神经胶质瘤患者外周血淋巴细胞亚群检测的临床分析

目的：分析神经胶质瘤患者检测其外周血淋巴细胞亚群的临床意义。方法选择2011-12-2013-08在我院接受诊治的神经胶质瘤患者65例作为观察组,选择同期在我院健康体检者43例作为对照组。分析2组患者采用流式细胞术检测CD3＋CD4＋、CD3＋CD8＋、CD4＋/CD8＋、CD3＋ T淋巴细胞及CD4＋CD25＋ T reg水平。结果观察组外周血中CD3＋ CD4＋T细胞与CD4＋/CD8＋比值较对照组显著降低（ P＜0.05）;A组外周血中CD3＋CD4＋百分比、CD4＋/CD8＋比值均较B组显著降低（ P＜0.05）;A组外周血中CD3＋CD8＋百分比较B组显著升高（ P＜0.05）;观察组CD4＋ CD25＋ T reg百分比较对照组显著升高（P＜0.05）;A组CD4＋CD25＋ Treg百分比较B组显著升高（P＜0.05）。结论恶性肿瘤存在免疫功能紊乱,对于神经胶质瘤外周血淋巴细胞的检测,尤其T reg细胞,有利于对神经胶质瘤患者恶性程度及疗效进行判断。     

神经梅毒患者淋巴细胞亚群分析

目的分析神经梅毒患者外周血中淋巴细胞亚群的特点,探讨其与神经梅毒发病的关系。方法选取神经梅毒患者52例为研究对象,分为无症状神经梅毒组、早期神经梅毒组与晚期神经梅毒组,选取同期健康体检者50例为健康对照组,采集外周血,运用流式细胞术分析神经梅毒组与健康对照组的外周血淋巴细胞的表达水平。结果神经梅毒患者的CD3~+CD8~+T淋巴细胞数量明显高于健康对照组,差异有统计学意义(P0.05),神经梅毒患者NK细胞数量显著低于健康对照组;无症状神经梅毒患者的CD3~+T细胞与CD3~+CD8~+T细胞数显著高于早期神经梅毒患者和晚期神经梅毒患者,差异均有统计学意义(P0.05)。结论神经梅毒患者的T淋巴细胞亚群失衡与NK细胞减少与神经梅毒的发生发展有一定关系。     

卒中T淋巴细胞亚群动态变化及其与卒中相关感染的关系

目的 观察卒中急性期患者外周血中C D 3 +T细胞、C D 3 + C D 4 +T细胞、C D 3 + C D 8 +T细胞及 CD4+CD25+FoxP3+调节性T细胞（regulatory T cells,Tregs）的动态变化,探讨卒中后机体免疫状态及其 对卒中后感染的影响。 方法 选取卒中急性期患者32例为卒中组,根据发病1周内是否发生感染,将患者分为卒中后非感 染组24例和卒中后感染组8例。另选取性别、年龄匹配的健康体检者23例为对照组。采用流式细 胞术分别于病程24 h内、3 d、7 d检测卒中患者和健康体检者外周血中CD3+T细胞、CD3+CD4+T细胞、 CD3+CD8+T细胞及Tregs水平。 结果 ① 与对照组比较,卒中后非感染组CD3+CD4+T细胞与Tregs于发病后7 d升高（P分别为0.02和 0.03）;CD3+CD8+T细胞在发病后24 h及3 d下降（P分别为0.01和0.03）,发病后7 d升至与健康对照组 无显著差异。卒中后感染组CD3+T细胞、CD3+CD4+T细胞、CD3+CD8+T细胞及Tregs在发病24 h内（P分别为 ＜0.001,＜0.001,0.03和＜0.001）、3 d（P均＜0.001）、7 d（P分别为＜0.001,0.01,0.01和0.01）均较健 康对照组明显下降;②卒中后感染组CD3+T细胞、CD3+CD4+T细胞及Tregs在发病后24 h内（P分别为0.01, ＜0.001和＜0.001）、3 d（P分别＜0.001,＜0.001和0.04）、7 d（P均＜0.001）均显著低于卒中后非感染 组;两组CD3+CD8+T细胞在发病后24 h内无明显差异,但3 d（P＜0.001）、7 d（P =0.02）卒中后感染组显 著低于卒中后非感染组。 结论 C D3+T淋巴细胞、CD3+CD4+T淋巴细胞、CD3+CD8+T淋巴细胞及Tregs参与卒中早期病理生理过 程,其动态变化可能导致卒中后免疫抑制,并参与卒中后感染的发生。     

