海默病(AD)患者外周血淋巴细胞蛋白磷酸酯酶-2A(PP-2A)的变化.方法 用放射性配体结合试验检测11名健康对照者、11例MCI患者及11例AD患者外周血淋巴细胞PP-2A的活性,并用Western bolt检测PP-2A蛋白的表达.结果 MCI组外周血淋巴细胞PP-2A活性(0.71±0.12)及蛋白表达(0.80±0.05)与健康对照组[分别为(1.01±0.09)和(0.96±0.07)]相比明显降低(均P＜0.01);AD组PP-2A活性(0.64±0.11)及蛋白表达(0.76±0.06)亦明显降低,与对照组相比差异有统计学意义(均P＜0.01);AD组PP-2A活性及蛋白表达比MCI组有降低的趋势,但差异无统计学意义.结论 MCI及AD患者外周血淋巴细胞PP-2A的活性及表达降低,在MCI的诊断中可能有参考价值,同时亦可能对AD的诊断有一定的潜在的意义.     

Aβ25-35刺激下人淋巴细胞中p53表达水平的变化

目的 探讨散发型阿尔茨海默病(sporadic Alzheimer's disease,sAD)患者外周血淋巴细胞中总p53蛋白水平增高,同时出现异常的“突变样”p53表达的可能机制.方法 采用诱导AD早期病理改变的重要物质即淀粉样蛋白Aβ25-35刺激健康人外周血淋巴细胞,通过Western blot检测总p53蛋白及其突变型的表达情况.结果 与对照组对比显示,在低浓度(10-8 mol/L) Aβ25-35作用下,不同时程的淋巴细胞中总p53表达即增高,差异有统计学意义(P＜0.01),并可见微量突变型p53的表达;随着Aβ25-35刺激浓度的提高,总p53水平有增高的趋势,但几乎无统计学意义;突变型p53表达无差异.结论 上述结果模拟了sAD患者外周血淋巴细胞中的变化,证实了可溶性A8在AD发病中的重要作用,以及对p53表达的影响.因而,这一模型将有望在探讨AD的发病机制以及筛选防治AD的药物中发挥一定的作用.     

miR-27a-3p在高血压性脑出血患者外周血中呈下调表达

目的探讨脑出血患者外周血血清miR-27a-3p的表达情况。方法选取31例脑出血患者(脑出血组)和11例体检健康对照者(对照组),收集临床资料,qRT-PCR法测定外周血血清中miR-27a-3p的表达水平。结果与健康对照组比较,脑出血组血清miR-27a-3p表达呈显著下调(miR-27a-3p相对表达量为0.29),脑出血组血清miR-27a-3p表达水平与血肿体积呈明显线性负相关(相关系数R=--0.502,P=0.004)。结论在脑出血患者外周血血清miR-27a-3p表达水平呈下调趋势,其表达量与血肿体积呈线性负相关关系,可能成为脑出血患者血肿体积的预测指标。     

Skp2和p27在人脑胶质瘤中的表达及其意义

目的探讨胶质瘤中Skp2的表达及其与p27蛋白的关系。方法用免疫组化方法检测33例胶质瘤和6例正常脑组织标本中Skp2和p27蛋白的表达。结果Skp2在正常脑组织中表达不明显,在低级别胶质瘤中表达较少,在高级别胶质瘤中表达较多,两两比较差别显著(P<0.01)。p27在正常脑组织中表达明显,在低级别胶质瘤中表达较少,在高级别胶质瘤中表达很少,两两比较差别显著(P<0.01)。胶质瘤中Skp2蛋白表达与p27蛋白表达呈负相关(r=-0.809,P<0.01)。结论Skp2的表达与胶质瘤的病理分级有关,它通过促进的降解参与胶质瘤细胞的恶性进展,可能成为胶质瘤治疗的新靶点。     

