DNA vaccination using bacillus Calmette-Guerin-DNA as an adjuvant to enhance immune response to three kinds of swine diseases

In order to enhance the immune efficacy of DNA vaccination, experiments were conducted to investigate the regulating effects of Bacillus Calmette-Guerin （BCG）-DNA as an adjuvant on immune responses of mice against foot-and-mouth disease （FMD）, Aujeszky＇s disease （AID） and classical swine fever （CSF）. BCG-DNA was purified from BCG by ion-exchange chromatography. Three DNA vaccines （pVSG, pVgD and pVE2） against the respective infection were constructed, and BCGDNA was coimmunized to mice by muscle injection. The results showed that titres of specific immunoglobulin （Ig）G to the vaccines mounted remarkably in the sera of the adjuvant covaccinated mice （P〈0.01）. Antibody isotype IgG2a and IgG1 also increased, respectively, in mice coimmunized with BCG-DNA compared with those of the control groups （P〈0.01）. Cellular immune cytokine interferon-gamma and cytotoxic T lymphocytes were detected in coimmunized BCG-DNA groups （P〈0. 05）. Whereas interleukin-4, humoral immune cytokine, was not significant （P〉 0. 05）. These results suggest that codelivery of BCG-DNA with DNA vaccines against FMD, AjD and CSF can enhance the induction of antigen-specific, especially, cell-mediated immunity.     

国产麻黄的形态组织学研究——Ⅱ.西南主产种及其它种

本文报道我国西南产麻黄——丽江麻黄Ephedra likiangensis Florin、匍枝丽江麻黄E.likiangensis f.mairei(Florin)C.Y.Cheng、藏麻黄E.saxatilis Royle ex Florin、山岭麻黄E.gerardiana Wall、垫状山岭麻黄E.gerardiana Var.congesta C.Y.Cheng、矮麻黄E.minuta Florin和异株矮麻黄E.minuta var.dioeca C.Y.Cheng,以及形态组织特征较特殊的宁夏产斑子麻黄E.lepidosperma C.Y.Cheng、新疆产窄膜麻黄E.lomatolepis Schrenk,西藏产西藏中麻黄E.intermedia var.tibetica Stapf的生药形态组织学研究结果。并根据对国产麻黄的生药形态组织学的系统研究结果,分别编写了各种国产麻黄(包括13种3变种1变型)的生药性状和生药显微特征检索表。     

临床感染病原菌菌种分布及其耐药性分析

目的了解笔者所在医院2008年度临床感染病原菌的菌种分布及其耐药现状。方法用VITEK微生物自动鉴定仪鉴定菌种,纸片扩散法检测临床分离菌对各种抗菌药物的敏感性,判读标准参照当年的CLSI M100文件。结果临床分离的1900株病原菌中革兰阴性杆菌、革兰阳性球菌和真菌所占比例分别为68.42%、18.16%和12.32%。分离率最高的五种病原菌:铜绿假单胞菌、大肠埃希菌、肺炎克雷伯菌、鲍曼不动杆菌和白色假丝酵母菌,占所有分离病原菌的59.26%,排前20位的病原菌则占到近90%(89.63%)。革兰阴性杆菌仅对亚胺培南、头孢哌酮/舒巴坦、美洛培南、哌拉西林/他唑巴坦的平均耐药率低于30%,铜绿假单胞菌和鲍曼不动杆菌对亚胺培南、美洛培南的耐药率分别28.9%、32.9%和11.6%、24.7%,大肠埃希菌、肺炎克雷伯菌和奇异变形杆菌的ESBL阳性率分别是52.8%、37.2%和34.6%,金黄色葡萄球和凝固酶阴性葡萄球菌中耐甲氧西林株各占44.0%和60.0%,未检出万古霉素中介或耐药的葡萄球菌,检出万古霉素耐药的粪肠球菌和屎肠球菌各1株。念珠菌对氟康唑耐药率接近5%。结论临床感染病原菌构成变化不大,但真菌检出率逐年升高。非发酵菌耐药率最低的是头孢哌酮/舒巴坦,肠杆菌科耐药率最低的是亚胺培南,ESBL的阳性率逐年升高。加强病原菌及其耐药性检测和监测,对病原学诊断及指导合理选用抗生素具重要参考价值。     

An evaluation of chemical photoreactivity and the relationship to phototoxicity

The existing regulatory guidance for photosafety testing of new drug products states that studies are warranted for those chemicals that both absorb light in the range of 290–700 nm, and that are either applied locally/topically, or “reach” (EMEA)/“significantly partition” (FDA) to the skin or eyes. The initial in vitro study recommended for the assessment of phototoxic potential is the 3T3 Neutral Red Uptake (NRU) Assay. The current study was undertaken to establish superior triggers for the initiation of biological photosafety testing. In this study, photophysical and photochemical parameters for 40 drug or drug-like molecules were studied. Principal Component Analysis (PCA), Partial Least Squares-Discriminant Analysis (PLS-DA), and a fivefold cross-validation PLS algorithm were used to evaluate the relationship between subsets of photophysical and photochemical parameters with the 3T3 NRU PIF/MPE (Photo Irritation Factor/Mean Photo Effect) results. The parameters most indicative of a 3T3 NRU positive PIF or MPE score were the extent of degradation in solution, the quantum yield of formation of singlet oxygen and the relative formation of superoxide anion. The results demonstrate that while absorption of light is critical to the induction of a light-induced process, it is the resultant events that may be used to predict the 3T3 NRU assay result. It is therefore proposed that the trigger for photosafety testing be revised to include a molecular basis for photoreactivity. From this limited investigation, estimated thresholds leading to 3T3 NRU positive results due to photodegradation, formation of singlet oxygen quantum yield or a relative superoxide anion formation value are proposed.     

