恶性疟原虫FCC1/HN株环子孢子蛋白的原核表达、纯化及其对肝细胞靶向作用的观察

目的从恶性疟原虫FCC1／HN株基因组中扩增得到环子孢子蛋白(circumsporozoite protein,CSP)的全长编码基因,克隆至原核表达载体pET-32c中进行表达纯化,观察重组环子孢子蛋白对肝细胞的结合能力,探讨其作为原发性肝癌基因治疗的靶向分子的可行性。方法根据恶性疟原虫3D7株环子孢子蛋白的编码序列设计一对引物,利用聚合酶链反应(PCR)技术从恶性疟原虫FCC1／HN株基因组中扩增出CSP的全长编码基因,将其克隆到载体pGEM-T中,通过基因测序加以证实;进一步亚克隆至原核表达载体pET-32c中,在大肠杆菌BL21中用IPTG诱导表达,表达产物用Ni2+螯合柱亲和纯化,采用免疫印迹技术(WB)对纯化的融合蛋白进行免疫反应性检测;采用免疫组化(IHC)技术观察重组环子孢子蛋白对不同组织细胞的结合能力。结果从恶性疟原虫FCC1／HN株基因组中成功扩增到1263 bp的全长CSP基因,该基因在原核系统中经诱导表达出一相对分子质量(Mr)约62×103大小的融合蛋白,表达产物以包涵体的形式存在;通过Ni2+亲和柱纯化获得重组CSP融合蛋白;WB表明,重组CSP融合蛋白能被疟原虫阳性血清特异性识别;免疫组化结果显示重组CSP融合蛋白能够与肝癌和正常肝细胞特异性地结合,与其他组织来源的细胞则未见反应。结论CSP是疟原虫子孢子表面主要的蛋白,重组环子孢子蛋白作为原发性肝癌基因治疗的靶向分子具有一定的潜在应用价值。     

外源性五羟色胺大鼠肺出血模型及其与浓度关系

目的　通过外源性五羟色胺 (5 -HT)气管滴入建立新生大鼠肺出血模型及其与浓度关系 .方法　模型制作 :日龄 4～ 5天Wistar二级大鼠 5 0只 ,随机分为 4组 :生理盐水对照组 (A组 )及 3种不同浓度外源性 5 -HT实验组(B、C、D组 ) :经气管导管分别滴入生理盐水和不同浓度 5 -HT ,4小时后处死 ,观察肺大体及组织病理改变 ,将肺出血程度分为 5级 :Ⅰ正常 ;Ⅱ肺水肿 ;Ⅲ点状肺出血 ;Ⅳ局灶性肺出血 ;Ⅴ弥漫性肺出血 ,选择出制作肺出血模型的最佳的浓度 .结果　不同浓度 5 -HT气管内滴入均能引起不同程度肺出血 ,但随着浓度增加 ,B、C、D三组间的肺出血程度无差异 (p >0 .0 5 ) ,其中D组死亡率 30 % ,对照组及B组和C组均无死亡 ,死亡鼠肺为弥漫性出血 .结论　 5 -HT可致大鼠肺出血 ,以 1× 10 -5mol/ml浓度为宜 ,随着 5 -HT浓度增加 ,大鼠死亡率增加 ,但肺出血发生率无差异     

冠状动脉血管直径测量方法的研究

心脏是人体生命活动的重要器官，能否正常工作主要是由心血管系和心传导系决定。冠状动脉是给心脏供血的唯一途径，心肌能否得到足够的营养、保持连续有节奏地运动，完全取决于冠状动脉是否能不断地供给其营养。冠状动脉直径的测量以及血管直径参数对于反映心脏功能和疾病的诊断具有重要意义。文中提出一种新型的、简便的血管直径测量方法，利用计算得到的血管直径，我们就可以判别心血管狭窄位置。     

Effects of interferons alpha and gamma on cytokine production and phenotypic pattern of human bronchial epithelial cells

Human bronchial epithelial cells are involved in airway immune mechanisms through secretion of cytokines and through cell-cell contacts with immunocompetent cells. The aim of our study was to assess the ability of interferon (IFN) alpha and gamma alone and in combination to modulate human bronchial epithelial cell (HBECs) release of the inflammatory cytokines IL-8 and IL-6 and fibronectin and to induce the surface expression of HLA-DR and ICAM-1 molecules involved in immune interactions with other cells. HBECs spontaneously secreted a limited amount of IL-8, which was significantly increased by IFN gamma. IFN alpha inhibited IFN gamma stimulated IL-8 secretion in a concentration-dependent manner. Further, IFN gamma induced IL-6 and fibronectin secretion, and this was also inhibited by IFN alpha. The expression of HLA-DR antigens was significantly increased by IFN gamma and partially inhibited by co-stimulation with IFN alpha. In contrast, IFN gamma also induced ICAM-1 expression by HBECs but co-stimulation with IFN alpha had no significant effect on the expression of this surface antigen. IFN alpha modulation of HBEC functions does not seem to be restricted to IFN gamma stimulation since either stimulatory or inhibitory effects of INF alpha on IL-8 production have been found in pilot experiments using IL-1 beta, TNF alpha, and TGF beta as stimuli. In summary, IFN-gamma induces a number of responses in HBECs including increased secretion of IL-6, IL-8 and fibronectin and increased expression of HLA-DR and ICAM-1. IFN alpha can inhibit all these except expression of ICAM-1 which is unaffected. IFN alpha can also interact with other inflammatory cytokines, but whether the effects are inhibitory or augmentive depends on the cytokines.     

