Age-related changes of neuron-specific enolase, S-100 protein, and myelin basic protein concentrations in cerebrospinal fluid.

Studies on cerebrospinal fluid (CSF) concentrations of neuron-specific enolase (NSE), S-100 protein, and myelin basic protein (MBP) in patients with neurological lesions indicate a quantitative relation between the degree of cell damage in the central nervous system (CNS) and the concentration of these CNS-specific proteins in CSF. Thus NSE, S-100, and MBP could be of use as markers for destructive processes in the CNS. We collected 937 specimens of CSF from children and adults (from newborns to age 91 years) who were undergoing a diagnostic lumbar puncture for several clinical indications. Of these, 79 samples from subjects ranging in age from 0.7 to 66 years could be used retrospectively to construct a reference interval according to our criteria. In these 79 samples no sex dependency existed. The relative increase of NSE, S-100, and MBP with age was similar (1% per year), suggesting a common underlying mechanism. These results emphasize the necessity of using age-matched reference values when the CNS-specific proteins are to be evaluated in neurological diseases. We also present three case histories to discuss the possible clinical relevance of the measurement of NSE, S-100, and MBP in children and adults.     

肠道病毒71型脑炎患儿血清和脑脊液神经元特异性烯醇化酶、S-100蛋白及髓鞘碱性蛋白的测定

目的 探讨肠道病毒71型（enterovirus A group type 71,EV71）脑炎患儿血清和脑脊液（Cerebrospinal fluid,CSF）中神经元特异性烯醇化酶（NSE）、S-100蛋白和髓磷脂碱性蛋白（MBP）的水平变化及其临床意义。方法 依据卫生部《肠道病毒71型（EV71）感染重症病例临床救治专家共识》的诊断标准,将患儿分为普通病例组、重症病例组（再分为重症重型组和重症危重型组）,同时设立对照组。应用双抗体夹心酶联免疫吸附测定（ELISA）法分别检测各组患儿不同时期血清、CSF中S-100和MBP的水平;采用电化学发光法检测各组患儿不同时期血清、CSF中NSE的水平。结果 重症危重型组和重症重型组中,血清和CSF中NSE以及S-100水平均显著高于普通病例组和对照组,差异有统计学意义;重症危重型组CSF中NSE、S-100水平也高于重症重型组,差异有统计学意义;普通病例组血清和CSF中NSE、S-100水平与对照组差异无统计学意义。重症病例组急性期血清和CSF中NSE、S-100含量均显著高于恢复期和对照组,差异有统计学意义,但恢复期血清和CSF中NSE、S-100含量与对照组差异无统计学意义;各组血清和CSF中MBP水平差异无统计学意义。重症病例组血清与CSF中NSE和S-100的含量均呈正相关（r=0.886,P0.01; r=0.875, P0.01）。结论 检测EV71脑炎患儿血清和CSF中NSE、S-100水平,可评估其脑损伤程度及预后。     

急性CO中毒后迟发性脑病神经元特异性烯醇化酶测定的临床意义

目的　探讨急性CO中毒后迟发性脑病 (DEACMP)患者脑脊液 (CSF)和血清神经元特异性烯酶化酶(NSE)的变化及临床意义。方法　对 49例DEACMP患者均在急性期入院 3d内抽取CSF和血标本进行NSE测定 ,其中 34例于治疗 30d后再次采取标本复查 ,对比分析治疗前后NSE的变化。结果　患者组急性期CSF和血清NSE含量较对照组明显增高 (P <0 .0 0 1) ;治疗后明显下降 (P <0 .0 0 1,P <0 .0 0 5 ) ,与对照组比较差异无显著性 (P >0 .0 5 ) ;患者组CSF和血清NSE呈明显正相关 (r =0 .46 14,P <0 .0 0 1)。结论　CSF和血清NSE测定可作为DEACMP诊断和病情判断的有效指标。     

