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Number of results: 24
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Abstract

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.
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Abstract

Osteocalcin is a major non-collagenous component of the bone extracellular matrix and is considered to be an indicative factor of osteoblast differentiation. In the present study, we detected osteocalcin expression in different antler areas and growth phases by immunohisto- chemistry. Osteocalcin was highly expressed in all areas during the mineralization period and in mesenchymal cell and chondrocyte areas during the rapid growth period. The nucleotide sequence of the osteocalcin gene in sika deer antler was determined. The open reading frame was 303 bp encoding a protein of 100 amino acids. The estimated molecular mass of osteocalcin was 10.38 kDa and the theoretical isoelectric point was 5.37. The osteocalcin gene with a 6× His-tag at the C-terminus was cloned into the pGEX-4T1 vector and expressed in Escherichia coli under optimal conditions. The recombinant soluble protein fused with GST was purified with Ni-NTA resin. The purified osteocalcin protein exhibited a significant increase in HA adhesion and promoted antler chondrocyte proliferation. Osteocalcin is an important factor in regulating the rapid growth and differentiation of deer antlers.
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Abstract

The Intrauterine fetal development process is complicated and affected by many regulating factors such as maternal nutritional status, transcription factors and adipokines. Adipokines are kinds of active substances secreted by adipose tissue, including more than 50 kinds of molecules. To explore the correlation between calf birth weights and adipokines including adiponectin, leptin, visfatin, and IGF-1 in cows venous and venous cord blood. Fifty-four healthy multiparous Chinese Holstein cows were used; in which, cows with a calf weight less than 40 kg were included in group A (n=9); those with a calf weight between 40 kg~45 kg were included in group B (n=25) and ≥45 kg were included in group C (n=20), venous blood and cord venous blood was collected. An ELISA kit was used to evaluate the concentration of adiponectin, leptin, visfatin, and IGF-1, correlations between index-index and index-calf birth weight were analysed. In both cows venous and cord venous blood, adiponectin, leptin, visfatin, and IGF-1 levels were significantly correlated with each other (p<0.01), and levels of these adipokines in venous blood were significantly higher than cord venous blood (p<0.01). Adiponectin, leptin, visfatin, and IGF-1 in venous cord blood were positively correlated with calf birth weights, and significantly correlated with calf birth weights respectively (p<0.01). Our study showed that adiponectin, leptin, and IGF-1 were found in venous blood and cord venous blood, and adiponectin, leptin, and IGF-1 in venous and cord venous blood potentially inter-regulated each other; adiponectin, leptin, and IGF-1 in venous blood were not significantly correlated with calf birth weights, while adiponectin, leptin, visfatin, and IGF-1 in venous cord blood were significantly correlated with calf birth weights, respectively.
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Abstract

Based on the mould temperature measured by thermocouples during slab continuous casting, a difference of temperature thermograph is developed to detect slab cracks. In order to detect abnormal temperature region caused by longitudinal crack, the suspicious regions are extracted and divided by virtue of computer image processing algorithms, such as threshold segmentation, connected region judgement and boundary tracing. The abnormal regions are then determined and labeled with the eight connected component labeling algorithm. The boundary of abnormal region is also extracted to depict characteristics of longitudinal crack. Based on above researches, longitudinal crack with abnormal temperature region can be detected and is different from other abnormalities. Four samples of temperature drop are picked up to compare with longitudinal crack on the abnormal region formation, length, width, shape, et al. The results show that the abnormal region caused by longitudinal crack has a linear and vertical shape. The height of abnormal region is more than the width obviously. The ratio of height to width is usually larger than that of other temperature drop regions. This method provides a visual and easy way to detect longitudinal crack and other abnormities. Meanwhile it has a positive meaning to the intelligent and visual mould monitoring system of continuous casting.
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Abstract