抽动障碍患儿免疫功能检测与其临床意义

目的　观察抽动障碍患儿外周血 T淋巴细胞亚群、自然杀伤细胞和体液免疫 ( Ig A,Ig G,Ig M)功能。方法　应用直接免疫荧光染色法 ,采用流式细胞术分别测定 3 0例抽动障碍患儿和 2 0名健康对照组儿童外周血 T淋巴细胞亚群、自然杀伤细胞。并用免疫透射比浊法测定患儿的体液免疫 ( Ig A、Ig G、Ig M)功能。比较两组的检测结果。结果　抽动障碍患儿外周血 CD4+细胞百分比、CD4+ /CD8+细胞比值和自然杀伤 ( NK)阳性细胞百分比分别为 ( 2 6.89± 9.0 8) %、( 0 .88± 0 .3 5 ) %、( 9.61± 6.45 ) % ,较对照组 ( 3 8.3 1± 6.95 ) %、( 1 .5 8± 0 .2 9) %、( 1 4.83±4.1 6) %明显降低 ,CD8+ 细胞百分比 [( 3 2 .5 8± 9.0 4) % ]较对照组 [( 2 4.82± 5 .5 4) % ]明显升高 ,而体液免疫Ig A、Ig G、Ig M含量与对照组比较则差异无显著性。结论　抽动障碍患儿存在细胞免疫功能紊乱 ,表现为 T淋巴细胞亚群平衡失调和自然杀伤细胞阳性率低。提示细胞免疫功能紊乱可能与某些儿童易患抽动障碍有关。     

异基因T淋巴细胞诱导血管内皮细胞组织因子表达的机制

目的 血管内皮细胞是移植物抗宿主病中异基因T淋巴细胞的重要靶组织，而血管内皮细胞损伤、活化后组织因子（TF）表达显著增高。因此血管内皮细胞TF表达可能在移植物抗宿主病引起的组织损伤中发挥重要作用。本研究的目的是探讨异基因T淋巴细胞能否诱导血管内皮细胞TF表达及其分子机制。方法 磁珠阳性分选异基因CD4+和CD8+T淋巴细胞。实时定量PCR和流式细胞术检测异基因T淋巴细胞作用后脐静脉内皮细胞（HUVEC）中TF在基因和蛋白水平的表达。Western blot方法检测HUVEC中MAPK的表达。观察MAPK阻滞剂对异基因T淋巴细胞诱导的HUVEC中TF表达的影响。结果 异基因CD4+ T淋巴细胞作用6、12、24h，HUVEC膜表面TF表达率分别为9.3±0.6%、34.5±1.1%、15.7±0.8%，较对照组（3.3±0.4%）显著增高（P<0.05）； HUVEC中TFmRNA表达水平亦显著增高，分别为对照组的14.4、8.9、5.3倍（P<0.05）。异基因CD8+T淋巴细胞作用12h，HUVEC膜表面TF表达率为14.6±0.7%，较对照组（2.8±0.3%）显著增高；HUVEC中TFmRNA表达水平显著增高，达对照组的5.7倍（P<0.05）。异基因T淋巴细胞作用后，HUVEC内磷酸化p38MAPK和JNK表达增强，而磷酸化ERK表达则无变化。P38MAPK阻滞剂(SB203580)和JNK阻滞剂(SP600125)可显著下调异基因T淋巴细胞诱导的TF表达。结论 异基因T淋巴细胞通过JNK和p38MAPK信号通路诱导血管内皮细胞TF表达。     

尼古丁抑制6-羟基多巴胺损伤诱导的CD8+T淋巴细胞浸润

目的:采用6-羟基多巴胺部分损伤大鼠帕金森病模型,探讨尼古丁保护多巴胺能神经元的机制。方法:动物分别接受尼古丁(0.2mg/kg或2.0mg/kg)或生理盐水腹膜腔内注射。注射7天后,尼古丁高、低剂量治疗组和模型对照组动物分别接受纹状体内6-OHDA20μg注射,制作部分损伤帕金森病模型。免疫组织化学及体视学方法定量分析黑质多巴胺能神经元和纹状体CD8+T淋巴细胞数量。结果:尼古丁低剂量治疗组和高剂量治疗组,6-OHDA注射侧的酪氨酸羟化酶免疫阳性细胞分别为相应对照侧的64.97±10.33%和67.24±12.67%;和模型对照组(25.27±11.79%)比较,差异有统计学意义(P<0.01)。此外,尼古丁也可明显减少纹状体CD8+T淋巴细胞浸润(P<0.05)。结论:尼古丁可能通过抑制CD8+T淋巴细胞浸润,保护多巴胺能神经元。     

