Publications by authors named "Guo-Qiang Zhu"

41 Publications

First report on the phylogenetic relationship, genetic variation of Echinococcus shiquicus isolates in Tibet Autonomous Region, China.

Parasit Vectors 2020 Nov 23;13(1):590. Epub 2020 Nov 23.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory for Animal Echinococcosis/Key Laboratory of Veterinary Parasitology of Gansu Province/Key Laboratory of Zoonoses of Agriculture Ministry, Lanzhou Veterinary Research Institute (CAAS), Lanzhou, 730046, Gansu, People's Republic of China.

Background: Cystic or alveolar echinococcosis caused by the larval stages of Echinococcus spp. is a very severe zoonotic helminth infection. Echinococcus shiquicus is a newly discovered species that has only been reported in the Qinghai and Sichuan provinces of the Qinghai-Tibet plateau, China where, to date, it has only been confirmed in Tibetan foxes and wild small mammal populations of the Tibetan plateau. Information on its genetic and evolutionary diversity is scanty. The aim of this study was to investigate the prevalence of E. shiquicus in plateau pikas (Ochotona curzoniae), a known intermediate host, and to determine the genetic variation and phylogenetic relationship of the E. shiquicus population in the Tibet region of China based on mitochondrial DNA.

Methods: Echinococcus shiquicus samples were collected from Damxung and Nyêmo counties (located in Tibet Autonomous Region, China). The mitochondrial cox1 and nad1 gene sequences were analyzed, and the genetic diversity and epidemiology of E. shiquicus in the region were discussed based on the results.

Results: The prevalence of E. shiquicus in pikas in Damxung and Nyêmo counties was 3.95% (6/152) and 6.98% (9/129), respectively. In combination with previous public sequence data, the haplotype analysis revealed 12 haplotypes (H) characterized by two distinct clusters (I and II), and a sequence distance of 99.1-99.9% from the reference haplotype (H1). The diversity and neutrality indices for the entire E. shiquicus populations were: haplotype diversity (Hd) ± standard deviation (SD) 0.862 ± 0.035; nucleotide diversity (Hd ± SD) 0.0056 ± 0.0003; Tajima's D 0.876 (P > 0.05); and Fu's F 6.000 (P > 0.05).

Conclusions: This was the first analysis of the newly discovered E. shiquicus in plateau pikas in the Tibet Autonomous Region of China. The neutrality indices suggest a deficiency of alleles, indicative of a recent population bottleneck.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13071-020-04456-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7686673PMC
November 2020

Regulation of LOX-1 on adhesion molecules and neutrophil infiltration in mouse keratitis.

Int J Ophthalmol 2020 18;13(6):870-878. Epub 2020 Jun 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To determine whether lectin-like ox-LDL receptor (LOX-1) regulates adhesion molecules expression and neutrophil infiltration in () keratitis of C57BL/6 mice.

Methods: C57BL/6 mice were pretreated with a neutralizing antibody to LOX-1 (5 µg/5 µL) or control nonspecific IgG (5 µg/5 µL), LOX-1 inhibitor Poly-I (2 µg/5 µL) or PBS by subconjunctival injection. Fungal keratitis (FK) mouse models of C57BL/6 mice were established by scraping corneal central epithelium, smearing on the corneal surface and covering the eye with contact lenses. The corneal response to infection was assessed clinical score. The mRNA levels of the adhesion molecules intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), P-selectin and E-selectin were tested in control and infected corneas by reverse transcription-polymerase chain reaction (RT-PCR). The protein levels of ICAM-1 were evaluated by immunofluorescence (IF) and Western blot. Neutrophils were extracted from the abdominal cavity of C57BL/6 mice followed by pretreatment using antibody to LOX-1 (10 µg/mL) or control nonspecific IgG (10 µg/mL), the Poly-I (4 µg/mL) or PBS. The cells were then stimulated with and tested mRNA and protein levels of lymphocyte function-associated antigen-1 (LFA-1) using RT-PCR and Western blot. IF and myeloperoxidase (MPO) assays were used to assess neutrophil infiltration in mice corneas.

Results: Pretreatment of LOX-1 antibody or the Poly-I reduced the degree of inflammation of cornea and decreased the clinical FK score compared with pretreatment of IgG or PBS (both <0.01). And these pretreatment also displayed an obvious decline in the mRNA levels of ICAM-1, VCAM-1, P-selectin, E-selectin and LFA-1 expression compared with control groups (all <0.01). Furthermore, pretreated with LOX-1 antibody or Poly-I, the protein levels of ICAM-1 and LFA-1 also decreased compared with control groups (all <0.05). Neutrophil infiltration in the cornea was significantly reduced after pretreatment of LOX-1 antibody or Poly-I compared with control groups by IF and MPO assays (both <0.01).