Isothermal hot compression experiments were carried out using the Gleeble-1500D thermal mechanical simulator. The flow stress of the Cu-1%Zr and Cu-1%Zr-0.15%Y alloys was studied at hot deformation temperature of 550°C, 650°C, 750°C, 850°C, 900°C and the strain rate of 0.001 s–1, 0.01 s–1, 0.1 s–1, 1 s–1, 10 s–1. Hot deformation activation energy and constitutive equations for two kinds of alloys with and without yttrium addition were obtained by correlating the flow stress, strain rate and deformation temperature. The reasons for the change of hot deformation activation energy of the two alloys were analyzed. Dynamic recrystallization microstructure evolution for the two kinds of alloys during hot compression deformation was analyzed by optical and transmission electron microscopy. Cu-1%Zr and Cu-1%Zr-0.15%Y alloys exhibit similar behavior of hot compression deformation. Typical dynamic recovery occurs during the 550-750°C deformation temperature, while dynamic recrystallization (DRX) occurs during the 850-900°C deformation temperature. High Zr content and the addition of Y significantly improved Cu-1%Zr alloy hot deformation activation energy. Compared with hot deformation activation energy of pure copper, hot deformation activation energy of the Cu-1%Zr and Cu-1%Zr-0.15%Y alloys is increased by 54% and 81%, respectively. Compared with hot deformation activation energy of the Cu-1%Zr alloy, it increased by 18% with the addition of Y. The addition of yttrium refines grain, advances the dynamic recrystallization critical strain point and improves dynamic recrystallization.
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Abstract

In order to compare the pathogenicity of different Tembusu virus (TMUV) strains from geese, ducks and chickens, 56 5-day-old Cherry Valley ducklings which were divided into 7 groups and infected intramuscularly with 7´105 PFU/ml per duck of six challenge virus stocks. The clinical signs, weight gain, mortality, macroscopic and microscopic lesions, virus loads in sera of 1, 3, 5, 7, 11 and 14 dpi and serum antibody titers were examined. The results showed that these viruses could make the young ducks sick, but the clinical signs differed with the different species-original strains. All the experimental groups lose markedly in weight gain compared to the control, but there were no obvious distinctions in weight gains, as well as macroscopic and microscopic lesions of dead ducks between the infected groups. However, the groups of waterfowl-derived strains (from geese and ducks) showed more serious clinical signs and higher relative expressions of virus loads in sera than those from chicken-derived. The mortality of waterfowl groups was 37.5%, and the greatest mortality of chicken groups was 12.5%. The serum antibodies of the geese-species group JS804 appeared earlier and were higher in the titers than others. Taken toghter, the pathogenicity of waterfowl-derived TMUV was more serious than chicken-derived TMUV and JS804 could be chosen as one TMUV vaccine strain to protect from the infection.
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Abstract

MDAP-2 is a new antibacterial peptide with a unique structure that was isolated from house- flies. However, its biological characteristics and antibacterial mechanisms against bacteria are still poorly understood. To study the biological characteristics, antibacterial activity, hemolytic activi- ty, cytotoxicity to mammalian cells, and the secondary structure of MDAP-2 were detected; the results showed that MDAP-2 displayed high antibacterial activity against all of the tested Gram-negative bacteria. MDAP-2 had lower hemolytic activity to rabbit red blood cells; only 3.4% hemolytic activity was observed at a concentration of 800μg/ml. MDAP-2 also had lower cytotoxicity to mammalian cells; IC50 values for HEK-293 cells, VERO cells, and IPEC-J2 cells were greater than 1000 μg/ml. The circular dichroism (CD) spectra showed that the peptide most- ly has α-helical properties and some β-fold structure in water and in membrane-like conditions. MDAP-2 is therefore a promising antibacterial agent against Gram-negative bacteria. To deter- mine the antibacterial mechanism(s) of action, fluorescent probes, flow cytometry, and transmis- sion electron microscopy (TEM) were used to study the effects of MDAP-2 on membrane perme- ability, polarization ability, and integrity of Gram-negative bacteria. The results indicated that the peptide caused membrane depolarization, increased membrane permeability, and destroyed membrane integrity. In conclusion, MDAP-2 is a broad-spectrum, lower hemolytic activity, and lower cytotoxicity antibacterial peptide, which is mainly effective on Gram-negative bacteria. It exerts its antimicrobial effects by causing bacterial cytoplasm membrane depolarization, increas- ing cell membrane permeability and disturbing the membrane integrity of Gram-negative bacte- ria. MDAP-2 may offer a new strategy to for defense against Gram-negative bacteria.
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Abstract

