Oxidative Stress in Cerebrospinal Fluid of Patients with Aseptic and Bacterial Meningitis

This study aimed to determine whether patients with aseptic and bacterial meningitis presented alterations in oxidative stress parameters of cerebrospinal fluid (CSF). A total of 30 patients were used in the research. The CSF oxidative stress status has been evaluated through many parameters, such as lipid peroxidation through thiobarbituric acid reactive substances (TBARS) and antioxidant defense systems such as superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH) and ascorbic acid. TBARS levels, SOD and GST activity increase in aseptic meningitis and in bacterial meningitis. The ascorbic acid concentration increased significantly in patients with both meningitis types. The reduced glutathione levels were reduced in CSF of patients with aseptic and bacterial meningitis. In present study we may conclude that oxidative stress contributes at least in part to the severe neurological dysfunction found in meningitis.     

脑脊液腺苷脱氨酶和干扰素-r水平在结核性脑膜炎中的诊断价值

目的：探讨脑脊液腺苷脱氨酶和干扰素-酌水平在结核性脑膜炎中的诊断价值。方法：选取2012 年6 月-2014 年6 月期间于 我院进行治疗的结核性脑膜炎患者50 例作为研究组,同期健康体检者50 例作为对照组1,50 例非结核性脑膜炎者作为对照组 2。检测脑脊液中腺苷脱氨酶和干扰素-酌水平,并进行比较。结果：研究组患者脑脊液中腺苷脱氨酶和干扰素-r水平分别为 （22.45± 4.23）U/L和（36.45± 13.56）ng/L;对照组1 患者脑脊液中腺苷脱氨酶和干扰素-r水平分别为（16.32± 3.24）U/L 和 （12.78± 2.67）ng/L,对照组2 患者脑脊液中腺苷脱氨酶和干扰素-r水平分别为（7.48± 4.01）U/L 和（13.25± 2.89）ng/L,三组比较 差异有统计学意义（P＜0.05）,其中研究组患者脑脊液中腺苷脱氨酶和干扰素-r水平显著高于对照组1 和对照组2,两两比较差 异均有统计学意义（均P＜0.05）。结论：脑脊液腺苷脱氨酶和干扰素-r在结核性脑膜炎患者中具有诊断价值,值得推广应用。     

Carbon-Mercaptooctadecane/Carboxylated Multi-walled Carbon Nanotubes Composite Based Genosensor for Detection of Bacterial Meningitis

Human brain bacterial meningitis is a life-threatening disease mainly caused by Neisseria meningitidis, lead to several complications including damage of brain or even death. The present available methods for diagnosis of meningitis have one or more limitations. A rmpM gene based genosensor was fabricated by immobilizing 5′-amino modified 19-mer single stranded DNA probe onto carbon-mercaptooctadecane/carboxylated multi-walled carbon nanotubes composite electrode and hybridized with 2.5–40 ng/6 μL of single stranded genomic DNA (ssG-DNA) of N. meningitidis from cerebrospinal fluid (CSF) of the suspected meningitis patients. The electrochemical response was measured by using cyclic voltammetry and differential pulse voltammetry (DPV) using 1 mM methylene blue as redox indicator in 30 min (including a response time of 1 min) at 25 °C. The sensitivity of the genosensor was 3.762 (μA/cm²)/ng and limit of detection was 2 ng of ssG-DNA of N. meningitidis with DPV. The genosensor has specificity only to N. meningitidis and does not hybridize with the genomic DNA of any other possible pathogen in human CSF. The immobilization of the probe and hybridization of the ssG-DNA were characterized by using electrochemical impedance in presence of 5 mM potassium ferricyanide and scanning electron microscopy. The genosensor loses only 12 % of its original DPV current on storage at 4 °C for 6 months. Carbon composite based electrochemical array can be constructed to detect multiple bacterial meningitis suspected patient CSF samples during an outbreak of the disease.     

