14 results on '"Xianjing, He"'
Search Results
2. Evaluation of Immunoprotective Effects of Fusobacterium necrophorum Outer Membrane Proteins 43K OMP, Leukotoxin and Hemolysin Multi-Component Recombinant Subunit Vaccine in Mice
- Author
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Jiawei Xiao, Jiancheng Jiang, Xianjing He, Siyao Zhang, Zhihui Wang, Fengfeng Wang, Lina Wang, and Donghua Guo
- Subjects
43K OMP ,hemolysin ,leukotoxin ,immunoprotective ,Fusobacterium necrophorum ,Veterinary medicine ,SF600-1100 - Abstract
We evaluated the efficacy of three vaccine formulations containing different combinations of proteins (43K OMP, leukotoxin recombinant protein PL4 and hemolysin recombinant protein H2) and killed whole cell Fusobacterium necrophorum in preventing liver abscess. Four subcutaneous vaccines were formulated: vaccine 1 (43K OMP), vaccine 2 (PL4 and H2), vaccine 3 (43K OMP, PL4 and H2), and vaccine 4 (killed whole bacterial cell). 43K OMP, PL4, and H2 proteins were produced by using recombinant protein expression. To evaluate vaccine efficacy, we randomly allocated 50 BALB/c female mice to one of five different treatment groups: PBS control group, vaccine 1, vaccine 2, vaccine 3, and vaccine 4. Mice were vaccinated three times, with 14 days between each immunization. After immunization, the mice were challenged with F. necrophorum. The three key findings of this study are as follows: (1) Vaccine 3 has enabled mice to produce higher antibody titer following bacterial challenge, (2) in the liver pathology of mice, the vaccine 3 liver showed the least pathology, and (3) all four vaccines produced high levels of antibodies and cytokines in mice, but the level of vaccine 3 was the highest. Based on our results, it has been demonstrated that a mixture of F. necrophorum 43K OMP, PL4, and H2 proteins inoculated with mice can achieve protection against liver abscess in mice. Our research may therefore provide the basis for the development of a vaccine against F. necrophorum bovine infections.
- Published
- 2021
- Full Text
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3. Supplementary material to 'The biogeography of relative abundance of soil fungi and bacteria in top surface soil'
- Author
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Kailiang Yu, Johan van den Hoogen, Zhiqiang Wang, Colin Averill, Devin Routh, Gabriel R. Smith, Rebecca E. Drenovsky, Kate M. Scow, Fei Mo, Mark P. Waldrop, Yuanhe Yang, Weize Tang, Franciska T. De Vries, Richard D. Bardgett, Peter Manning, Felipe Bastida, Sara G. Baer, Elizabeth M. Bach, Carlos García, Qingkui Wang, Linna Ma, Baodong Chen, Xianjing He, Sven Teurlincx, Amber Heijboer, James A. Bradley, and Thomas W. Crowther
- Published
- 2022
4. The biogeography of relative abundance of soil fungi and bacteria in top surface soil
- Author
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Kailiang Yu, Johan van den Hoogen, Zhiqiang Wang, Colin Averill, Devin Routh, Gabriel R. Smith, Rebecca E. Drenovsky, Kate M. Scow, Fei Mo, Mark P. Waldrop, Yuanhe Yang, Weize Tang, Franciska T. De Vries, Richard D. Bardgett, Peter Manning, Felipe Bastida, Sara G. Baer, Elizabeth M. Bach, Carlos García, Qingkui Wang, Linna Ma, Baodong Chen, Xianjing He, Sven Teurlincx, Amber Heijboer, James A. Bradley, and Thomas W. Crowther
- Abstract
Fungi and bacteria are the two dominant groups of soil microbial communities worldwide. By controlling the turnover of soil organic matter, these organisms directly regulate the exchange of carbon between the soil and the atmosphere. Fundamental differences in the physiology and life history of bacteria and fungi suggest that variation in the biogeography of soil fungal and bacterial relative abundance could drive striking differences in carbon decomposition and soil organic matter formation across different biomes. However, a lack of global and predictive information on the distribution of these organisms in terrestrial ecosystems has prevented the inclusion of soil fungal and bacterial relative abundance and the associated processes into global biogeochemical models. Here, we used a global scale dataset in the top soil surface (>3000 distinct observations of soil fungal and bacterial abundance) to generate the first quantitative and spatially high resolution (1 km) explicit map of soil fungal proportion, defined as fungi/fungi + bacteria, across terrestrial ecosystems. We reveal striking latitudinal trends where fungal dominance increases in cold and high latitude environments with large soil carbon stocks. There was strong non-linear response of fungal dominance to environmental gradient, i.e., mean annual temperature (MAT) and net primary productivity (NPP). Fungi and bacteria dominated in regions with low and high MAT and NPP, respectively, thus representing slow vs. fast soil energy channels, a concept with a long history in soil ecology. These high-resolution models provide the first steps towards representing the major soil microbial groups and their functional differences in global biogeochemical models to improve predictions of soil organic matter turnover under current and future climate scenarios. Raw datasets and global maps generated in this study are available at https://doi.org/10.6084/m9.figshare.19556419 (Yu, 2022).
