Structural models and surface equation of state for pulmonary surfactant monolayers

A simple surface equation of state is proposed to describe pi-A isotherms of pulmonary surfactant monolayers. The monolayer is considered as undergoing three characteristic states during the compression: the disordered liquid-expanded (LE) state, the ordered liquid-condensed (LC) state and the collapse state. Structural models of pure protein (SP-B and SP-C) monolayer are proposed to interpret the behavior characteristics of monolayer in the states. The area, ALC, is defined as an instantaneous LC-state area when the monolayer is under the complete LC state. The area, At, is defined as a transition area from the ordered LC state to the collapse state. And the collapse pressure, pi(max), is defined as the maximum surface pressure that the monolayer can bear before collapse. The ideal equation of state is revised by ALC, At and pi(max), and a new equation of state is obtained, which is applicable for pure components of pulmonary surfactant. The theoretical pi-A isotherms described by the equation of state are compared with the experimental ones for SP-B, SP-C, DPPC and DPPG, and good agreements are obtained. The equation of state is generalized to protein-lipid binary mixtures by introducing mixing rules. The predicted pi-A isotherms agree with the experimental ones for various pulmonary surfactant components and the average deviation is about 9.2%.     

多孔淀粉负载青蒿素微球的溶出、生物利用度和组织分布研究

探讨多孔淀粉负载青蒿素微球(ART-PS)在体外溶出实验中,相比于青蒿素原药的溶出效果以及在大鼠体内的生物利用度和组织分布规律。在体外溶出实验中,分别在水、人工胃液和人工肠液三种溶出介质中,与青蒿素原药的溶出效果进行比较分析。在体内生物利用度实验中,通过对18只大鼠分别灌胃青蒿素原药与多孔淀粉负载青蒿素微球后,检测不同时间点的血药浓度,考察药物在大鼠体内的吸收和代谢差异。在组织分布规律的研究中,对98只大鼠分别灌胃多孔淀粉负载青蒿素微球和青蒿素原药,在不同时间点检测大鼠心、肝、脾、肺、肾、脑,共6个组织器官中的药物浓度。多孔淀粉负载青蒿素微球的体外溶出率在水、人工胃液、人工肠液中分别是青蒿素原药的4.04、3.59和3.82倍。多孔淀粉负载青蒿素微球在大鼠体内的血药浓度明显高于青蒿素原药,生物利用度提高为青蒿素原药的2.90倍。在组织分布的结果中,多孔淀粉负载青蒿素微球和青蒿素原药都主要分布在心脏和肝脏中,其中多孔淀粉负载青蒿素微球在不同时间各个组织中的相应含量都比原药高。多孔淀粉负载青蒿素微球相比于青蒿素原药,在体外的溶出效果更好,在体内的吸收释放效果更佳,在各组织器官中的药物含量明显高于原药,为解决青蒿素因难溶于水而在实际应用中受限提供了重要的理论依据。     

Effects of BmCPV Infection on Silkworm Bombyx mori Intestinal Bacteria

The gut microbiota has a crucial role in the growth, development and environmental adaptation in the host insect. The objective of our work was to investigate the microbiota of the healthy silkworm Bombyx mori gut and changes after the infection of B. mori cypovirus (BmCPV). Intestinal contents of the infected and healthy larvae of B. mori of fifth instar were collected at 24, 72 and 144 h post infection with BmCPV. The gut bacteria were analyzed by pyrosequencing of the 16S rRNA gene. 147(135) and 113(103) genera were found in the gut content of the healthy control female (male) larvae and BmCPV-infected female (male) larvae, respectively. In general, the microbial communities in the gut content of healthy larvae were dominated by Enterococcus, Delftia, Pelomonas, Ralstonia and Staphylococcus, however the abundance change of each genus was depended on the developmental stage and gender. Microbial diversity reached minimum at 144 h of fifth instar larvae. The abundance of Enterococcus in the females was substantially lower and the abundance of Delftia, Aurantimonas and Staphylococcus was substantially higher compared to the males. Bacterial diversity in the intestinal contents decreased after post infection with BmCPV, whereas the abundance of both Enterococcus and Staphylococcus which belongs to Gram-positive were increased. Therefore, our findings suggested that observed changes in relative abundance was related to the immune response of silkworm to BmCPV infection. Relevance analysis of plenty of the predominant genera showed the abundance of the Enterococcus genus was in negative correlation with the abundance of the most predominant genera. These results provided insight into the relationship between the gut microbiota and development of the BmCPV-infected silkworm.     