脑脊液检查在早产儿及足月儿细菌性脑膜炎的诊断价值

目的:评价脑脊液检查在早产儿及足月儿细菌性脑膜炎诊断中的价值。方法:选取2014年6月1日至2016年12月31日上海市儿童医院新生儿科收治的行腰椎穿刺检查的447例新生儿,回顾性分析新生儿的一般资料、脑脊液常规生化、培养等指标,根据胎龄将患儿分为早产儿167例与足月儿280例,再根据有无患发细菌性脑膜炎分为早产儿细菌性脑膜炎27例(早产儿观察组)、早产儿非细菌性脑膜炎140例(早产儿对照组)、足月儿细菌性脑膜炎38例(足月儿观察组)、足月儿非细菌性脑膜炎242例(足月儿对照组),采用受试者工作特征(ROC)曲线评估蛋白定量、白细胞计数、葡萄糖对早产儿及足月儿细菌性脑膜炎的诊断价值。结果:与同组对照组相比,足月儿观察组和早产儿观察组蛋白定量和白细胞计数均明显升高,而葡萄糖含量显著下降,且差异均具有统计学意义(P0.05);本研究65例细菌性脑膜炎患儿脑脊液培养分离出11株细菌(16.9%)。足月儿脑脊液白细胞计数、蛋白定量以及葡萄糖诊断细菌性脑膜炎的ROC曲线下面积分别为0.995、0.846、0.703。早产儿脑脊液白细胞计数、蛋白定量以及葡萄糖诊断细菌性脑膜炎ROC曲线下面积分别为0.970、0.711、0.705。结论:脑脊液白细胞计数、蛋白定量在足月儿和早产儿细菌性脑膜炎中具有较高的诊断价值。     

Rapid Diagnosis and Simultaneous Identification of Tuberculous and Bacterial Meningitis by a Newly Developed Duplex Polymerase Chain Reaction

The present study describes the development and evaluation of a duplex polymerase chain reaction (D-PCR) for diagnosis and simultaneous identification of tuberculous meningitis (TBM) and bacterial meningitis (BM) in a single reaction. A D-PCR with primers amplifying portions of the Mycobacterium tuberculosis IS6110 and the eubacteria 16SrDNA sequence in a same reaction mix was developed and tested on DNA extracted from 150 clinical CSF samples from different categories (TBM = 39, BM = 26, control infectious and non-infectious category = 85). The results indicate a clear differentiation between bands for eubacteria and M. tuberculosis with an analytical sensitivity of 10³ cfu/ml for eubacteria and 10² cfu/ml for M. tuberculosis. When evaluated in clinical samples, D-PCR overall diagnosed 100 % confirmed TBM and 100 % confirmed BM cases with overall specificity of 96.5 %. D-PCR can be an effective tool for diagnosis and simultaneous identification of TBM or BM in a single PCR reaction. It saves time, cost, labour and sample amount and help in administration of appropriate antimicrobial therapy. The proposed diagnostic assay would be helpful in correct and rapid management of TBM and BM patients.

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-015-0517-9) contains supplementary material, which is available to authorized users.     

Nociceptive Response and Adenine Nucleotide Hydrolysis in Synaptosomes Isolated from Spinal Cord of Hypothyroid Rats

Purinergic system exerts a significant influence on the modulation of pain pathways at the spinal site. Adenosine has antinociceptive properties in experimental and clinical situations, while ATP exerts pronociceptive actions in different pain models. In this study we investigated the hydrolysis of ATP to adenosine in synaptosomes from spinal cord in parallel with the nociceptive response of rats at different ages after hypothyroidism induction. Hypothyroidism elicited a significant increase in AMP hydrolysis to adenosine in synaptosomes from spinal cord of rats subjected to neonatal hypothyroidism and in 420-day-old rats submitted to thyroidectomy. Accordingly, these rats presented an analgesic response as a consequence of hypothyroidism. In contrast, the ATP hydrolysis was decreased in the spinal cord of 60-day-old hypothyroid rats in parallel with a significant increase in nociceptive response. These results indicate the involvement of adenine nucleotides in the control of the hypothyroidism-induced nociceptive response during development.     