- Published
- 2022
5. Selenium Deficiency in Chickens Induces Intestinal Mucosal Injury by Affecting the Mucosa Morphology, SIgA Secretion, and GSH-Px Activity
- Author
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Jianfa Wang, Donghua Guo, Yucai Lin, Rui Wu, Dongbo Sun, Shuai Lian, and Xianjing He
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medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,Ileum ,010501 environmental sciences ,digestive system ,01 natural sciences ,Biochemistry ,Antioxidants ,Inorganic Chemistry ,Jejunum ,Selenium ,03 medical and health sciences ,Intestinal mucosa ,Selenium deficiency ,Internal medicine ,medicine ,Animals ,Intestinal Mucosa ,0105 earth and related environmental sciences ,chemistry.chemical_classification ,0303 health sciences ,Glutathione peroxidase ,030302 biochemistry & molecular biology ,Biochemistry (medical) ,General Medicine ,medicine.disease ,Small intestine ,medicine.anatomical_structure ,Endocrinology ,chemistry ,Immunoglobulin A, Secretory ,Duodenum ,Intraepithelial lymphocyte ,Chickens - Abstract
The small intestine is one of the target organs of dietary selenium (Se) deficiency. Our objective was to investigate the effects of Se deficiency on small intestinal mucosa morphology and function in chickens. In the present study, 1-day (d)-old chickens were fed either a commercial diet with 0.15 mg/kg Se (control group) or a Se-deficient diet with 0.016 mg/kg Se (Se-group). The average daily weight gain, Se content in the blood, secretory immunoglobulin A (SIgA) secretion, and glutathione peroxidase (GSH-Px) activity in the small intestine in chickens were examined after 10, 20, 30, and 40 days of feeding. We also observed the morphology of the small intestine and recorded the number of intraepithelial lymphocytes (IELs). The average daily weight gain decreased; the level of Se in the blood decreased significantly; and SIgA secretion and GSH-Px activity in the duodenum, jejunum, and ileum decreased to different degrees. Histological analysis showed that the villus length, crypt depth, mucosal thickness, and number of IELs in the small intestine decreased to different extents in different periods. In the Se-group, longer feeding times were associated with more severe injury to physiological structure and function in the intestinal mucosa in chickens. In conclusion, Se deficiency induced injury of the mucosal immune barrier and physical barrier of the small intestine, and decreased the growth performance and antioxidant capacity in chickens.
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- 2020
6. The biogeography of relative abundance of soil fungi versus bacteria in surface topsoil
- Author
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Kailiang Yu, Johan van den Hoogen, Zhiqiang Wang, Colin Averill, Devin Routh, Gabriel Reuben Smith, Rebecca E. Drenovsky, Kate M. Scow, Fei Mo, Mark P. Waldrop, Yuanhe Yang, Weize Tang, Franciska T. De Vries, Richard D. Bardgett, Peter Manning, Felipe Bastida, Sara G. Baer, Elizabeth M. Bach, Carlos García, Qingkui Wang, Linna Ma, Baodong Chen, Xianjing He, Sven Teurlincx, Amber Heijboer, James A. Bradley, Thomas W. Crowther, Microbial Ecology (ME), AKWA, Aquatic Ecology (AqE), Terrestrial Ecology (TE), Ecosystem and Landscape Dynamics (IBED, FNWI), and IBED Other Research (FNWI)
- Subjects
General Earth and Planetary Sciences - Abstract