Clinical relevance of increased neuron-specific enolase concentration in cerebrospinal fluid

Neuron-specific enolase (NSE) concentrations in cerebrospinal fluid (CSF) and serum have been studied by an EIA-method using monoclonal antibodies against human NSE. In a control group (n = 24) the mean NSE value (+/- SD) in CSF was 10.8 (+/- 4.5) ng/ml. Increased NSE values in CSF (greater than or equal to 20 ng/ml, ie greater than or equal to means + 2s) have been detected in 33/172 patients with the following neurological diseases: CNS tumors (6/30), infarctions (6/36), cerebral ischemias (5/25), inflammatory diseases (7/33), epilepsias (3/10) and miscellaneous neurological diseases (6/38). The NSE assay in CSF was not specific for a single neurological disease. In 9% of all patients with an organic neurological disease the increased NSE concentration was the only abnormal result in the CNS out of variables routinely determined in CSF. The discrimination between an organic and psychogenic origin of epilepsy may be possible by an NSE analysis in CSF. The NSE assay in CSF can be recommended as an unspecific screening parameter for pathological organic CNS processes.     

标本溶血对电化学发光免疫法结果的影响

目的探讨标本不同程度溶血对电化学发光免疫法(ECLIA)测定结果的影响。方法将血液标本人为干预成不同程度的溶血标本,使用ECLIA检测各组标本中的甲状腺功能、肿瘤标志物、内分泌激素等共28项指标的浓度并做分析。结果标本溶血可使胰岛素(INS)浓度降低,使叶酸(FA)、神经元特异性烯醇化酶(NSE)浓度升高,重度溶血有使铁蛋白测定结果偏高的趋势,但差异无统计学意义,其余指标测定结果不受溶血的影响。结论除某些项目外大部分溶血标本的ECLIA测定结果没有受到影响。测定INS、FA、NSE时应绝对避免使用溶血标本,以确保结果准确。     

Cerebrospinal fluid and serum neuron-specific enolase in acute benign headache

We determined the cerebrospinal fluid (CSF) and serum neuron-specific enolase (NSE) concentrations in 19 patients with acute benign headache. All patients had normal neurological examination, CSF and head computed tomography scan. The final diagnoses were: primary thunderclap headache ( n  = 7), primary exertional headache ( n  = 3), primary cough headache ( n  = 1), migraine without aura ( n  = 4), headache unspecified ( n  = 2), probable infrequent episodic tension-type headache ( n  = 1), headache attributed to hypertensive crisis without hypertensive encephalopathy ( n  = 1). A group of 108 healthy subjects served as controls. CSF NSE concentration was 14.16 ng/ml [95% confidence interval (CI) 11.86, 16.47)] in the headache sample (controls 17.19 ng/ml, 95% CI 16.23, 18.15). Serum NSE concentration was 7.50 ng/ml (95% CI 5.20, 9.80) in the headache sample (controls 8.45 ng/ml, 95% CI 7.67, 9.23). CSF/serum ratio was 2.81 (95% CI 2.21, 3.40) in the headache sample (controls 2.23, 95% CI 2.03, 2.42). Acute benign headache is not associated with neuronal damage as estimated by means of CSF and serum NSE concentration.     

Factors affecting the accuracy and reliability of the measurement of anti-Müllerian hormone concentration in the clinic

ObjectiveWe aimed to identify the factors that influence serum anti-Müllerian hormone (AMH) concentration measurements.MethodsWe collected serum samples between May and September 2018 and compared the effect on AMH concentration measured by ELISA of conditions including venepuncture, storage time, storage temperature, locations of the reaction microplate, and the use of the oral contraceptive pill and gonadotrophin-releasing hormone (GnRH).ResultsAMH concentration was not affected by food intake but was affected by haemolysis. It was also much higher in samples on the edge of the ELISA microtitre plate. AMH concentration increased after incubation at room temperature for 1 day, 4°C for 3 days, −20°C for 1 month and −40°C for 4 months, but no change occurred during storage at −80°C for 9 months. AMH concentration was high in patients following GnRH agonist treatment but was not affected by oral contraceptives.ConclusionsNo fasting is required prior to AMH measurement. Placement of serum samples on the edge of microtitre plates affects the results of the AMH ELISA. If serum samples cannot be assayed immediately, it is best to store them at −80°C. Basal AMH concentration cannot be used as a measure of ovarian reserve after GnRH agonist treatment.     

