经颅磁刺激安全性的实验研究

目的 :探讨重复经颅磁刺激的安全性。方法 :选用 3 6只健康Wistar大鼠 ,随机分为对照组 ,低频刺激组和高频刺激组 ,对低频组和高频组分别给予不同频率和强度的经颅磁刺激 ,然后对各组大鼠的行为、组织病理形态学、血清髓鞘碱性蛋白 (MBP)及神经元特异性烯醇化酶 (NSE)含量进行观察。结果 :低频刺激组 (5Hz)和高频刺激组 (2 0Hz)在刺激过程中均未出现异常活动 ,无肢体强直、阵挛等 ,脑组织形态学包括大体观察、普通光镜及电镜改变不明显 ,其血清MBP和NSE含量与正常对照组比较 ,差异无显著性意义 (P >0 0 5 )。结论 :在一定强度和频率内经颅磁刺激是一种比较安全的方法     

中枢神经特异蛋白和神经元特异性烯醇化酶水平对急诊高血压脑出血患者预后预测的价值

目的:探讨中枢神经特异蛋白(Soluble protein-100β,S100β)和神经元特异性烯醇化酶(Neuron specific enolase,NSE)水平预测急诊高血压脑出血患者预后的价值。方法:回顾性分析我院2018年04月至2019年07月期间急诊入院的190例高血压脑出血患者血清S100β蛋白和NSE水平,统计患者入院后30天的病死率。采用Pearson分析血清S100β蛋白、NSE水平与预后的相关性;以ROC曲线评估血清S100β蛋白和NSE水平单独和联合预测高血压脑出血患者的预后。结果:Pearson相关性分析结果显示患者血清S100β蛋白与NSE含量呈正相关(r=0.16,P0.05)。ROC曲线显示:S100β、NSE均可单独预测患者预后,而预测的准确性依次为:S100β(S100β+NSE)=NSE。结论:S100β、NSE作为生物学指标可以预测患者预后,S100β准确性更高。     

急性颅脑损伤患者血清BNP和NSE含量的变化及临床意义

目的：探讨急性脑损伤患者血清脑钠肽（BNP）和神经元特异性烯醇化酶（NSE）含量的变化及其机理和临床意义。方法：采用放射免疫分析对134例急性颅脑损伤患者和40例健康对照者血清BNP和NSE进行测定,并对结果进行统计学分析。结果：颅脑损伤后血清BNP含量显著低于正常对照组,且伤情越重,BNP浓度越低;NSE含量的变化正好与之相反,且两者之间存在明显的负相关（r=-0.674,P〈0.01）。结论：血清BNP和NSE有助于估计急性颅脑损伤患者病情轻重及预后。     

S-100B、NSE和MBP对急性脑出血预后的临床评估

目的:研究脑出血后血清S-100B蛋白、神经特异性烯醇化酶(NSE)和碱性髓鞘蛋白 (MBP)浓度在预后评估中的价值。方法:对40例急性脑出血住院病人在病后72 h内进行血清S- 100B、NSE和MBP浓度检测,并结合格拉斯哥昏迷量表(GOS)评分进行比较分析。结果:本组40例病人脑出血后血清S-100B、NSE和MBP浓度均显著高于正常对照组,而且病人中不同预后组之间S- 100B、NSE和MBP浓度存在显著差异。分别以脑出血后72 h血清S-100B浓度2．0μg／L,NSE浓度 30μg／ml和MBP浓度10 μg／ml为临界标准评估顶后,S-100B的特异度为91％,敏感度为72％;NSE 特异度为77％,敏感度为67％;MBP特异度为63％,敏感度为61％。结论:脑出血后血清S-100B蛋白、NSE和MBP浓度对评估脑出血病人预后具有较高的特异性和敏感性。     

