SAG1和ROP1混合基因真核表达质粒接种小鼠诱导产生拮抗致死性弓形虫感染的保护性免疫

目的检测混合SAG1和ROP1编码基因真核表达质粒疫苗诱导小鼠的免疫应答,评价其抗弓形虫感染的保护性免疫效果。方法将SAG1和ROP1编码基因片段克隆入pEGFP-N3表达载体,构建重组质粒;RT-PCR体外验证重组质粒在NIH3T3细胞中的表达;通过检测抗体、抗体分型及细胞因子来评价体液免疫和细胞免疫;流式细胞仪测定脾脏淋巴细胞亚群;腹腔内注射毒性株弓性虫速殖子攻击免疫小鼠。结果 RT-PCR结果显示重组质粒能在哺乳动物细胞内表达;SAG1和ROP1混合重组质粒疫苗诱导小鼠产生很强体液免疫和细胞免疫,对于毒性株弓形虫感染攻击具有免疫保护作用。结论不同候选抗原编码基因混合重组质粒能够诱导小鼠产生抗弓形虫感染保护性免疫,提示含有多种成份的混合DNA疫苗的研制可作为核酸免疫研究的策略之一。

葡糖酸醋杆菌内源复制片段的克隆

采用改进的碱裂解法提取Gluconacetobacter hansenii ATCC23769自发不产膜突变体的内源隐蔽质粒。用不同的限制性内切酶对混合质粒直接进行酶切,酶切后的片段混合物与pUC18载体连接构建重组载体。重组载体回转入G.hansenii ATCC23769获得隐蔽质粒上具有复制能力的片段,序列结果分析表明:该片段上没有某些其他质粒所具有的Rep蛋白。利用该片段,构建了能同时在大肠杆菌和葡糖酸醋杆菌中复制的质粒载体,体内的抗生素抗性实验证明该载体具有良好的稳定性。

Dependence of Mixed Aerosol Light Scattering Extinction on Relative Humidity in Beijing and Hong Kong

The hygroscopic properties of mixed aerosol particles are crucial for the application of remote sensing products of aerosol optical parameters in the study of air quality and climate at multiple scales. In this study, the authors investigated aerosol optical properties as a func tion of relative humidity （RH） for two representative me tropolises： Beijing and Hong Kong. In addition to the RH data, mass concentrations of PM10 （particulate matter up to 10 utm in diameter） and aerosol scattering extinction coefficient （aext） data were used. The relationship between the mass scattering extinction efficiency （MEE, defined as O＇ext/PMl0） and RH can be expressed by regression func tions asf= 1.52x ＋ 0.29 （re= 0.77）,f= 1.42x ＋ 1.53 （re= 0.58）,f= 1.19x ＋ 0.65 （re= 0.59）, andf= 1.58x ＋ 1.30 （re = 0.61） for spring, summer, autumn, and winter, respec tively, in Beijing. Here, f represents MEE, x represents I/（1-RH）, and the coefficients of determination are pre sented in parentheses. Conversely, in Hong Kong, the corresponding functions are f= 1.98x- 1.40 （r^2= 0.55）,f = 1.32x - 0.36 （r^2 = 0.26）,f= 1.87x - 0.65 （r^2 = 0.64）, and f= 2.39x - 1.47 （r^2 = 0.72）, respectively. The MEE values for Hong Kong at high RHs （RH 〉 70%） are higher than those for Beijing, except in summer; this suggests that aerosols in Hong Kong are more hygroscopic than those in Beijing for the other three seasons, but the aerosol hy groscopicity is similarly high in summer over both cities. This study describes the effects of moisture on aerosol scattering extinction coefficients and provides a potential method of studying atmospheric visibility and ground level air quality using some of the optical remote sensing products of satellites.

Influence of water content and shear rate on the mechanical behavior of soil-rock mixtures

Soil-rock mixtures(S-RMs) are widely distributed in the nature. The mesoscopic deformation and failure mechanisms as well as the macro-mechanical behaviors of the S-RMs depend largely upon the rate of deformation, water content and particle sizes. In this research, a series of large-scale direct shear tests with different water contents and different grain-size distributions were conducted to study the influence of the aforementioned factors on the mechanical properties of the S-RMs. Due to the effect of the rock blocks' breakage in the S-RMs, the relationship between the shear strength and the vertical stress of S-RM follows a power law instead of a linear one. It is found that there exists a threshold value for the vertical stress during the shearing process,below which the soil strength is mainly determined by the inter-locking of particles and the re-arrangement of meso-structure,and otherwise large-sized rock blocks are gradually broken into smaller fragments, resulting in a decrease in the soil strength.The shear rate can also significantly influence the degree of particle breakage and the meso-structural rearrangement of the SRMs, namely, under low shear rate, the particles of the samples are fully broken resulting in enhanced macro-strength. As a result, the lower the shear rate, the higher the macroscopic strength. So under unsaturated conditions, the water content will affect the strength of the S-RMs by reducing the strength of rock blocks. As the water content increases, the soil strength decreases gradually, and assumes a moderate value when the water content reaches 8%. At the same water content, the soil strength increases with the sizes of large rock blocks. For the occlusion, breakage and structure re-arrangement of the oversized rock blocks inside S-RM, which have a huge influence on the mechanical characteristics of the samples.

SERS imaging for label-free detection of the phospholipids distribution in hybrid lipid membrane

A label-free and low-cost mapping method based on SERS imaging was reported for illustrating the distribution of phospho- lipids with similar structures in binary lipid membranes on Ag nanoparticles （Ag NPs） films. The Ag NPs films exhibited strong SERS activity and good reproducibility which were investigated with p-aminothiopbenol （p-ATP） as probe molecules. Atomic force microscope （AFM） measurement proved that compact lipid membranes formed on the Ag NPs films. Basing on the Ag NPs films, the SERS spectra of phospholipids in the mixed lipid membranes were achieved and the inherent vibration of 1,2-dimyristoyl-sn-lycero-3-phosphoglycerol, sodium salt （DMPG）, 1482 cm-1, was used to distinguish between DMPG and dimyristoylphosphatidylcholine （DMPC）. The proportions of phospholipids in the mixed lipid membranes were represented by the intensity ratio of peaks at 1482 cm-1 and 1650 cm1 （R1482/1650） simultaneously： increasing R1482/1650 indicated higher propor- tion of DMPG and lower proportion of DMPC. SERS imaging of the lipid membranes was constructed as a combination of spacial information and the semiquantitative detection of phospholipids according to R1482/1650, which showed that the charged phospholipids, DMPG, aggregated in the hybrid lipid membranes. The presented mapping strategy based on SERS imaging carried out on Ag NPs films supplied a facile, label-free and inexpensive way for potential applications in the research on the structure of the lipid membrane, such as lipid domains and rafts.