多梗死性痴呆患者脑脊液肿瘤坏死因子和神经元特异性烯醇化酶的测定及临床意义

InAsia，theincidencerateofvasculardementiaishigherthanthatofAlzheimer＇sdisease．Vasculardementiaisoneofthemostcommondiseasesofseniledementia．Multi－infarctdementia（MID）isthemostcommontypeinvasculardementia．Inthestudywereporthere，wemeasuredtheconcentrationsofTNF－αandNSEincere－brospinalfluid（CSF）of７２patientswithcerebralinfarction（CI）and５５patientswithMID，soastoevaluatetheroleofthechangesofTNF－αandNSEinthepathogenesisofMID．１Subjectandmethod１．１SubjectThesubjects…     

Factors influencing analysis of prolyl endopeptidase in human blood and cerebrospinal fluid: increase in assay sensitivity

Prolyl endopeptidase (EC 3.4.21.26) (PEP) is present in nearly all investigated mammalian cells and biological fluids and might be involved in the degradation of physiologically important neuropeptides. To be able to investigate the variation of PEP in blood and cerebrospinal fluid (CSF) in human disease, the factors influencing analysis of PEP in these body fluids must be determined. The purpose of the present work was to study the influence of storage conditions, anticoagulation additives, freezing and thawing and substrate solvent on determination of PEP in blood plasma/serum and CSF. It was found that the PEP activity was about 10% higher in plasma (with EDTA and heparinate for anticoagulation) than in serum. Storage at room temperature (20 degrees C) caused a rapid decline in enzyme activity, which was smaller but still considerable at 4 degrees C. Storage at -20 degrees C and -70 degrees C did not decrease the PEP activity. Freezing and thawing of plasma/serum samples showed that the first freeze-thawing cycle produced a 20% reduction in enzyme activity but little further decrease was observed during subsequent cycles of freeze-thawing. In conclusion, PEP activity should preferably be measured within one hour after sampling using EDTA- or heparinate plasma. For long-term storage, samples should be immediately frozen and stored at -20 degrees C or colder. The selection and amount of the organic solvent used to dissolve the fluorogenic substrate strongly influenced the sensitivity of the assay. By developing an optimal solvent system an increase in assay sensitivity of about 400% could be obtained, which for the first time allowed measurement of the PEP activity in CSF.     

The automated processing algorithm to correct the test result of serum neuron‐specific enolase affected by specimen hemolysis

IntroductionSerum neuron‐specific enolase (NSE) is an important tumor marker for small cell lung cancer and neuroblastoma. However, the test of serum NSE compromised by specimen hemolysis is presented as a falsely higher result, which seriously disturbs clinical decision. This study aimed to establish a solution integrated with laboratory information system to clear the bias from hemolysis on serum NSE test.MethodsThe reference range of serum hemolysis index (HI) was first established, and specimen hemolysis rate was compared between HI test and visual observation. NSE concentration in serum pool with normal HI was spiked with serial diluted lysates from red blood cells to deduce individual corrective equation. The agreement between individual corrective equation and original NSE test was assayed by Bland and Altman plots.ResultsThe high HI existed in 32.6% of specimens from patients. The NSE median of hemolyzed specimens was significant higher than the baseline (p = 0.038), while the corrected NSE median had no difference compared with the baseline (p = 0.757). The mean difference of corrected NSE and initial NSE was 1.92%, the SD of difference was 5.23%, and furthermore, the difference was independent of tendency of HI (Spearman r = −0.069, p = 0.640). The 95% confidence interval of mean difference (from −8.33% to 12.17%) was less than the acceptable bias range (±20%).ConclusionThe agreement between individual correction equation and NSE assay was satisfied. Our automated processing algorithm for serum NSE could provide efficient management of posttest data and correct positive bias from specimen hemolysis.     

