新型冠状病毒肺炎患者4S呼吸康复指引

2019新型冠状病毒(2019-nCoV)感染的肺炎具有高度传染性,本文结合4S(simple,safe,satisfy,save)呼吸康复内容和2019新型冠状病毒肺炎的诊治标准,对2019-nCoV所致的肺炎患者提供可行的呼吸康复指引.     

新型冠状病毒肺炎疫情防控期间开展肺功能检查的专家共识

新型冠状病毒肺炎(COVID-19)主要通过呼吸道飞沫传播及密切接触传播,也可能会通过气溶胶传播。肺功能检查可增加医务人员和受检者发生COVID-19传播的风险,必须严格执行有效的预防和控制措施以防止院内感染。为了指导肺功能检查室医务人员做好防控工作,本文结合肺功能检查的特点,总结了当前疫情下肺功能检查在医务人员管理、检查流程管理和检查环境物品管理3个方面的要求及注意事项。文中强调在疫情流行期间,必须严格掌握肺功能检查的适应证,强烈建议COVID-19确诊病例或疑似病例在传染期内暂缓检查,其他病患如非病情急需也暂缓检查;肺功能室医务人员应严格执行标准分级防护措施;受试者应在单独区域进行隔离检查;检查时必须使用一次性呼吸过滤器;并重视肺功能检查环境和设备的清洁消毒。     

利用LMS法建立全年龄段FEV1正常参考值

目的探讨一种建立肺功能全年龄段的正常参考值的方法。方法利用LMS法对4个中心共12829名正常健康人FEV1建立全年龄段正常参考值曲线和正常值下限,并对该模型进行评价。结果FEV1与身高、年龄之间存在密切关联,在校正身高影响之后,FEV1在全年龄段与年龄呈非线性关系,而且变异随着年龄变化而变化。LMS方法考虑个体间变异,适用于不服从正态分布的偏峰数据,而且还可以校正身高等其他因素的影响,计算得到的不同年龄的正常参考值最低下限更符合实际。结论LMS法是一种建立全年龄肺功能正常参考值的有效统计方法。     

螨制剂舌下免疫治疗对单一和多重致敏变应性鼻炎的疗效评估

目的观察单一和多重变应性鼻炎患者舌下脱敏的疗效异同并分析原因。方法回顾性分析舌下脱敏治疗1年以上单一和多重变应性鼻炎患者终止治疗1年后的疗效,并对粉尘螨变应原组分与其他点刺变应原作同源进化树分析。结果纳入患者50例,舌下免疫治疗显效22例,有效15例,无效13例,总有效率为74%。50例患者中30例(60%)为多重变态反应患者,阳性率最高的前3种变应原依次为蟑螂(83%)、冬季花粉(70%)和蚕丝(63%)。用有序多分类资料比较的秩和检验分析舌下脱敏治疗1年以上单一尘螨过敏患者与多重变态反应患者的疗效,两组总体分布相同。结论使用舌下含服粉尘螨滴剂治疗变应性鼻炎,多重与单一尘螨变态反应患者治疗效果相当。粉尘螨滴剂亦可用于多重变态反应患者常规治疗。     