固相法合成心肌兴奋肽及其类似物

用Boc-和Tos-基团分别保护氨基和侧链胍基,以1%交联度聚苯乙烯二苯甲氨基树脂为载体,用DCC固相法合成肽,HF断裂肽树脂键和去除侧链保护基团,粗产物经高效液相层析纯化,合成了心肌兴奋肽Phe-Met-Arg-Phe-NH_2及其类似物Phe-Pro-Arg-Phe-NH_2,并观察了此二种肽对大鼠血压和心率的影响。     

羧乙基锗倍半氧化物对正常和异丙基肾上腺素损伤培养乳鼠心肌细胞的影响

羧乙基锗倍半氧化物(Ge—132)能促进培养乳鼠心肌细胞DNA和RNA的合成,提高超氧化物歧化酶(SOD)的活性,维持细胞膜结构的稳定。异丙基肾上腺素可使心肌细胞乳酸脱氢酶(LDH)释放增加,细胞搏动停止,超微结构显示肌膜和线粒体严重受损。Ge—132可显著减少异两基肾上腺素引起心肌细胞释放LDH,维持细胞的搏动功能和超微结构的完整。     

苦楝化学成份的研究

从苦楝(Melia azedarach L.)果中分得苦楝新醇(Ⅰ),苦楝醇(Ⅱ)、苦楝酮(Ⅲ)、苦楝二醇(Ⅳ)、香草醛(Ⅴ)和香草酸(Ⅵ)。根据波谱(IR,MS,~1HNMR,~(13)CNMR)分析和理化常数测定,确定了它们的结构。其中苦楝新醇(Ⅰ)为新化合物,对菜青小虫有一定的拒食活性。     

6－甲氧基正丁苯酞在大鼠肝微粒体中的代谢研究

报道了用ＧＣ／ＭＳ方法及衍生化技术研究６甲氧基正丁苯酞（ＭＢＰ）在大鼠肝微粒体中的代谢转化结果。６甲氧基正丁苯酞在苯巴比妥（ＰＢ）诱导的大鼠肝微粒体中主要转化为３羟基、γ羟基取代物，６羟基正丁苯酞与一个环氧化代谢产物     

冻存复苏条件对髓性白血病细胞株生物学特性的影响

目的:采用不同方法对白血病HL60细胞株进行冻存和复苏,观察生物学特性改变,筛选最佳冻存复苏方案。方法:实验于2006-04/06在吉林医药学院临床检验实验室(国家三级标准)进行。①HL60细胞株由中国科学院上海生物细胞研究所提供。将欲冷冻保存的HL60细胞调整到良好的生长状态(对数生长期),随机数字表法分成4组,离心收集后在含体积分数为0.1小牛血清的RPMI-1640培养基中分别加入二甲基亚砜,使其终浓度依次为50,100,150,200g/L。冻存15d后,每组取1支冻存管复苏并接种于细胞培养板,培养12h后用锥虫蓝拒染法计算细胞复苏率。②选择冻存条件和冻存细胞密度相同的两支冻存管,按不同方法进行HL60细胞复苏。37℃水浴传统复苏法:立即将冻存管置于37℃水浴中,待融化后800r/min离心5min,吸去上清液,加入含体积分数为0.1小牛血清的RPMI-1640培养基,混匀后接种于细胞培养板,1mL/孔,9孔/组,置于体积分数为0.05的CO2培养箱37℃继续培养。40℃溶解后再37℃恒温改良复苏法:将冻存细胞于40℃水浴中迅速溶解,转入37℃水浴箱,融化后离心、加培养基等操作同传统复苏法。复苏细胞培养12h后采用锥虫蓝染色计算细胞存活率。③用无血清RPMI-1640培养基制备HL60细胞悬液,按3×107L-1密度接种于细胞培养板中,1mL/孔,然后分别加入体积分数为0.05,0.1,0.12,0.15,0.2的灭活小牛血清,每种血清浓度设置8个试验孔,并分别于培养12,24,36,48,60,72,84,96h后计数每孔细胞数量,并绘制细胞生长曲线。结果:①不同浓度二甲基亚砜冻存后HL60细胞复苏率的比较:二甲基亚砜200g/L组的细胞复苏率明显低于二甲基亚砜50,100,150g/L组[(64.6±2.8)%,(87.0±1.4)%,(86.4±2.1)%,(85.7±2.8)%;t=25.44,P<0.01],二甲基亚砜50,100,150g/L组间差异无显著性意义(t=0.82～1.44,P>0.05)。②不同复苏方法HL60细胞存活率的比较:与37℃水浴传统复苏法比较,40℃溶解后再37℃恒温改良复苏法的细胞存活率显著升高[(69.5±1.5)%,(87.4±1.8)%,t=23.24,P<0.01]。③RPMI-1640培养基不同血清含量对HL60细胞生长情况的影响:在体积分数为0.05的血清含量培养基中,HL60细胞基本不生长,且有逐渐死亡的趋势;在体积分数为0.1的血清含量培养基中,HL60细胞生长趋势明显,但生长速度相对较慢;在体积分数为0.12,0.15,0.2的血清含量培养基中,HL60细胞生长迅速,3种血清浓度间细胞生长趋势无明显差异。结论:以50g/L二甲基亚砜作为HL60细胞冻存保护剂、联合40℃溶解后再37℃恒温改良复苏法可使细胞保持最佳生物学特性,体积分数为0.12的血清含量为HL60细胞常规培养的最适浓度。