Publications by authors named "Jun-Ling Hou"

38 Publications

[Main chemical constituents in aerial parts of Glycyrrhiza uralensis by UPLC-Q-Exactive Orbitrap-MS].

Zhongguo Zhong Yao Za Zhi 2021 Mar;46(6):1449-1459

School of Chinese Materia Medica,Beijing University of Chinese Medicine Beijing 102488,China Engineering Research Center of Good Agricultural Practice for Chinese Crude Drugs,Ministry of Education Beijing 102488,China.

Chemical constituents from aerial parts of Glycyrrhiza uralensis were analyzed and identified using ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS). The chromatographic column of Waters Acquity UPLC BEH-C_(18)(2.1 mm×100 mm, 1.7 μm) was adopted, with acetonitrile-water(0.5% formic acid) as mobile phase at a flow rate of 0.2 mL·min~(-1). Data was collected in positive and negative modes of electrospray ionization(ESI). A total of 55 compounds, including 42 flavonoids, 9 stilbenes, 2 coumarins, 1 lignin and 1 phenolic acid, which were characterized in the aerial parts of G. uralensis based on accurate molecular mass information of molecular and product ions provided by UPLC-Q-Exactive Orbitrap-MS based on comparison with standard substances and references. It is an effective and accurate method to provide chemical information of constituents in aerial parts of G. uralensis, and can provide a reference for further study on pharmacodynamic material basis and resources development and utilization.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20201225.301DOI Listing
March 2021

Molecular detection and subtype distribution of Blastocystis in farmed pigs in southern China.

Microb Pathog 2021 Feb 19;151:104751. Epub 2021 Jan 19.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, PR China. Electronic address:

Blastocystis is one of the most common causative agents of intestinal diseases, which can cause enteric diseases in animals and humans. However, limited data is available on the prevalence or subtypes of Blastocystis infections in farmed pigs in southern China. In this study, a total of 396 fecal samples were collected from farmed pigs in three provinces in southern China in 2016, and screened for Blastocystis by PCR amplification of the small subunit rRNA (SSU rRNA) gene fragment. One hundred and seventy (42.93%) of the examined fecal samples were detected Blastocystis-positive, and two known zoonotic subtypes ST1 and ST5 were identified, with ST5 being the predominate subtype. Moreover, gender, age and region were considered as risk factors that associated with Blastocystis infection in farmed pigs. The present study revealed the prevalence and subtypes of Blastocystis infections in farmed pigs in southern China, which provided essential data for the control of Blastocystis infections in pigs, other animals and humans in China.
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http://dx.doi.org/10.1016/j.micpath.2021.104751DOI Listing
February 2021

Advances in the Development of Anti- Vaccines: Challenges, Opportunities, and Perspectives.

Vaccines (Basel) 2020 Sep 22;8(3). Epub 2020 Sep 22.

MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.

The gastrointestinal nematode parasite () is a resident of tropical and subtropical regions worldwide that imposes significant production losses, economic losses, and animal health issues in the small ruminant industry, particularly sheep and goats. Considerable efforts have been made to understand how immunity is elicited against infection. Various potential vaccine antigens have been tested by different methods and strategies applied in animal models, and significant progress has been made in the development of vaccines against . This review highlighted and shared the knowledge about the current understanding of host immune responses to and ongoing challenges in the development of a protective, effective, and long-lasting vaccine against infection. We have also pinpointed some achievements and failures in the development and testing of vaccines, which will establish a road map for future research directions to explore new effective vaccine candidates for controlling and preventing infection.
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http://dx.doi.org/10.3390/vaccines8030555DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7565421PMC
September 2020

Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry-Based Metabolomics Reveals Metabolic Alterations in the Mouse Cerebellum During Infection.

Front Microbiol 2020 10;11:1555. Epub 2020 Jul 10.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

is a protozoan parasite with a remarkable neurotropism. We recently showed that infection can alter the global metabolism of the cerebral cortex of mice. However, the impact of infection on the metabolism of the cerebellum remains unknown. Here we apply metabolomic profiling to discover metabolic changes associated with infection of the mouse cerebellum using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Multivariate statistics revealed differences in the metabolic profiles between the infected and control mouse groups and between the infected mouse groups as infection advanced. We also detected 10, 22, and 42 significantly altered metabolites (SAMs) in the infected cerebellum at 7, 14, and 21 days post infection (dpi), respectively. Four metabolites [tabersonine, arachidonic acid (AA), docosahexaenoic acid, and oleic acid] were identified as potential biomarker or responsive metabolites to infection in the mouse cerebellum. Three of these metabolites (AA, docosahexaenoic acid, and oleic acid) play roles in the regulation of host behavior and immune response. Pathway analysis showed that infection of the cerebellum involves reprogramming of amino acid and lipid metabolism. These results showcase temporal metabolomic changes during cerebellar infection by in mice. The study provides new insight into the neuropathogenesis of infection and reveals new metabolites and pathways that mediate the interplay between and the mouse cerebellum.
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http://dx.doi.org/10.3389/fmicb.2020.01555DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7381283PMC
July 2020

First report of Cryptosporidium spp. infection and risk factors in black-boned goats and black-boned sheep in China.

Parasitol Res 2020 Sep 25;119(9):2813-2819. Epub 2020 Jun 25.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, Gansu Province, People's Republic of China.