单次癫痫发作后患者血清和脑脊液中神经元特异性烯醇化酶及髓鞘碱性蛋白的变化

目的探讨单次癫痫发作是否会引起脑损伤。方法 2007年6月-2009年11月,采用电化学发光法检测癫痫发作后24h内40例和对照组40例患者血清和脑脊液中神经元特异性烯醇化酶（neuron-specific enolase,NSE）水平,采用ELISA法测定其血清和脑脊液中髓鞘碱性蛋白（myebin bosic protein,MBP）水平。结果癫痫组血清和脑脊液中NSE水平明显高于对照组（P〈0.01）;癫痫组血清MBP水平与对照组比较差异无统计学意义（P〉0.05）;癫痫组脑脊液中MBP水平高于对照组（P〈0.05）。结论单次癫痫患者血清和脑脊液中NSE明显升高,脑脊液中MBP升高,提示单次癫痫发作可导致神经元损伤。     

Acyclovir concentrations in serum and cerebrospinal fluid at steady state

A long-term clinical trial of acyclovir, 800 mg tid, as a therapeutic agent in multiple sclerosis (MS) is in progress. In three patients paired serum and cerebrospinal fluid (CSF) specimens were sampled after one, four, eight and twelve months of continuous treatment. These samples were collected 1.5 h before or 1.5 h after an oral dose. Acyclovir concentrations were assessed by radioimmunoassay. In the CSF, the acyclovir concentration was relatively stable, with a mean of 0.83 microM, while the serum acyclovir concentration was variable with mean peak and trough concentrations of 4.08 and 2.47 microM, respectively. In two other MS patients the acyclovir concentration time profile in serum and CSF was studied at steady state during the 8 h dose interval. In this study the acyclovir concentration in the CSF was only slightly affected by the fluctuations in serum and the acyclovir CSF/acyclovir serum ratio was apparently not influenced by the blood-brain barrier function. We found no indication of an accumulation of acyclovir in cerebrospinal fluid after one to twelve months of oral treatment.     

Factors influencing analysis of prolyl endopeptidase in human blood and cerebrospinal fluid: increase in assay sensitivity

Prolyl endopeptidase (EC 3.4.21.26) (PEP) is present in nearly all investigated mammalian cells and biological fluids and might be involved in the degradation of physiologically important neuropeptides. To be able to investigate the variation of PEP in blood and cerebrospinal fluid (CSF) in human disease, the factors influencing analysis of PEP in these body fluids must be determined. The purpose of the present work was to study the influence of storage conditions, anticoagulation additives, freezing and thawing and substrate solvent on determination of PEP in blood plasma/serum and CSF. It was found that the PEP activity was about 10% higher in plasma (with EDTA and heparinate for anticoagulation) than in serum. Storage at room temperature (20°C) caused a rapid decline in enzyme activity, which was smaller but still considerable at 4°C. Storage at ?20°C and ?70°C did not decrease the PEP activity. Freezing and thawing of plasma/serum samples showed that the first freeze‐thawing cycle produced a 20% reduction in enzyme activity but little further decrease was observed during subsequent cycles of freeze‐thawing. In conclusion, PEP activity should preferably be measured within one hour after sampling using EDTA‐ or heparinate plasma. For long‐term storage, samples should be immediately frozen and stored at ?20°C or colder. The selection and amount of the organic solvent used to dissolve the fluorogenic substrate strongly influenced the sensitivity of the assay. By developing an optimal solvent system an increase in assay sensitivity of about 400% could be obtained, which for the first time allowed measurement of the PEP activity in CSF.     