人髓母细胞瘤中细胞周期素依赖激酶抑制物p27KIP1的表达及预后关系

目的检测p27蛋白在人髓母细胞瘤中的表达及定位情况,探讨p27与髓母细胞瘤预后的关系。方法应用免疫组化SP法和激光共聚焦显微术检测57例髓母细胞瘤标本中p27蛋白的表达及其定位情况,结合随访资料进行生存分析。结果在全部57例髓母细胞瘤标本中,p27阳性表达为32.41%～96.53%,平均为64.34%±18.10%,Kaplan-Meier生存曲线分析示p27高表达组≥80%较低表达组(<80%)有显著性差异(P<0.0001);以p27阳性表达率60%为截取值,两组间生存曲线有显著性差异(P<0.0001)。激光共聚焦显微术检测显示p27在肿瘤细胞质中有表达,生存曲线分析显示在全部病例及儿童病例组中有显著性差异(P<0.05),而在成人病例组中则无显著性差异P>0.05。Cox比例风险模型分析显示p27表达降低和p27在细胞质中呈阳性表达在髓母细胞瘤中的相对危险度分别为4.5(95%CI2.3～6.7)、3.4(95%CI1.6～4.1)。结论在髓母细胞瘤中p27表达有不同水平的减低,p27表达的降低及p27在细胞质呈阳性表达是人髓母细胞瘤的危险因素,p27在人髓母细胞瘤中具有预后判断意义的最佳截取值为阳性表达率60%。     

高血压脑出血患者外周血miR-155、miR-27a-3p表达水平及其临床意义

目的 研究高血压脑出血(HICH)患者外周血微小microRNA-155(miR-155)、microRNA-27a-3p(miR-27a-3p)表达水平及其临床意义.方法 选取2018年01月-2020年01月四川绵阳四0四医院神经外科收治的100例HICH患者作为研究对象,设为观察组,根据格拉斯哥昏迷(GCS)评分...     

雌激素在阿尔茨海默病患者的改变及其意义

目的分析AD患者血清雌激素水平和内源性雌激素的改变,探讨雌激素水平与AD严重程度、病程及年龄的关系。方法采用1∶1病例配对的研究方法,通过面对面问卷调查对全部研究对象进行神经心理测验,包括MMSE、ADL、POD、FOM、WISE、HMT抑郁量表等;同时调查女性研究对象的初潮年龄、绝经年龄、怀孕次数及母乳喂养等;用放射免疫法测定血清性激素水平。结果AD组雌二醇水平(E2)明显低于对照组,有显著性差异(P<0.05);AD组血清雌二醇水平与痴呆严重程度和MMSE分值呈正相关,(P<0.05);AD组初潮年龄明显大于对照组,而绝经年龄明显早于对照组,有显著性差异(P<0.05);怀孕次数尽管在两组比较无统计学差异(P>0.05),但AD组仍少于对照组。结论AD组雌激素水平明显低于对照组,雌激素缺乏是AD的危险因素之一。     

阿尔茨海默病患者外周血OSTM1表达的变化

目的观察阿尔茨海默病(AD)患者与健康对照组外周血单核粒细胞中骨硬化症相关蛋白1(OSTM1)表达的差异并探讨其意义。方法 AD患者与对照组各30例,采用Ficoll-Hypaque密度梯度离心法分离人外周血单核粒细胞,培养24h收集细胞。细胞爬片后,采用免疫荧光细胞化学技术,观察OSTM1在单核粒细胞中分布并比较两组荧光信号强弱;蛋白印迹法(Western blotting)检测OSTM1蛋白表达水平;实时定量多聚链式反应(RT-PCR)法检测OSTM1mRNA表达水平。结果 1OSTM1表达情况:人外周血中表达OSTM1主要集中在胞浆,与对照组比较,AD组细胞荧光信号明显增强且分布范围扩大;2两组OSTM1在蛋白表达水上的平差异:AD组OSTM1表达(0.53!0.04)较对照组(0.36!0.03)比较差异有统计学意义(P<0.05);3两组在OSTM1基因水平表达情况:AD组与对照组OSTM1 mRNA分别为0.75!0.13和0.32!0.11,AD组OSTM1 mRNA表达水平与对照组相比,差异有统计学意义(P<0.05)。结论 AD组OSTM1在蛋白、基因水平较对照组均明显升高,推测OSTM1在AD发病机制中具有关键作用。     