吉兰-巴雷综合征患者免疫球蛋白治疗前后T淋巴细胞亚群的变化及其意义

目的 研究吉兰-巴雷综合征(GBS)患者应用静脉大剂量注射免疫球蛋白(IVIG)治疗前后T淋巴细胞亚群的变化,并进一步探讨IVIG治疗GBS的可能机制.方法 选择31例临床确诊的GBS患者,以治疗前后作为自身对照,根据治疗效果评定为效优组和效劣组,应用流式细胞分析仪检测GBS患者外周血中T淋巴细胞亚群相对计数.结果 ①GBS患者急性期治疗后CD8+T细胞(28.77%±11.02%)和CD4+CD29+T细胞百分率(56.71%±12.44%)较治疗前(分别为31.84%±12.35%、62.40%±12.72%)显著降低(t=2.995、3.919,P＜0.05),CD4+/CD8+值和CD4+CD45RA+T细胞百分率较治疗前显著升高(t=2.368、3.860,P＜0.05),但治疗前后CD3+T细胞和CD4+T细胞百分率差异无统计学意义.②效优组CD8+T细胞和CD4+CD29+T细胞百分率较治疗前显著降低(t=2.144、3.343,P＜0.05),CD4+/CD8+值和CD4+CD45RA+T细胞百分率较治疗前显著升高(t=2.159、3.277,P＜0.05),效劣组T淋巴细胞亚群分布在治疗前后未发生明显变化.③在本组资料中,IVIG治疗急性期GBS 2周内的显效率为61.29%(19/31),无死亡病例.结论 急性期GBS患者在IVIG治疗前后发生了不同程度T淋巴细胞亚群分布的改变,为探讨GBS的发病机制及IVIG治疗GBS的机制提供了免疫学基础;IVIG治疗效果好,能够有效抑制病情进展,促进神经功能恢复.     

早期肠内营养联合合生元制剂对高血压脑出血患者细胞免疫功能的影响

目的 观察早期肠内营养联合合生元制剂对高血压脑出血患者细胞免疫功能和感染性并发症的影响. 方法 将53例高血压脑出血患者按照随机数字表法分为早期肠内营养组(对照组,26例)和早期肠内营养联合合生元制剂组(研究组,27例),对照组于伤后24～48 h采用肠内营养制剂(瑞素)开始营养支持;研究组则在对照组的基础上联合应用合生元制剂(金双歧,2 g,3次/d)14 d.2组患者在肠内营养营养支持0d、4d、8d和15 d分别检测外周血T淋巴细胞CD3+,T淋巴细胞亚群CD4+、CD8+和CD4+/CD8+,同时观察2组患者发生感染性并发症的差异. 结果研究组患者在营养支持的第8天,CD3+、CD4+、CD4+/CD8+的表达明显比对照组高,差异均有统计学意义(P＜0.05).在营养支持第15天,研究组CD3+、CD4+的表达仍比对照组明显升高,差异均有统计学意义(P＜0.05).研究组术后的感染性并发症的发生率低于对照组[33.33%(9/27)vs46.15%(12/26)],但差异无统计学意义(P＞0.05). 结论 与普通早期肠内营养相比,添加合生元制剂的早期肠内营养有利于促进高血压脑出血患者细胞免疫功能恢复和降低感染性并发症的发生率.     