Conclusion: Inhibition of LOX-1 can decrease the expression of adhesion molecules and reduce neutrophil infiltration in infected corneas of C57BL/6 mice.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2020.06.03DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7270261PMC
June 2020

Expression and role of aryl hydrocarbon receptor in keratitis.

Int J Ophthalmol 2020 18;13(2):199-205. Epub 2020 Feb 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To observe the expression and role of aryl hydrocarbon receptor (AhR) in the immune response of mouse cornea infected with .

Methods: Murine models of keratitis were established by scraping the central epithelium of mouse cornea, daubing on the cornea and covering with a contact lens. The mice were randomly divided into the control group and the -infected (A.F.) group for 1, 3 and 5d respectively, which corneas were daily monitored by a slit lamp microscope and the clinical scores were also recorded timely after infection. In this study, immunofluorescence staining was used to detect the expression and localization of AhR in mouse corneas, and the mRNA and protein of AhR were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. In addition, mouse peritoneal macrophages were stimulated by with or without the pretreatment of AhR antagonist CH223191 and AhR agonist FICZ, and the tumor necrosis factor alpha (TNF-α), inducible nitric oxide synthase (iNOS), interleukin-10 (IL-10) and Arg-1 mRNA were detected by RT-PCR.

Results: According to the results of the slit light photography, it was clearly indicated that the corneal inflammation were the most severe and the clinical score became the highest as well on the 3 day after the infection of . Contrasted with the control group, the expression of AhR in the corneal epithelial cells infected with was significantly increased detected by immunofluorescence staining. AhR mainly expressed in the nucleus and cytoplasm of corneal epithelial cells. Consistent with the transcriptional level of AhR mRNA, the expression level of AhR protein reached the peak on the 3 day after infection which was detected by Western blot. Furthermore, RT-PCR showed that CH223191 up-regulated the expression of TNF-α and iNOS and down-regulated the expression of IL-10 and Arg-1 in peritoneal macrophages; inversely, FICZ reduced the expression of TNF-α and iNOS while elevated the expression of IL-10 and Arg-1.

Conclusion: AhR is involved in the pathogenesis of keratitis and induced immune protection in anti- immune response by inhibiting M1 and increasing M2 phenotype macrophage-related inflammatory factors.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2020.02.01DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013794PMC
February 2020

Changes in corneal innervation and pain responses in fungal keratitis.

Int J Ophthalmol 2020 18;13(1):1-6. Epub 2020 Jan 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 26600, Shandong Province, China.

Aim: To characterize changes in the cornea nerve and pain responses in fungal keratitis (FK).

Methods: A retrospective analysis of confocal microscopy images of 11 FK corneas was performed, and the results were compared with those for 11 normal corneas. Subbasal corneal nerves were analyzed for total nerve number, main nerve trunk number, branching patterns and tortuosity. C57BL/6 mice were infected with . Disease severity was determined through clinical scoring and slit lamp photography. Corneas were harvested at 1, 3, 5, and 7d post infection (p.i.) and assessed for β III tubulin. Corneal mechanical sensitivity thresholds were detected by von Frey test. β-endorphin (β-EP) and µ receptor protein expression was detected through Western blotting.

Results: Total nerve number, main nerve trunk number, and nerve branching were significantly lower in FK patients than in controls, but tortuosity was not significantly different. In infected mice, subbasal nerve density decreased from 1d p.i., reaching a minimum at 5d p.i. Clinical scores rose at 1d p.i., peaked at 3d p.i., and decreased at 5d p.i. Mechanical sensitivity thresholds showed the same trends. β-EP and µ receptor protein expression increased after infection.

Conclusion: Corneal nerve density is lower in FK patients and -infected mice than in controls. Pain sensitivity decreases with postinfection corneal ulcer aggravation. β-EP and µ receptor proteins are both upregulated in infected mouse corneas.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2020.01.01DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6942951PMC
January 2020

Genetic variation of Echinococcus spp. in yaks and sheep in the Tibet Autonomous Region of China based on mitochondrial DNA.

Parasit Vectors 2019 Dec 27;12(1):608. Epub 2019 Dec 27.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis, Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, CAAS, Lanzhou, 730046, Gansu, People's Republic of China.

Background: Cystic echinococcosis (CE) in humans and livestock is caused by Echinococcus granulosus (sensu lato). In China where CE is endemic, a number of studies have shown that Echinococcus granulosus (sensu stricto) is majorly responsible for CE. However, E. canadensis (G6) which is the second leading cause of CE is now being detected in most parts of the country. In this study, the species diversity and genetic variation of Echinococcus granulosus (s.l.) in four counties in Tibet Autonomous Region of China were investigated.