结核性脑膜炎与化脓性脑膜炎脑脊液与血浆生化指标比值的比较研究

目的:比较结核性脑膜炎与化脓性脑膜炎脑脊液与血浆生化指标比值。方法:选择2010年2月~2014年12月我院结核性脑膜炎患者82例,化脓性脑膜炎98例,检测脑脊液与血浆中的蛋白、糖及氯化物含量,并计算比值。结果:两组患者脑脊液蛋白、糖含量的差异无统计学意义(P0.05),化脓性脑膜炎组的氯化物含量高于结核性脑膜炎组(P0.05);两组血浆糖含量的差异无统计学意义(P0.05),化脓性脑膜炎组蛋白和氯化物含量明显高于结核性脑膜炎组(P0.05);化脓性脑膜炎组蛋白比值低于结核性脑膜炎组,氯化物比值则高于化脓性脑膜炎组,差异均有统计学意义(P0.05);两组间糖比值比较,差异无统计学意义(P0.05)。结论:脑脊液与血浆生化指标比值对鉴别诊断结核性脑膜炎与化脓性脑膜炎有重要意义。     

颅内感染患者血清和脑脊液降钙素原水平的观察

目的 研究降钙素原(PCT)在脑膜炎患者血清和脑脊液(CSF)中的水平,探讨其在脑膜炎诊断中的临床意义.方法 用免疫透射比浊法测定42例脑膜炎患者(细菌性脑膜炎18例、病毒性脑膜炎24例)急性期内血清和脑脊液PCT,并与CSF的常规生化指标作相关性分析;20例神经系统非感染性疾病为对照组.结果 细菌性脑膜炎患者血清和CSF中的PCT含量均显著高于病毒性脑膜炎和对照组(P＜0.01);但病毒性脑膜炎患者与对照组之间的PCT水平差异无统计学意义(P＞0.05).相关性分析显示,在CSF中PCT与白细胞数(r=0.161,P＞0.05)无明显相关性,但与CSF蛋白含量和血清PCT水平呈弱相关性(r=0.465和0.570,P＜0.05).结论 PCT升高是颅内细菌感染的标志之一;PCT可作为一项CSF的常规生化指标,有助于指导临床对脑膜炎的诊治.     

脑脊液乳酸和血清降钙素原及C反应蛋白对小儿细菌性脑膜炎的诊断价值

目的 探讨脑脊液乳酸、血清降钙素原及C反应蛋白对小儿细菌性脑膜炎的诊断价值。方法 选取我院2016年4月至2017年6月收治的50例细菌性脑膜炎患儿以及50例病毒性脑膜炎患儿进行作为研究对象,比较2类患儿脑脊液乳酸（LA）、血清降钙素原（PCT）及C反应蛋白（CRP）的水平,并分析其诊断价值。结果 细菌性脑膜炎组患儿脑脊液LA、血清PCT及CRP水平显著高于病毒性脑膜炎患儿（均P<0.05）。血清PCT诊断的灵敏度和特异度最高（96.4%、90.9%,P<0.05）。3项指标联合检测的灵敏度（100.0%）和特异度（95.5%）明显高于任一单项指标（均P<0.05）。经过Pearson相关性分析,脑脊液LA、血清PCT及CRP与小儿细菌性脑膜炎均呈显著正相关关系（均P<0.05）。结论 脑脊液乳酸、血清PCT及CRP对小儿细菌性脑膜炎的诊断和治疗效果监测有重要应用价值。     

Isolation and Characterization of Mumps Virus Strains in a Mumps Outbreak with a High Incidence of Aseptic Meningitis

In 1993, mumps with a high incidence of aseptic meningitis became prevalent in Akita prefecture, Japan. Three mumps virus isolates obtained from the nonvaccine-associated cases lacked the BamHI restriction cleavage site of the P gene, like the Urabe strain (Yamada, A. et al, Vaccine 8: 553-557). However, four additional nucleotide substitutions were found in the determined region of 157 bp. Fourteen of 19 cases from which mumps virus showing the Urabe-like RFLP profile was detected were complicated with symptomatic meningitis, whereas there were only four cases of meningitis among 23 individuals infected with the wild type showing no Urabe-like RFLP profile (non-“Urabe-like” wild-type). The incidence of meningitis was over 70% among patients infected with the “Urabe-like” wild-type virus. The “Urabe-like” wild-type disappeared after February 1994 in the epidemic area and was replaced by the non-“Urabe-like” wild-type. Patients infected with the “Urabe-like” wild-type lived in a closed colony, in which there were two instances of transmission between siblings. Thus this outbreak was transient and narrowly localized.     

