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Number of results: 8
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Abstract

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.
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Authors and Affiliations

H. Xu
P. Zhang
R. Li
W. Wu
S. Wang
Y. Xu
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Abstract

Taraxacum Officinale, commonly called dandelion, is herbaceous perennial belonging to the family of Asteraceae, having good antibacterial effects which are related to its phenolic substances. In this study, the effect of phenolic contents as well as the antibiofilm activity against Staphylococcus aureus of phenolic extract from T. Officinale were evaluated in vitro. With 70% metha- nol-water (v/v) as a solvent, the dandelion was extracted by ultrasonic assisted extraction method. Subsequent identification and quantification of phenol in extract was carried out using High Performance Liquid Chromatography (HPLC). The minimum inhibitory concentration and anti- bacterial kinetic curve of dandelion phenolic extract were analyzed by spectrophotometry. Changes in extracellular alkaline phosphatase (AKP) contents, electrical conductivity, intracellular protein contents, and DNA of S. aureus after the action of dandelion phenolic extract were determined to study its effect on the permeability of S. aureus cell wall and cell membrane. The results showed that chlorogenic acid (1.34 mg/g) was present in higher concentration, followed by luteolin (1.08 mg/g), ferulic acid (0.22 mg/g), caffeic acid (0.21 mg/g), and rutin (0.19 mg/g) in the dandelion phenolic extract. The minimum inhibitory concentration (MIC) of dandelion phenolic extract against S. aureus was 12.5 mg/mL. The antibacterial kinetic curve analysis showed that the inhibitory effect of dandelion phenolic extract on S. aureus was mainly in the exponential growth phase. After applying the dandelion phenolic extract, the growth of S. aureus was signifi- cantly inhibited entering into the decay phase early. Furthermore, after the action of dandelion, the extracellular AKP contents of S. aureus, the electrical conductivity and the extracellular protein contents were all increased. The phenolic extract also affected the normal reproduction of S. aureus. These results suggest that dandelion has an inhibitory effect on S. aureus, and the mechanism of its action was to destroy the integrity of the cell walls and cell membranes.
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Authors and Affiliations

P. Xu
1 2 3
X.B. Xu
1
A. Khan
4
T. Fotina
3
S.H. Wang
2

  1. School of Life Science and Basic Medicine, Xinxiang University, Jinsui St. 191, 453003 Xinxiang, China
  2. College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Eastern Hua Lan Street, 453003 Xinxiang, China
  3. Department of Veterinary Medicine, Sumy National Agrarian University, Herasima Kondratieva St. 160, 40021 Sumy, Ukraine
  4. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi 030801, P.R. China
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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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Authors and Affiliations

X. Li
M. Liu
X. Bai
Y. Li
Y. Zhao
S. Wang
J. Wang
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Abstract

To explore the role of Toll-like receptors (TLRs) and interferon (IFN) in the innate immunity against porcine epidemic diarrhea virus (PEDV), we detected the expression of TLR genes in PEDV-infected IPEC-J2 cells by real-time PCR. We also detected the level of interferon α (IFN-α) and interferon γ (IFN-γ) by enzyme-linked immunosorbent assay (ELISA). Results showed that IPEC-J2 cells exhibited a clear pathological change after PEDV infection at 24 h. In addition, TLR7, TLR9 and TLR10 expressions were significantly upregulated in PEDV-infected IPEC-J2 cells at 24 h. Interestingly, the expression patterns of TLR2 and TLR4 were consistent at different stages of PEDV infection. The expression level of TLR3 decreased significantly with the increase of infection time, but the expression levels of TLR5 and TLR8 genes at 6 h and 12 h were significantly lower than those in the control group (p<0.01). There were significant correlations among the expression levels of TLR genes (p<0.05). Cytokine detection showed that the secretion level of IFN-α in the PEDV-infected group was significantly higher than that in the control group (p<0.01), and IFN-γ at 6 h and 12 h after PEDV infection was significantly higher than that in control group (p<0.01). Therefore, our results suggest that PEDV infection can induce innate immune responses in intestinal porcine jejunum epithelial cells, leading to changes in the expression of Toll-like receptors, and can regulate the resistance to virus infection by affecting the release levels of downstream cytokines.