Cryptosporidium is an opportunistic protozoan parasite that can inhabit in the gastrointestinal tract of various hosts. Cryptosporidium infection in black-boned goats and black-boned sheep may pose a threat to the survival and productivity, causing considerable economic losses to the livestock industry. However, it is yet to know whether black-boned goats and black-boned sheep in China are infected with Cryptosporidium. Thus, the objective of the present study was to investigate the prevalence and associated risk factors of Cryptosporidium infection in black-boned goats and black-boned sheep in Yunnan province, China. A total of 590 fecal samples were obtained from black-boned goats and black-boned sheep from five counties in Yunnan province, and the prevalence and species distribution of Cryptosporidium were determined by amplification of the 18S rDNA fragment using the nested PCR. The overall Cryptosporidium prevalence was 13.2% (78/590), with 18.0% (55/305) in black-boned goats and 8.1% (23/285) in black-boned sheep. The age and sampling site were identified as main factors that result in significant differences in Cryptosporidium prevalence. Three species, namely C. muris, C. xiaoi, and C. ubiquitum, were identified in black-boned goats and black-boned sheep in the present study, with C. muris (46/78) as the predominant species. This is the first report of Cryptosporidium infection in black-boned goats and black-boned sheep in China, and the findings will facilitate better understanding, prevention, and control of Cryptosporidium infection in black-boned goats and black-boned sheep in China.
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http://dx.doi.org/10.1007/s00436-020-06781-6DOI Listing
September 2020

Prevalence and genetic characterization of Enterocytozoon bieneusi and Giardia duodenalis in Tibetan pigs in Tibet, China.

Infect Genet Evol 2019 11 27;75:104019. Epub 2019 Aug 27.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China; Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu Province 225009, PR China. Electronic address:

Enterocytozoon bieneusi and Giardia duodenalis are important opportunistic enteric zoonotic pathogens that cause diarrhoea and intestinal diseases in animals and humans. China is the largest producer of pigs, but whether Tibetan pigs, a unique pig breed in Tibet, are infected with E. bieneusi and G. duodenalis is unknown. Therefore, we conducted a molecular epidemiological survey to determine the prevalence of E. bieneusi and G. duodenalis in Tibetan pigs in Tibet, China, and identified the genotypes of these causative agents. A total of 345 faecal specimens were collected from Tibetan pigs from three Tibet counties (Milin, Cuona and Gongbujiangda), examined by nested PCR and sequenced utilizing genetic markers in the ribosomal internal transcribed spacer (ITS) region of the rRNA and glutamate dehydrogenase (gdh) gene for E. bieneusi and G. duodenalis, respectively. Moreover, using multilocus sequence typing, the subtypes of E. bieneusi were identified based on four loci (MS1, MS3, MS4 and MS7). A total of 41 (11.88%) faecal samples from Tibetan pigs were E. bieneusi-positive, and 2 (0.58%) were G. duodenalis-positive. The multivariate logistic regression analysis showed that age was considered a risk factor for Tibetan pig infection of E. bieneusi. Two novel (GB11, GB31) and four known E. bieneusi genotypes (EbpC, EbpD, PigEBITS5 and CHS12) were identified and were all classified as zoonotic group 1 according to the phylogenetic analysis. Two MLGs (MLGI and MLGII) were further identified in the E. bieneusi EbpC genotype by multilocus sequence typing analysis. In addition, two G. duodenalis assemblages (D and E) were found in the present study. To our knowledge, the current study is the first to detect the prevalence and perform genetic characterization of G. duodenalis in Tibetan pigs in Tibet, China. The results could provide essential data for controlling E. bieneusi and G. duodenalis infections in Tibetan pigs that are in contact with other animals and humans, as Tibetan pigs could be a potential source for human infection by these pathogens.
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http://dx.doi.org/10.1016/j.meegid.2019.104019DOI Listing
November 2019

The Multitasking Cathepsin B Interferes With Various Functions of Goat Peripheral Blood Mononuclear Cells .

Front Immunol 2019 23;10:1707. Epub 2019 Jul 23.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

Cathepsin B, a lysosomal cysteine protease, is thought to be involved in the pathogenesis of infection, but its exact role remains unclear. In the present study, a recombinant cathepsin B (rFgCatB) protein was expressed in the methylotrophic yeast . Western blot analysis confirmed the reactivity of the purified rFgCatB protein to serum from -infected goats. The effects of serial concentrations (10, 20, 40, 80, and 160 μg/ml) of rFgCatB on various functions of goat peripheral blood mononuclear cells (PBMCs) were examined. We demonstrated that rFgCatB protein can specifically bind to the surface of PBMCs. In addition, rFgCatB increased the expression of cytokines (IL-2, IL-4, IL-10, IL-17, TGF-β, and IFN-γ), and increased nitric oxide production and cell apoptosis, but reduced cell viability. These data show that rFgCatB can influence cellular and immunological functions of goat PBMCs. Further characterization of the posttranslational modification and assessment of rFgCatB in immunogenicity studies is warranted.
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http://dx.doi.org/10.3389/fimmu.2019.01707DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6664072PMC
November 2020

Metabolomic signature of mouse cerebral cortex following Toxoplasma gondii infection.

Parasit Vectors 2019 Jul 29;12(1):373. Epub 2019 Jul 29.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, Gansu, People's Republic of China.

Background: The protozoan parasite Toxoplasma gondii infects and alters the neurotransmission in cerebral cortex and other brain regions, leading to neurobehavioral and neuropathologic changes in humans and animals. However, the molecules that contribute to these changes remain largely unknown.

Methods: We have investigated the impact of T. gondii infection on the overall metabolism of mouse cerebral cortex. Mass-spectrometry-based metabolomics and multivariate statistical analysis were employed to discover metabolomic signatures that discriminate between cerebral cortex of T. gondii-infected and uninfected control mice.

Results: Our results identified 73, 67 and 276 differentially abundant metabolites, which were involved in 25, 37 and 64 pathways at 7, 14 and 21 days post-infection (dpi), respectively. Metabolites in the unsaturated fatty acid biosynthesis pathway were upregulated as the infection progressed, indicating that T. gondii induces the biosynthesis of unsaturated fatty acids to promote its own growth and survival. Some of the downregulated metabolites were related to pathways, such as steroid hormone biosynthesis and arachidonic acid metabolism. Nine metabolites were identified as T. gondii responsive metabolites, namely galactosylsphingosine, arachidonic acid, LysoSM(d18:1), L-palmitoylcarnitine, calcitetrol, 27-Deoxy-5b-cyprinol, L-homophenylalanine, oleic acid and ceramide (d18:1/16:0).