脑白质损伤早产儿血清IL-6及NSE的变化及其临床意义

目的了解血清白细胞介素-6（IL-6）及神经元特异性烯醇化酶（NSE）的变化对脑白质损伤早产儿早期诊断的价值。方法观察组为脑白质损伤早产儿35例,对照组为正常早产儿35名。采用ELISA分别于生后1、7、14 d检测两组血清IL-6、NSE水平。结果对照组患儿生后第1、7及14天血清IL-6水平分别为（19.14±1.18）pg/ml、（19.14±1.14）pg/ml及（19.11±1.34）pg/ml,观察组患儿分别为（25.19±3.03）pg/ml、（24.48±2.97）pg/ml及（23.74±2.95）pg/ml,观察组显著高于对照组,差异有统计学意义（P均=0.000）。对照组患儿生后第1、7及14天血清NSE水平分别为（4.70±0.36）ng/ml、（4.31±0.29）ng/ml及（4.14±0.30）ng/ml,观察组患儿分别为（6.30±0.89）ng/ml、（6.05±0.86）ng/ml及（5.64±0.75）ng/ml,观察组显著高于对照组,差异有统计学意义（P均=0.000）。结论脑白质损伤早产儿血清IL-6和NSE的浓度明显高于对照组。监测早产儿血清IL-6和NSE水平,对脑白质损伤的诊断和治疗效果的评价具有一定临床价值。     

急性颅脑外伤患者血清NSE含量变化及临床意义

目的：探讨急性颅脑外伤患者血清神经元特异性烯醇化酶（Neuron-specificenolase,NSE）的含量及动态变化对颅脑损伤的诊断和评估伤情、观察疗效、预后判断的临床价值。方法：采用发光免疫分析,对60例急性颅脑外伤患者及62例正常人血清NSE含量进行同步检测。并于治疗前和治疗后3d、1周、2周、3周NSE的含量进行动态观察。对38例不同预后的急性颅脑外伤患者治疗前后的血清NSE浓度及GCS评分进行回顾分析。结果：急性颅脑外伤患者血清NSE含量显著增高,与正常对照组比较差异具有极显著意义（P〈0.001）,其NSE增高的程度,重型组〉中型组〉轻型组（P〈0.01）,与GCS评分密切负相关（r=-0.9140,P〈0.01）。动态观察显示56例不同程度的颅脑外伤患者治疗后血清NSE浓度随着治程的延续而逐渐下降,还发现急性颅脑外伤患者的血清NSE浓度如果高滴度持续增高,则预后不佳,反之则预后相对较好。而且NSE浓度下降越迅速,恢复正常的时间越早,其预后越佳。结论：血清NSE含量反映脑细胞损伤的程度,是一项灵敏的量化指标。进行NSE含量的检测不仅对脑损伤的诊断和评估伤情富有临床价值,而且对于疗效观察及预后的早期判断均有帮助。     

ACI患者治疗前后血清NSE和hs-CRP检测的临床意义探讨

目的：探讨急性脑梗死（ACI）患者治疗前后血清神经元特异性烯醇化酶（NSE）和超敏C-反应蛋白（hs-CRP）水平的变化及临床意义。方法：应用发光免疫分析和免疫荧光定量技术对51例ACI患者治疗前后血清NSE和hs-CRP水平进行动态检测,并与98例正常人的检测结果作比较。结果：治疗前ACI患者血清NSE和hs-CRP水平均显著高于正常对照组（P〈0.001）,发现大面积脑梗死组治疗前血清NSE和hs-CRP水平〉中面积组（P〈0.01）〉小面积组（P〈0.01）。且ACI患者治疗前血清NSE和hs-CRP的含量与梗死灶面积密切相关（r（1）=0.8811,P〈0.01;r（2）=0.7913,P〈0.01）。治疗1周血清NSE和hs-CRP水平均显著下降（P1〈0.05,P2〈0.01）,治疗后2周hs-CRP水平与正常对照组差异无统计学意义（P〉0.05）,NSE水平仍明显高于正常对照组（P〈0.01）。结论：检测ACI患者血清NSE和hs-CRP水平的变化不仅有助于脑梗死的诊断和病情及预后的判断,而且对于疗效的评估、指导临床实践亦有价值。     