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Authors and Affiliations

F. Wang
S.Q. Wang
H.F. Wang
Z.C. Wu
W.B. Bao
S.L. Wu
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Abstract

Information regarding the correct pedigree of and relationship between animals is useful for managing dairy breeding, reducing inbreeding, estimating breeding value, and establishing correct breeding programs. Additionally, the successful implementation of progeny testing is crucial for improving the genetics of dairy cattle, which depends on the availability of correct pedigree information. Incorrect pedigree information leads to bias in bull evaluation. In this study, Neogen GeneSeek Genomic Profiler (GGP) 50K SNP chips were used to identify and verify the sire of Taiwanese Holstein dairy cattle and analyze the reasons that lead to incorrect sire records. Samples were collected from 2,059 cows of 36 dairy farms, and the pedigree information was provided by breeders. The results of sire verification can be divided into three categories: submitted unconfirmed sire, submitted confirmed sire, and incorrectly submitted verified sire. Data on the sires of 1,323 (64.25%) and 572 (27.78%) dairy cows were verified and discovered, respectively. Sires of 1,895 (92.03%) dairy cattle were identified, which showed that the paternal pedigree of dairy cattle could be discovered and verified through genetic testing. An error-like analysis revealed that the data of 37 sires were incorrectly recorded because the bull’s NAAB code number was incorrectly entered into the insemination records: for 19 sires, the wrong bull was recorded because the frozen semen of a bull placed in the wrong storage tank was used, 6 had no sire records, and for 12 sires, the NAAB code of the correct bull was recorded but with a wrong stud code, marketing code, or unique number for the stud or breed. To reduce recorded sire error rates by at least 27.78%, automated identification of the mated bull must be adopted to reduce human error and improve dairy breeding management on dairy farms.
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Bibliography


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Authors and Affiliations

C.H. Chao
1
Y.H. Yeh
1
Y.M. Chen
1
K.H. Lee
1
S.H. Wang
1
T.Y. Lin
1

  1. Hsinchu Branch, Livestock Research Institute, Council of Agriculture, Executive Yuan, 207-5, Bi-tou-mian, Wu-hoo village, Si-hoo Township, Miaoli County, Taiwan
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Abstract

The interface characteristics, bending and impact behavior, as well as fracture characteristics of stainless steel clad plates fabricated by vacuum hot rolling at different rolling temperatures of 1100°C, 1200°C and 1300°C are investigated in detail. The interface bonding strength is gradually increased with the increasing rolling temperature due to the sufficient diffusion behavior of alloy element. The bending toughness and impact toughness are gradually decreased, while the bending strength increase with the increase of the rolling temperature, which is attributed to mechanisms of matrix softening and interface strengthening at high rolling temperature. Due to the weak interface at 1100°C, the bending and impact crack propagation path was displaced by delamination cracks, which in turn lead to reduction in stress intensity of the main crack, playing an effective role in toughening the stainless steel clad plates. Moreover, the impact fracture morphologies of clad plates show a typical ductile-brittle transition phenomenon, which is attributed to the matrix softening behavior with the increasing rolling temperature.

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Authors and Affiliations

Q. An
K.Y. Fan
Y.F. Ge
B.X. Liu
J. He
S. Wang
C.X. Chen
P.G. Ji
O. Tolochko
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Abstract

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a major public health concern. Nucleocapsid (N) protein is the most abundant structural protein on SARS-CoV-2 virions and induces the production of antibodies at the early stage of infection. Large-scale preparation of N protein is essential for the development of immunoassays to detect antibodies to SARS-CoV-2 and the control of virus transmission. In this study, expression of water-soluble N protein was achieved through inducing protein expression at 25°C with 0.5 mM IPTG for 12 h. Western blot and ELISA showed that recombinant N protein could be recognized by sera collected from subjects immunized with Sinovac inactivated SARS-CoV-2 vaccine. Four monoclonal antibodies namely 2B1B1, 4D3A3, 5G1F8, and 7C6F5 were produced using hybridoma technology. Titers of all four monoclonal antibodies in ELISA reached more than 1.28×10 6.0. Moreover, all monoclonal antibodies could react specifically with N protein expressed by transfection of pcDNA3.1-N into BHK-21 cells in IPMA and IFA. These results indicated that water-soluble N protein retained high immunogenicity and possessed the same epitopes as that of native N protein on virions. In addition, the preparation of water-soluble N protein and its monoclonal antibodies laid the basis for the development of immunoassays for COVID-19 detection.
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Authors and Affiliations

Y.B. Wang
1
S.W. Wang
2
Q.Y. Jin
3
L.P. Chen
4
F.Q. Zhang
1
J.J. Shi
1
Y. Yin
5
Z.X. Fan
1
X.Y. Liu
6
L.P. Wang
6
P. Li
6

  1. School of Public Health, Xinxiang Medical University, Xinxiang 453003, P.R. China
  2. School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453003, P.R. China
  3. Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, P.R. China
  4. Gushi County Center for Animal Disease Control and Prevention, Xinyang 465200, P.R. China
  5. Mingde College of Xinxiang Medical University, Xinxiang 453003, P.R. China
  6. School of Biological Engineering, Xinxiang University, Xinxiang 453003, P.R. China

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