药理剂量维生素C杀灭流感病毒的体外研究

目的 探讨药理剂量维生素C体外杀灭流感病毒A/CA/7/09 (H1N12009)的作用及其机制.方法 将流感病毒A/CA/7/09感染人类正常呼吸道上皮细胞(NHBE)后,立即或8h后分别加入2.5～20 mmol/L维生素C、维生素C+过氧化氢酶和不含维生素C的NHBE培养基,或直接在流感病毒A/CA/7/09中分别加入上述培养基.培养4～12 h后,分别收集培养基上清液,通过半数组织培养感染剂量( TCID50)法对病毒进行定量检测.利用高压液相层析法(HPLC)测量样品中维生索C含量和Clark型氧电极法测量样品中过氧化氢浓度.结果 药理剂量的维生素C可杀灭体外游离的流感病毒,也能杀灭体外培养细胞内的流感病毒;维生素C杀灭病毒的效应与剂量相关:2.5 mmol/L的维生素C可杀灭约90％的流感病毒,20 mmol/L的维生素C可杀灭所有病毒;维生素C在病毒感染不同时间窗的杀灭病毒效应不同,在病毒感染8～12h内,维生素C杀灭病毒的效果最好;4 h内可清除感染NHBE细胞的所有流感病毒;感染后期维生素C杀灭病毒的效果最差.维生素C杀灭病毒的效应可被过氧化氧酶完全抑制.结论 药理剂量的维生素C作为一种强氧化剂,可能通过生成过氧化氢发挥杀灭细胞内外流感病毒的作用.     

钙池操纵性钙内流在非小细胞肺癌中的作用

目的探讨钙池操纵性钙通道（store-operated calcium channel,SOCC）介导的钙池操纵性钙内流（storeoperated calcium entry,SOCE）在非小细胞肺癌中的作用。方法应用实时定量PCR方法和Western blot方法检测非小细胞肺癌组织和A549肺腺癌细胞中SOCC的mRNA和蛋白表达。应用Fura-2Am荧光素方法检测A549细胞的SOCE;经过不同浓度SOCE抑制剂处理A549细胞后,应用WST-1比色法和Transwell方法检测A549细胞的增殖和侵袭能力改变。结果在非小细胞肺癌组织和A549细胞中有STIM1、ORAI1～3的mRNA和STIM1、ORAI1的蛋白表达,分化差的肿瘤组织比分化好的高表达ORAI1 mRNA（P=0.028）;A549细胞中存在SOCE,并且SOCE抑制剂SKF-96365或Ni Cl2可抑制A549细胞的增殖和侵袭。结论非小细胞肺癌组织中表达STIM1和ORAI1～3。A549肺腺癌细胞存在SOCE,并且由SOCC介导的SOCE在肺癌细胞生长和转移过程中起着重要的作用。     

人工外泌体共递送siRNA和蛋白的递送系统的设计及体外评价

本文制备了人工外泌体,共传递蛋白和核酸,实现多组分药物高效安全共传递。采用阳离子脂质赋形剂二油酰基三甲基铵丙烷(dioleyl trimethylammonium propane, DOTAP)修饰聚乳酸-羟基乙酸共聚物(polylactic acidglycolic acid copolymer, PLGA)基质来设计优化的制剂,双乳化法制备包裹蛋白和核酸的PLGA/DOTAP纳米粒,再用逆相蒸发法制备最外层的膜结构,此膜结构由二棕榈酰磷脂酰胆碱(1, 2-dipalmitoyl-sn-glycero-3-phosphocholine,DPPC)、二油酰磷脂酰胆碱(1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC)、二硬脂酰磷脂酰胆碱(1,2-distearoylsn-glycero-3-phosphocholine, DSPC)、胆固醇及膜蛋白组成,通过超声分散和挤出的方式形成人工外泌体结构,并分析其物理特性和传递效果性质。结果表明,人工外泌体粒径约为156.13 nm,带负电荷(-18.23±0.57 mV),能高效共传递蛋白和siRNA,且siRNA能高效抑制目标基因Trim 28的表达。这说明人工外泌体模拟了外泌体结构,实现了多组分药物安全高效的共递送。     

A bivalent outer membrane vesicle-based intranasal vaccine to prevent infection of periodontopathic bacteria