Methods: Infection with Echinococcus granulosus (s.s.) in yaks and sheep was identified using NADH dehydrogenase subunit 1 and 5 (nad1 and nad5) mitochondrial genes while the genotype G6 of E. canadensis initially diagnosed with NADH dehydrogenase subunit 1 (nad1) was further confirmed by analysis of the complete mitochondrial genome and a phylogenetic network constructed based on the nad2 and nad5 genes.

Results: Out of 85 hydatid cyst samples collected from slaughtered sheep (n = 54) and yaks (n = 31), 83 were identified as E. granulosus (s.s.) G1 (n = 77), G3 (n = 6) and 2 were identified as E. canadensis G6. Analysis of the nad1/nad5 genes revealed 16/17 mutations with 9/14 parsimony informative sites resulting in 15/14 haplotypes, respectively. Haplotype diversity (Hd) and nucleotide diversity (π) of E. granulosus (s.s.) population were 0.650 and 0.00127 for nad1 and 0.782 and 0.00306 for nad5, respectively, with an overall negative Tajima's D and Fu's Fs. A low F indicated no genetic difference between isolates from sheep and yaks.

Conclusion: Pockets of infection with E. canadensis (G6, G7, G8 and G10) have been previously reported in sheep, goats, yaks and/or humans in different parts of China. While the G6 genotype has been previously reported in sheep in the Tibet Autonomous Region, the detection in a yak in the present study represents the first to the best of our knowledge. Therefore, we recommend future surveys and control efforts to comprehensively investigate other potential intermediate hosts for the prevalence and genetic diversity of the E. canadensis group (G6, G7, G8 and G10) across the country and their inclusion into the existing CE control programme.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13071-019-3857-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6935104PMC
December 2019

Correction to: First molecular description, phylogeny and genetic variation of Taenia hydatigena from Nigerian sheep and goats based on three mitochondrial genes.

Parasit Vectors 2019 11 21;12(1):547. Epub 2019 Nov 21.

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

Following publication of the original article [1], the have authors flagged that the information in the legend of Fig. 1 is detailed in the wrong order.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13071-019-3807-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6873671PMC
November 2019

First molecular description, phylogeny and genetic variation of Taenia hydatigena from Nigerian sheep and goats based on three mitochondrial genes.

Parasit Vectors 2019 Nov 5;12(1):520. Epub 2019 Nov 5.

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

Background: Cysticercosis caused by the metacestode larval stage of Taenia hydatigena is a disease of veterinary and economic importance. A considerable level of genetic variation among isolates of different intermediate hosts and locations has been documented. Generally, data on the genetic population structure of T. hydatigena is scanty and lacking in Nigeria. Meanwhile, similar findings in other cestodes like Echinococcus spp. have been found to be of epidemiological importance. Our aim, therefore, was to characterize and compare the genetic diversity of T. hydatigena population in Nigeria based on three mitochondrial DNA markers as well as to assess the phylogenetic relationship with populations from other geographical regions.

Methods: In the present study, we described the genetic variation and diversity of T. hydatigena isolates from Nigerian sheep and goats using three full-length mitochondrial genes: the cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 1 (nad1), and NADH dehydrogenase subunit 5 (nad5).

Results: The median-joining network of concatenated cox1-nad1-nad5 sequences indicated that T. hydatigena metacestodes of sheep origin were genetically distinct from those obtained in goats and this was supported by high F values of nad1, cox1, and concatenated cox1-nad1-nad5 sequences. Genetic variation was also found to be higher in isolates from goats than from sheep.

Conclusions: To the best of our knowledge, the present study described the genetic variation of T. hydatigena population for the first time in Nigeria using full-length mitochondrial genes and suggests the existence of host-specific variants. The population indices of the different DNA markers suggest that analysis of long mitochondrial DNA fragments may provide more information on the molecular ecology of T. hydatigena. We recommend that future studies employ long mitochondrial DNA sequence in order to provide reliable data that would explain the extent of genetic variation in different hosts/locations and the biological and epidemiological significance.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13071-019-3780-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6833231PMC
November 2019

A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections.

BMC Infect Dis 2019 Oct 16;19(1):854. Epub 2019 Oct 16.

State Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAAS, Lanzhou, 730046, Gansu Province, People's Republic of China.

Background: Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction.

Methods: Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay.

Results: A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously.

Conclusions: This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s12879-019-4512-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6796438PMC
October 2019

Review of Cystic Echinococcosis in Nigeria: A Story of Neglect.

Acta Parasitol 2020 Mar 24;65(1):1-10. Epub 2019 Sep 24.

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.