Alzheimer病患者外周血淋巴细胞周期分布及细胞外信号调节激酶1/2/哺乳动物雷帕霉素靶蛋白通路的改变及意义

目的研究Alzheimer病(AD)患者外周血淋巴细胞周期分布及上游调控通路细胞外信号调节激酶1/2(ERK1/2)/哺乳动物雷帕霉素靶蛋白(m TOR)的改变及意义。方法对30例AD患者(AD组)用流式细胞分析仪检测外周血淋巴细胞周期分布,用Western Blot法检测淋巴细胞ERK1/2/m TOR通路和周期素依赖性蛋白激酶抑制剂p21的表达,并与帕金森病(PD)患者(PD组)及正常对照者(NC组)进行比较。分析AD组细胞周期相关蛋白与简易精神状态检查(MMSE)量表评分、病程的关系。结果与NC组相比,AD组外周淋巴细胞分布于G1期的比例(G1%)显著下降而S%显著升高(均P0.01);淋巴细胞中p-ERK1/2、pm TOR和p21水平显著下降(均P0.01)。与NC组相比,PD组ERK1/2、p-ERK1/2、m TOR、p-m TOR、p21的差异均无统计学意义。AD组外周血淋巴细胞p-m TOR水平与MMSE评分呈正相关(r=0.592,P=0.018),与病程呈负相关(r=-0.558,P=0.025)。结论 AD患者外周血淋巴细胞周期分布存在异常,淋巴细胞的pERK1/2、p-m TOR和p21水平显著下降,并且病情越重、病程越长者p-m TOR水平下降越明显。外周血淋巴细胞p-ERK1/2、p-m TOR和p21有可能成为AD诊断及病情评价的外周标志物。     

miR-27-3p、miR-128-3p及miR-140-3p表达在急性脑梗死患者诊疗中的价值研究

目的 探讨miR-27-3p、miR-128-3p及miR-140-3p表达在急性脑梗死(ACI)患者诊疗中的应用价值。方法 选取2020年1月至2022年12月南通市第三人民医院全科医学科收治的ACI患者100例作为研究对象,定义为ACI组,根据ACI患者3个月改良Rankin评分量表(mRS评分)将其分为预后良好组(mRS评分≤2分)68例与预后不良组(mRS>2分)32例,另选同期于南通市第三人民医院全科医学科体检的健康者100例作为对照组。记录各组miR-27-3p、miR-128-3p及miR-140-3p表达水平并进行比较,采用Pearson相关分析探讨两变量的相关性,通过绘制受试者工作特征(ROC)曲线评价miR-27-3p、miR-128-3p及miR-140-3p联合检测在ACI诊断中的效能,采用多因素Logistic回归分析探讨ACI预后的危险因素。结果 ACI组平均年龄、吸烟史比例、合并高血压比例、miR-140-3p与miR-128-3p及miR-27-3p表达水平均明显高于对照组(^均P<0.05)。miR-27-3p、miR-...     

Alzheimer病患者外周血淋巴细胞Akt及其活化水平的改变及意义

目的 研究Alzheimer病(AD)患者外周血淋巴细胞Akt及其活化水平的改变及意义.方法 对20例AD患者(AD组)进行简易精神状态检查(MMSE)量表评分,用免疫印迹法检测其外周血淋巴细胞的Akt及磷酸化Akt (p-Akt)水平,并与帕金森病(PD)患者(PD组)和正常对照者(正常对照组)进行比较.分析AD患者外周血淋巴细胞的Akt水平与病程和MMSE量表评分的关系.结果 与正常对照组相比,AD组外周血淋巴细胞中Akt及p-Akt水平明显升高(均P＜0.01);PD组p-Akt水平显著增高(P＜0.05).与PD组相比,AD组Akt水平升高明显(P＜0.05).AD患者外周血淋巴细胞中Akt水平与病程呈正相关(r=0.602,P=0.014),与MMSE量表评分呈负相关(r=-0.560,P=0.024).结论 AD患者外周血淋巴细胞的Akt及其活化水平显著增高,并且病程越长、病情越重者增高的越明显.提示Akt有可能成为AD诊断及病情评价的外周标志物.     