十字花科黑腐病菌中转录调控因子HpaR1调控一个纤维素酶基因的表达

十字花科黑腐病菌（Xcc8004）中的一个转录调控因子XC₂736（HpaR1）在致病过程中具有重要的作用。前期研究发现该转录调控因子可能调控胞外纤维素酶的合成。为了解HpaR1对纤维素酶的转录调控机理,本研究对HpaR1进行原核表达纯化,并与488bp的包含XC₀639的启动子区DNA片段进行凝胶电泳迁移率试验,发现HpaR1与XC₀639启动子可以发生结合。将488bp的XC₀639的启动子DNA片段与报告基因gus融合,构建XC₀639的报告质粒pGUS0639r,分别导入野生型8004菌株和缺失突变体DM2736中,分析发现在突变体背景下GUS的表达水平比野生型背景明显降低。表明HpaR1正调控XC₀639的表达。构建XC₀639的极性整合突变体PK0639,检测发现PK0639几乎丧失胞外纤维素酶的活力;通过功能反式互补构建的互补菌株CPK0639可以恢复纤维素酶活性。研究结果表明HpaR1通过调控纤维素酶基因XC₀639的表达来调控细胞的纤维素酶活性。本研究为更深入地了解HpaR1如何调控细菌生理生化功能奠定了基础。     

Development and Evaluation of a DAS-ELISA for Rapid Detection of Tembusu Virus Using Monoclonal Antibodies against the Envelope Protein

Since April 2010, Tembusu virus (TMUV) which is a contagious pathogen of waterfowls, causing symptoms of high fever, loss of appetite and fall in egg production, has been reported in east of China. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) which detects for TMUV was developed, using two monoclonal antibodies (mAbs) against the TMUV envelope (E) protein. BALB/c mice were immunized with purified recombinant E protein expressed in E. coli. Three hybridoma cell lines designated as 12B1, 10C6 and 2D2, were screened by cell fusion and indirect ELISA for their ability to recognize different linear epitopes on the E protein, and were characterized subsequently. High-affinity mAbs 12B1 and 2D2 were used as capture and detection antibodies, respectively. The reaction conditions for the DAS-ELISA were optimized for TMUV detection. The cross-reactivity of the DAS-ELISA was determined using TMUV, duck plague virus, avian influenza virus subtype H9, Newcastle disease virus, duck hepatitis A virus type 1 and duck reovirus samples. A total of 191 homogenized tissues of field samples were simultaneously detected by DAS-ELISA and by RT-PCR. The former was found to have a high specificity of 99.1% and a sensitivity of 93.1%. These results reveal a positive coincidence between DAS-ELISA and RT-PCR at a coincidence rate of 95.8%. The method developed in this study can be used for the diagnosis of TMUV infection of duck origin.     

Rad9 plays an important role in DNA mismatch repair through physical interaction with MLH1

Rad9 is conserved from yeast to humans and plays roles in DNA repair (homologous recombination repair, and base-pair excision repair) and cell cycle checkpoint controls. It has not previously been reported whether Rad9 is involved in DNA mismatch repair (MMR). In this study, we have demonstrated that both human and mouse Rad9 interacts physically with the MMR protein MLH1. Disruption of the interaction by a single-point mutation in Rad9 leads to significantly reduced MMR activity. This disruption does not affect S/M checkpoint control and the first round of G₂/M checkpoint control, nor does it alter cell sensitivity to UV light, gamma rays or hydroxyurea. Our data indicate that Rad9 is an important factor in MMR and carries out its MMR function specifically through interaction with MLH1.