Purpose: Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu lato is a widespread zoonotic disease of global concern. In Nigeria, the exact picture/status of CE is unclear, as most of the states are largely uninvestigated. Yet, as with every parasitic zoonosis, the first step towards planning a comprehensive management and control programme involves assessment of available national/regional prevalence data, host range, and risk factors at play in the transmission dynamics.

Methods: Published articles on echinococcosis were searched on PubMed and Africa Journal Online (AJOL) databases. Inclusion criteria were based on studies reporting prevalence of echinococcosis in animals and humans (including case reports) from 1970 to 2018.

Results: In this study, we evaluated and summarized cystic echinococcosis reports in Nigeria and found that post 1970-80s, studies on cystic echinococcosis have remained sparse regardless of the high prevalence recorded in the early years of CE investigation. In addition, information on the genetic population structure and the role of wildlife in CE transmission is still lacking.

Conclusions: This study appraises the prevalence and distribution of CE in Nigeria and identified areas where surveillance and control efforts should be focused and intensified.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2478/s11686-019-00124-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7223537PMC
March 2020

Cystic echinococcosis in Nigeria: first insight into the genotypes of Echinococcus granulosus in animals.

Parasit Vectors 2019 Aug 7;12(1):392. Epub 2019 Aug 7.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, 730046, Gansu, P. R. China.

Background: Cystic echinococcosis (CE) is a zoonosis caused by cestodes of Echinococcus granulosus (sensu lato) complex. In Nigeria, reports on the prevalence of CE, although limited, have been found to vary with location and host with higher prevalence and fertility rate observed in camels than other livestock. Until now, information regarding the molecular characteristics, genetic population structure, and genotypes of Echinococcus is lacking. Therefore, this study was aimed at addressing these gaps in knowledge.

Methods: We describe the genetic status of 31 Echinococcus isolates collected from slaughtered livestock (camels, cattle and goats) based on the full-length mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes.

Results: The resulting nucleotide sequences via the NCBI BLAST algorithm and Bayesian phylogeny of cox1 and cox1-nad1 genes using MrBayes v.3.1.2 showed that all isolates were clearly E. canadensis (G6/G7) and were 99-100% identical to previously reported G6/G7 haplotypes across Europe, Asia, North and East Africa.

Conclusions: Although, the G1 genotype is believed to be responsible for the majority of global CE burden, reports from a number of West African countries including Nigeria suggest that E. canadensis G6/G7 genotype could be the major causative agent of CE in the subregion. This study provides for the first time insight into the genetic population structure of Echinococcus species as well as implications for CE control in Nigeria.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13071-019-3644-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6686243PMC
August 2019

Inhibition of LOX-1 alleviates the proinflammatory effects of high-mobility group box 1 in keratitis.

Int J Ophthalmol 2019 18;12(6):898-903. Epub 2019 Jun 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To investigate the inflammatory amplification effect of high-mobility group box 1 (HMGB1) in () keratitis and the relationship between lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) and HMGB1 in keratitis immune responses.

Methods: Phosphate buffer saline (PBS), and Boxb were injected into BALB/c mice subconjunctivally before the corneas were infected with RAW264.7 macrophages and neutrophils were pretreated with PBS and Boxb to determine the HMGB1 inflammatory amplification effects. Abdominal cavity extracted macrophages were pretreated with Boxb and Poly (I) (a LOX-1 inhibitor) before hyphae stimulation to prove the the relationship between the two molecules. LOX-1, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), macrophage inflammatory protein-2 (MIP-2) and IL-10 were assessed by polymerase chain reaction and Western blot.

Results: Pretreatment with Boxb exacerbated corneal inflammation. In macrophages and neutrophils, induced LOX-1, IL-1β, TNF-α and MIP-2 expression in Boxb group was higher than those in PBS group. Poly (I) treatments before infection alleviated the proinflammatory effects of Boxb in abdominal cavity extracted macrophages. Pretreatment with Boxb did not influence Dectin-1 mRNA levels in macrophages and neutrophils.

Conclusion: In fungal keratitis, HMGB1 is a proinflammatory factor in the first line of immune response. HMGB1 mainly stimulates neutrophils and macrophages to produce inflammatory cytokines and chemokines during the immune response. LOX-1 participates in HMGB1 induced inflammatory exacerbation in keratitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2019.06.03DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6580210PMC
June 2019

Disparate expression of autophagy in corneas of C57BL/6 mice and BALB/c mice after infection.

Int J Ophthalmol 2019 18;12(5):705-710. Epub 2019 May 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To determine the disparate expression of autophagy in the () keratitis between susceptible C57BL/6 mice and resistant BALB/c mice.