颅脑损伤患者血清S-100B和神经元特异性烯醇化酶的检测

目的探讨颅脑损伤患者血清S鄄100B和神经元特异性烯醇化酶(NSE)的变化及其临床意义。方法用双抗体夹心ELISA法检测30名健康人和108例颅脑损伤患者血清S鄄100B和NSE水平。结果108例颅脑损伤患者在伤后12h血清S鄄100B水平[(1.24±0.32)滋g/L]较对照组[(0.38±0.12)滋g/L]升高,重型、中型和轻型颅脑外伤患者的血清NSE浓度均明显高于对照组的NSE浓度(P<0.01);而不同组别颅脑外伤患者血清NSE水平差异亦有显著性(P<0.05)。结论血清S鄄100B和NSE检测可作为诊断早期脑损伤的指标之一。     

创伤性脑损伤后大鼠外周血CD8的变化

目的:探讨外周血CD8在创伤性脑损伤后的变化特点。方法:改良Feeney's自由落体打击法制备创伤性脑损伤模型;ELISA检测大鼠血清中神经元特异性烯醇化酶(NSE)和CD8的含量,双变量分析两者的相关性;Western blot法检测大鼠血液淋巴细胞FasL蛋白的表达。结果:颅脑外伤后大鼠血清CD8升高滞后于NSE。大鼠伤后1 d血清NSE水平即显著升高,伤后3 d达到峰值,随后逐渐下降;大鼠血清CD8在伤后3 d开始升高,峰值出现在伤后7 d,随后逐渐下降。统计分析显示伤后1 d、3 d、7 d的血清的NSE与伤后3 d、7 d、14 d的血清CD8含量变化呈现正相关。但伤后不同时间大鼠血液淋巴细胞FasL蛋白表达的差异均无统计学显著性。结论:创伤性脑损伤后释放入血的NSE刺激机体免疫应答,引起外周血CD8增多,推测其可能与创伤性脑损伤后继发性损伤的免疫应答有关。     

The development of immunoreactivity for neuron-specific enolase of preoptic and septal neurons in dissociated cultures

The immunocytochemical visualization of neuron-specific enolase, which is a marker protein for differentiated neurons, was applied to follow the differentiation of preoptic and septal neurons in dissociated cultures. From 4 to 24 daysin vitro, the relative numbers of stained neurons were counted and the staining intensity of individual neurons determined by absorbancy measurements using a television-based densitometer. Whereas few stained cells could be observed at 4 DIV, 80% of the neurons were neuron-specific enolase-positive at 13 daysin vitro. This value remained constant up to 24 daysin vitro. The density of the immunoreaction product increased dramatically from 13 to 17 daysin vitro and was still higher at 24 daysin vitro. The glial and ependymal cells of the carpet, as well as neuroblasts, remained unstained.Comparison with morphological observations and immunocytochemical demonstration of neuronal peptides made earlier shows that expression of neuron-specific enolase closely parallels neuronal differentiation. These observations indicate that cultures derived from preoptic and septal neurons represent a viable model system for the study of neuronal maturationin vitro.     

The contribution of postnatal steroid administration to early brain damage in preterm babies with bronchopulmonary dysplasia

Background/aimPostnatal corticosteroids are commonly used to treat bronchopulmonary dysplasia (BPD). We aimed to show whether S100 calcium-binding B (S100B), neuron-specific enolase (NSE), Tau protein or microtubule-associated protein tau (MAPT), and glial fibrillary acid protein (GFAP) levels would provide any evidence of early neurological damage in premature infants receiving postnatal low dose dexamethasone therapy for BPD treatment.Materials and methods In this cohort study, 136 preterm infants diagnosed with BPD at ≤32 weeks of gestation formed the study group, and 64 preterm infants formed the control group. NSE, S100B, GFAP, and MAPT levels were first measured before the postnatal corticosteroid treatment in both the patient and the control group on the 28th day and, for a second time, after treatment termination in the patient group.Results There were significant differences between the measured GFAP, MAPT, and NSE values of the BPD and control groups on the 28th day, whereas there was no significant difference between the measured S100B values of the two groups. There were a statistically significant difference between the NSE values measured on the 28th day and after the treatment within the BPD group, whereas no significant difference existed between the GFAP, MAPT, and S100B values.Conclusion NSE levels, which indicate brain damage in the early period, increased in preterm babies with BPD who had been administered postnatal dexamethasone.     