Fungi and bacteria are the two dominant groups of soil microbial communities worldwide. By controlling the turnover of soil organic matter, these organisms directly regulate the cycling of carbon between the soil and the atmosphere. Fundamental differences in the physiology and life history of bacteria and fungi suggest that variation in the biogeography of relative abundance of soil fungi versus bacteria could drive striking differences in carbon decomposition and soil organic matter formation between different biomes. However, a lack of global and predictive information on the distribution of these organisms in terrestrial ecosystems has prevented the inclusion of relative abundance of soil fungi versus bacteria and the associated processes in global biogeochemical models. Here, we used a global-scale dataset of >3000 distinct observations of abundance of soil fungi versus bacteria in the surface topsoil (up to 15 cm) to generate the first quantitative and high-spatial-resolution (1 km2) explicit map of soil fungal proportion, defined as fungi/fungi + bacteria, across terrestrial ecosystems. We reveal striking latitudinal trends where fungal dominance increases in cold and high-latitude environments with large soil carbon stocks. There was a strong nonlinear response of fungal dominance to the environmental gradient, i.e., mean annual temperature (MAT) and net primary productivity (NPP). Fungi dominated in regions with low MAT and NPP and bacteria dominated in regions with high MAT and NPP, thus representing slow vs. fast soil energy channels, respectively, a concept with a long history in soil ecology. These high-resolution models provide the first steps towards representing the major soil microbial groups and their functional differences in global biogeochemical models to improve predictions of soil organic matter turnover under current and future climate scenarios. Raw datasets and global maps generated in this study are available at https://doi.org/10.6084/m9.figshare.19556419 (Yu, 2022)., Earth System Science Data, 14 (9), ISSN:1866-3516, ISSN:1866-3508
- Published
- 2022
7. Liver X receptor α participates in LPS-induced reduction of triglyceride synthesis in bovine mammary epithelial cells
- Author
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Shuai Lian, Jianfa Wang, Jun Song, Dandan Hao, Xianjing He, Hai Wang, Xu Zhang, and Rui Wu
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Lipopolysaccharides ,medicine.medical_specialty ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Mammary Glands, Animal ,Internal medicine ,medicine ,Animals ,Liver X receptor ,Fatty acid synthesis ,Cells, Cultured ,Triglycerides ,030304 developmental biology ,Liver X Receptors ,chemistry.chemical_classification ,0303 health sciences ,Triglyceride ,Fatty acid ,Epithelial Cells ,General Medicine ,Metabolism ,Sterol regulatory element-binding protein ,Endocrinology ,Enzyme ,chemistry ,Gene Expression Regulation ,030220 oncology & carcinogenesis ,Sterol Regulatory Element Binding Protein 1 ,Animal Science and Zoology ,Cattle ,Female ,Food Science - Abstract
Lipopolysaccharides (LPS) could induce milk fat depression via regulating the body and blood fat metabolism. However, it is not completely clear how LPS might regulate triglyceride synthesis in dairy cow mammary epithelial cells (DCMECs). DCMECs were isolated and purified from dairy cow mammary tissue and treated with LPS. The level of triglyceride synthesis, the expression and activity of the liver X receptor α (LXRα), enzymes related to de novo fatty acid synthesis, and the expression of the fatty acid transporters were investigated. We found that LPS decreased the level of triglyceride synthesis via a down-regulation of the transcription, translation, and nuclear translocation level of the LXRα. The results also indicated that the transcription level of the LXRα target genes, sterol regulatory element binding protein 1 (SREBP1), fatty acid synthetase (FAS), acetyl-CoA carboxylase-1 (ACC1), were significantly down-regulated in DCMECs after LPS treatment. Our data may provide new insight into the mechanisms of milk fat depression caused by LPS.
