The humoral immune response and protective efficacy of vaccination with inactivated split and whole influenza virus vaccines in BALB/c mice

Recently the urgency of developing a pandemic influenza vaccine has lead to the re-evaluation of the use of whole virus vaccine. We have compared the humoral immune response and the protective efficacy of whole and split influenza virus vaccines in mice. Whole virus vaccine was more immunogenic particularly after the first dose of vaccine, generally eliciting higher numbers of systemic antibody secreting cells and an earlier and higher neutralising antibody response. Immunisation with one dose of whole virus vaccine more effectively reduced viral shedding upon non-lethal homologous viral challenge, but two doses of split virus vaccine was most effective at limiting viral replication and this was correlated with high influenza specific serum IgG concentrations. The two vaccine formulations induced different T helper profiles particularly after one dose of vaccine; split virus vaccine induced a type 2 bias response, whereas whole virus vaccine elicited a dominant type 1 response.     

肺纤维化模型大鼠肺组织中转化生长因子β1及转化生长因子β1mRNA的表达及黄芪莪术合剂的干预效应

目的:分析中药黄芪莪术合剂对博莱霉素所致大鼠肺间质纤维化的干预作用。方法:实验于2002-10/2003-02在天津中医药大学动物实验室完成。①健康SD大白鼠60只,正常饲养1周后,随机分为正常对照组6只、模型组18只、黄芪莪术合剂组18只、强的松组18只。②经气管滴入博莱霉素复制肺纤维化大鼠模型,黄芪莪术合剂组给予40.5mg/L黄芪莪术合剂,强的松组给予0.63g/L强的松盐水溶液,均采用灌胃方式给药,每次1.5mL,1次/d。正常对照组、模型组给予等量生理盐水。③模型组、黄芪莪术合剂组、强的松组分别于造模第3,7,21天每组6只麻醉下取右肺下叶组织后处死,正常对照组于第3天麻醉下取右肺下叶组织后处死。④采用免疫组织化学方法和原位杂交方法检测肺组织转化生长因子β1及转化生长因子β1mRNA的表达。结果:进入结果分析大鼠55只,中途脱落5只。①转化生长因子β1表达:造模第3,7,21天模型组高于正常对照组(P<0.01),黄芪莪术合剂组和强的松组明显低于模型组(P<0.01～0.05)。造模第3,21天强的松治疗组低于黄芪莪术合剂治疗组(P<0.05)。②转化生长因子β1mRNA表达:造模第3,7,21天模型组高于正常对照组,黄芪莪术合剂组和强的松组明显低于模型组(P<0.01);第7天时,强的松治疗组低于黄芪莪术合剂治疗组,第21天时黄芪莪术合剂治疗组低于强的松治疗组,差异均有显著性意义(P<0.05)。结论:中药黄芪莪术合剂具有显著抑制博莱霉素致大鼠肺纤维化的作用,抑制转化生长因子β1及转化生长因子β1mRNA的表达是其可能的机制之一。     

10种中药有效成分抑制口腔主要致病菌比较

目的：为评价常见中药对口腔主要致病菌作用及其作为口腔保健用药的可能性。方法：从文献报导中有抗致龋作用的中草药中选择出１０种中药或天然植物进行１０种代表性的口腔致病菌的药敏试验。结果：茶多酚和鞣酸具有较强的抑菌和杀菌作用。结论：初步认为茶多酚和鞣酸更适合作为预防和治疗口腔疾病的保健用药。     

常见角膜致病真菌基因芯片检测方法的实验

目的:建立一种根据基因芯片原理,对国内真菌性角膜病常见致病菌种进行快速检测和准确检测的实验方法。方法:实验于2004-11/2006-04在河南省生物工程中心分子生物学实验室和河南省角膜病重点实验室完成。将针对国内临床上常见的角膜致病真菌6属12种,包括腐皮镰孢菌、串珠镰孢菌、梨状镰孢菌、尖孢镰孢菌、烟曲霉菌、黄曲霉菌、土曲霉菌、黑曲霉菌、新月弯孢菌、牵连青霉菌、链格孢霉菌、白色念珠菌,并经修饰的特异性寡核苷酸探针固定在两种材料的片基上。用一对通用引物,对上述真菌的DNA基因片断进行扩增,将带有特殊标记的扩增产物与片基上排列的探针进行杂交,对12株标准菌株和82株临床分离株样本采用双盲法原则进行检测,从而对两种不同片基芯片的特异性和灵敏度进行判断和评估。结果:①在两种不同材料的片基上,均可以高效率结合经过修饰和加尾的寡核苷酸探针,玻片约为35mer,尼龙膜约为400mer,可靠性较好。②通过琼脂糖凝胶电泳观察,12种真菌均产生了530～630bp的聚合酶链反应扩增产物,在相同的条件下对其进行杂交检测,得到了具有各自特征的杂交后显色图谱,玻片为荧光显色,尼龙膜为生物素-亲和素显色,可直接从杂交显色图谱上对不同菌种区分判断。③两种芯片均可与低于琼脂糖凝胶电泳显色浓度一个数量级的聚合酶链反应扩增产物反应,敏感性高于聚合酶链反应检测;在对82株临床分离株样本检测中,79株经聚合酶链反应扩增琼脂糖凝胶电泳均显示扩增产物条带,无非特异性杂交反应发生;玻片灵敏度为92.4%,尼龙膜灵敏度87.3%,χ2检验差异无显着性(P>0.05)。结论:实验建立了以ITS区基因为靶标的寡核苷酸探针芯片检测系统,具有较好的特异性和灵敏度,能够在三四个小时内完成对临床上常见的12种角膜致病真菌的菌种检测。     