银杏内酯K通过PI3K/Akt/mTOR通路促进血管生成改善缺血性卒中

目的 探究银杏内酯 K（Ginkgolide K ，GK）通过磷脂酰肌醇 -3- 激酶（PI3K）/ 蛋白激酶 B （Akt）/ 哺乳动物雷帕霉素靶蛋白（mTOR）信号通路对缺血性卒中小鼠血管内皮生长因子（VEGF）表达 和脑血管生成的作用。方法 将 40 只 C57BL/6 小鼠随机分为对照组、大脑中动脉闭塞（MCAO）组、低 剂量 GK（GK-L）组（3.5 mg/kg）、中剂量 GK（GK-M）组（7 mg/kg）和高剂量（GK-H）组（14 mg/kg），每组各 8 只。建立 MCAO 模型和氧葡萄糖剥夺发（OGD）体外模型；采用改良后的神经系统严重程度评分（mNSS） 评估法检测小鼠神经功能缺损；2，3，5- 三苯基氯化四氮唑（TTC）染色检测小鼠脑缺血面积；免疫荧 光染色检测小鼠梗死灶周围皮质血管生成和星形胶质细胞覆盖。培养 hCMEC/D3 细胞，分为对照组， OGD 组、OGD+CK 组、OGD+LY 组（LY 为 PI3K 信号通路抑制剂）和 OGD+GK+LY 组，采用细胞计数试 剂盒 -8（CCK-8）检测内皮细胞活性；Western blot 检测内皮细胞缺氧诱导因子 1α（HIF-1α）、VEGF 和 PI3K/Akt/mTOR 通路相关基因蛋白表达；细胞划痕实验检测内皮细胞迁移能力；血管形成实验检测内 皮细胞管腔形成能力。结果 与 MCAO 组［（5.37±1.25）分、（11.99±1.72）%］比较，GK-M 组和 GK-H 组 小鼠 mNSS 评分［分别为（3.37±1.32）、（2.23±0.38）分］和脑缺血面积［（4.75±0.89）%、（2.42±0.42）%］ 均 显 著 降 低（P＜ 0.05）。 与 MCAO 组 EdU+ /CD31+ 细 胞 数、EdU+ /GFAP+ 细胞数和星形胶质细胞覆盖 率［分 别 为（3.33±0.58）个、（4.33±1.53）个、（69.20±5.60）%］比 较，GK 组 小 鼠 EdU+ /CD31+ 细胞数 ［（13.67±2.08）个，t=3.576］、EdU+ /GFAP+ 细 胞 数［（8.33±1.53）个，t=6.008］和 星 形 胶 质 细 胞 覆 盖 率 ［（82.26±7.77）%］显著升高（均P＜ 0.05）。与对照组细胞活力、HIF-1α、VEGF 蛋白表达、Akt 和 mTOR 磷酸化水平［分别为（100.31±3.01）%、（0.09±0.03）、（0.13±0.03）、（0.20±0.04）、（0.18±0.03）］比较，OGD 组细胞活力［（52.37±9.06）%］、HIF-1α（0.17±0.02）和 VEGF 蛋白表达（0.18±0.03）、Akt 和 mTOR 磷酸 化水平［（0.28±0.06），（0.38±0.05）］均显著升高（均P＜ 0.05）；与 OGD 组比较，OGD+GK 组细胞 HIF-1α （0.22±0.03）和 VEGF 蛋白表达（0.23±0.03）、Akt 和 mTOR 磷酸化水平［（0.48±0.09），（0.52±0.05）］、细胞 活力［（61.07±3.48）%］、迁移率［（85.26±11.03）%］和管状结构数量［（81.97±5.79）%］均显著升高（均 P＜0.05），OGD+LY组则表现出相反变化；PI3K信号通路抑制剂LY294002可逆转GK对OGD细胞的影响。 结论 GK 通过 PI3K/Akt/mTOR 信号通路上调 VEGF 表达促进脑血管生成，改善小鼠缺血性卒中。     

动脉瘤蛛网膜下腔出血患者外周血淋巴细胞程序性死亡受体1表达特性及意义的研究

目的通过观察动脉瘤蛛网膜下腔出血(aSAH)患者外周血淋巴细胞程序性死亡受体1(PD-1)的表达,探索其在aSAH患者中的表达特性及意义。方法选取aSAH患者共计21例,根据Glasgow昏迷量表(GCS)评分分为有神经症状组(GCS15分,10例)和无神经症状组(GCS=15分,11例)。另选取6名健康志愿者作为正常对照组。使用流式细胞仪检测PD-1在各组患者外周血CD4~+和CD8~+ T淋巴细胞的表达情况,分析其特性及意义。结果无神经症状组PD-1~+CD4~+T淋巴细胞百分率和PD-1~+CD8~+ T淋巴细胞百分率均明显高于正常对照组,明显低于有神经症状组(均P0.05)。aSAH患者PD-1~+CD4~+ T淋巴细胞百分率和PD-1~+CD8~+ T淋巴细胞百分率与GCS评分密切相关,Spearman系数分别为0.87和0.83(均P0.05)。aSAH患者中,有神经症状组肺部感染发病率明显高于无神经症状组(P0.05),且有肺部感染组PD-1~+CD4~+ T淋巴细胞百分率和PD-1~+CD8~+ T淋巴细胞百分率明显高于无肺部感染组(均P0.05)。结论aSAH患者,特别是有神经症状的患者,CD4~+和CD8~+ T淋巴细胞PD-1表达强度明显增加,其表达强度增加与aSAH后神经症状严重程度和出血后肺部感染有相关性。     