Conclusions: Our data provide novel insight into the dysregulation of the metabolism of the mouse cerebral cortex during T. gondii infection and have important implications for studies of T. gondii pathogenesis.
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http://dx.doi.org/10.1186/s13071-019-3623-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6664753PMC
July 2019

[Investigation and analysis on difference of cultivation technique situation of Salvia miltiorrhiza].

Zhongguo Zhong Yao Za Zhi 2019 Apr;44(7):1314-1320

Institute of Medicinal Plant Development, Chinese Academy of Medicinal Sciences and Peking Union Medical Collage Beijing 100193, China School of Chinese Pharmacy, Beijing University of Chinese Medicine Beijing 100102, China Engineering Research Center of GAP for Chinese Crude Drugs, Ministry of Education Beijing 100102, China.

Salvia miltiorrhiza is one of the commonly used bulk medicinal materials, which has significant effect on cardiovascular disease, and are heavy demanded in Asia, Europe, North America, Russia and Africa. Consequently, increasing the yield and quality of S. miltiorrhiza has become a major concern worldwide. With the current wild resources of S. miltiorrhiza gradually decreasing, cultivated products occupy most of the markets. However, the cultivation area is widely distributed and the cultivation techniques is different, which lead to the quality and yield of S. miltiorrhiza in consistent. This paper combined visiting survey with document analysis to carry out the cultivation situation of S. miltiorrhiza in main cultivation areas of Shandong, Henan, Sichuan, Shanxi and Hebei provinces. There exist big differences of the ecological environment, mode of cultivation, fertilization, pest control, harvesting processing among the producing areas. We should carry on the ecological suitability zoning analysis and suitable cultivation of each area study to form a pattern of high quality and high yield for the sustainable development of S. miltiorrhiza cultivation.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20190129.010DOI Listing
April 2019

Serological evidence of hepatitis E virus and influenza A virus infection in farmed wild boars in China.

Acta Trop 2019 Apr 6;192:87-90. Epub 2019 Feb 6.

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, People's Republic of China; College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province 130118, People's Republic of China; Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu Province 225009, People's Republic of China. Electronic address:

Hepatitis E virus (HEV) and influenza A virus (IAV) are two important pathogens which can infect humans and various animals causing public health problems. In this study, the seroprevalence and risk factors associated with HEV and IAV infection in farmed wild boars were investigated in China. A total of 758 serum samples were collected from farmed wild boars between 2015 and 2016, and antibodies against HEV and IAV were examined by enzyme-linked immunosorbent assay (ELISA) using commercially available kits. The overall prevalence of anti-HEV antibodies was 24.54% (186/758, 95% CI 21.48-27.60) in farmed wild boars. There were statistically significant differences in the HEV seroprevalence in farmed wild boars of different ages (<22 days: 8.33%; 22-66 days: 18.89%; >66 days: 26.36%) (P < 0.05) and different genders (50.00% in male and 23.49% in female) (P <  0.01). However, there was no statistically significant difference in the HEV seroprevalence in farmed wild boars of different regions and different years. The overall IAV seroprevalence was 5.80% (44/758, 95% CI 4.14-7.46), and there was no statistically significant difference in the IAV seroprevalence in farmed wild boars of different ages and genders, collected from different regions and different years. Our results indicate that HEV and IAV infections in farmed wild boars may pose a potential risk for human infection. To our knowledge, this is the first report of HEV and IAV seroprevalence in farmed wild boars in China, which provides baseline data for further studies and for control of HEV and IAV infection in farmed wild boars.
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http://dx.doi.org/10.1016/j.actatropica.2019.02.004DOI Listing
April 2019

Proteomic analysis of Fasciola gigantica excretory and secretory products (FgESPs) interacting with buffalo serum of different infection periods by shotgun LC-MS/MS.

Parasitol Res 2019 Feb 18;118(2):453-460. Epub 2018 Dec 18.

Institute of Comparative Medicine, Yangzhou University, and Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis, and Jiangsu Key Laboratory of Zoonosis, Yangzhou, 225009, Jiangsu Province, People's Republic of China.

Fasciolosis, caused by Fasciola hepatica and Fasciola gigantica, is an important zoonotic disease in the world. It affects livestock, especially for sheep and cattle, causing major economic loss due to morbidity and mortality. Although the excretory and secretory products (ESPs) of F. hepatica have been relatively well studied, little is known about the interaction between the ESP and host, and the mechanism of the key proteins involved in interaction. In this study, buffaloes were infected by Fasciola gigantica, and infection serum was collected at three different periods (42dpi, 70dpi, and 98dpi). The interaction proteins were pulled down with three different period serum by Co-IP assay, respectively, and then identified by LC-MS/MS analysis. A number of proteins were identified; some of them related to the biological function of the parasite, while most of them the functions were unknown. For the annotated proteins, 13, 5, and 7 proteins were pulled down by the infected serum in 42dpi, 70dpi, and 98dpi, respectively, and 18 proteins could be detected in all three periods. Among them, 13 belong to the cathepsin family, 4 proteins related to glutathione S-transferase, and 3 proteins are calcium-binding protein; other proteins related to catalytic activity and cellular process. This study could provide new insights into the central role played by ESPs in the protection of F. gigantica from the host immune response. At the same time, our research provided material for further studies about the interaction between F. gigantica and host.
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http://dx.doi.org/10.1007/s00436-018-6169-zDOI Listing
February 2019

Protective Efficacy Against Acute and Chronic Infection Induced by Immunization With the DNA Vaccine TgDOC2C.