- Published
- 2020
8. Interaction of 43K OMP of Fusobacterium necrophorum with fibronectin mediates adhesion to bovine epithelial cells
- Author
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Xianjing, He, Kai, Jiang, Jiawei, Xiao, Shuai, Lian, Yaping, Chen, Rui, Wu, Lina, Wang, Dongbo, Sun, and Donghua, Guo
- Subjects
Fusobacterium necrophorum ,General Veterinary ,Fusobacterium Infections ,Animals ,Cattle Diseases ,Cattle ,Epithelial Cells ,Female ,General Medicine ,Foot Rot ,Microbiology ,Fibronectins - Abstract
Fusobacterium necrophorum, a Gram-negative anaerobe, is an important bovine pathogen that causes hepatic abscesses, foot rot, mastitis and endometritis. We have previously shown that the 43 kDa outer membrane protein (43 K OMP) of F. necrophorum is a porin protein that plays an important role in bacterial infections; however, the molecular mechanisms by which this protein mediates adhesion remain unclear. In this study, we investigated the role of 43 K OMP in F. necrophorum adhesion to bovine epithelial cells using 43 K OMP-deficient mutants, and identified the protein that interacts with 43 K OMP by immunoprecipitation-mass spectrometry. Our results indicated that the native 43 K OMP and recombinant 43 K OMP could bind to the cell membrane of MAC-T or bovine endometrial epithelial cells (BEECs). When F. necrophorum was preincubated with antibodies against the recombinant 43 K OMP or bovine epithelial cells were preincubated with 43 K OMP, the adhesion of F. necrophorum to MAC-T or BEECs decreased significantly (P0.01). We successfully constructed a 43 K OMP-deficient strain (A25Δ43 K OMP) and bacterial attachment to MAC-T or BEECs was significantly higher with the F. necrophorum A25 strain than with mutant strain A25Δ43 K OMP (P0.01). The deficiency of 43 K OMP reduced the binding of F. necrophorum to bovine epithelial cells by 90.5 %-94.9 %. Among the 39 potential differential proteins, fibronectin, collagen and myosin were selected as the target proteins, and direct interaction between 43 K OMP of F. necrophorum and fibronectin was demonstrated. Taken together, these results suggest that 43 K OMP plays a key role in adhesion of F. necrophorum to bovine epithelial cells through its interaction with fibronectin. These findings provide a theoretical basis for the pathogenic mechanism of F. necrophorum.
- Published
- 2022
9. LPS-induced reduction of triglyceride synthesis and secretion in dairy cow mammary epithelial cells via decreased SREBP1 expression and activity
- Author
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Chunqiu Li, Dongbo Sun, Bin Yang, Xufei Shan, Jianfa Wang, Xianjing He, Hai Wang, Xu Zhang, and Rui Wu
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Lipopolysaccharides ,0301 basic medicine ,medicine.medical_specialty ,03 medical and health sciences ,Mammary Glands, Animal ,Transcription (biology) ,Internal medicine ,medicine ,Animals ,Secretion ,Transcription factor ,Triglycerides ,chemistry.chemical_classification ,Lipogenesis ,Triglyceride synthesis ,food and beverages ,Fatty acid ,Epithelial Cells ,General Medicine ,medicine.disease ,Mastitis ,Sterol regulatory element-binding protein ,030104 developmental biology ,Endocrinology ,Gene Expression Regulation ,chemistry ,Sterol Regulatory Element Binding Protein 1 ,Cattle ,Female ,Animal Science and Zoology ,Food Science - Abstract
Sterol regulatory element binding protein 1 (SREBP1) has a central regulatory effect on milk fat synthesis. Lipopolysaccharides (LPS) can induce mastitis and cause milk fat depression in cows. SREBP1 is also known to be associated with inflammatory regulation. Thus, in the current study, we hypothesized that LPS-induced milk fat depression in dairy cow mammary epithelial cells (DCMECs) operates via decreased SREBP1 expression and activity. To examine the hypothesis, DCMECs were isolated and purified from dairy cow mammary tissue and treated with LPS (10 µg/ml). LPS treatment of DCMECs suppressed lipid-metabolism-related transcription factor SREBP1 mRNA expression, nuclear translocation and protein expression, leading to reduced triglyceride content. The transcription levels of acetyl-CoA carboxylase-1 and fatty acid synthetase were significantly down-regulated in DCMECs after LPS treatment, suggesting that acetyl-CoA carboxylase-1 and fatty acid synthetase involved in de novo milk fat synthesis was regulated by SREBP1. In summary, these results suggest that LPS induces milk fat depression in dairy cow mammary epithelial cells via decreased expression of SREBP1 in a time-dependent manner.