Methods: C57BL/6 and BALB/c mice were used to establish fungal keratitis models. Disease severity and inflammatory response were observed by slit lamp microscopy in -infected corneas of C57BL/6 and BALB/c mice at 1, 3 and 5d. Hematoxylin-eosin (H&E) staining was used to detect pathological changes of corneas. The expression of autophagy-related proteins Beclin-1, LC3, SQSTM1/p62, and LAMP-1 was assessed by Western blot in C57BL/6 and BALB/c mice at 1, 3 and 5d post infection (p.i.). Immunofluorescent staining was used to test the expression of LC3 in corneas after infection.

Results: Keratitis severity was higher in C57BL/6 mice versus BALB/c mice at 1, 3 and 5d p.i. H&E staining showed that the number of inflammatory cells was larger and the severity of ulcer was higher in C57BL/6 mice than in BALB/c mice after stimulation with Higher expression of LAMP-1, Beclin-1, and LC3 was shown in C57BL/6 mice corneas than in BALB/c mice corneas at 1, 3 and 5d p.i., while the expression of p62 was lower in C57BL/6 mice. The fluorescence of LC3 was significantly increased in corneas of C57BL/6 mice compared with BALB/c mice after infection.

Conclusion: The expression of autophagy is higher in corneas of C57BL/6 mice than in BALB/c mice after infection. Autophagy may be positively correlated with keratitis severity and pathological changes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2019.05.02DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6520259PMC
May 2019

Role of the IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to infection.

Int J Ophthalmol 2019 18;12(4):549-556. Epub 2019 Apr 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To investigate the expression of interleukin (IL)-33 in the cornea and human corneal epithelial cells (HCECs) exposed to (), and to determine the function of IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to infection.

Methods: The mRNA and protein expression of IL-33 in HCECs and mice corneas were examined by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analysis, respectively. IL-33 expression was also detected in cornea samples from healthy donors and patients with fungal keratitis with immunohistochemistry. The cultured HCECs were treated with inactive hyphae at various concentrations with or without recombinant human IL-33 protein, soluble recombinant ST2 protein, specific ST2 neutralizing antibody, or the mitogen-activated protein kinase (MAPK) p38 inhibitor SB203580 for evaluation of the expression and activation of IL-33/ST2/p38 signaling in the regulation of proinflammatory cytokines. The production levels of IL-6 and IL-1β were determined by qRT-PCR and enzyme-linked immunosorbent assay (ELISA). The proliferation of HCECs was determined by a Cell Counting Kit-8 (CCK8) assay and cell count.

Results: IL-33 expression levels increased in the corneal tissues of patients with fungal keratitis and in mice corneas of experimental infection, as well as in HCECs with infection of strongly stimulated HCECs-generated proinflammatory cytokine (IL-6 and IL-1β) production at both the mRNA and protein levels. This production of pro-inflammatory mediators stimulated by was further stimulated by IL-33 and was prevented by soluble ST2 protein or ST2 neutralizing antibody. Moreover, IL-33 naturally promoted the p38 phosphorylation induced by , which was suppressed by soluble ST2 protein. The MAPK p38 inhibitor SB203580 also inhibited the -induced proinflammatory cytokine production. IL-33 administration for 48h and 72h promoted the proliferation of HCECs, which was attenuated by treatment with soluble recombinant human ST2 protein.

Conclusion: elevates IL-33 expression in human and mice corneas and HCECs. Thus, IL-33/ST2/p38 signaling may play an important role in amplifying the immune response of corneal epithelial cells to infection. Besides, IL-33 promotes the cell proliferation of HCECs its receptor ST2. These findings suggest a novel autocrine mechanism of amplification of the fungal-induced inflammatory response in the corneal epithelium, highlighting a potential therapeutic target for fungal keratitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2019.04.04DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6469567PMC
April 2019

Comparative transcriptome analysis reveals PERP upregulated during Salmonella Enteritidis challenge in laying ducks.

J Cell Physiol 2019 07 27;234(7):11330-11347. Epub 2018 Nov 27.

Joint International Research Laboratory of Agriculture & Agri-Product Safety of Ministry of Education, Yangzhou University, Yangzhou, China.

Salmonella Enteritidis (SE) can be transmitted to eggs through cecum or the ovary from infected layers and causes food poisoning in humans. The mechanism of cecal transmission has been extensively studied. However, the mechanism and route of transovarian transmission of SE remain unclear. In this study, the ducks were orally inoculated with SE, and the ovarian follicles and stroma were collected to detect SE infection. The immune responses were triggered and the innate and adaptive immune genes (TLR4, NOD1, AvβD7, and IL-1β) were upregulated significantly during the SE challenge. Moreover, the ovary tissues (small follicle and stroma) of susceptible and resistant-laying ducks were performed by RNA sequencing. We obtained and identified 23 differentially expressed genes (DEGs) between susceptible and resistant-laying ducks in both small follicle and stroma tissues ( p < 0.05). The DEGs were predominately identified in the p53 signaling pathway. The expression of key genes (p53, MDM2, PERP, caspase-3, and Bcl-2) involved in the signaling pathway was significantly higher in granulosa cells (dGCs) from SE-infected ducks than those from uninfected ducks. Moreover, the overexpression of PERP resulted in further induction of p53, MDM2, caspase-3, and Bcl-2 during SE infection in dGCs. Whereas, an opposite trend was observed with the knockdown of PERP. Besides, it is further revealed that the PERP could enhance cell apoptosis, SE adhesion, and SE invasion in SE-infected dGCs overexpression. Altogether, our results demonstrate the duck PERP involved in the ovarian local immune niche through p53 signaling pathway in dGCs challenged with SE.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jcp.27790DOI Listing
July 2019