Front Microbiol 2018 4;9:2965. Epub 2018 Dec 4.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

is a ubiquitous intracellular apicomplexan parasite that can cause zoonotic toxoplasmosis. Effective vaccines against infection are necessary to prevent and control the spread of toxoplasmosis. The present study analyzed the B-linear epitopes of DOC2 (TgDOC2) protein and then cloned the C-terminus of the TgDOC2 gene (TgDOC2C) to construct the pVAX-TgDOC2C eukaryotic vector. After intramuscular injection of pVAX-TgDOC2C, immune responses were monitored. Two weeks after the last immunization, the protective effects of pVAX-TgDOC2C against acute and chronic toxoplasmosis were evaluated by challenges with RH tachyzoites (genotype I) and PRU cysts (genotype II). The DNA vaccine elicited strong humoral and cellular immune responses with high levels of IgG antibody, IL-2 and IFN-γ production compared to those of the controls. The percentage of CD4+ and CD8+ T cells in mice immunized with pVAX-TgDOC2C was significantly increased compared to that of mice injected with empty pVAX I or PBS. After acute infection with 10 lethal tachyzoites, mice immunized with pVAX-TgDOC2C survived longer (12.5 days) than mice treated with pVAX I (8 days) and PBS (7.5 days). Mice immunized with pVAX-TgDOC2C had significantly less brain cysts (1600.83 ± 284.61) compared to mice immunized with pVAX I (3016.67 ± 153.84) or PBS (3100 ± 246.98). Together, these results demonstrated that TgDOC2C confers protective immunity against infection and may be a promising candidate antigen for further development of an effective multicomponent vaccine for veterinary use against toxoplasmosis in livestock animals.
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http://dx.doi.org/10.3389/fmicb.2018.02965DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6288300PMC
December 2018

First report of Neospora caninum seroprevalence in farmed raccoon dogs in China.

Acta Trop 2019 Feb 5;190:80-82. Epub 2018 Nov 5.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, PR China; Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu Province, 225009, PR China. Electronic address:

Neospora caninum is an intracellular protozoan parasite with a worldwide distribution, which can cause abortion and stillbirth in ruminants. However, little is known of N. caninum infection in raccoon dogs in China. Thus, a total of 1181 serum samples of farmed raccoon dogs were collected from the major production areas of Jilin, Changchun in Jilin province, and Rizhao, Weihai and Yantai in Shandong province in China. The antibodies to N. caninum were examined by a competitive-inhibition enzyme-linked immunoassay (cELISA). While the N. caninum seroprevalence in farmed raccoon dogs from different regions (cities) were not statistically significant, farmed raccoon dogs in Shandong province had a significantly higher N. caninum seroprevalence (8.99%, 95% CI 7.13-10.85) than those in Jilin province (4.46%, 95% CI 1.99-6.93) (P <  0.05). N. caninum seroprevalence in male and female raccoon dogs were 7.91% (95% CI 5.99-9.83) and 8.06% (95% CI 5.46-10.66), respectively. In addition, the seroprevalence of N. caninum infection in infancy, youth and adult raccoon dogs was 7.32% (95% CI 4.31-10.33), 7.93% (95% CI 5.44-10.42) and 8.41% (95% CI 5.82-11.00), respectively. This is the first report of N. caninum seroprevalence in farmed raccoon dogs in China, which provided baseline data for the prevention of N. caninum infection in raccoon dogs in China.
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http://dx.doi.org/10.1016/j.actatropica.2018.11.002DOI Listing
February 2019

Prevalence and genotypes of Enterocytozoon bieneusi in pigs in southern China.

Infect Genet Evol 2018 12 13;66:52-56. Epub 2018 Sep 13.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China; College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province 130118, PR China. Electronic address:

Enterocytozoon bieneusi is one of the most important causative agents of microsporidiosis, causing diarrhoea the symptoms of enteric disease in humans and animals. Although there is some information on the prevalence and genotypes of E. bieneusi in China, there is still a lack of data in pigs in southern China. In the present study, a total of 396 faecal specimens were collected from pigs in Zhejiang, Guangdong and Yunnan provinces in southern China, and were examined by nested PCR amplification of the ribosomal internal transcribed spacer (ITS) for the prevalence and genotypes of E. bieneusi. The overall prevalence of E. bieneusi in pigs was 31.57% (125/396), forming 15 genotypes, including 9 known genotypes (EbpC, EbpA, D, G, H, PigEBITS5, Henan-IV, KIN-1, CHS5) and 6 novel genotypes (GD1, ZJ1, ZJ2, YN1, YN2 and YN3), which were all clustered into Group 1. Moreover, multilocus sequence typing (MLST) showed that 6, 3, 4 and 5 types were identified in MS1, MS3, MS7 and MS4 loci, respectively, representing four multilocus genotypes (MLGs), designated as MLGs novel-1 to novel-4 in the present study. This is the first detailed study of E. bieneusi using MLST in pigs in southern China, which extended information about the distribution of E. bieneusi genotypes in China.
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http://dx.doi.org/10.1016/j.meegid.2018.09.006DOI Listing
December 2018

Immunization with plasmid DNA expressing Heat Shock Protein 40 confers prophylactic protection against chronic Toxoplasma gondii infection in Kunming mice.

Parasite 2018 23;25:37. Epub 2018 Jul 23.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu 730046, PR China.

Toxoplasma gondii causes one of the most common protozoal diseases of humans and animals worldwide. With the aim of designing an effective vaccine against T. gondii infection, we examined the immunogenicity of a DNA vaccine expressing heat shock protein 40 (HSP40) against challenge with T. gondii (type I RH and type II Pru) strains in Kunming mice. The plasmid pVAX1-HSP40 was constructed and used to immunize mice by intramuscular injection for three sequential immunizations with two-week intervals. This immunization regimen significantly reduced parasite cyst burden in pVAX1-HSP40-immunized mice (1871.9 ± 142.3) compared with control mouse groups immunized with pVAX1 (3479.2 ± 204.4), phosphate buffered saline (3024.4 ± 212.8), or left untreated (3275.0 ± 179.8) as healthy controls (p < 0.01). However, immunization failed to protect mice against challenge with the virulent RH strain. There was a significant increase in T lymphocyte subclasses (CD3eCD4 T and CD3eCD8a T lymphocytes) in splenic tissues in immunized mice compared with controls (p < 0.05). However, the level of antibodies, lymphocyte proliferation and concentration of cytokines (IFN-γ, IL-2, IL-4, IL-10 and IL-12p70) were not significantly different between immunized and control mouse groups (p < 0.05). These data indicate that pVAX1-HSP40 induced specific immune responses and achieved a significant reduction in the number of brain cysts in Pru-infected mice, and thus can be tested in future immunization studies along with plasmids containing other immunogenic proteins as a cocktail vaccine to fully abolish chronic toxoplasmosis.
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http://dx.doi.org/10.1051/parasite/2018040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6057741PMC
October 2018

[Effects of different concentrations of nitrogen,phosphorus and potash on Glycyrrhiza uralensis and its relationship with soil element content].

