Duck viral hepatitis (DVH) is an acute and fatal disease of young ducklings characterized by rapid transmission and damages. The most important agent of DVH is duck hepatitis virus 1 (DHV-1). The effective control of DVH was achieved by active immunization of 1-day-old duck- lings with an attenuated DHV-1 virus vaccine. However, the attenuated virus might reverse to virulence. In this study, a DHV-1 strain, Du/CH/LBJ/090809, was identified and its genomic se- quences were determined. The genome of Du/CH/LBJ/090809 is composed of 7,692 nt excluding poly A and the virus was clustered into genotype A by comparing with other referenced DHV-1 strains. Du/CH/LBJ/090809 could lead to 30% mortality of 10-day-old specific pathogen free (SPF) ducklings. The virus was passaged serially in SPF chicken embryonated eggs and three vi- ruses, passage 16 (P16), P29 and P40, were selected for genomic analysis. P29 and P40 were used to evaluate the attenuation in duckling by inoculating the virus to 10-day-old SPF ducklings. Re- sults of vaccination-challenge assay showed that the inactivated virus P40 could evoke protection against the pathogenic parent virus. Nucleotide and amino acid sequences of the genomes of Du/ CH/LBJ/090809, P16, P29 and P40 were compared. Changes both in nucleotides and amino acids, which might be contributed to the decreasing in virulence by chicken embryo-passaging of DHV- 1, were observed. We speculated that these changes might be important in the adaption and at- tenuation of the virulent virus. Additionally, strains obtained in this study will provide potential candidate in the development of vaccines against DHV-1.
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.
Insulin receptor substrate 2 (IRS-2) modulates the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, which controls the suppression of gluconeogenic genes; IRS-2 is also a critical node of insulin signaling. Because of the high homology between pig and human IRS-2, we investigated the expression pattern and function of porcine IRS-2. QPCR and immunoblotting were used to detect the IRS-2 expression level in different tissues. There were high IRS-2 levels in the cerebral cortex, hypothalamus, and cerebellum in the central nervous system. In peripheral tissues, IRS-2 was expressed at relatively high levels in the liver. Immunohistochemistry analysis revealed that IRS-2 was mainly distributed in the hypothalamus and cerebral cortex. Furthermore, IRS-2 knockdown porcine hepatocytes and porcine aortic endothelial cells (PAECs) were generated. The IRS-2 knockdown induced abnormal expression of genes involved in glycolipid metabolism in hepatocytes and reduced the antiatherosclerosis ability in PAECs. In addition, we disrupted IRS-2 in porcine embryonic fibroblasts (PEFs) using the CRISPR/Cas9 genome editing system, before finally generating IRS-2 knockout embryos by somatic cell nuclear transfer (SCNT). Taken together, our results indicate that IRS-2 might be a valuable target to establish diabetes and vascular disease models in the pig.
Cu–4.7 wt. % Sn alloy wire with Ø10 mm was prepared by two-phase zone continuous casting technology, and the temperature field, heat and fluid flow were investigated by the numerical simulated method. As the melting temperature, mold temperature, continuous casting speed and cooling water temperature is 1200 °C, 1040 °C, 20 mm/min and 18 °C, respectively, the alloy temperature in the mold is in the range of 720 °C–1081 °C, and the solid/liquid interface is in the mold. In the center of the mold, the heat flow direction is vertically downward. At the upper wall of the mold, the heat flow direction is obliquely downward and deflects toward the mold, and at the lower wall of the mold, the heat flow deflects toward the alloy. There is a complex circular flow in the mold. Liquid alloy flows downward along the wall of the mold and flows upward in the center.
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.
Ludwigite is the main available boron-bearing resource in China. In order to enrich the theory system and optimize its utilization processes, this paper study the mechanism and kinetics on non-isothermal decomposition of ludwigite in inert atmosphere by means of thermal analysis. Results show that, the decomposition of serpentine and szajbelyite is the main cause of mass loss in the process. At the end of decomposition, hortonolite and ludwigite are the two main phases in the sample. The average E value of structural water decomposition is 277.97 kJ/mol based on FWO method (277.17 kJ/mol based on KAS method). The results is proved to be accurate and reliable. The mechanism model function of structural water decomposition is confirmed by Satava method and Popescu method. The form of the most probable model function is G(α) = (1 – α)–1 – 1 (integral form) and f (α) = (1 – α)2 (differential form), and its mechanism is chemical reaction. This is verified by the criterion based on activation energy of model-free kinetics analysis.
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.
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.
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.
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.
In vitro embryogenic callus is a critical factor for genetic transformation of rice, especially for indica varieties. In this study, we investigated the relationship between polyamines, including putrescine (Put), spermidine (Spd) and spermine (Spm), and callus browning, and we studied the effect of exogenous Put on callus regeneration and on the content of endogenous polyamines. In addition, the expression levels of arginine decarboxylase gene (Adcl) and S-adenosylmethionine decarboxylase gene (Samdc) in embryogenic callus were studied by quantitative Real-time PCR analysis. The results showed that the contents of endogenous Put and Spd in the browning callus were significantly lower than those in normal callus. Exogenous Put could effectively improve the growing state of callus of indica rice and enhance the development of embryogenic callus. The content of endogenous polyamines in embryogenic callus, especially Spd and Spm, was increased after addition of exogenous Put. Additionally, exogenous Put also had an obvious impact on the expression levels of Adcl but partial effect on the expression levels of Samdc gene. This study could increase the knowledge of both embryogenic callus induction and polyamine catabolism in callus in indica rice.
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.