Regulation of lipoxygenase-1 and Dectin-1 on interleukin-10 in mouse keratitis.

Int J Ophthalmol 2018 18;11(6):905-909. Epub 2018 Jun 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To investigate the regulation of lipoxygenase (LOX)-1 and Dectin-1 on interleukin-10 (IL-10) production in mice with () keratitis.

Methods: The corneas of C57BL/6 mice were pretreated with LOX-1 inhibitor Poly(I) or Dectin-1 siRNA separately before the infection of . Polymerase chain reaction (PCR) and Western blot were used to detect the expression of IL-10.

Results: The mRNA and protein expressions of IL-10 were significantly increased in mice with keratitis. Compared with the group pretreated with sterile water before infection, Poly(I) pretreatment suppressed IL-10 expression significantly. Compared with the group pretreated with scrambled siRNA before infection, Dectin-1 siRNA pretreatment significantly reduced IL-10 expression in response to infection.

Conclusion: LOX-1 and Dectin-1 regulate IL-10 production in mouse keratitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2018.06.02DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6010385PMC
June 2018

Mincle in the innate immune response of mice fungal keratitis.

Int J Ophthalmol 2018 18;11(4):539-547. Epub 2018 Apr 18.

Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China.

Aim: To investigate how macrophage inducible C-type lectin (Mincle) influences inflammation in mice fungal keratitis induced by ().

Methods: C57BL/6 mice were infected with after pretreated with Mincle agonist TDB or Mincle neutralizing antibody (MincleAb), taking DMSO or IgG as control group respectively. The cornea lesions were monitored with slit-lamp microscope and evaluated by clinical score. Mincle expression was assessed using reverse transcription-ploymerase chain reaction (RT-PCR) and immunostaining. The expression of cytokines (IL-1β, TNF-α and IL-6) chemokines (CXCL-1 and MIP-2) was determined by RT-PCR and ELISA. Neutrophil infiltration was observed by immunostaining. The levels of nitric oxide (NO) generated by corneas were tested by Griess reaction.

Results: Mincle mRNA and protein levels were higher in infected corneas than normal corneas of C57BL/6 mice, saving clinical scores revealed differences. When pretreated with Mincle agonist TDB, the mRNA and protein levels of IL-1β, TNF-α and IL-6 in infected corneas were significantly increased compared with the control group (<0.01). Results of the counterpart in corneas pretreated with Mincle neutralizing antibody was decreased consistently (<0.01). Expression of CXCL1 and MIP-2 mRNA levels were up-regulated in TDB group and down-regulated in MincleAb group (<0.01), coincide with neutrophil aggregation degree in corneas showed by immunostaining. As for the concentration of NO, it was promoted in TDB group compared with DMSO control group, and decreased in MincleAb group compared with IgG control group.

Conclusion: Mincle plays a dual role in mice fungal keratitis. It participates in the innate immune system by enhancing inflammation. What's more, Mincle can mediate cytotoxic effects by regulating the formation of NO.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.18240/ijo.2018.04.01DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5902354PMC
April 2018

Insight into the molecular mechanism of miR-192 regulating resistance in piglets.

Biosci Rep 2018 02 21;38(1). Epub 2018 Feb 21.

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

MicroRNAs (miRNAs) have important roles in many cellular processes, including cell proliferation, growth and development, and disease control. Previous study demonstrated that the expression of two highly homologous miRNAs (miR-192 and miR-215) was up-regulated in weaned piglets with F18 infection. However, the potential molecular mechanism of miR-192 in regulating infection remains unclear in pigs. In the present study, we analyzed the relationship between level of miR-192 and degree of resistance using transcription activator-like effector nuclease (TALEN), bacterial adhesion assays, and target genes research. A TALEN expression vector that specifically recognizes the pig miR-192 was constructed and then monoclonal epithelial cells defective in miR-192 were established. We found that miR-192 knockout led to enhance the adhesion ability of the strains F18ab, F18ac and K88ac, meanwhile increase the expression of target genes ( and ) by qPCR and Western blotting analysis. The results suggested that miR-192 and its key target genes ( and ) could have a key role in infection. Based on our findings, we propose that further investigation of miR-192 function is likely to lead to insights into the molecular mechanisms of infection.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1042/BSR20171160DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5821941PMC
February 2018

ROS-Mediated Cell Cycle Arrest and Apoptosis Induced by Zearalenone in Mouse Sertoli Cells via ER Stress and the ATP/AMPK Pathway.