脑死亡状态巴马小型猪肺水含量及肺超微结构变化

目的:观察脑死亡状态下巴马小型猪肺水含量及肺组织常规及超微结构改变。方法:实验于2003-08/2004-12在河南省实验动物中心及河南省病理学重点实验室完成。选用健康巴马小型猪11只,采用随机数字表法随机分为2组,对照组5只,脑死亡组6只。对照组持续麻醉维持24h,不建立脑死亡模型,24h后撤除麻醉,动物苏醒后处死;脑死亡组6只,应用改进的缓慢间断颅内加压法建立脑死亡模型,判定脑死亡后通过呼吸、循环支持维持动物脑死亡状态24h,24h后撤除呼吸、循环支持,动物心肺死亡。维持脑死亡状态下巴马小型猪的生命体征达到脑死亡供体要求,即平均动脉压≥60mmHg(1mmHg=0.133kPa),血氧饱和度≥95%,血氧分压≥100mmHg,体温≥35℃,中心静脉压≤10~12mmHg,血流动力学稳定。观察脑死亡组动物脑死亡24h后肺组织肺水含量、肺脏常规组织形态(光镜,苏木精-伊红染色)及超微结构变化(电镜),并与对照组比较。结果:脑死亡组1只动物在脑死亡造模过程中因心肺死亡脱失。①两组动物肺水含量比较:脑死亡组肺水含量显著高于对照组[(0.83±0.00,0.76±0.02)g/g(P<0.05)]。②两组动物肺脏常规组织形态及超微结构变化:脑死亡组动物肺组织光镜下见肺间质弥漫性水肿,肺泡间隔增宽,肺泡腔有渗出液,局灶性肺不张,肺毛细血管充血;电镜下可见肺泡Ⅱ型上皮细胞线粒体肿胀,线粒体嵴存在排列紊乱、消失,微绒毛缺失,板层小体数量减少、脱颗粒,结构有破坏。对照组光镜及电镜下未见明显的损伤性变化。结论:脑死亡状态可导致肺水含量增加,肺组织出现损伤性改变,可能对移植后肺组织功能造成一定影响。     

Intramuscular Matrix-M-adjuvanted virosomal H5N1 vaccine induces high frequencies of multifunctional Th1 CD4 cells and strong antibody responses in mice

Ideally, a candidate pandemic influenza vaccine should elicit rapid and strong cell-mediated and humoral immune responses, which are long-lasting and exhibit broad cross-reactivity against drifted strains. The present study investigated the detailed humoral and cellular immune responses in mice vaccinated intranasally or intramuscularly with inactivated influenza H5N1 (NIBRG-14) virosomal vaccine alone or formulated with Matrix-M adjuvant. The intramuscular Matrix-M-adjuvanted vaccine induced a strong immediate and long-term humoral immune response with high cross-reactivity against drifted H5N1 viruses and showed a dose-sparing potential. Additionally, the vaccine induced a balanced Th1/Th2 cytokine profile and most importantly high frequencies of multifunctional Th1 CD4⁺ cells. Our results highlight that Matrix-M adjuvant is a promising parenteral adjuvant for formulating pandemic candidate vaccines.     

Intranasal c-di-GMP-adjuvanted plant-derived H5 influenza vaccine induces multifunctional Th1 CD4+ cells and strong mucosal and systemic antibody responses in mice

Vaccination is the best available measure of limiting the impact of the next influenza pandemic. Ideally, a candidate pandemic influenza vaccine should be easy to administer and should elicit strong mucosal and systemic immune responses. Production of influenza subunit antigen in transient plant expression systems is an alternative to overcome the bottleneck in vaccine supply during influenza pandemic. Furthermore, a needle-free intranasal influenza vaccine is an attractive approach, which may provide immunity at the portal of virus entry.The present study investigated the detailed humoral and cellular immune responses in mice vaccinated intranasally or intramuscularly with plant-derived influenza H5N1 (A/Anhui/1/05) antigen alone or formulated with bis-(3′,5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) as adjuvant. The use of c-di-GMP as intramuscular adjuvant did not enhance the immune response to plant-derived influenza H5 antigen. However, intranasal c-di-GMP-adjuvanted vaccine induced strong mucosal and systemic humoral immune responses. Additionally, the intranasal vaccine elicited a balanced Th1/Th2 profile and, most importantly, high frequencies of multifunctional Th1 CD4⁺ cells. Our results highlight that c-di-GMP is a promising mucosal adjuvant for pandemic influenza vaccine development.     

蛇床子中一新的角型呋喃香豆素

从辽宁新民产蛇床Cnidium monnieri 果实中分离鉴定8个化合物,分别为欧山芹素(oroselone,Ⅰ),β谷舀醇(Ⅱ),哥伦比亚内酯(columbianadin,Ⅲ),佛手柑内醋(bergapten,Ⅳ),O-乙酰哥伦比亚甙贰元(O-acetycolumbianetin,v),O-乙酰异蛇床素(cniforinA,Ⅵ),爱得尔庭(edultin,Ⅶ)和'-乙酰白芷素(2'-acetylangelicin,Ⅷ)。其中化合物Ⅷ为一新化合物,化合物Ⅰ系首次从该植物中分得。