Observations of development of resistance to detachment of cultured bovine bronchial epithelial cells in response to protease treatment.

Protease injury of the bronchial epithelium may play an important role in the pathogenesis of many airway diseases including asthma and chronic bronchitis. We hypothesized that neutrophil elastase can cause significant injury to the bronchial epithelium leading to detachment of bronchial epithelial cells. This detachment might be prevented if elastase is released into the airway lumen and the bronchial epithelium forms a barrier preventing access to the basal attachment sites. To assess this, the detachment of bronchial epithelial cells by elastase from extracellular matrix was measured. An increase in the resistance to detachment with time in culture was observed. The resistance to detachment was confirmed in bronchial epithelial cells, which were grown to electrically resistant monolayers on millipore filters and exposed to trypsin and elastase applied to both the apical and basal surfaces. Significantly greater detachment occurred when the proteases were applied at the basal surface versus the apical surface. Injury to the bronchial epithelium may enhance the proteolytic effect on the epithelium by disrupting epithelial barrier function. This was tested by exposing bronchial epithelial cell cultures to cigarette smoke extract before exposure to trypsin and elastase. The detachment of the bronchial epithelial cells exposed on the apical surface was increased greatly after smoke exposure. These data suggest that an intact bronchial epithelium can act as a barrier against proteolytic injury. Such a mechanism might protect the airway epithelium during intraluminal inflammation, and, if defective, might potentiate cigarette smoke-induced airway injury.     

品管圈对精神分裂症患者约束率及护理满意度的影响

目的总结和分析精神分裂症患者的护理中采用品管圈活动的临床效果。方法以本院于2017年8月—2019年6月收治的122例精神分裂症患者为研究对象,随机分为品管圈组和常规护理组,各61例。其中常规护理组患者选择传统的常规护理方法,而品管圈组患者则采取品管圈活动。比较两组患者的护理效果,对比指标主要有患者的约束率和护理满意度。结果品管圈组患者各项评价指标均优于常规护理组,差异具有统计学意义(P<0.05)。结论精神分裂症患者采用品管圈活动的总体临床效果较好,可提高患者的约束率,总体满意度高。     

2013年-2017年我国血站收入的结构变动情况分析△

目的：对我国不同类型血站的收入状况进行问卷调查和深度访谈,分析近年来我国血站收入结构变动情况及特征,找到影响血站收入结构变动的主要项目,合理优化血站收入结构。方法：2018 年8月采用普查的方法对全国353家（32 家血液中心、321 家中心血站）的收入状况进行问卷调查,同时选取山东省6地市中心血站的负责人进行深度访谈。对收集的血站收入数据,运用结构变动方法进行分析。 结果：2013年-2017年公益一类血站收入的结构变动度为24.22%,财政补助收入、事业收入是构成公益一类血站收入变动的主要来源,其中财政补助收入的贡献率53.20%;2013年-2017年我国公益二类血站收入的结构变动度为16.79%,财政补助收入、事业收入是构成公益二类血站收入变动的主要来源,其中贡献率最大的是事业收入49.99%。结论：财政补助收入、事业收入是影响血站收入结构变动的主要项目,两类血站的日常运营与建设对财政补助收入高度依赖,公益二类经营收入增幅不明显,发展不足。     

夜间最低血氧饱和度联合睡眠相关量表筛查儿童阻塞性睡眠呼吸暂停的应用

目的 评价夜间最低血氧饱和度(LSaO₂)联合OSA-18量表和针对儿童的改良版Epworth嗜睡量表(ESS-CHAD)初筛儿童阻塞性睡眠呼吸暂停(OSA)的准确性,以探讨更简便、快捷、可行的基层医院初筛疑似儿童OSA的方法。方法 纳入2020年8月13日—2021年2月8日就诊于本院耳鼻咽喉头颈外科睡眠监测中心完成整夜多导睡眠监测的儿童139例作回顾性分析研究,其家长在医生协助下完成OSA-18量表和ESS-CHAD。分析LSaO₂和两种量表及三者联合早期初筛疑似儿童OSA的诊断价值。结果 LSaO₂预测儿童是否患OSA的最佳界值为90.5%(P<0.00, AUC=0.82)。OSA-18和ESS-CHAD依据不同界值分别诊断儿童OSA的灵敏度最高,分别为74.8%、22.7%。LSaO₂/OSA-18,LSaO₂/ESS-CHAD、OSA-18/ESS-CHAD诊断儿童OSA的灵敏度最高,分别为88.2%、68.1%、77.3%。 结论 LSaO₂有诊断儿童OSA的临床价值,OSA-18量表较ESS-CHAD初筛疑似儿童OSA有更高的灵敏度,但特异度低,LSaO₂联合OSA-18量表诊断儿童OSA的灵敏度、特异度均较其他联合诊断方式高。在无法进行睡眠监测的基层医院可考虑应用LSaO₂ 与OSA-18量表结合的方式初步筛查儿童OSA。