Toxins (Basel) 2018 01 1;10(1). Epub 2018 Jan 1.

College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.

Zearalenone (ZEA) can perturb the differentiation of cells, reduce the generation of reproductive cells and induce a death of germ cells, but the molecular mechanism remains unclear. In order to investigate the potential mechanism of ZEA-induced cell cycle arrest and apoptosis, we studied the effects of ZEA on cell proliferation, cell-cycle distribution, cell-cycle-related proteins, cell death, cell apoptosis, ROS generation and the ATP/AMPK pathway in Sertoli cells. The role of ROS, ER stress and the ATP/AMPK pathway in ZEA-induced cell-cycle arrest and cell apoptosis was explored by using the antioxidant NAC, ER stress inhibitor 4-PBA and the AMPK inhibitor dorsomorphin, respectively. The results revealed that ZEA inhibited the cell proliferation, influenced the distribution of the cell cycle and induced cell apoptosis through the ATP/AMPK pathway. The ATP/AMPK pathway was regulated by ER stress that was induced by ROS generation after exposure to ZEA. Taking these together, this study provided evidence that ROS regulated the process of ZEA-induced cell cycle arrest and cell apoptosis through ER stress and the ATP/AMPK signal ways.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/toxins10010024DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793111PMC
January 2018

New insights into the codon usage patterns of the bactericidal/permeability-increasing (BPI) gene across nine species.

Gene 2017 Jun 21;616:45-51. Epub 2017 Mar 21.

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, PR China; Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, Yangzhou, Jiangsu, PR China. Electronic address:

Bactericidal/permeability-increasing (BPI) protein is a member of a new generation of proteins known as super-antibiotics that are implicated as endotoxin neutralising agents. Non-uniform usage of synonymous codons for a specific amino acid during translation of a protein is known as codon usage bias (CUB). Analysis of CUB and compositional dynamics of coding sequences could contribute to a better understanding of the molecular mechanism and the evolution of a particular gene. In this study, we performed CUB analysis of the complete coding sequences of the BPI gene from nine different species. The codon usage patterns of BPI across different species were found to be influenced by GC bias, particularly GC3s, with a moderate bias in the codon usage of BPI. We found significant similarities in the codon usage patterns in BPI gene among closely related species, such as Sus_scrofa and Bos_taurus. Moreover, we observed evolutionary conservation of the most over-represented codon CUG for the amino acid leucine in the BPI gene across all species. In conclusion, our analysis provides a novel insight into the codon usage patterns of BPI. This information facilitates an improved understanding of the structural, functional and evolutionary significance of BPI gene among species, and provides a theoretical reference for developing antiseptic drug proteins with high efficiency across species.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.gene.2017.03.016DOI Listing
June 2017

Direct Involvement of the Master Nitrogen Metabolism Regulator GlnR in Antibiotic Biosynthesis in Streptomyces.

J Biol Chem 2016 Dec 8;291(51):26443-26454. Epub 2016 Nov 8.

From the Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032,

GlnR, an OmpR-like orphan two-component system response regulator, is a master regulator of nitrogen metabolism in the genus Streptomyces In this work, evidence that GlnR is also directly involved in the regulation of antibiotic biosynthesis is provided. In the model strain Streptomyces coelicolor M145, an in-frame deletion of glnR resulted in markedly increased actinorhodin (ACT) production but reduced undecylprodigiosin (RED) biosynthesis when exposed to R2YE culture medium. Transcriptional analysis coupled with DNA binding studies revealed that GlnR represses ACT but activates RED production directly via the pathway-specific activator genes actII-ORF4 and redZ, respectively. The precise GlnR-binding sites upstream of these two target genes were defined. In addition, the direct involvement of GlnR in antibiotic biosynthesis was further identified in Streptomyces avermitilis, which produces the important anthelmintic agent avermectin. We found that S. avermitilis GlnR (GlnRsav) could stimulate avermectin but repress oligomycin production directly through the respective pathway-specific activator genes, aveR and olmRI/RII To the best of our knowledge, this report describes the first experimental evidence demonstrating that GlnR regulates antibiotic biosynthesis directly through pathway-specific regulators in Streptomyces Our results suggest that GlnR-mediated regulation of antibiotic biosynthesis is likely to be universal in streptomycetes. These findings also indicate that GlnR is not only a master nitrogen regulator but also an important controller of secondary metabolism, which may help to balance nitrogen metabolism and antibiotic biosynthesis in streptomycetes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1074/jbc.M116.762476DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5159505PMC
December 2016

CD14 in the TLRs signaling pathway is associated with the resistance to E. coli F18 in Chinese domestic weaned piglets.