Immunohistochemical expression of neural tissue markers (neuron-specific enolase, glial fibrillary acidic protein, S100 protein) in ameloblastic fibrodentinoma: a comparative study with ameloblastic fibroma

Formalin-fixed paraffin-embedded sections of three cases of ameloblastic fibrodentinoma (AFD) were studied by the avidin-biotin-peroxidase complex method using antibodies against neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and S100 protein and the results were compared with those in ameloblastic fibroma (AF). A striking histopathological characteristic of AFD was the formation of abortive dentin with various degrees of maturation at the epithelial-mesenchymal tissue interface. Central cells of enamel organ-like epithelia with various stages of abortive dentin induction in AFD were generally positive for NSE. Dental lamina-like epithelial cells also showed positive staining in some areas. No cells were positive for NSE in AF. Positive staining for GFAP was observed in the juxta-epithelial mesenchymal tissue of the formation stage of immature dentin with various numbers of entrapped cells in AFD, but GFAP staining was negative in other mesenchymal and epithelial tissues at other stages. In AF, no GFAP-positive cells were found. There were a few S100 protein-positive cells found in the foci of epithelial components in both AFD and AF. Mesenchymal cells showing a dendritic or spindle shape were positive for S100 protein in some areas of AFD and AF. Although such cells in the mesenchymal component of pigmented AFD were more numerous than in non-pigmented AFD and AF, their distribution pattern in the former condition was basically similar to that in the latter. Although the present results, obtained from conventional immunohistochemical procedures, do not directly reflect the expression of neural crest-derived cells in the dentinogenesis of AFD, such results do not disprove the possibility of the expression of neural proteins probably related to neural crest-derived cells in dentinogenesis under certain pathologic conditions in odontogenic mixed tumors. Such a phenomenon may also occur during dentinogenesis in other odontogenic mixed tumors and in normal tooth differentiation, but at an undetectable level.     

NSE在慢性砷中毒大鼠海马CA3区的表达

目的:利用神经元特异性烯醇化酶(NSE)免疫组化染色,光镜下观察慢性砷中毒对成年大鼠海马CA3区神经元形态学影响,以及慢性砷中毒对成年大鼠脑的神经毒性。方法:取3～4月龄SD大鼠60只,随机分为正常对照组(自由饮用蒸馏水)、低剂量组(As2O3按4mg/l的比例用蒸馏水配制,自由饮用)和高剂量组(As2O3按100mg/l的比例用蒸馏水配制,自由饮用)。每组20只,各组均用普通饲料喂养,连续喂养4月后取大鼠海马组织,常规石蜡切片尼式染色(Nissl),并利用神经元特异性烯醇化酶(NSE)抗体进行SABC法免疫组化染色,观察大鼠海马CA3区神经元的形态学改变,进行细胞形态学计数及测量NSE阳性反应产物的平均光密度。结果:砷中毒组神经元数量减少,细胞形态不规则,可见大量坏死的神经细胞,NSE免疫组化染色阳性细胞减少(P0.01),阳性反应产物平均光密度降低(P0.01)。结论:慢性砷中毒可以导致大鼠海马CA3区神经元结构损伤。     

Subepithelial neuroendocrine cells and carcinoid tumours of the human small intestine and appendix. A comparative immunohistochemical study with regard to serotonin, neuron-specific enolase and S-100 protein reactivity