Periodontal disease has become a serious public health problem, not only causing tooth loss, but also inducing chronic disorders of extra-oral organs. The present study assessed an intranasal vaccine strategy to prevent periodontal disease using outer membrane vesicles (OMVs) of two major periodontopathic bacteria, Porphyromonas gingivalis (Pg) and Aggregatibacter actinomycetemcomitans (Aa). We compared the morphology, composition, and immune activity between OMVs of Pg strain ATCC 33277 and Aa strain Y4. Aa OMVs had a smoother surface and stronger lipid A activity compared to Pg OMVs. The in vitro immune activity elicited by Aa OMVs in macrophage-like cells was remarkably stronger than that of Pg OMVs. Intranasal immunization of mice with Aa OMVs alone resulted in robust, humoral immune responses in blood and saliva. Despites the intrinsically low mucosal immunogenicity of Pg OMVs alone, using Aa OMVs as a mucosal adjuvant strongly enhanced Pg-specific immune responses, resulting in both serum IgG and salivary IgA, both of which aggregated Pg and Aa cells. Furthermore, Aa OMVs were found to be a more potent mucosal adjuvant than Poly(I:C) in the context of enhancing the production of Pg-specific IgG (especially IgG2a) and IgA. In addition, in a randomized, blinded study, mice oral challenged with Pg and Aa after intranasal immunization with Pg OMVs and Aa OMVs had significantly decreased numbers of both microorganisms compared to mock-immunized mice. Furthermore, in an intracerebral injection mouse model, there were no serious adverse effects on the brain even after administrating a dose of OMVs as same as that used for intranasal administration. Taken together, the bivalent OMV intranasal vaccine may be effective in preventing colonization of periodontopathic bacteria in the oral cavity and related systemic disorders associated with periodontal diseases.     

Recent advances in the development of vaccines for chronic inflammatory autoimmune diseases

Chronic inflammatory autoimmune diseases leading to target tissue destruction and disability are not only causing increase in patients’ suffering but also contribute to huge economic burden for the society. General increase in life expectancy and high prevalence of these diseases both in elderly and younger population emphasize the importance of developing safe and effective vaccines. In this review, at first the possible mechanisms and risk factors associated with chronic inflammatory autoimmune diseases, such as rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE) and type 1 diabetes (T1D) are discussed. Current advances in the development of vaccines for such autoimmune diseases, particularly those based on DNA, altered peptide ligands and peptide loaded MHC II complexes are discussed in detail. Finally, strategies for improving the efficacy of potential vaccines are explored.     

Addition of C3d-P28 adjuvant to a rabies DNA vaccine encoding the G5 linear epitope enhances the humoral immune response and confers protection

Rabies DNA vaccines based on full-length glycoprotein (G) induce virus neutralizing antibody (VNA) responses and protect against the virus challenge. Although conformational epitopes of G are the main target of VNAs, some studies have shown that a polypeptide linear epitope G5 is also able to induce VNAs. However, a G5 DNA vaccine has not been explored. While multiple doses of DNA vaccines are required in order to confer a protective immune response, this could be overcome by the inclusion of C3d-P28, a molecular adjuvant is know to improve the antibody response in several anti-viral vaccine models. To induce and enhance the immune response against rabies in mice, we evaluated two DNA vaccines based on the linear epitope G5 of Rabies Virus (RABV) glycoprotein (pVaxG5 vaccine) and another vaccine consisting of G5 fused to the molecular adjuvant C3d-P28 (pVaxF1 vaccine). VNA responses were measured in mice immunized with both vaccines. The VNA levels from the group immunized with pVaxG5 decreased gradually, while those from the group vaccinated with pVaxF1 remained high throughout the experimental study. After challenge with 22 LD50 of the Challenge Virus Strain (CVS), the survival rate of mice immunized with pVaxG5 and pVaxF1 was increased by 27% and 50% respectively, in comparison to the PBS group. Furthermore, the in vitro proliferation of anti-rabies specific spleen CD4+ and CD8+ T cells from mice immunized with pVaxF1 was observed. Collectively, these results suggest that the linear G5 epitope is a potential candidate vaccine. Furthermore, the addition of a C3d-P28 adjuvant contributed to enhanced protection, the sustained production of VNAs, and a specific T-cell proliferative response.