Zhongguo Zhong Yao Za Zhi 2018 Jun;43(12):2474-2479

School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 102488, China.

With annual Glycyrrhiza uralensis seedlings as experimental material, using "3414" optimal regression design and applied fertilizer, through the sampling of G. uralensis at harvest, root fresh weight and content of active components were measured in Lanzhou, Bayan Nur city, Chifeng, Jiuquan. Combined with NPK content in soil, potted experiments were used to study the effects of different nitrogen and phosphorus ratios on the dry matter accumulation and accumulation of active components of G. uralensis. The results reported as follows: the optimum fertilizer treatment in Lanzhou, Bayan Nur city, Chifeng, Jiuquan was N₁P₂K₁,N₂P₂K₁,N₁P₁K₂ and N₂P₁K₂, respectively. The efforts of single fertilizer on the fresh root weight acted as parabolic type.There was no significant effect of fertilizer treatment on the accumulation of active components of G. uralensis. Furthermore, in terms of nitrogen and phosphorus, the type of fertilizers that restricted the growth of the region was the type of elements with lower content in the soil. The optimal fertilizer usage was in inverse proportion to content of elements in soil. When the content of phosphorus in soil was low, nitrogen fertilizer and potash fertilizer showed positive interaction with phosphorus fertilizer, whereas, they showed negative interaction.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.2018.0078DOI Listing
June 2018

A recombinant Fasciola gigantica 14-3-3 epsilon protein (rFg14-3-3e) modulates various functions of goat peripheral blood mononuclear cells.

Parasit Vectors 2018 03 6;11(1):152. Epub 2018 Mar 6.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.

Background: The molecular structure of Fasciola gigantica 14-3-3 protein has been characterized. However, the involvement of this protein in parasite pathogenesis remains elusive and its effect on the functions of innate immune cells is unknown. We report on the cloning and expression of a recombinant F. gigantica 14-3-3 epsilon protein (rFg14-3-3e), and testing its effects on specific functions of goat peripheral blood mononuclear cells (PBMCs).

Methods: rFg14-3-3e protein was expressed in Pichia pastoris. Western blot and immunofluorescence assay (IFA) were used to examine the reactivity of rFg14-3-3e protein to anti-F. gigantica and anti-rFg14-3-3e antibodies, respectively. Various assays were used to investigate the stimulatory effects of the purified rFg14-3-3e protein on specific functions of goat PBMCs, including cytokine secretion, proliferation, migration, nitric oxide (NO) production, phagocytosis, and apoptotic capabilities. Potential protein interactors of rFg14-3-3e were identified by querying the databases Intact, String, BioPlex and BioGrid. A Total Energy analysis of each of the identified interaction was performed. Gene Ontology (GO) enrichment analysis was conducted using Funcassociate 3.0.

Results: Sequence analysis revealed that rFg14-3-3e protein had 100% identity to 14-3-3 protein from Fasciola hepatica. Western blot analysis showed that rFg14-3-3e protein is recognized by sera from goats experimentally infected with F. gigantica and immunofluorescence staining using rat anti-rFg14-3-3e antibodies demonstrated the specific binding of rFg14-3-3e protein to the surface of goat PBMCs. rFg14-3-3e protein stimulated goat PBMCs to produce interleukin-10 (IL-10) and transforming growth factor beta (TGF-β), corresponding with low levels of IL-4 and interferon gamma (IFN-γ). Also, this recombinant protein promoted the release of NO and cell apoptosis, and inhibited the proliferation and migration of goat PBMCs and suppressed monocyte phagocytosis. Homology modelling revealed 65% identity between rFg14-3-3e and human 14-3-3 protein YWHAE. GO enrichment analysis of the interacting proteins identified terms related to apoptosis, protein binding, locomotion, hippo signalling and leukocyte and lymphocyte differentiation, supporting the experimental findings.

Conclusions: Our data suggest that rFg14-3-3e protein can influence various cellular and immunological functions of goat PBMCs in vitro and may be involved in mediating F. gigantica pathogenesis. Because of its involvement in F. gigantica recognition by innate immune cells, rFg14-3-3e protein may have applications for development of diagnostics and therapeutic interventions.
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http://dx.doi.org/10.1186/s13071-018-2745-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5840819PMC
March 2018

Identification of two novel host proteins interacting with Toxoplasma gondii 14-3-3 protein by yeast two-hybrid system.

Parasitol Res 2018 Apr 3;117(4):1291-1296. Epub 2018 Mar 3.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

Toxoplasma gondii deploys many effector proteins in order to hijack and manipulate host cell signaling pathways, allowing parasite colonization, subversion of immune responses, and disease progression. T. gondii effector protein 14-3-3 (Tg14-3-3) promotes parasite dissemination inside the body, by enhancing the migratory ability of infected microglia and dendritic cells. Understanding both the mechanism of action and the host targets of Tg14-3-3 effector is important because of their importance to the parasite's virulence. The aim of the present study was to explore the function of Tg14-3-3 by utilizing the yeast two-hybrid system (Y2HS) to identify novel Tg14-3-3 interactors/substrates in host cells. A human cDNA library was screened using Tg14-3-3 as the bait. Tg14-3-3 (RH strain, Type I) was cloned into the pGBKT7 vector and expressed in the Y2HGold yeast strain. The bait protein expression was validated by Western blotting analysis, auto-activation, and toxicity investigation compared with control (Y2HGold yeast strain transformed with empty pGBKT7 vector). Two positive Tg14-3-3 interactors identified by this screening, hCG1821272 and eIF5B (eukaryotic translation initiation factor 5B), were isolated and characterized. This approach made it possible to gain a better understanding of the function of Tg14-3-3 in regulating host proteins involved in key cellular processes, such as translational initiation and cell migration.
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http://dx.doi.org/10.1007/s00436-018-5812-zDOI Listing
April 2018

Expression profiles of genes involved in TLRs and NLRs signaling pathways of water buffaloes infected with Fasciola gigantica.