高迁移率族蛋白1拮抗剂BoxA对细菌性脑膜炎大鼠的治疗作用

目的:探究高迁移率族蛋白1(HMGB1)拮抗剂BoxA尾静脉注射对细菌性脑膜炎(BM)大鼠模型的临床体征改善和炎症抑制作用。方法:除外正常对照的雄性Sprague-Dawley(SD)大鼠设为对照组(n=20),另取60只大鼠行脑室立体定向注射20μL大肠杆菌Escherichia coli (DH5α1×10~7CFU/m L)建立BM模型,之后随机分为两组(各组n=30),一组尾静脉注射HMGB1拮抗剂BoxA,即BoxA组;一组麻醉后进行尾静脉注射无菌磷酸盐(PBS),即Vehicle组。造模3 d后,对各组大鼠的临床指标以及病理生理参数(颅内压和脑脊液白细胞(WBC)计数)进行评估,使用酶联免疫吸附实验(ELISA)法检测血清中HMGB1的相对含量,使用伊文思蓝染色观察血脑屏障(BBB)通透性,使用免疫荧光染色检测大脑皮层炎症因子(IL-1β和TNF-α)的表达水平。结果:相比Control组,Vehicle组临床指标,颅内压,WBC计数以及血清HMGB1含量明显提升(P0.05);而BoxA组相比Vehicle组,以上改变有部分减少(P0.05)。另外,Vehicle组较Control组EB渗漏增加且炎症因子(IL-1β和TNF-α)表达水平增高(P0.05)。与Vehicle组相比,BoxA组的这些变化亦被部分调节(P0.05)。结论:HMGB1抑制剂BoxA尾静脉注射能够下调HMGB1表达水平并同时缓解细菌性脑膜炎大鼠的临床症状和炎症反应。     

成人化脓性脑膜炎患者脑脊液变化

分析成人化脓性脑膜炎患者脑脊液细胞、病原菌等的变化及其关系, 为该病的有效诊断、病情监测和针对性治疗提供参考。

对我院32例成人化脓性脑膜炎患者用药前后不同时期的脑脊液分别进行常规、生化与细胞学检测, 对检测结果进行统计分析。

与恢复期比较, 患者急性期时脑压、脑脊液白细胞总数与乳酸脱氢酶水平显著升高(均P < 0.05)。32例患者中, 15例脑脊液细菌培养呈阳性。典型菌与非典型菌感染者急性期脑脊液白细胞总数、小淋巴细胞、单核吞噬细胞和嗜中性粒细胞比例差异均有统计学意义(均P < 0.05)。

成人化脓性脑膜炎患者脑脊液变化显著, 脑脊液细胞学变化与感染类型相关, 可为个性化诊疗提供参考。

     

细菌性脑膜炎患儿脑脊液Caveolin-1、VE-cadherin、CXCl-5水平与炎性因子的关系及其诊断价值分析

摘要 目的：探讨细菌性脑膜炎（BM）患儿脑脊液陷窝蛋白-1（Caveolin-1）、血管内皮钙黏素（VE-cadherin）、CXC类趋化因子配体5（CXCl-5）水平与炎性因子的相关性,分析Caveolin-1、VE-cadherin、CXCl-5诊断BM的价值。方法：选取2016年1月1日至2020年5月1日期间我院收治的106例BM患儿（BM组）、76例病毒性脑炎（VM）患儿（VM组）、72例无神经系统损伤患儿（对照组）。检测BM组、VM组患儿脑脊液中Caveolin-1、VE-cadherin、CXCl-5水平,检测三组血清肿瘤坏死因子-α（TNF-α）、降钙素原（PCT）和C-反应蛋白（CRP）、白介素-6（IL-6）水平。Pearson相关性分析Caveolin-1、VE-cadherin、CXCl-5与TNF-α、PCT、CRP、IL-6之间相关性,受试者工作特征（ROC）曲线分析Caveolin-1、VE-cadherin、CXCl-5诊断BM的价值。结果：BM组脑脊液Caveolin-1、VE-cadherin、CXCl-5水平高于VM组（P＜0.05）,血清TNF-α、PCT、CRP、IL-6水平高于VM组和对照组（P＜0.05）,VM组血清TNF-α、PCT、CRP、IL-6水平高于对照组（P＜0.05）。BM组患儿脑脊液中Caveolin-1、VE-cadherin、CXCl-5水平与血清TNF-α、PCT、CRP、IL-6水平均呈正相关（P＜0.05）。ROC曲线分析结果显示联合Caveolin-1、VE-cadherin、CXCl-5诊断BM的曲线下面积（AUC）为0.888,高于以上指标单独诊断的0.690、0.661、0.639。结论：BM患儿脑脊液中Caveolin-1、VE-cadherin、CXCL5水平较VM患儿增高,其可能通过介导神经系统炎症反应参与BM发病,联合三项指标在BM鉴别诊断中具有较高价值。     

