儿童病毒性脑膜炎、脑炎中肠道病毒感染的分子生物学分型检测及临床研究

目的探讨儿童病毒性脑膜炎、脑炎中肠道病毒感染的分子生物学分型。方法采用逆转录聚合酶链反应(RT-PCR)及简并引物扩增并测序,利用Genbank核对分型,检测东营地区2008年1月-2012年2月63例临床诊断为病毒性脑膜炎、脑炎患儿脑脊液中的肠道病毒(EV)RNA,并结合临床特点进行分析。结果 63例病毒性脑膜炎、脑炎患儿的脑脊液经RT-PCR检测,EV感染阳性率为74.6%(47/63)。利用通用引物筛选出的阳性标本,并采用简并引物经两次PCR共获得阳性结果 31例(66.0%)。随机抽取23条阳性片段进行克隆测序,测序结果与Gen Bank标准EV毒株进行同源性对比分析,相关序列同源性均在97%以上。肠道病毒引起中枢神经系统感染的临床特征在各个年龄组有所不同,33例急性期EV患儿脑脊液中白细胞数不同程度增高(均数76×106/L)。结论 EV是儿童无菌性脑膜炎、脑炎的最主要的病原体。采用RT-PCR可以快速准确的检测出中枢神经系统感染的肠道病毒;采用分型引物扩增并测序,将EV的VP1部分序列与GENBANK中的EV标准毒株进行对比,根据基因序列的同源性进行EV型别鉴定是一种切实可行的方案。EV引起的病毒性脑膜炎临床症状一般较轻、无特异性,多数患儿CSF中白细胞高于正常范围。     

新生儿化脓性脑膜炎脑脊液酸碱平衡与电解质变化的研究

目的 分析化脓性脑膜炎新生儿脑脊液电解质、酸碱平衡的临床意义。方法 收集521医院新生儿科2013年1月—2017年6月,确诊为化脓性脑膜炎的患儿44例作为观察组,以同时期收治的未患颅内感染性疾病的患儿40例为对照组,对所有患儿进行脑脊液电解质、气体分析测定。观察组临床治愈后再复测。结果 1)观察组治疗前脑脊液PCO₂、HCO^-₃、BE、乳酸(Lac)、Cl^-与对照组相比差异有统计学意义(P＜0.05),而K⁺、Na⁺、Ca²⁺差异无统计学意义(P＞0.05);2)观察组治疗前与临床治愈后脑脊液pH、PCO₂、HCO^-₃、BE、Lac、Cl^-差异有统计学意义(P＜0.05),而K⁺、Na⁺、Ca²⁺差异无统计学意义(P＞0.05);3)观察组脑脊液的pH、PCO₂、Lac同动脉血上述酸碱变量之间存在相关性。结论 化脓性脑膜炎患儿存在脑脊液酸碱失衡及Cl^-的减低,脑脊液气体分析、Cl^-测定可作为诊断、评价治疗效果的直接客观指标。     

单核细胞增生性李斯特菌脑膜炎免疫应答机制的研究进展

李斯特菌脑膜炎是单核细胞增生性李斯特菌感染引起的脑膜炎，具有极高的致死率。对李斯特菌病的发病机制的研究目前主要集中在细菌如何跨肠道屏障、胎盘屏障方面。而免疫学机制及炎症通路有待表征。研究发现在感染早期宿主就能识别细菌DNA启动先天性免疫反应，随后引起相应靶细胞分泌炎症因子启动适应性免疫反应。该文从固有性免疫、适应性免疫反应、补体系统等方面综述了单核细胞增生性李斯特脑膜炎发病机制的研究进展。     

儿童单核细胞增生性李斯特菌侵袭性感染22例分析

目的 了解儿童单核细胞增生性李斯特菌(LM)感染的临床特点，为本病的经验诊治提供参考。 方法 通过检索电子病历系统，回顾性分析2010年1月至2018年12月我院无菌来源标本培养结果为LM的患儿临床特点。结果 确诊为LM侵袭性感染患儿22例，男11例。年龄范围为2h~14y8m，中位数为3d，新生儿占68.2%。血培养阳性16例，脑脊液培养阳性11例，其中5例血液脑脊液培养同时分离到LM。16例诊断为化脓性脑膜炎，其中9例出现颅内并发症(颅内出血4例、硬膜下积液4例、脑积水3例、脑室管膜炎1例)。临床表现新生儿主要为气促发绀、发热、状态反应差、末梢循环差、呻吟及黄疸等；非新生儿以发热、呕吐、精神状态改变、头痛、抽搐及颈强直常见。脑脊液检查白细胞数、蛋白、乳酸脱氢酶升高14例，糖降低12例，氯化物降低11例。药敏试验显示，LM对氨苄西林、万古霉素和利奈唑胺敏感率为100%。本组资料治愈好转率77.3%，病死率18.2%。结论     

Immune reconstitution inflammatory syndrome (IRIS) associated with Cryptococcus neoformans infection in AIDS patients