溶血对时间分辨免疫荧光分析法测定胰岛素的影响

目的：探讨溶血对时间分辨免疫荧光分析法测定胰岛素的影响。方法收集30例未发生溶血的血清标本,利用低渗溶血法分别制备轻度溶血、中度溶血、重度溶血标本各10例;采用时间分辨免疫荧光分析法分别测定各组标本溶血前（0 h）和溶血后0．5、2、18 h的胰岛素浓度;通过配对t检验、重复测量数据的方差分析对溶血程度和时间对胰岛素浓度检测结果的影响进行分析。结果轻度溶血、中度溶血、重度溶血标本之间胰岛素浓度检测结果比较差异无统计学意义（P＞0．05）;随着溶血时间的延长,胰岛素浓度呈明显下降趋势,0．5、2、18 h检测结果与0 h检测结果相比,比较差异均有统计学意义（P＜0．05）。结论溶血程度对时间分辨免疫荧光分析法测定胰岛素浓度的影响不明显,但溶血时间则对测定结果具有较为明显的影响;标本溶血和溶血时间均为影响胰岛素浓度检测结果的重要因素。     

Acyclovir levels in serum and cerebrospinal fluid after oral administration of valacyclovir

The possible involvement of herpesviruses in the pathogenesis of multiple sclerosis (MS) was recently investigated in a clinical trial of valacyclovir in patients with MS. As an important part of that survey we performed an independent pharmacokinetic study in order to determine the concentration of acyclovir in cerebrospinal fluid (CSF). The concentrations of acyclovir in serum and CSF were measured at steady state after 6 days of oral treatment with 1,000 mg of valacyclovir three times a day. Samples were obtained from 10 patients with MS. All patients had normal renal function, and none had signs of a damaged blood-CSF barrier. The maximum concentration of acyclovir in serum was reached after 1 to 3 h (mean +/- standard deviation [SD], 27.1 +/- 5.6 micro M), and the minimum concentration in serum was 3.1 +/- 1.1 micro M (mean +/- SD). The acyclovir concentrations in CSF at 2 and 8 h were essentially stable, with the mean +/- SD levels being 2.5 +/- 0.9 and 2.3 +/- 0.7 micro M, respectively. Similar levels were recorded in serum and CSF samples from five other MS patients after 6 months of oral treatment with valacyclovir at identical dosages. The area under the concentration-time curve (AUC) for acyclovir in CSF to the AUC for acyclovir in serum (CSF/serum AUC ratio) was approximately 20%. We conclude that the improved bioavailability previously reported for valacyclovir in plasma results in higher concentrations in CSF, while the CSF/serum AUC ratio remains constant.     

神经元特异性烯醇化酶在中枢神经疾病中的作用

烯醇化酶是参与无氧酵解中的关键酶,它是由α,β,γ组成的5种二聚体同工酶,其中γγ型因其特异性地存在于神经元和神经内分泌细胞中,被称作为神经元特异性烯醇化酶(NSE)。分子量为78000。NSE在神经系统含量亦不同,表现为中枢神经系统含量大于周围神经系统。在许多中枢神经系统疾病,如:脑卒中、脑外伤、脑肿瘤,特别是脑胶质瘤和成神经细胞瘤患者中,因NSE从病变神经元中溢出进入外周血和脑脊液,使血清和脑脊液中NSE水平上升,故广泛用于中枢神经系统疾病的诊断和预后估计。另外,因NSE存在于神经内分泌细胞中,故被用作恶性肿瘤,特别是肺部肿瘤的检测,NSE是敏感性与特异性强的肿瘤标志物。在其他一些疾病,如:肝硬化、肝性脑病以及晕厥等疾病中含量也有变化,可能有助于疾病诊断与预后判断。     