Geomechnical model testing has been widely applied as a kind of research technique in underground engineering problems. However, during the practical application process, due to the influence of many factors, the desired results cannot be obtained. In order to solve this problem, based on the measurement requirements of the model test, combined with FBG(Fiber Bragg Grating) sensor technology and traditional measurement methods, an FBG monitoring system, Micro-multi-point displacement test system, resistance strain test system and surrounding rock pressure monitoring system are developed. Applying the systems to a model test of the tunnel construction process, the displacement in advance laws of tunnel face, radial displacement distribution laws and surrounding rock pressure laws are obtained. Test results show that a multivariate information monitoring system has the advantage of high precision, stability and strong anti-jamming capability. It lays a solid foundation for the real-time data monitoring of the tunnel construction process model test.
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Abstract

Based on the theory of heat transfer, the influence of expansion joints on the temperature and stress distribution of ladle lining is discussed. In view of the current expansion joint, the mathematical model of heat transfer and the three dimensional finite element model of ladle lining brick are established. By analyzing the temperature and stress distribution of ladle lining brick when the expansion joints are in different sizes, the thermal mechanical stress caused by the severe temperature difference can be reduced by the suitable expansion joint of the lining brick during the ladle baking and working process. The analysis results showed that the thermal mechanical stress which is caused by thermal expansion can be released through the 2 mm expansion joint, which is set in the building process. So we can effectively reduce the thermal mechanical stress of the ladle lining, and there is no risk of steel leakage, thus the service life of ladle can be effectively prolonged.
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Abstract

The friction and wear properties of 201HT aluminum alloys and the corresponding competitive coupons were tested on an electrohydraulic servo face friction and wear testing machine (MM-U10G). The microstructures of the competitive coupons were investigated by scanning electron microscopy (SEM) and consequently the corresponding friction and wear mechanisms were studied. The results demonstrated that: (1) the best competitive material of friction and wear performance of the 201HT was the 201HTC. (2) the 201HTC modified by carbon following the initial mill for oil storage of the micro-groove to be produced, increased the corresponding lubrication performance reduced the friction coefficient and wear rate effectively. (3) the 201HT-201HTC could obtain both better friction and wear mainly due to the initial process of grinding following the 201HT plastic deformation occurred in the surface and the formation of a series of re-melting welding points, whereas the 201HT material hardness would be similar to the 201HTC material hardness, which led into the competitive material friction and wear performance improvement.
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Abstract

Self-biting disease occurs in most farmed fur animals in the world. The mechanism and rapid detection method of this disease has not been reported. We applied bulked sergeant analysis (BSA) in combination with RAPD method to analyze a molecular genetic marker linked with self-biting trait in mink group. The molecular marker was converted into SCAR and loop-mediated isothermal amplification (LAMP) marker for rapid detection of this disease. A single RAPD marker A10 amplified a specific band of 1000bp in self-biting minks. The sequences of the bands exhibited 73% similarity to the Canis Brucella. SCAR and LAMP marker were designed for the specific fragment of RAPD marker A10 and validated in 30 self-biting minks and 30 healthy minks. c2 test showed difference (p<0.05) with SCAR and significant difference (p<0.01) with LAMP in the detection rate between the two groups, but LAMP method was more accurate than SCAR method. This indicated that LAMP can be used as a positive marker to detect self-biting disease in minks.
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