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.
Senecavirus A (SVA) the only member of the Senecavirus genus within the Picornaviridae family, is an emerging pathogen causing swine idiopathic vesicular disease and epidemic transient neonatal losses. Here, SVA strain (CH-HNKZ-2017) was isolated from a swine farm exhibiting vesicular disease in Henan Province of Central China. A phylogenetic analysis based on complete genome sequence indicated that CH-HNKZ-2017 was closely related to US-15-40381IA, indica- ting that a new SVA isolate had emerged in China.
The study was aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA), which can detect specifically Feline herpesvirus type 1 (FHV-1). The primers were designed based on the conserved sequence of FHV-1 glycoprotein B gene. The recombinant protein with reactogenicity was purified as coating antigen of the assay. The indirect ELISA, characterized by high sensitivity showed no cross-reaction with two types of feline virus, had detection limit at 1:2000 dilution. The positive rate of the assay, according to the determined cutoff value (0.25), was basically consistent with Feline Herpes Virus Antibody ELISA kit. In conclusion, the indirect ELISA with high repeatability and reproducibility can be used for detecting FHV-1, and can provide necessary support to related research.
In order to overcome the shortcomings of the dolphin algorithm, which is prone to falling into local optimum and premature convergence, an improved dolphin swarm algorithm, based on the standard dolphin algorithm, was proposed. As a measure of uncertainty, information entropy was used to measure the search stage in the dolphin swarm algorithm. Adaptive step size parameters and dynamic balance factors were introduced to correlate the search step size with the number of iterations and fitness, and to perform adaptive adjustment of the algorithm. Simulation experiments show that, comparing with the basic algorithm and other algorithms, the improved dolphin swarm algorithm is feasible and effective.
In the current study, twenty lambs, aged 4 months, half male and half female, were classified into four groups, with five in each group. The experimental three groups of lambs were given intravenous (IV), intramuscular (IM) and subcutaneous (SC) administrations of recombinant ovine interferon-τ (roIFN-τ). The fourth group (normal control) of lambs was given normal saline injections in the same way. After administrations, blood samples were collected from the tested animals at different time points post injection, and the serum titers of roIFN-τ were measured using cytopathic effect (CPE) inhibition bioassay. The results of calculating pharmacokinetic (PK) parameters using DAS software showed that the PK characteristics of roIFN-τ through IV injection conformed to the two-compartment open model, whose half-life of distribution phases (T1/2α) was 0.33±0.034 h and the elimination half-life(T1/2β) was 5.01±0.24 h. However, the PK features of IM injection and SC injection of roIFN-τ conformed to the one compartment open model, whose Tmax were 3.11±0.26 h and 4.83±0.43 h, respectively, together with an elimination half life(T1/2β) of 9.11±0.76 h and 7. 43±0.58 h, and an absorption half-life (T1/2k(a)) of 1.13±0.31 h and 1.85±0.40 h, respectively. The bioavailability of roIFN-τ after IM administration reaches 73.57%, which is greater than that of SC administration (53.43%). These results indicate that the drug administration effect can be preferably obtained following a single dose IM administration of the roIFN-τ aqueous preparation. This study will facilitate the clinical application of roIFN-τ as a potential antiviral agent in future work.
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.
Vaccination is a common routine for prevention and control of human and animal diseases by inducing antibody responses and cell-mediated immunity in the body. Through vaccinations, smallpox and some other diseases have been eradicated in the past few years. The use of a patho- gen itself or a subunit domain of a protein antigen as immunogens lays the basis for traditional vaccine development. But there are more and more newly emerged pathogens which have expe- rienced antigenic drift or shift under antibody selective pressures, rendering vaccine-induced im- munity ineffective. In addition, vaccine development has been hampered due to problems includ- ing difficulties in isolation and culture of certain pathogens and the antibody-dependent enhancement of viral infection (ADE). How to induce strong antibody responses, especially neu- tralizing antibody responses, and robust cell-mediated immune responses is tricky. Here we re- view the progress in vaccine development from traditional vaccine design to reverse vaccinology and structural vaccinology and present with some helpful perspectives on developing novel vac- cines.