Denervation study of synapses of organ of corti of old world monkeys

Fine structural characteristics of synapses in the spiral organ of Corti were examined, with reference to differences between inner and outer haircell systems, and to location of neurons of origin of efferent axons. Surgical interruption of crossed olivocochlear bundle, of vestibular nerve, of facial nerve, and excision of superior cervical ganglia were used to determine the pathways of efferent axons. Interruption of the vestibular nerve near the brainstem results in degeneration of all efferent terminals on outer hair cells. Mid-line lesions at, and caudal to, the facial colliculus result in degeneration of about half of these efferent terminals. Efferent synaptic bulbs to the inner hair-cell system are small, of the order of one micron, and form type 2 junctions with afferent dendrites. They tend to have more large dense-core vesicles (about 80 nm) than the large efferent terminals of the outer hair-cell system, and appear to be the terminals of axons in the habenula perforata, which exhibit varicosities laden with large dense core vesicles. The varicosities are unaffected by excision of the superior cervical ganglia. So far as our material can reveal, it appears that the varicosities in the habenula perforata do not survive vestibular root interruption, nor do the efferent processes in the internal spiral bundle or at the base of inner hair cells. Most interestingly, the afferent processes of the inner hair-cell system, as identified for example by their relation to pre-synaptic bodies in the inner hair cells, are subject to a trans-synaptic reaction after severance of the vestibular root. They undergo a dramatic cytological transformation, characterized by increase of volume, engorgement with microtubules, microfilaments, microvesicles of various sizes, and clusters of lysosomes. Thus, both the efferent and afferent terminals of the inner hair-cell system show marked cytological differences from the corresponding terminals of the outer hair cell system.     

Mechanism of action of tubocurarine on nicotinic cholinoreceptors of neurons of the sympathetic ganglia of rats

Tubocurarine (Tc) effect on membrane currents elicited by acetylcholine (ACh) was studied in isolated superior cervical ganglion neurons of rat using patch-clamp method in the whole-cell recording mode. The "use-dependent" block of ACh current by Tc was revealed in the experiments with ACh applications, indicating that Tc blocked the channels opened by ACh. Mean lifetime of Tc-open channel complex, tau, was found to be 9.8 +/- 0.5 s (n = 7) at -50 mV and 20-24 degrees C. tau exponentially increased with membrane hyperpolarization (e-fold change in tau corresponded to the membrane potential shift by 61 mV). Inhibition of the ACh-induced current by Tc (3-30 microM/1) was completely abolished by membrane depolarization to the level of 80-100 mV. Inhibition of ACh-induced current was augmented at increased ACh doses. It is concluded that the open channel block produced by Tc is likely to be the only mechanism for Tc action on nicotinic acetylcholine receptors in superior cervical ganglion neurons of rat.     

The effect of age of onset of PD on risk of dementia

Background Dementia occurs in the majority of patients with Parkinson’s disease (PD). Late onset of PD has been reported to be associated with a higher risk for dementia. However, age at onset (AAO) and age at baseline assessment are often correlated. The aim of this study was to explore whether AAO of PD symptoms is a risk factor for dementia independent of the general effect of age. Methods Two community-based studies of PD in New York (n = 281) and Rogaland county, Norway (n = 227) and two population-based groups of healthy elderly from New York (n = 180) and Odense, Denmark (n = 2414) were followed prospectively for 3–4 years and assessed for dementia according to DSM-IIIR. All PD and control cases underwent neurological examination and were followed with neurological and neuropsychological assessments. We used Cox proportional hazards regression based on three different time scales to explore the effect of AAO of PD on risk of dementia, adjusting for age at baseline and other demographic and clinical variables. Findings In both PD groups and in the pooled analyses, there was a significant effect of age at baseline assessment on the time to develop dementia, but there was no effect of AAO independent of age itself. Consistent with these results, there was no increased relative effect of age on the time to develop dementia in PD cases compared with controls. Interpretation This study shows that it is the general effect of age, rather than AAO that is associated with incident dementia in subjects with PD. Received in revised form: 22 December 2005     

Limits of deinstitutionalization--perspectives of relatives of psychiatric patients

After a hopeful beginning, the social process of the reintegration of those with severe mental illness has come to a standstill. I am led to wonder whether "the community" really wants to live together with people suffering from severe mental illness, and if so, how closely? As long as the medical treatment of mental illness provided by the general practitioners is fundamentally deficient, as they are not able to prescribe the necessary interventions--such as out-patient psychiatric nursing, and service providers in the out-patient sector are content with offering increasingly intensive forms of care for the less seriously ill at the cost of the Social Welfare System--the reintegration of those with serious mental illness remains an illusion--which is mainly to the benefit of providers of residential care in homes and hostels.