A comparative immunocytochemical study was performed of subepithelial neuroendocrine cells of the human small intestine and appendix and carcinoid tumours of these sites, using a monoclonal antibody to serotonin and polyclonal antisera against neuron-specific enolase (NSE) and S-100 protein. Subepithelial neuroendocrine cells were easily identified in the lamina propria of the appendix. These cells, which sometimes occurred in aggregates, displayed serotonin and NSE immunoreactivity and were surrounded by S-100 protein immunoreactive cells, presumably of Schwann cell origin. In the appendix scattered cells with corresponding morphological features and immunoreactivity were also observed deep in the submucosa. In addition, subepithelial neuroendocrine cells were sparsely present in the lamina propria of the small intestine, occurring only as single cells in the deeper part of the mucosa below or between the epithelial crypts. Most appendiceal carcinoid tumours (11 of 12 examined cases) were biphasic and consisted of neuroendocrine tumour cells with intermingled S-100 protein immunoreactive cells (Schwann cells) with long cytoplasmic extensions. However, small intestinal (11 cases) and caecal (10 cases) carcinoids lacked S-100 protein immunoreactive cells as an integral component. The results indicate that the appendiceal carcinoids are mostly closely related structurally to the subepithelial neuroendocrine and Schwann cell aggregates of the lamina propria and are thus presumed to be histogenetically related to this cell system, while the histogenesis of small-intestinal and caecal carcinoids remains less clear.     

Time course of serum neuron-specific enolase levels from infancy to early adulthood in a female patient with beta-propeller protein-associated neurodegeneration

Beta-propeller protein-associated neurodegeneration (BPAN), a subgroup of neurodegeneration with brain iron accumulation, is typically characterized by non-progressive global developmental delay and seizures in childhood, followed by progressive neurological decline with parkinsonism and dementia in adolescence or early adulthood. It is difficult to clinically identify a patient with BPAN in childhood. Recent studies reported that serum levels of neuron-specific enolase (NSE) were elevated in children with BPAN. We reviewed the time course of serum NSE levels in a 21-year-old female patient genetically diagnosed (a de novo WDR45 variant c.268A > T) with BPAN, which was suspected based on prolonged elevation of serum NSE. There was an overall tendency for serum NSE levels to decrease in a stepwise fashion. The peak serum NSE level was observed during the first 2 years of age and then decreased rapidly in 1 year. High serum NSE levels persisted between 3 and 11 years of age. Subsequently, serum NSE levels decreased and plateaued after 13 years of age. There were tendencies for both blood AST and LDH levels to decrease over time in parallel with serum NSE levels. Serum NSE levels may be a diagnostic biomarker of BPAN in children but becomes of less value in identifying a patient with BPAN after childhood.     

流式荧光法检测NSE的应用评价

建立流式荧光法定量检测神经元特异性烯醇化酶(NSE)的正常参考值,评估其在肺癌诊断中的应用价值。采用流式荧光法检测176例临床血清样本的NSE水平,并与罗氏Elecsys电化学发光法检测结果进行比较,确定两种方法之间的可比性。进一步采用流式荧光法检测668例健康者血清NSE水平,建立健康人群的NSE正常参考值。最后,以流式荧光法检测242例肺癌患者、108例肺部良性疾病者和147例健康者血清NSE水平,评价流式荧光法检测NSE的应用。结果显示,176例临床样本的检测结果表明流式荧光法和电化学发光法相关性良好(Y=0.9746X-0.2075,R2=0.9653),二者具有可比性。根据668例健康者检测结果的第95%百分位数(17.07ng/ml)确定流式荧光法的NSE正常参考值,与罗氏Elec-sys 2010电化学发光法的相似。以NSE>17.07ng/ml判断为阳性,NSE在小细胞肺癌患者中的阳性率显著高于肺鳞癌、肺腺癌患者(P<0.01)。建立了流式荧光法检测NSE的正常参考值,并初步评估了流式荧光法检测NSE在肺癌诊断中的应用,为流式荧光法检测NSE的合理应用和广泛推广奠定了基础。     

Immunohistochemical and Ultrastructural Analysis of Bronchopulmonary Neuroendocrine Neoplasms. I. Carcinoids

Twenty-five strictly defined bronchopulmonary carcinoids were studied by light microscopic immunohistochemistry by the peroxidase technique for NSE (neuronspecific enolase), serotonin, and a broad spectrum of neuropeptides. Eighteen cases were also studied by electron microscopy.