颅脑创伤患者血浆和脑脊液血管升压素的动态变化及意义

目的 探讨颅脑创伤患者血浆和脑脊液血管升压素(AVP)的动态变化及其临床意义。方法 将36例中重度颅脑创伤患者按GCS和GOS评分分组，采用放射免疫RIA方法和渗透压测定仪对患者血浆和脑脊液中AVP、血浆和脑脊液渗透压和电解质进行动态检测，同时选择30例外科手术患者作为对照。结果 颅脑创伤组血浆和脑脊液AVP明显高于对照组并与GCS评分有关。对照组血浆AVP术前与术后有显著性差异，GOS各分组间脑脊液AVP有显著性差异，AVP水平与脑脊液渗透压和颅内压相关；血浆AVP与血浆渗透压相关；Spearman相关性检验提示，相对低钠血症，血浆AVP变化较脑脊液AVP敏感；相对颅内压变化血浆渗透压较脑脊液渗透压敏感。结论 血浆和脑脊液中AVP可以通过不同环节和不同因素参与继发性脑损害的过程。脑脊液或血浆中的AVP可以通过不同环节和不同因素参与继发性脑损害的过程，脑脊液或血浆中的AVP与颅脑创伤损伤严重程度密切相关，可以作为判断伤情程度的客观指标；创伤患者预后越差，脑脊液中AVP浓度越高，测定颅脑创伤患者CSF中AVP水平比血浆中AVP更合适作为预后的判断指标，用来评估脑水肿严重程度和颅内压也较血浆中AVP敏感、准确。     

头颅局部亚低温对颅内感染患者近期疗效的观察

目的观察头颅局部亚低温对颅内感染患者的近期疗效并探讨其可能的机制。方法 40例颅内感染患者,其中病毒性脑炎12例,结核性脑膜炎16例,化脓性脑膜炎12例,以上患者随机分为两组,亚低温治疗组在常规药物治疗的基础上加用局部亚低温治疗,常温治疗组为单纯常规药物治疗。分别应用双抗体夹心酶联免疫吸附测定（ABC-ELISA）法和放射免疫法测定患者血清和脑脊液中基质金属蛋白酶9（MMP-9）、神经元特异性烯醇化酶（NSE）含量,进行脑脊液的常规和生化检测。结果亚低温治疗7天后,亚低温治疗组脑脊液中白细胞、蛋白、颅内压明显较常温组下降,糖升高（P〈0.01）;亚低温治疗组的NSE、MMP-9较常温组明显下降（P〈0.01）;各观察组MMP-9和NSE的含量均显著高于对照组（P〈0.01）。结论颅内感染过程中存在着血脑屏障受损和神经元损伤,亚低温可以同时减少神经细胞的损伤及改善血脑屏障,从而发挥良好的治疗作用。     

脑梗死及多梗死性痴呆患者脑脊液S-100、NSE的测定

目的　研究脑梗死 (CI)及多梗死性痴呆 (MID)患者脑脊液 (CSF)S - 10 0、神经元特异性烯醇化酶 (NSE)含量的变化及意义。方法　采用放免法及ELISA法对 72例CI及 2 1例MID患者CSFS - 10 0、NSE含量进行测定。MID患者的智能水平采用简易智能量表 (MMSE)测定 ,并参考Hachinski缺血评分及DSM -Ⅳ诊断标准进行确诊。结果　CI急性期组患者CSFS- 10 0、NSE浓度明显高于对照组 (P <0 0 1) ;而CI恢复期组患者CSFS - 10 0、NSE的浓度与对照组无显著差异 (P >0 0 5 )。MID患者CSFS - 10 0浓度高于对照组及CI恢复期组 (P <0 0 5 ) ;而NSE浓度低于对照组及CI恢复期组 (P <0 0 5 )。结论　NSE、S - 10 0蛋白可反映MID患者脑神经细胞、神经胶质细胞的损害情况和功能状态 ,对MID患者的病情及预后判断有重要意义。