结核性脑膜炎颅神经损害与脑脊液特点的关系研究

目的:探讨结核性脑膜炎颅神经损害与脑脊液特点的关系。方法:回顾性收集我院神经内科确诊的173例结核性脑膜炎患者临床资料,将所有患者分为伴颅神经损害和不伴颅神经损害两组,对其临床特点及脑脊液外观、压力及细胞学、生化、免疫球蛋白结果进行统计学分析。结果:颅神经损害者占所有病人的22.5%(39/173),视神经损害占61.5%(24/39),外展神经损害占53.8%(21/39),动眼神经损害占15.4%(6/39),听神经损害占15.4%(6/39);颅神经损伤组较无颅神经损伤组脑脊液压力、蛋白质及Ig G、Ig M、Alb明显升高(P0.01或0.001),且颅神经损害者头颅MRI脑膜强化较无颅神经损害者差异有统计学意义(P0.001),而脑脊液外观、白细胞计数及比例两组间并无发现显著差异。结论:结核性脑膜炎患者伴颅神经损害时,脑脊液压力、蛋白质及部分免疫球蛋白较无颅神经损害者明显升高,这将有助于结核性脑膜炎合并颅神经损害的临床诊断,对于有颅神经损害的结核性脑膜膜炎患者加强并早期启动抗结核治疗,以减少蛋白质沉积,从而降低颅神经损害的发生率。     

Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Streptococcus agalactiae (Group B Streptococcus) is a commensal of the human intestine and vagina of adult women but is the leading cause of invasive infection in neonates. This Gram-positive bacterium displays a set of virulence-associated surface proteins involved in the interaction with the host, such as adhesion to host cells, invasion of tissues, or subversion of the immune system. In this study, we characterized a cell wall-localized protein as an ecto-5′-nucleoside diphosphate phosphohydrolase (NudP) involved in the degradation of extracellular nucleotides which are central mediators of the immune response. Biochemical characterization of recombinant NudP revealed a Mn²⁺-dependent ecto-5′-nucleotidase activity on ribo- and deoxyribonucleoside 5′-mono- and 5′-diphosphates with a substrate specificity different from that of known orthologous enzymes. Deletion of the gene coding the housekeeping enzyme sortase A led to the release of NudP into the culture supernatant, confirming that this enzyme is anchored to the cell wall by its non-canonical LPXTN motif. The NudP ecto-5′-nucleotidase activity is reminiscent of the reactions performed by the mammalian ectonucleotidases CD39 and CD73 involved in regulating the extracellular level of ATP and adenosine. We further demonstrated that the absence of NudP activity decreases bacterial survival in mouse blood, a process dependent on extracellular adenosine. In vivo assays in animal models of infection showed that NudP activity is critical for virulence. These results demonstrate that Group B Streptococcus expresses a specific ecto-5′-nucleotidase necessary for its pathogenicity and highlight the diversity of reactions performed by this enzyme family. These results suggest that bacterial pathogens have developed specialized strategies to subvert the mammalian immune response controlled by the extracellular nucleotide signaling pathways.     