Twenty-three of the twenty-five cases showed immunostaining for NSE; 24 cases displayed immuno-staining for at least two of the hormones tested for; the single case that failed to show hormonal immuno-reactivity was however positive for NSE and had granules by electron microscopy. Serotonin was the most frequently demonstrated hormone followed by bombesin, vasoactive intestinal peptide, gastrin, leuenkephalin, alphamelanocyte stimulating hormone, somatostatin, substance P, and calcitonin. In several cases, adjacent-step sections stained for different hormones strongly indicated immunoreactivity for more than one hormone in single neoplastic cells.

By electron microscopy, all 18 cases studied showed generally abundant neurosecretory granules, which, however, displayed considerable heterogeneity in their size, shape, and density. Twelve of these eighteen cases displayed evidence of squamous differentiation and 10 showed characteristic exocrine lumina.

The capability of single neuroendocrine tumors and single neuroendocrine tumor cells to produce more than one immunoreactive hormone is hereby amply confirmed; these broad capabilities are certainly reflected in the heterogeneous granule populations seen by electron microscopy. The synchronous presence of squamous and exocrine features in bronchopulmonary carcinoids indicates that they too are capable of multidirectional differentiation, which should not detract from their being regarded basically as well-differentiated neuroendocrine neoplasms. The clinical significance of strictly defining bronchial carcinoids is underscored by the fact that of 25 cases followed for 2-13 years, only one developed metastases 9 years postoperatively.     

Paragangliomas of the Head and Neck: Ultrastructural and Immunohistochemical Analysis

Eighteen head and neck paragangliomas were studied by light microscopy and light microscopic immunohistochemistry by the peroxidase technique for the presence of NSE (neuron-specific enolase), serotonin, and a battery of neuropeptides. Seven of these tumors were also studied by electron microscopy.

All 18 cases demonstrated immunostaining for NSE; 10 of the 11 carotid body tumors had immunostaining for multiple hormones. Considering all 18 cases, the most frequently demonstrated hormonal substances were in order: serotonin, leuenkephalin, gastrin, substance P, vasoactive intestinal polypeptide (VIP), somatostatin, bombesin, calcitonin, and alpha MSH. In several tumors, adjacent-step sections stained for different hormonal substances strongly suggested reactivity for more than one hormone in given tumor cells.

By electron microscopy, all 7 cases studied displayed considerable heterogeneity of the neurosecretory granules with respect to size, shape, and electron density. This demonstrated that branchiomeric paragangliomas are capable of producing a spectrum of neuropeptides in addition to their known amine content. The presence of immunoreactive serotonin in most of these neoplasms was confirmed. In addition to these findings, neurofibrils within the substance of carotid body paragangliomas demonstrated immunoreactivity for somatostatin and a gastrinlike neuropeptide. The significance of the neuropeptides in these neoplasms and their possible presence and role in normal and hyperplastic paraganglia remain to be defined.     

冠状动脉搭桥术患者围术期血浆神经元特异性烯醇化酶浓度与术后认知功能

目的研究冠状动脉搭桥术患者围术期血浆神经元特异性烯醇化酶（NSE）浓度及术后认知功能。方法40例接受冠状动脉搭桥术的男性患者，分为体外循环组和非体外循环组，每组20例。在术前、术后6、12、18和24h采血，用放射免疫方法测定血浆NSE浓度。在术前1天和术后7～10天进行神经心理测试。结果体外循环组患者术后6h血浆NSE浓度比术前显著升高，同时高于非体外循环组；非体外循环组患者各时点无差别。非体外循环组患者在数字广度测验（逆向）、斯特鲁字色干扰测验改正反应和阻塞反应的结果显著优于体外循环组患者。体外循环组患者术后6hNSE水平与连线测验A型相关，术后12hNSE浓度与数字广度测验（顺向）、连线测验A型、斯特鲁字色干扰测验时间分项相关。结论心脏手术后血浆NSE浓度的升高可在某种程度上反映术后认知功能障碍，但在具体应用时应考虑NSE的组织多源性及体外循环因素的影响。