- Published
- 2018
10. Selenium deficiency induces duodenal villi cell apoptosis via an oxidative stress-induced mitochondrial apoptosis pathway and an inflammatory signaling-induced death receptor pathway
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Debin Yu, Jianfa Wang, Shuai Lian, Dongbo Sun, Xianjing He, Rui Wu, Jianbin Liang, and Zhe Liu
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Male ,0301 basic medicine ,Antioxidant ,Proteome ,Duodenum ,medicine.medical_treatment ,Biophysics ,Apoptosis ,medicine.disease_cause ,Biochemistry ,Antioxidants ,Biomaterials ,Selenium ,03 medical and health sciences ,chemistry.chemical_compound ,Selenium deficiency ,medicine ,Animals ,chemistry.chemical_classification ,Reactive oxygen species ,Glutathione peroxidase ,Metals and Alloys ,Glutathione ,medicine.disease ,Molecular biology ,Mitochondria ,Oxidative Stress ,030104 developmental biology ,chemistry ,Chemistry (miscellaneous) ,Cytokines ,Inflammation Mediators ,Signal transduction ,Reactive Oxygen Species ,Chickens ,Oxidation-Reduction ,Oxidative stress ,Signal Transduction - Abstract
Selenium (Se) is an important nutritional trace element possessing antioxidant properties. Our goal was to elucidate the effect and mechanism of Se deficiency on the intestinal cell fate. One-day-old three-yellow chickens were fed a low Se diet for 1, 3, and 5 weeks. Histologic characteristics, protein expression profiles, antioxidant activities, inflammatory signaling, and the apoptosis status in duodenum mucosa were investigated. Histological results showed that Se deficiency could increase inflammatory cell infiltration, karyopyknosis of the epithelial cells, cytoplasm vacuolization and dissolution of goblet cells. The proteomics results indicated that Se deficiency could induce apoptosis of cells in duodenal villi via inhibition of antioxidant redox signaling and activation of NF-κB signaling. Further analysis results showed that Se deficiency decreased the total antioxidant capacity of duodenum mucosa via down-regulating the transcription level and activities of glutathione peroxidase (GPX), reduced glutathione (GSH), and thioredoxin reductase (TrxR). The NF-κB signaling pathway was activated by Se deficiency-induced reactive oxygen species (ROS). TUNEL, DNA ladder, immunohistochemical assay, and western blotting proved that selenium deficiency could induce duodenal villi cell apoptosis. The results also indicated that Se deficiency can cause duodenal villi cell apoptosis via an oxidative stress-induced mitochondrial apoptosis pathway (intrinsic pathway) and an inflammatory signaling-induced death receptor pathway (extrinsic pathway). Our data may provide new insight into the prevention and treatment of chronic diarrhea caused by Se deficiency.
- Published
- 2018
11. 17β-Estradiol and progesterone decrease MDP induced NOD2 expression in bovine mammary epithelial cells
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Xu Zhang, Dan-dan Xu, Rui Wu, Qian-yu He, Xianjing He, Bin Yang, Jianfa Wang, Gang Wang, Dongbo Sun, and Zhi-peng Sun
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Blotting, Western ,Immunology ,Nod2 Signaling Adaptor Protein ,Biology ,Real-Time Polymerase Chain Reaction ,03 medical and health sciences ,chemistry.chemical_compound ,Mammary Glands, Animal ,Adjuvants, Immunologic ,Internal medicine ,NOD2 ,medicine ,Animals ,Receptor ,Protein kinase A ,Progesterone ,Innate immune system ,Estradiol ,General Veterinary ,0402 animal and dairy science ,Interleukin ,Epithelial Cells ,04 agricultural and veterinary sciences ,040201 dairy & animal science ,digestive system diseases ,030104 developmental biology ,Real-time polymerase chain reaction ,Endocrinology ,chemistry ,Cattle ,Female ,Tumor necrosis factor alpha ,Acetylmuramyl-Alanyl-Isoglutamine ,Muramyl dipeptide - Abstract
During the periparturient period, many neuroendocrine changes develop in cows. Periparturient hormone fluxes may adversely affect mammary gland immunity and mastitis susceptibility. 17β-Estradiol (E2) and progesterone (P4) have been reported to function on immune regulation, and their concentration fluctuates dramatically during the perinatal period. Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) mediate numerous aspects of innate immunity in humans and experimental animals. This study aimed to explore the effects of E2 and P4 on NOD2 expression in bovine mammary epithelial cells (BMECs). BMECs were isolated and purified from bovine mammary tissue and treated with E2/P4 and muramyl dipeptide (MDP). After these treatments, the mRNA levels of NOD2, receptor-interacting protein kinase (RIP) 2, interleukin (IL) 1β, IL-6, IL-8 and tumor necrosis factor (TNF) α were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) respectively, and the protein levels of NOD2 were analyzed by western blotting. The results showed that E2 and P4 decreased MDP-induced transcriptional expression of NOD2 and the downstream molecules. Moreover, E2 reduced MDP-induced NOD2 protein expression levels. Our study suggests that down-regulation of NOD2 by E2 and P4 may be one of the reasons for mastitis susceptibility in periparturient dairy cows.
