Sperm-mediated gene transfer (SMGT) is based on the ability of spermatozoa to bind exoge- nous DNA and transfer it into oocytes by fertilization. However, SMGT is still undergoing opti- mization to improve its efficiency to produce transgenic animals. The acrosome reaction is neces- sary for spermatozoa to carry the exogenous DNA into oocytes. In this study, the effect of the acrosome reaction on the efficiency of spermatozoa carrying exogenous DNA was evalua- ted. The results showed that the efficiency of the acrosome reaction was significantly higher (p<0.05) after incubation with 50 μmol/L progesterone compared to incubation without proges- terone. It was significantly higher (p<0.05) in the 20, 40, and 60 min of progesterone treatment groups than in the 0 min treatment group. The spermatozoa were further incubated with cyanine dye Cy5 labeled DNA (Cy5-DNA) for 30 min at 37°C, and positive fluorescence signals were detected after the acrosome reaction was induced by progesterone at concentrations of 0 and 50 μmol/L for 40 min. The percentage of positive Cy5-DNA signals in spermatozoa was 96.61±2.06% and 97.51±2.03% following exposure to 0 and 50 μmol/L progesterone, respective- ly. The percentage of partial spermatozoa heads observed following combination with Cy5-DNA was 39.73±3.03% and 56.88±3.12% following exposure to 0 and 50 μmol/L progesterone, respec- tively. The ratio of positively stained spermatozoa combined with exogenous DNA showed no reduction after the acrosome reaction. These results suggest that the acrosome reaction might not be the key factor affecting the efficiency of SMGT.
The microstructures and mechanical properties of T92 martensitic steel/Super304 austenitic steel weld joints with three welding consumables were investigated. Three types of welding materials ERNiCr-3, ERNiCrCoMo-1and T-304H were utilized to obtain dissimilar welds by using gas tungsten arc weld (GTAW). The results show that heat affect zone (HAZ) of T92 steel consists of coarse-grained and fine-grained tempered martensites. The microstructures of joints produced from ERNiCrCoMo-1 consist of equiaxed dendrite and columnar dendrite grains, which are more complicated than that of ERNiCr-3. In the tensile tests, joints constructed from ERNiCrCoMo-1 and T-304H met the ASME standard. The highest fracture energy was observed in specimens with the welding material ERNiCrCoMo-1. Ni content in weld seam of ERNiCrCoMo-1 was highest, which was above 40%. In conclusion, the nickel alloy ERNiCrCoMo-1 was the most suitable welding material for joints produced from T92 martensitic steel/Super304 austenitic steel.
A novel phase shift full bridge (PSFB) converter with voltage-doubler and decoupling integrated magnetics in photovoltaic (PV) systems is proposed. Considering the demand that the output voltage is higher than the input voltage in PV systems, the voltage-doubler is added to achieve higher voltage gain compared with the traditional PSFB. In order to avoid current oscillation caused by the voltage-doubler and obtain the wide zero voltage switching (ZVS) ranges, an external inductor is imposed on the circuit. Especially, to obtain much higher power density, the external inductor and transformer are integrated into one magnetic core. The operation and voltage gain of proposed converter are analyzed. Also, in order to reveal the effects the integrated magnetics gives to the converter, the decoupling condition and the expression of leakage inductor of integrated magnetics are obtained in detail. Finally a 100 W prototype converter is made and the experimental results are given to verify the analysis.
Sapelovirus A (SV-A) is a positive-sense single-stranded RNA virus which is associated with acute diarrhea, pneumonia and reproductive disorders. The virus capsid is composed of four proteins, and the functions of the structural proteins are unclear. In this study, we expressed SV-A structural protein VP1 and studied its antigenicity and immunogenicity. SDS-PAGE analysis revealed that the target gene was expressed at high levels at 0.6 mM concentration of IPTG for 24 h. The mouse polyclonal antibody against SV-A VP1 protein was produced and reached a high antiserum titer (1: 2,048,000). Immunized mice sera with the recombinant SV-A VP1 protein showed specific recognition of purified VP1 protein by western blot assay and could recognize native SV-A VP1 protein in PK-15 cells infected with SV-A by indirect immunofluorescence assay. The successfully purified recombinant protein was able to preserve its antigenic determinants and the generated mouse anti-SV-A VP1 antibodies could recognize native SV-A, which may have the potential to be used to detect SV-A infection in pigs.