Canine parvovirus (CPV) causes acute gastroenteritis in domestic dogs, cats, and several wild carnivore species. In this study, the full-length VP2 gene of 36 CPV isolates from dogs and cats infected between 2016 and 2017 in Beijing was sequenced and analyzed. The results showed that, in dogs, the new CPV-2a strain was the predominant variant (n = 18; 50%), followed by the new CPV-2b (n = 6; 16.7%) and CPV-2c (n = 3; 8.3%) strains, whereas, among cats, the predominant strain was still CPV-2 (n = 9; 25%). One new CPV-2a strain, 20170320-BJ-11, and two CPV-2c strains, 20160810-BJ-81 and 20170322-BJ-26, were isolated and used to perform experimental infections. Multiple organs of beagles that died tested PCR positive for CPV, and characteristic histopathological lesions were observed in organs, including the liver, spleen, lungs, kidneys, small intestines, and lymph nodes. Experimental infections showed that the isolates from the epidemic caused high morbidity in beagles, indicating their virulence in animals and suggesting the need to further monitor evolution of CPV in China.
In order to understand infection of avian influenza A virus (AIV) and canine distemper virus (CDV) in the Siberian Tiger in Northeast China, 75 Siberian Tiger serum samples from three cap- tive facilities in northeastern China were collected. AIV and CDV antibody surveillance was test- ed by using hemagglutination inhibition and serum neutralization methods. The results showed that the seroprevalence of H5 AIV, H9 AIV and CDV was respectively 9.33% (7/75), 61.33% (46/75) and 16% (12/75). In the 1<years <2 and > 5 year-old group, the seroprevalence of the H9 AIV was 24% and 80% (P < 0.01), and the CDV seroprevalence was 6% and 36% (P < 0.01), respectively. It was demonstrated that 3 (4%) out of 75 serum samples were AIV+CDV seropos- itive, with 2.67% (2/75) in H9+AIV and 1.33% (1/75) in H5+H9+AIV. To our knowledge, this is the first report of AIV and CDV seroprevalence in Siberian Tigers in China, which will provide base-line data for the control of AIV and CDV infection in Siberian Tigers in China.
In order to predict the distribution of shrinkage porosity in steel ingot efficiently and accurately, a criterion R√L and a method to obtain its threshold value were proposed. The criterion R√L was derived based on the solidification characteristics of steel ingot and pressure gradient in the mushy zone, in which the physical properties, the thermal parameters, the structure of the mushy zone and the secondary dendrite arm spacing were all taken into consideration. The threshold value of the criterion R√L was obtained with combination of numerical simulation of ingot solidification and total solidification shrinkage rate. Prediction of the shrinkage porosity in a 5.5 ton ingot of 2Cr13 steel with criterion R√L>0.21 m･℃1/2･s -3/2 agreed well with the results of experimental sectioning. Based on this criterion, optimization of the ingot was carried out by decreasing the height-to-diameter ratio and increasing the taper, which successfully eliminated the centreline porosity and further proved the applicability of this criterion.
The drainage consolidation method has been efficiently used to deal with soft ground improvement. Nowadays, it has been suggested to use a new sand soil which is a composite of sand and recycled glass waste. The permeability performance of glass-sand soil was explored to judge the feasibility of glass-sand soil backfilled in the drainage consolidation of sand-drained ground. For comparison purposes, different mix proportions of recycled glass waste, fineness modulus, and glass particle size were analyzed to certify the impact on the permeability coefficient and the degree of consolidation. The numerical results show that adding a proper amount of recycled glass waste could promote the permeability performance of glass-sand soil, and the glasssand soil drain could be consolidated more quickly than a sand drain. Experiments showed that glass-sand soil with the a 20% mix of recycled glass waste reveals the optimum performance of permeability.
Heterogeneous nuclear ribonucleoprotein K (hnRNP K), is a multifunctional protein that participates in a variety of regulatory processes of signal transduction and gene expression. To further characterize the significance of hnRNP K in different male germ cells, we investigated the expression profiles of hnRNP K at different developmental stages in pig and rat testes, and conducted a comparative analysis of expression patterns between these two species. In porcine testis development, both the mRNA and protein level of hnRNP K were down-regulated from 3 months to 8 months. However, the expression level of hnRNP K was abundant across the embryonic period in rats, and decreased gradually from 0 day post partum (dpp) to 14 dpp, then increased with the highest level presenting at 90 dpp. Immunolocalization analysis further confirmed the differential expression and localization of hnRNP K protein during testis development in pigs and rats. The results showed that hnRNP K was widely distributed in gonocytes, spermatogonia, sertoli cells and Leydig cells. The dynamic expression profile of hnRNP K may imply its crucial and potential roles in the development of the testis, which will provide a theoretical basis for the future study of molecular mechanism regulation of spermatogenesis.