AD患者外周血单核细胞中TLR-4表达 水平变化的临床意义

目的 探讨外周血单核细胞Toll样受体-4(Toll-like receptor 4,TLR-4)的表达水平变化与AD发病及进展的关系。方法 检测阿尔茨海默病(Alzheimer's disease,AD)及轻度认知功能障碍(Mild cognitive impairment,MCI)患者及同年龄正常对照者(Healthy Controls,HC)外周血单核细胞Toll样受体-4(Toll-like receptor 4,TLR-4)的表达水平; 采集并分离AD(n=28)、MCI(n=26)及NC组(n=20)外周血单核细胞; 用实时荧光定量PCR(Quantitative Real-Time,Q-PCR)、流式细胞术(Flow Cytometry)检测TLR-4的表达水平; 双抗体夹心法(Double antibody sandwich method,ELISA)检测各组血清中肿瘤坏死因子(Tumor necrosis factor-α(TNF-α)、白介素6(Interleukin 6,IL-6)的水平。结果 与HC组比较,AD患者外周血单核细胞TLR-4表达水平均明显上调(P均<0.05)。MCI组略有上升,但无统计学意义; AD患者外周血单核细胞TLR-4表达水平与血清TNF-a、IL-6水平呈正相关(r=0.3885和0.3270,P均<0.05)。结论 外周血单个核细胞TLR-4表达水平上调提示炎性机制在AD的发病中发挥了作用,TLR4可作为评估疾病进展的潜在生物学标志物。     

Interleukin-2 receptor in peripheral blood lymphocytes of Alzheimer's disease patients.

In this study, we investigated the differences in the incidence rate of human leukocyte antigen-DR (HLA-DR) and interleukin 2 receptor (IL-2R) in the peripheral blood lymphocytes of 13 women who were diagnosed clinically as having Alzheimer's disease (AD group), and 13 healthy women with no mental disorders (control group). As a result, the AD group showed a markedly higher rate of lymphocyte subsets of CD4+HLA-DR+, CD4+IL-2R+, CD8+HLA-DR+, CD8-HLA-DR+ and CD8+IL-2R+. This finding indicates a possible immune reaction occurring in the peripheral blood of AD patients. In addition, these higher rates correlate well with the changes in the immune system reported in the postmortem brains of AD patients. Our findings indicate that immunological interactions exist between the central nervous system and the peripheral blood lymphocytes of AD patients, and AD might be induced by an immune reaction occurring in the brain.     

Immune system-associated antigens on the surface of peripheral blood lymphocytes in patients with Alzheimer's disease.

We investigated the immune-associated antigens of peripheral lymphocytes from 13 patients with Alzheimer's disease (AD) and 13 age-matched healthy control subjects using two-color analysis with flow cytometry. Four ratios of immune-related antigens, T/B lymphocytes, CD4/CD8, CD4/CD45R and CD4/HLA-DR, were compared for the AD and control groups. The T/B and CD4/CD8 ratios did not differ between the groups, the ratio of CD4+CD45R+ subset in the AD group was lower than the ratio in the control group, and the ratios of CD4+ CD45R- and CD4+HLA-DR+ subsets in the AD group were significantly higher. Further, in the AD group, the CD4+ CD45R+/CD4+ ratio was lower and the CD4+ CD45R- CD4+ ratio was higher than in the control group.     