Mol Immunol 2018 02 11;94:18-26. Epub 2017 Dec 11.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu Province 225009, PR China. Electronic address:

Infection of ruminants and humans with Fasciola gigantica is attracting increasing attention due to its economic impact and public health significance. However, little is known of innate immune responses during F. gigantica infection. Here, we investigated the expression profiles of genes involved in Toll-like receptors (TLRs) and NOD-like receptors (NLRs) signaling pathways in buffaloes infected with 500F. gigantica metacercariae. Serum, liver and peripheral blood mononuclear cell (PBMC) samples were collected from infected and control buffaloes at 3, 10, 28, and 70days post infection (dpi). Then, the levels of 12 cytokines in serum samples were evaluated by ELISA. Also, the levels of expression of 42 genes, related to TLRs and NLRs signaling, in liver and PBMCs were determined using custom RT Profiler PCR Arrays. At 3 dpi, modest activation of TLR4 and TLR8 and the adaptor protein (TICAM1) was detected. At 10 dpi, NF-κB1 and Interferon Regulatory Factor signaling pathways were upregulated along with activation of TLR1, TLR2, TLR6, TLR10, TRAF6, IRF3, TBK1, CASP1, CD80, and IFNA1 in the liver, and inflammatory response with activated TLR4, TLR9, TICAM1, NF-κB1, NLRP3, CD86, IL-1B, IL-6, and IL-8 in PBMCs. At 28 dpi, there was increase in the levels of cytokines along with induction of NLRP1 and NLRP3 inflammasomes-dependent immune responses in the liver and PBMCs. At 70 dpi, F. gigantica activated TLRs and NLRs, and their downstream interacting molecules. The activation of TLR7/9 signaling (perhaps due to increased B-cell maturation and activation) and upregulation of NLRP3 gene were also detected. These findings indicate that F. gigantica alters the expression of TLRs and NLRs genes to evade host immune defenses. Elucidation of the roles of the downstream effectors interacting with these genes may aid in the development of new interventions to control disease caused by F. gigantica infection.
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http://dx.doi.org/10.1016/j.molimm.2017.12.007DOI Listing
February 2018

[Effects of different concentrations of nitrogen and phosphorus on growth and active components of Salvia miltiorrhiza].

Zhongguo Zhong Yao Za Zhi 2016 Nov;41(22):4175-4182

School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China.

With annual Salvia miltiorrhiza seedlings as experimental material, using "3414" optimal regression design recommended by the Ministry of Agriculture and regularly watered with nutrient solution, through the dynamic sampling of S. miltiorrhiza in different growing stages, and the growth index, dry weight of plant root and content of active components were measured. The potted experiments were applied to study the effects of different nitrogen and phosphorus ratios on the growth, dry matter accumulation and accumulation of active components of S. miltiorrhiza, in order to explore a compatible fertilization method of nitrogen and phosphorus ratio that are suitable for production and quality of S. miltiorrhiza. The results reported as follows:①High concentrations of nitrogen fertilizer was beneficial to dry matter accumulation of S. miltiorrhiza aerial parts, and low concentration of nitrogen fertilizer transferred the dry matter accumulation to underground, and N1P1 could make the transfer ahead of time;②Regression analysis showed that in the early growth stage (before early July), we could use the nitrogen and phosphorus as basic fertilizer at a concentration of 1.521,0.355 g•L⁻¹ respectively to promote the growth of S. miltiorrhiza and at a concentration of 2.281,0.710 g•L⁻¹ respectively to promote the dry matter accumulation of root (after mid-August);③Five kinds of active components of S. miltiorrhiza decreased with the increase of nitrogen concentration, and increased with the increase of the concentration of phosphate fertilizer. Nitrogenous fertilizer, phosphate fertilizer in N-P=2∶3 ratio was more suitable for the accumulation of salvianolic acids, in N-P=1∶2 ratio was more suitable for the accumulation of tanshinone.
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http://dx.doi.org/10.4268/cjcmm20162215DOI Listing
November 2016

Serum levels of cytokines in water buffaloes experimentally infected with Fasciola gigantica.

Vet Parasitol 2017 Sep 25;244:97-101. Epub 2017 Jul 25.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu Province, 225009, PR China. Electronic address:

Fasciola gigantica infection in water buffaloes causes significant economic losses especially in developing countries. Although modulation of the host immune response by cytokine neutralization or vaccination is a promising approach to control infection with this parasite, our understanding of cytokine's dynamic during F. gigantica infection is limited. To address this, we quantified the levels of serum cytokines produced in water buffaloes following experimental infection with F. gigantica. Five buffaloes were infected via oral gavage with 500 viable F. gigantica metacercariae and blood samples were collected from buffaloes one week before infection and for 13 consecutive weeks thereafter. The levels of 10 cytokines in serum samples were simultaneously determined using ELISA. F. gigantica failed to elicit the production of various pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-2, IL-6, IL-12, and IFN-γ. On the other hand, evidence of a Th2 type response was detected, but only early in the course of parasite colonization and included modest increase in the levels of IL-10 and IL-13. The results also revealed suppression of the immune responses as a feature of chronic F. gigantica infection in buffaloes. Taken together, F. gigantica seems to elicit a modest Th2 response at early stage of infection in order to downregulate harmful Th1- and Th17-type inflammatory responses in experimentally infected buffaloes. The full extent of anti-F. gigantica immune response and its relation to pathogenesis requires further study.
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http://dx.doi.org/10.1016/j.vetpar.2017.07.028DOI Listing
September 2017

First detection and genetic characterization of Toxoplasma gondii in market-sold oysters in China.