Cloning and Characterization of the Genomic RNA Sequence of the Mumps Virus Strain Associated with a High Incidence of Aseptic Meningitis

cDNA clones of the mumps virus wild-type strain, associated with a high incidence of aseptic meningitis (ODATE-1 strain), were isolated and analyzed from genomic nucleotide position 22 to 8520 containing the NP, P, M., F, SH and HN protein coding region. The ODATE-1 strain exhibited a RFLP profile identical to that of the Urabe vaccine strain in spite of the fact that the virus was isolated from non-vaccinated cases. However, a comparison of nucleotide and amino acid sequences among the ODATE-1 strain, Urabe strain and Miyahara strain revealed that the ODATE-1 strain was not related to the Urabe strain.     

脑脊液PCT、VEGF、S-100B、NSE、MMP及CGRP水平在病毒性脑膜炎患儿中的临床研究

目的:探讨脑脊液降钙素原(PCT)、血管内皮生长因子(VEGF)、S100B蛋白(S-100B)、神经元特异性烯醇化酶(NSE)、基质金属蛋白酶(MMP)及降钙素基因相关肽(CGRP)水平联合检测在病毒性脑炎患儿中的诊断价值和意义。方法:选取2012年6月至2014年6月收治的病毒性脑炎患儿106例作为研究组,选取同时期来我院进行健康体检的60例儿童作为对照组,比较两组脑脊液PCT、VEGF、S-100B、NSE、MMP及CGRP水平,并分析研究组依照不同层次分组后以上各指标的差异。结果:研究组脑脊液中PCT、VEGF、S-100B、NSE、MMP及CGRP水平明显高于对照组,差异具有统计学意义(P0.05);研究组重症患儿高于轻症患者,急性期患者高于非急性期患儿,差异均具有统计学意义(P0.05)。以上各指标中任意二种、三种、四种及五种联合检测,其诊断效能均在0.923-0.967之间,六种指标联合检测的效能最高为0.975。结论:病毒性脑膜炎患儿血清及脑脊液PCT、VEGF、S-100B、NSE、MMP及CGRP水平均呈现升高的趋势,且不同严重程度及分期患儿的以上指标均有一定的差异,六种指标联合检测诊断病毒性脑炎患儿的效能最高。     

Epidemiological Profile of Cryptococcal Meningitis Patients in Rio Grande do Sul,Brazil

Cryptococcosis is a major opportunistic mycosis which has meningitis as its most frequent clinical presentation and can be fatal in the absence of antifungal therapy. The aetiological agents are Cryptococcus neoformans, which affects mainly immunocompromised subjects, and C. gattii, the aetiologic agent for cryptococcosis in healthy individuals. A recent outbreak of cryptococcosis on Vancouver Island, Canada, raised the level of concern about the epidemiology of this disease. In Brazil, between 1980 and 2002, six per cent of AIDS patients had cryptococcosis in course at the time of diagnosis. To identify the profile of cryptococcal meningitis patients in Rio Grande do Sul (RS), Brazil, a retroactive study was realized using data from patients registered at Laboratório Central de Saúde Pública IPB-LACEN/RS from 2000 to 2005. Most of the patients were men (77.12%), Caucasian (83.5%), median age between thirty and thirty-nine years old (46.24%) and HIV positive (95%).     

Increase of Adenine Nucleotide Hydrolysis in Rat Hippocampal Slices after Seizures Induced by Quinolinic Acid

Quinolinic acid (QUIN), an endogenous convulsant compound, overstimulates the glutamatergic system stimulating N-methyl-D-aspartate receptors, enhancing glutamate release and inhibiting glutamate uptake. Glutamate releases the neuroprotector adenosine, which in turn reduces glutamate release and depresses the neuronal activity. Additionally, adenine nucleotides are an important source of adenosine, by action of ecto-nucleotidases. Here we evaluated the adenine nucleotide hydrolysis in hippocampal slices of adult rats in different times after seizures induced by QUIN. After 45 min, there was an increase of ATP and ADP hydrolysis. After 5 h, there was an increase of ATP, ADP and AMP hydrolysis. After 12 h, there was an increase only of ATP hydrolysis. After 24 h, all hydrolysis returned to control levels. As slice preparations maintain tissue integrity, this study indicates, more than previously observed with synaptosomal preparations, that the extracellular production of the neuroprotector adenosine may be involved in brain responses to seizures.