阿尔茨海默病患者外周血白细胞介素23和白细胞介素17测定及意义

目的探讨阿尔茨海默病(Alzheimer’s disease,AD)患者外周血中炎症因子白细胞介素23(Interleukin-23,IL-23)和IL-17水平及其与认知功能损害的相关性。方法纳入AD患者102例(研究组),以及健康老年志愿者100例(对照组)。搜集受试者的一般资料和疾病信息。采用简易精神状态量表(MMSE)和临床痴呆评定量表(CDR)对受试者认知功能和日常行为能力进行评估。应用实时荧光定量逆转录-聚合酶链式反应(RTPCR)检测各组入选者外周血淋巴细胞中IL-23和IL-17的mRNA表达,应用酶联免疫吸附试验(ELISA)检测入选者血清IL-23和IL-17蛋白的表达。比较两组入选者血清炎症因子mRNA及蛋白水平的差异,分析外周血IL-23和IL-17表达的相关性。以全部入选者为研究对象,分析两组IL-17和IL-23表达差异与MMSE评分的相关性。以AD组为研究对象,分析患者IL-17和IL-23表达水平与CDR评分的相关性。结果 AD组外周血淋巴细胞中IL-23和IL-17的mRNA表达水平及血清IL-23和IL-17蛋白的表达水平均显著高于对照组,差异均有统计学意义(P<0.05),且两者具有显著正相关性(P<0.01)。整体人群中,IL-23和IL-17水平均与MMSE评分呈负相关(P<0.05)。AD患者血IL-23和IL-17水平与CDR评分均无相关性。结论外周血IL-23和IL-17水平与整体人群的认知功能相关,但与AD患者的痴呆严重度无明显相关性。     

Impairment of mitogenic activation of peripheral blood lymphocytes in Alzheimer's disease.

Cell-cycle dysregulation might be critically involved in the process of neurodegeneration in Alzheimer's disease (AD). We now provide evidence for a dysfunction of the cell division cycle as a more general cellular phenomenon of the disease. Peripheral blood lymphocytes, stimulated with mitogenic compounds, were less able to express CD69, an early proliferation marker, in AD patients than in age-matched controls. Expression levels of CD69 of both T-cells and B-cells correlated inversely with the Mini-mental Scale. The results suggest that a systemic failure of cellular proliferation control might be of critical importance for the pathomechanism of AD.     

Drug treatment of Alzheimer's disease patients leads to expression changes in peripheral blood cells

BackgroundIncreasing cholinergic activity has been the primary mechanism for treating dementia due to Alzheimer's disease. However, the effectiveness of cholinesterase inhibitors (ChEIs) is still widely debated. The identification of specific biomarkers capable of identifying patients more likely to respond to these treatments could potentially provide specific evidence to clearly address this controversy through patient stratification. The goal of this study was to determine the feasibility of discovering biomarkers specific for the treatment of Alzheimer's disease.MethodsPeripheral blood was collected from a cohort of patients treated with different ChEIs. Total RNA was isolated and profiled on the human Genome-Wide SpliceArray (GWSA) to test the feasibility of discriminating the different treatment subgroups of subjects based on the expression patterns generated from the Genome-Wide SpliceArray.ResultsSpecific expression differences were identified for the various treatment groups that lead to a clear separation between patients treated with ChEIs versus naïve patients when Principal Component Analysis was performed on probe sets selected for differential expression. In addition, specific probe sets were identified to be dependent on the inhibitor used among the treated patients.ConclusionsDistinct separation between non-treated, galantamine, donepezil, and rivastigmine-treated patients was clearly identified based on small sets of expression probes. The ability to identify drug-specific treatment expression differences strengthens the potential for using peripheral gene signatures for the identification of individuals responding to drug treatment.     

T lymphocytes in peripheral blood from patients with neurological diseases

Absolute and relative numbers of T lymphocytes in peripheral blood were determined in 19 different groups of patients with neurological diseases, using the E-rosette test. A significantly decreased total number of T lymphocytes was found in the following groups: acute Guillain-Barré syndrome (GBS), active multiple sclerosis (MS), and malignant cerebral tumor. A less pronounced and not significant reduction was observed in patients with amyotrophic lateral sclerosis (ALS). In all other groups of patients normal total numbers of T lymphocytes were found, thus indicating that the decrease in T lymphocyte counts in GBS, MS and malignant cerebral tumors is not due to nonspecific injury of nerve tissue.