Infect Genet Evol 2017 10 12;54:276-278. Epub 2017 Jul 12.

College of Marine Science, Shandong University at Weihai, Weihai, Shandong Province 264209, PR China. Electronic address:

Toxoplasma gondii oocysts in the water might be filtered through the gills of shellfish in the process of feeding, and can be concentrated in the digestive glands. Consumers might become infected through ingestion of the shellfish unless they are discharged or inactivated by the shellfish. Thus, the purpose of this study was to evaluate the presence of T. gondii in market-sold oysters in China under natural conditions using a molecular approach. A total of 998 oysters were collected from markets in four cities (Weihai, Qingdao, Yantai and Rizhao) of Shandong province, eastern China. Of these, 26 samples (2.61%) were tested positive by nested PCR amplification of T. gondii B1 gene. Only one of the 26 positive samples was typed completely, and was identified as ToxoDB Genotype #9. This is first report of T. gondii in market-sold oysters in China, suggesting that oysters have the ability to filter and retain T. gondii oocysts in their tissues, which represents a risk to public health because oysters are frequently ingested in nature.
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http://dx.doi.org/10.1016/j.meegid.2017.07.014DOI Listing
October 2017

Detection of in White Yaks in Gansu Province, China.

Biomed Res Int 2017 6;2017:5790181. Epub 2017 Jun 6.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, China.

, the most common zoonotic pathogen of microsporidiosis, has been found in various animals and humans, but no information is available concerning the prevalence and genotypes of in white yaks . In the present study, 353 faecal samples from white yaks in Tianzhu Tibetan Autonomous County, Gansu Province, Northwestern China, were collected and examined by PCR amplification of the internal transcribed spacer gene to estimate prevalence and identify their genotypes. Of the 353 faecal samples, 4 (1.13%) were tested -positive. Sequences analysis revealed that two known genotypes, namely, I ( = 1) and BEB4 ( = 2), and a novel genotype, namely, WCY1 ( = 1), were found in this study. Among them, genotype WCY1 was clustered into Group 1, and genotypes I and BEB4 belonged to Group 2. The present study firstly indicates the existence of in yaks in Gansu Province, Northwestern China. This is also the first record of in white yaks. Effective measures should be taken to control infection in white yaks, other animals, and humans.
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http://dx.doi.org/10.1155/2017/5790181DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5476876PMC
March 2018

Molecular detection and genetic characterization of Toxoplasma gondii in farmed raccoon dogs (Nyctereutes procyonoides) in Shandong province, eastern China.

Acta Trop 2017 Aug 2;172:143-146. Epub 2017 May 2.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China; College of Marine Science, Shandong University at Weihai, Weihai, Shandong Province 264209, PR China. Electronic address:

Toxoplasma gondii is a successful opportunistic parasite, affecting a wide range of vertebrate animals and humans. Genetic diversity of T. gondii in raccoon dogs (Nyctereutes procyonoides) is of great importance to understand the transmission of T. gondii in the environment. However, no information is available about the distribution of genetic diversity of T. gondii infection in raccoon dogs. This study was conducted to estimate the prevalence and genetic characterization of T. gondii from raccoon dogs in Shandong province, eastern China. A total of 314 brain tissue samples of raccoon dogs were collected and genomic DNA was extracted and assayed for T. gondii infection using semi-nested PCR targeting B1 gene. The positive DNA samples were typed at 10 genetic markers (SAG1, SAG2(5'+3' SAG2, alter.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) by multiplex multilocus nested polymerase chain reaction-restriction fragment length polymorphism (Mn- PCR-RFLP) technology. Thirty-five (11.15%) of 314 DNA samples were detected positive. Only six samples were completely typed at all genetic loci, and these samples represented ToxoDB genotype#9. Two sample were typed at 9 genetic loci and one was typed at 8 genetic loci, all of them represented Type I. To our knowledge, this is the first report of genetic characterization of T. gondii in raccoon dogs in China. These results revealed the existence of genetic diversity of T. gondii in raccoon dogs in China. These data provided base-line information for controlling T. gondii infection in raccoon dogs.
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http://dx.doi.org/10.1016/j.actatropica.2017.05.001DOI Listing
August 2017

Occurrence of in Donkeys () in China: A Public Health Concern.

Front Microbiol 2017 31;8:565. Epub 2017 Mar 31.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesLanzhou, China.

is an important zoonotic parasite. It can infect virtually all animal species and has a global distribution. However, the prevalence of in donkeys () has only been reported in Algeria and Spain, and no information is available concerning genotypes of in donkeys worldwide. In the present study, a total of 301 donkey fecal samples (48 from Jilin Province, 224 from Shandong Province and 29 from Liaoning Province) were collected and examined by PCR amplification of the internal transcribed spacer (ITS) region. The overall . prevalence was 5.3% (16/301), with 6.3% (3/48) in Jilin Province, 4.9% (11/224) in Shandong Province, and 6.9% (2/29) in Liaoning Province. Prevalence in different age groups ranged from 4.2 to 5.5%. . prevalence in donkeys sampled in different seasons varied from 4.2 to 6.5%. Altogether, four genotypes were identified in this study, with two known genotypes (J and D) and two novel genotypes (NCD-1and NCD-2). Phylogenetic analysis revealed that genotypes D, NCD-1 and NCD-2 belonged to group 1, while the remaining genotype J was clustered into group 2. These findings revealed the occurrence of in donkeys in China for the first time. Moreover, the present study also firstly genotyped the . in donkeys worldwide. These findings extend the distribution of genotypes and provide baseline data for controlling infection in donkeys, other animals and humans.
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http://dx.doi.org/10.3389/fmicb.2017.00565DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374738PMC
March 2017

The presence of Giardia intestinalis in donkeys, Equus asinus, in China.

Parasit Vectors 2017 01 3;10(1). Epub 2017 Jan 3.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.

Background: Giardia intestinalis is one of the most important zoonotic enteric parasites. As no information regarding prevalence and genotype of G. intestinalis in donkeys (Equus asinus) in China is available, 181 faecal samples from 48 donkeys from Jilin Province, from 104 from Shandong Province and from 29 from Liaoning Province were examined between May and December 2015.