- Published
- 2017
12. A Highly Sensitive and Selective Fluorescence Chemosensor for Cu2+and Zn2+Based on Solvent Effect
- Author
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Xiaobo Huang, Xianjing He, Yixiang Cheng, Qian Miao, Jieming Jiao, and Lu Wang
- Subjects
Circular dichroism ,Chemistry ,General Chemistry ,Solvent effects ,Photochemistry ,Fluorescence ,Highly sensitive - Abstract
A novel fluorescence chemosensor 1 based on (R)-binaphthyl-salen can exhibit highly sensitive and selective recognition responses toward Cu2+ by "turn-off" fluorescence quench type in THF/H2O, and Zn2+ by "turn-on" fluorescence enhancement type in CHCl3/CH3CN, respectively, suggesting that solvents can dramatically affect the responsive properties of salen-based chemosensor. In addition, Cu2+ can lead to the most pronounced changes of CD spectra without the influence of solvents, which indicates this kind chemosensor can also be used as a sole Cu2+ probe based on CD spectra.
- Published
- 2012
13. Development of a novel LAMP diagnostic method for visible detection of swine Pasteurella multocida
- Author
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Huanmin Yang, Jiasan Zheng, Jianfa Wang, Xianjing He, Chunren Wang, Dongbo Sun, and Rui Wu
- Subjects
DNA, Bacterial ,Pasteurella multocida ,Diagnostic methods ,genetic structures ,Swine ,animal diseases ,Pasteurella Infections ,Loop-mediated isothermal amplification ,Biology ,Sensitivity and Specificity ,Animals ,Swine Diseases ,Detection limit ,General Veterinary ,Reproducibility of Results ,General Medicine ,biology.organism_classification ,Molecular biology ,eye diseases ,Respiratory pathogens ,genomic DNA ,Specific primers ,sense organs ,Pcr method ,Nucleic Acid Amplification Techniques - Abstract
A set of four specific primers for six regions of kmt1 gene from a species specific region was designed for developing the loop-mediated isothermal amplification diagnostic method of swine Pasteurella multocida (Pm-LAMP). After the Pm-LAMP was carried out at 63°C for 1 h, the LAMP products could be visually confirmed using fluorescent dyes as detection reagent under UV-illumination. In sensitivity, the detection limit of the Pm-LAMP was 10 cfu/mL, and was 1 log less than that of the PCR method. In specificity, the Pm-LAMP did not amplify genomic DNA of swine common respiratory pathogens. Furthermore, based on results for clinical swab samples (n = 31) using PCR detection as golden standard, relative sensitivity of the Pm-LAMP was 100%, relative specificity of the Pm-LAMP was 90.9%, and percentage of observation agreement was 93.5% (Kappa = 0.85). The Pm-LAMP method should be a useful diagnostic tool for rapid and visible detection of swine Pasteurella multocida.
- Published
- 2010
14. A polyclonal antibody against the C subunit of porcine aminopeptidase N expressed in Escherichia coli
- Author
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Hong Zhang, Qinghe Zhu, Rui Wu, Wang Yueqiang, Xianjing He, Dongbo Sun, Donghua Guo, and Wu Guojun
- Subjects
Immunogen ,medicine.drug_class ,Swine ,Protein subunit ,Recombinant Fusion Proteins ,Immunology ,Genetic Vectors ,Enzyme-Linked Immunosorbent Assay ,CD13 Antigens ,medicine.disease_cause ,Monoclonal antibody ,Polymerase Chain Reaction ,law.invention ,Plasmid ,law ,Antibody Specificity ,medicine ,Escherichia coli ,Immunology and Allergy ,Animals ,Cloning, Molecular ,biology ,Gastroenteritis, Transmissible, of Swine ,Immune Sera ,Molecular biology ,Protein Subunits ,Polyclonal antibodies ,biology.protein ,Recombinant DNA ,Rabbits ,Antibody ,Coronavirus Infections - Abstract
The entire pig aminopeptidase N (pAPN) gene was amplified by RT-PCR using total RNA extracted from intestinal brush border membrane of a newborn piglet. The amplified products of the pAPN gene were cloned into the vector pMD18-T, generating a recombinant plasmid pMD18-T-pAPN. The C subunit of pAPN (pAPN-C) produced by PCR from the plasmid pMD18-T-pAPN was expressed in Escherichia coli using vector pET-32a with His tag. After confirming reactivity of the recombinant protein pAPN-C to antibody against native pAPN, polyclonal antibody against the recombinant protein pAPN-C was prepared in rabbit using purified protein as immunogen. In Western blot analysis, the antibody elicited by the recombinant protein pAPN-C could recognize the native pAPN. These data demonstrate that the pAPN-C recombinant protein and its polyclonal antibody can provide some basis for further receptor antagonist.
- Published
- 2011
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