活菌制剂活菌数急剧衰降的原因与减缓衰降的方法

市售活菌制剂口服液活菌数急剧衰降的原因是营养缺乏,特别是缺乏部分氨基酸及某些维生素。增加动物性蛋白胨及强化酵母膏,添加适量维生素可使活菌数一年后保持在１０^６／ｍｌ以上。     

Quantitative regulation of the thermal stability of enveloped virus vaccines by surface charge engineering to prevent the self-aggregation of attachment glycoproteins

The development of thermostable vaccines can relieve the bottleneck of existing vaccines caused by thermal instability and subsequent poor efficacy, which is one of the predominant reasons for the millions of deaths caused by vaccine-preventable diseases. Research into the mechanism of viral thermostability may provide strategies for developing thermostable vaccines. Using Newcastle disease virus (NDV) as model, we identified the negative surface charge of attachment glycoprotein as a novel determinant of viral thermostability. It prevented the temperature-induced aggregation of glycoprotein and subsequent detachment from virion surface. Then structural stability of virion surface was improved and virus could bind to and infect cells efficiently after heat-treatment. Employing the approach of surface charge engineering, thermal stability of NDV and influenza A virus (IAV) vaccines was successfully improved. The increase in the level of vaccine thermal stability was determined by the value-added in the negative surface charge of the attachment glycoprotein. The engineered live and inactivated vaccines could be used efficiently after storage at 37°C for at least 10 and 60 days, respectively. Thus, our results revealed a novel surface-charge-mediated link between HN protein and NDV thermostability, which could be used to design thermal stable NDV and IAV vaccines rationally.     

用指纹图谱评价环境样本DNA的提取效果

研究了DNA指纹图谱技术在评价环境样本DNA提取效果方面的应用。采用 3种DNA提取方法提取垃圾渗滤水和活性污泥中的微生物DNA ,并用ARDRA和RISA图谱加以评价。实验结果表明 ,RISA图谱分析法可以作为评估DNA提取效果的有效手段。     

同型半胱氨酸对大鼠血管平滑肌细胞增殖的作用

血中同型半胱氨酸（ｈｏｍｏｃｙｓｔｅｉｎｅ,ＨＣＹ）浓度的升高已成为动脉粥样硬化发生的一个独立危险因子．为进一步阐明ＨＣＹ促进血管平滑肌细胞（ｖａｓｃｕｌａｒｓｍｏｏｔｈｍｕｓｃｌｅｃｅｌｓ,ＶＳＭＣｓ）增殖,从而引起动脉粥样硬化发生的机理．本实验采用细胞计数、３Ｈ－ＴｄＲ参入、细胞周期分析、Ｎｏｒｔｈｅｒｎ杂交方法证明,一定剂量的ＨＣＹ可促进离体培养的ＷＫＹ大鼠血管平滑肌细胞增殖,使其ＤＮＡ合成增加,细胞周期中Ｓ期细胞所占比例增加４３％,并促进ｃ－ｍｙｃ与ｃ－ｆｏｓ原癌基因ｍＲＮＡ表达增加．提示ＨＣＹ可能通过促进ＶＳＭＣｓ增殖而诱发动脉粥样硬化     

Determination of cefazedone in human plasma by high performance liquid chromatography–tandem mass spectrometry: Application to a pharmacokinetic study on Chinese volunteers

A rapid, sensitive and simple high performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) method was developed for determination of cefazedone in human plasma using metronidazole as internal standard (IS). The chromatographic separation was achieved on an Ultimate XB-CN column (2.1 mm × 150 mm, 5 μm) with an isocratic mobile phase of acetonitrile and 20 mM ammonium acetate in 0.1% formic acid in water (15:85, v/v). Detection was performed using electrospray ionization in positive ion multiple reaction-monitoring mode (SRM), monitoring the transitions m/z 548.2 → 344.1 for cefazedone and m/z 172.2 → 128.1 for IS. Calibration curves were linear over a wide range of 0.20–401.12 μg/mL for cefazedone in plasma. The lower limit of quantification (LLOQ) was 0.20 μg/mL. The intra- and inter-day precisions were less than 7.2%. The average recovery of cefazedone was 90.8–91.0%. The validated method was successfully applied to the pharmacokinetic study of cefazedone in Chinese healthy volunteers following intravenous (IV) administration of 500, 1000 and 2000 mg cefazedone injection.     

鼎突多刺蚁群体结构和生活史的研究

本文报告了鼎突多刺蚁(Polyrhachis vicina Roger)的群体结构和生活史。通过研究表明,鼎突多刺蚁一年发生一代,以蚁后、雄蚁、工蚁、幼虫和卵越冬。卵和幼虫在冬天发育停滞(或极慢)。到了春天,随着气温的上升,卵和幼虫又恢复正常发育。据室内人工饲养观察,卵的发育历期为23.8±2.5天(平均温度26℃)幼虫为20.4±4.4天(26℃),工蚁蛹为19.8±5.5天(27℃)。成长工蚁在5—11月出现;8—11月,雄蚁从蛹中羽化;10月,雌蚁从蛹中羽化。雌蚁分飞交尾后,进入邻近蚁巢或回到原巢,脱翅成为蚁后。蚁巢内存在着多后现象。     

Phosphorylation of a 25 kDa protein is induced by thermostable direct hemolysin of Vibrio parahaemolyticus

Thermostable direct hemolysin (TDH) is a possible virulence factor produced by Vibrio parahaemolyticus. Although TDH has a variety of biological activities, including hemolytic activity, the biochemical mechanism of action remains uncertain. Here we analysed biochemical events, especially phosphorylation, caused by TDH in erythrocytes, and found that TDH caused significant phosphorylations of proteins on erythrocyte membrane. Phosphorylation of proteins was studied using γ-³²P ATP and SDS-PAGE. A number of protein kinase inhibitors were tested, to determine which types of kinases were involved in the phosphorylation events. TDH induced the phosphorylation of two proteins on membranes of human erythrocyte that are sensitive to TDH. The estimated molecular weight of these proteins was 25 and 22.5 kDa. Interestingly, the 22.5 kDa, but not the 25 kDa protein, was phosphorylated on the membrane of TDH-insensitive (resistant) horse erythrocytes. Moreover, a mutant TDH (R7), which retained binding ability but lost hemolytic activity, also phosphorylated only the 22.5 kDa protein on human erythrocyte membranes. Among the protein kinase inhibitors used the protein kinase C inhibitors, (staurosporine and calphostin C) showed marked inhibition of phosphorylation of 25 kDa protein. In addition to phosphorylation, these protein kinase C inhibitors suppresssed hemolysis by TDH. These results indicate that the phosphorylation of the 25 kDa protein seems to be essential for the hemolysis by TDH after it binds to erythrocyte membranes.