Cryptococcosis is frequently associated to the immune reconstitution inflammatory syndrome (IRIS) in AIDS patients on highly active antiretroviral therapy (HAART). This study aimed to evaluate clinical and evolutive features of IRIS associated cryptococcosis patients in Uberaba, Brazil. Patients: Eighty‐one AIDS individuals admitted at the teaching hospital with cryptococcal meningitis were evaluated and from these, 40 were prospectively followed. Of 40 patients with cryptococcosis, nine (22.5%) presented clinical and laboratory features of IRIS. Six (66.6%) were male, with a mean age of 37.2. Five (55.5%) presented cryptococcosis as first AIDS defining condition. In seven (77.9%) IRIS was characterised as a relapse of meningeal symptoms after 10 weeks, mean time of 72 days, of starting HAART whereas, two asymptomatic patients developed the syndrome as an unmasked cryptococcosis after 10 and 12 weeks on HAART. Lymphadenitis as isolated finding associated with IRIS was evidenced in three cases. All patients presented low CD4⁺ and high RNA viral load baseline values. Cultures of cerebrospinal fluid and lymph‐node fragments tissues of these cases were negative. Six of nine individuals developed high intracranial pressure requiring a daily relief lumbar puncture. No deaths occurred during the evolution of these patients. The incidence and clinical evolutive profile observed in this case series are in accordance with other reports elsewhere.     

结核性脑膜炎脑脊液细胞学动态观察的临床意义

目的探讨结核性脑膜炎(tuberculous meningitis,TBM)患者脑脊液(cerebrospinal fluid,CSF)细胞学动态观察的临床意义。方法选取我院TBM患者20例,对其应用CSF细胞学检测方法,分析不同时期的TBM的临床特点以及持续时间。结果 TBM患者在临床经抗结核治疗前后CSF的中性粒细胞、激活单核细胞、小淋巴细胞、单核细胞、转移淋巴细胞以及淋巴样细胞所占比例具有统计学意义(χ2=2.055、1.583、2.054、3.512、2.025、2.050,P<0.05)。细胞学诊断结果治疗前后比较具有统计学意义(χ2=2.157、3.145,P<0.05)。结论 CSF细胞学动态检测对判断患者的治疗情况具有一定的指导意义。     

氯霉素联合青霉素治疗小儿难治性化脓性脑膜炎

目的探讨氯霉素联合青霉素对小儿难治性化脓性脑膜炎的治疗效果。方法对我科近5a来收治的27例难治性化脓性脑膜炎患儿应用氯霉素联合青霉素治疗,并对结果进行分析。结果所有27例患儿均临床治愈,达到出院标准,未出现明显不良反应。结论对小儿难治性化脓性脑膜炎可选择氯霉素联合青霉素进行治疗,安全有效。     

Antibody-induced neutrophil depletion prior to the onset of pneumococcal meningitis influences long-term neurological complications in mice

During pneumococcal meningitis, clearance of bacteria by recruited neutrophils is crucial for host protection. However, these innate immune mechanisms are often insufficient and treatment with antibiotics is necessary to prevent death. Despite this antibiotic treatment, approximately half of all survivors suffer lifelong neurological problems. There is growing evidence indicating the harmful effects of neutrophils on CNS integrity. Therefore, the present study investigated the roles of neutrophils in the acute inflammatory response and the resulting long-term neuropsychological effects in murine pneumococcal meningitis. Long-term behavioural and cognitive functions in mice were measured using an automated IntelliCage system. Neutrophil depletion with antibody 1A8 as adjunctive therapy was shown to remarkably impair survival in meningitic C57BL/6J mice despite antibiotic (ceftriaxone) treatment. This was accompanied by increased bacterial load in the cerebrospinal fluid (CSF) and an increase in IL-1β, but decrease in TNF, within the CSF at 20 h after bacterial inoculation. In the longer term, the surviving neutrophil-depleted post-meningitic (PM) mice displayed reduced diurnal hypolocomotion compared to PM mice treated with an isotype antibody. However, they showed nocturnal hyperactivity, and greater learning impairment in a patrolling task that is believed to depend upon an intact hippocampus. The data thus demonstrate two important mechanisms: 1. Neutrophil extravasation into the CNS during pneumococcal meningitis influences the pro-inflammatory response and is central to control of the bacterial load, an increase in which may lead to death. 2. Neutrophil-mediated changes in the acute inflammatory response modulate the neuropsychological sequelae in mice that survive pneumococcal meningitis.     

重症化脓性脑膜炎儿童血CD4~+CD25~+调节性T细胞的变化

目的初步探讨CD4~+CD25~+调节性T细胞(Tregs)在儿童重症化脓性脑膜炎病程早期的变化及其可能的作用。方法回顾性研究2014年8月至2015年12月入住PICU的39例严重化脓性脑膜炎患儿的临床资料,并根据患儿入院后12h内血Tregs降低与否分为无Tregs降低组和Tregs降低组(Tregs计数410个/mm3为降低),分析早期Tregs改变与临床表现、实验室指标和转归的关系。结果 39例患儿中,13例(33%)Tregs比例明显下降(31%),18例(46%)Tregs绝对计数下降(410个/mm~3)。4例病死患儿均来自于Tregs降低组。与无Tregs降低组比较,Tregs降低组患儿外周血白细胞低于正常的比例更高,血清降钙素原升高更显著(P0.05)。结论儿童重症化脓性脑膜炎患儿早期Tregs可能受到抑制,这与患儿炎症反应更重、病死率更高有关。