Findings: Twenty-eight (15.47%) out of 181 donkey samples were tested G. intestinalis-positive by nested amplification of the triosephosphate isomerase (tpi) gene. The prevalence in different regional groups varied from 10.42 to 18.27%. The prevalence in adult and young donkeys was 14.29 and 22.92%, respectively. Otherwise, the prevalence was 11.69% in summer and 18.27% in winter. However, no statistically significant differences were found in relation to region or age group. Sequence analysis of the tpi, glutamate dehydrogenase (gdh) and beta giardin (bg) loci identified 4, 1 and 3 subtypes of assemblage B, respectively. Moreover, four novel multilocus genotypes (MLGs novel-1 to novel-4) were identified in assemblage B.

Conclusions: This first report of G. intestinalis in donkeys in China indicates that further studies of nation-wide molecular epidemiology and geographical distribution of Giardia in donkeys are warranted. Effective strategies should be implemented to control G. intestinalis infection in donkeys, other animals and humans.
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http://dx.doi.org/10.1186/s13071-016-1936-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5209919PMC
January 2017

[Identification, Expression and Antigenicity Analysis of Serpin B6 of Taenia solium].

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 2016 Aug;34(4):334-8

Objective: To identify and express serpin B6 of Taenia solium (Tsserpin B6) and explore its possible use as a diagnostic antigen.

Methods: Primers for Tsserpin B6 were designed according to T. solium genome and transcriptome data. The Tsserpin B6 gene was amplified from the total RNA of T. solium cysticercus and subsequently analyzed by bioinformatics. Multiple amino acid sequence alignments of Tsserpin B6 and other parasites serpins were created using the Clustal X1.83. Phylogenetic analyses were performed using the MEGA 6.0. The recombinant expression vector pET-30a-Tsserpin B6 was constructed and expressed in E. coli strain BL21 (DE3). The expressed proteins were purified, isolated by SDS-PAGE, and analyzed by Western blotting using pig serum infected with T. solium cysticerci.

Results: The complete reading frame of Tsserpin B6 was 1 131 bp and encoded a protein of 376 amino acids. The encoded protein had a conservative reactive center loop and distinctive domains of NEEGAE and FTVDHPFLF, and harbored 9 potential linear B cell epitopes. The expressed products of Tsserpin B6 mainly existed as an inclusion body, and reacted with pig serum infected with T. solium, resulting in a specific band at the Mr 53 000.

Conclusion: The Tsserpin B6 gene was successfully cloned, and its expressed products can be recognized by pig serum infected with T. solium.
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August 2016

[Identification of Taenia solium Insulin Receptor TsIR-1316 and Expression of Its Ligand Binding Domain].

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 2016 04;34(2):93-8

Objective: To characterize the structure of insulin receptor of Taenia solium(TsIR-1316) and express its ligand binding domain (LBD).

Methods: Primers for TsIR-1316 were designed according to the genomic data of T. solium, and the TsIR-1316 gene was amplified by PCR. The nucleotide and amino acid sequences of TsIR-1316 were aligned using BLASTN and BLASTP, and the putative signal peptide and structure domains were predicted. The LBD fragment of TsIR-1316 was cloned into the pET-30a(+) vector and expressed. The expressed proteins were purified, separated by SDS-PAGE and analyzed with Western blotting using cysticercus cellulosae-positive serum and TsIR-LBD-immunized rabbit serum.

Results: The open reading frame of TsIR-1316 was 5 196 bp, encoded a protein of 1 732 amino acids which had a typical conserved domain of tyrosine kinase family, was 84% homologous with Echinococcus multilocularis, and had a “V”-shaped tertiary structure. As expected, SDS-PAGE showed that the expressed protein had a band at Mr 59 000. Western blotting showed that the recombinant protein had specific reactions with cysticercus cellulosae positive serum and TsIR-LBD immunized rabbit serum, resulting in a specific band at M(r) 59 000.

Conclusion: The TsIR-1316 gene was successfully cloned and identified. The expressed protein of TsIR-1316 LBD can be recognized by cysticercus cellulosae positive serum, which suggests a good antigenicity of this protein.
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April 2016

[Influence of Different Growing Years on Accumulation of Flavonoids and Saponins in Astragali Radix].

Zhong Yao Cai 2015 Jul;38(7):1366-9

Objective: To study the effects of different growing years on the accumulation of flavonoids and saponins in Astragali Radix, in order to offer evidence to determine the most appropriate harvest time.

Methods: HPLC method was developed for the determination of the contents of flavonoids and saponins.

Results: The contents of flavonoids and saponins in different growing years of Astragali Radix existed obvious differences, the longer growing, the content was relatively higher.

Conclusion: With the increase of growing years, the contents of flavonoids and saponins are roughly rising year by year, the harvest time has a great influence on the accumulation of the compositions.
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July 2015

[Effects of plant growth regulator uniconazole on plant morphology and biomass allocation of Salvia miltiorrhiza].

Zhongguo Zhong Yao Za Zhi 2015 May;40(10):1925-9

In this study, we use pot experiment to evaluate the effect of plant growth regulator on plant morphology and biomass allocation of Salvia miltiorrhiza. Different concentrations of uniconazole were supplied to S. miltioohiza by means of foliar spray. Height, breadth and stem diameter were measured dynamically, the biomass of leaf, stem, flower and fruit, root biomass and biomass ratio were also examined at the harvest time. Owing to the treatment, plant morphology showed significant changes, the height had been greatly reduced and the breadth decreased largely. Meanwhile, the biomass allocation changed too. The biomass ratio of leaf and stem had been notably reduced while the biomass ratio of root had been increased remarkably. It appears that foliar application of uniconazole during vigorous growth period in S. miltioohiza has dramatic effect on dwarfing plant and improving resistant to lodging. This measure could also be applied to condensed cultivation of S. miltioohiza to increase production.
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May 2015