目的 探讨肠道病毒相关性脑炎患儿的临床特点;以期能为临床医师对该病的认识提供一定的帮助。方法 回顾性分析2018年1月—2019年12月广州市妇女儿童医疗中心感染科收治的37例肠道病毒相关性脑炎患儿的临床相关资料。结果 37例患儿男28例,女9例,男女比例3.11:1。主要临床症状体征发热(97.30%)、口腔疱疹和(或)皮疹(54.05%);常见神经系统症状呕吐(56.76%)、头痛(56.76%)、惊厥(29.72%),其中惊厥及呕吐头痛症状≤1岁组与其他年龄组差异有统计学意义;脑脊液检查白细胞升高为主;所有患儿均康复出院且无神经系统后遗症。结论 儿童肠道病毒相关性脑炎近一半患儿临床无咽部疱疹或皮疹表现,1岁以内患儿主要以发热及惊厥为主要表现,大于1岁尤其学龄前期及学龄期患者以发热呕吐伴头疼为主要表现;早期诊治预后良好。

Objective To explore the clinical characteristics of children with enterovirus associated encephalitis, in order to provide some help for clinicians to understand the disease. Methods The clinical data of 37 children with enterovirus related encephalitis treated in the infection department of Guangzhou Women and Children's Medical Center from January 2018 to December 2019 were analyzed retrospectively. Results There were 28 males and 9 females, with a male to female ratio of 3.11:1. The main clinical symptoms and signs were fever (97.30%), oral herpes and/or rash (54.05%); the common nervous system symptoms were vomiting (56.76%), headache (56.76%) and convulsion (29.72%). There were significant differences in convulsion, vomiting and headache symptoms between ≤ 1 year old group and other age groups. The leukocytes level in cerebrospinal fluid was elevated. All children recovered and discharged without neurological sequelae. Conclusions Nearly half of children with enterovirus associated encephalitis had no clinical manifestations of pharyngeal herpes or rash. The main manifestations of children under 1 year old were fever and convulsion. The main manifestations of children over 1 year old, especially preschool and school-age patients, were fever and vomiting with headache. Early diagnosis and treatment had good prognosis.

目的 利用大肠杆菌原核表达系统优化表达纯化肠道病毒71型(EV71)VP3结构蛋白,为后续单克隆抗体制备及检测试剂盒研发提供前期基础。方法 采用PCR方法扩增EV71病毒VP3基因,将其插入表达载体pET28a(+),构建pET28a-VP3重组质粒,转化大肠杆菌BL21(DE3)菌株,分别在25 ℃、37 ℃下经IPTG诱导表达,重组表达的蛋白产物经凝胶电泳初步分析,比较不同温度诱导表达的蛋白产物。结果 成功构建pET28a-VP3重组质粒,不同温度下诱导表达的蛋白产物在30.5 kDa左右位置均出现目的条带;37 ℃下诱导表达的蛋白超声破碎并离心后,目的蛋白基本位于沉淀中,而25 ℃诱导表达的蛋白产物有少量目的蛋白溶解于上清液中。结论 在25 ℃或37 ℃下均能利用大肠杆菌原核表达系统有效表达EV71病毒VP3蛋白;37 ℃诱导时蛋白可融性表达低,目的蛋白获取效率较高。

Objective To express VP3 capsid protein of enterovirus 71 by using Escherichia coli prokaryotic expression system. Methods VP3 gene was amplified by PCR before inserted into pET28a(+) plasmid. Then the plasmid pET28a-VP3 was transformed and expressed in the Escherichia coli BL21 strain at 25 ℃ or 37 ℃. Finally the protein was analyzed by SDS-PAGE gel electrophoresis. Results The pET28a-VP3 plasmid was successfully constructed, and the EV71 VP3 protein was expressed. Supernatant of the production after ultrasonication and centrifugation got a little VP3 protein. Conclusion The EV71 VP3 protein was expressed. Expression at 25 ℃ may lead to the dissolution of the recombinant protein.