Preferential flow characteristics of reclaimed mine soils in a surface coal mine dump.

There are a large number of macropores/tubular channels of a few centimeters and plant roots in reclaimed dump soils, which are the main reasons for the formation of soil macropores and soil preferential flow. To systematically study the morphological characteristics and variation of soil preferential flow for different reclaimed vegetations in a dump, a dye-staining experiment and physical and chemical analysis were carried out to investigate the formation mechanism and influencing factors of soil preferential flow in the vegetation restoration process. The results indicate that there were differences in the soil water breakthrough curves for different plots. The macropore effluent rate generally increased at first and then tended to stabilize. The soil steady effluent rate decreased with increasing soil depth, which reached the maximum and minimum values at the depths of 0∼5 cm (0.0193∼0.0315 mm s-1) and 50∼60 cm (0.0028∼0.0035 mm s-1), respectively. Furthermore, the radius of soil macropores under different types of reclaimed vegetation ranged from 0.03 to 4.71 mm, most of which ranged from 0.11 to 2.36 mm. The soil macroporosity of different reclaimed vegetation types ranged from 0.03 to 16.58%, which was significantly greater than 5%. The soil macroporosity determined 65% of the variation in the steady effluent rate and 42% of the variation in the saturated hydraulic conductivity. Furthermore, the dye coverage ratio decreased as the soil layer depth increased in different plots, and there were some differences in each plot. The maximum dye coverage ratio occurred in the 0∼5 cm soil layer, which reached 90.37%. The dye coverage ratio at a depth of 0∼60 cm in six plots followed the order of Robinia pseudoacacia (26.48%) > Ulmus pumila (20.12%) > mixed forest (17.32%) > farmland (15.06%) > shrub (13.97%) > weeds (10.07%). The soil preferential flow mostly occurred in the 0∼40 cm soil depth layer, which occupied more than 93% of the total soil profile (0∼60 cm). Moreover, a Pearson correlation was used to analyze the relationship between environmental factors (soil, water, and plant factors) and the dye coverage ratio. The dye coverage ratio of soil preferential flow under different reclamation vegetations was very significantly or significantly positively correlated with the gravel content, mean radius of soil macropores, soil saturated hydraulic conductivity, root weight density, and root length density, which promoted the formation and development of soil preferential flow. This study will provide a scientific basis for understanding the formation mechanism and perfecting the research system of soil preferential flow, vegetation restoration, and reconstruction in a dump; furthermore, this research offers significance guidance in the construction of green mines and the development of regional economics.

Real time ex vivo detection and dynamic monitoring of in vivo expression of secreted luciferase gene injected by hydrodynamic method / 生物工程学报

We chose Gaussia luciferase (Gluc), a secreted luciferase gene as reporter to real-time detect and dynamically monitor hydrodynamic injection gene expression. First, we constructed an expression vector pAAV2neo-Gluc. Then Huh7 and HepG2 cells were transfected with pAAV2neo-Gluc and the activity of Gluc in the supernatant and cell lysates were assayed. Results showed that the Gluc activity in the supernatant was about 100 higher than that in cell lysates, indicating the expressed Gluc existing mainly as a secreted form as reported. Live bioluminescence imaging of mice hydrodynamic injected pAAV2neo-Gluc showed whole body distribution, while the pAAV2neo-Fluc primarily located in the liver. Then we injected different doses of pAAV2neo-Gluc into mice by tail-vein hydrodynamic injection, took minor amount of blood from mice tails at different time points and measured the luciferase activity to investigate dynamic changes of Gluc expression and secretion in vivo. The results suggested that the time courses of Gluc expression were highly consistent among each dose groups. The luciferase activity in blood could be detected as early as 2 h after injection, reached the peak at about 10 h and gradually decreased from then on. The expression level of Glue was positively correlated with the dose of injected plasmid DNA. To further detect the assay sensitivity of the ex vivo Gluc measurement method, we investigated three additional groups of mice injected with lower doses of 0.001 microg, 0.01 microg and 0.1 microg pAAV2neo-Gluc respectively. Results revealed that activity of Gluc in blood could be detected even at dose as low as 0.001 microg DNA, suggesting the assay sensitivity was extremely high. In conclusion, a real-time ex vivo detection method of dynamically monitoring of gene expression in vivo by hydrodynamic injection can be a valuable means for the study of gene expression regulation in vivo.