Sci Rep 2016 Apr 21;6:24611. Epub 2016 Apr 21.

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P. R. China.

Escherichia coli F18 (E. coli F18) is mainly responsible for post-weaning diarrhea (PWD) in piglets. The genetic basis and regulatory mechanism of E. coli F18 resistance in Chinese domestic weaned piglets remain unclear. Meishan piglets were used as model animals to test their susceptibility to E. coli F18. By performing a comparative transcriptome study on duodenum tissues of sensitive and resistant pigs, we identified 198 differentially expressed genes (DEGs; 125 upregulated and 73 downregulated) in the resistant pigs. DEGs were predominately involved in immune system pathways, including the Toll-like receptor (TLR) signaling pathway. qPCR and western blot showed CD14, IFN-α, TLR4 and IL-1β, etc. in the TLR signaling pathway had significantly higher expression levels in lipopolysaccharide (LPS)-induced small intestinal epithelial cell lines (IPEC-J2) than those in normal IPEC-J2 cells. Immunohistochemical analysis showed the increased expression of CD14 gene in the E. coli F18-resistant individuals. After CD14 knockdown, the levels of cytokines IL-6 and IL-12 were significantly reduced in IPEC-J2 cell supernatants. The adhesion ability of F18ab strain with IPEC-J2 cells was significantly increased (p < 0.01). This study revealed the TLR signaling pathway, and especially CD14, probably plays an important role in resistance to E. coli F18 infection in Chinese domestic piglets.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/srep24611DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4838916PMC
April 2016

[AFLP analysis on genetic diversity of Haloxylon ammodendron in China].

Zhongguo Zhong Yao Za Zhi 2014 Mar;39(6):959-64

To determine the genetic diversity of Haloxylon ammodendron collected from 14 sites in 5 provinces, 103 H. ammodendron samples of 12 wild populations and 2 cultivated which collected from 14 sites in 5 provinces were analyzed by amplified fragment length polymorphism (AFLP) DNA markers. PopGen32 and NTSYSpc2.1 was applied to evaluate genetic diversity of H. ammodendron populations. The average percentage of polymorphic loci (PPL) of total H. ammodendron populations was 94.13%, the average Nei's gene diversity index (H(e)) from 14 populations was 0.308 0, and the Shannon's genetic diversity index (I) was 0.467 6. The results indicated that the genetic diversity of H. ammodendron populations was high. Genetic differentiation index (G(st)) was 0.313 8, and the gene flow (N(m)) was 1.093 5 at the population level. The level of gene flow of H. ammodendron showed it possessed the feature of wind-pollinated outcrossing plants. AMOVA analysis indicated that genetic variation of H. ammodendron was much higher within groups (89.34%) than that among groups (10.66%), moreover genetic variation within groups mainly occurred among populations in different producing areas (84.80%). Cluster analysis (UPGMA) was applied to generate dendrogram based on Nei's genetic distances of 14 populations. Samples from Xinjiang and Qinghai were clustered respectively as a clade for their distant genetic relationship, while Samples from Gansu, Inner Mongolia and Ningxia were clustered together for their close genetic relationship. Genetic diversity of H. ammodendron populations is high in China, and genetic differentiation among regions is small, thus abundance within this specie is high at this stage. Therefore, wild nursery and artificial cultivating in different areas are effective measures for the conservation and sustainable utilization of H. ammodendron resources.
View Article and Find Full Text PDF

Download full-text PDF

Source
March 2014

Simple and rapid detection of Salmonella by direct PCR amplification of gene fimW.

Curr Microbiol 2014 Oct 17;69(4):429-35. Epub 2014 May 17.

College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China.

This study established a simple method of specifically detecting Salmonella species by amplifying fimW gene, which was involved in regulating Salmonella type I fimbriae expression. A pair of primers was designed to target and discriminate the 68 Salmonella strains of 23 Salmonella serovars available to us from 12 non-Salmonella strains of five different kinds of bacteria by polymerase chain reaction (PCR) amplification. Results showed that specific DNA fragment with an expected size of 477 bp was successfully amplified from all Salmonella serovars, while no target band was detected in non-Salmonella species. The sensitivity of this PCR-amplifying system reached to 1 pg DNA chromosome and 10(2) cfu of Salmonella enteritis strain CMCC(B) 50336. The above results demonstrated the method as a simple, sensitive, and specific way for Salmonella detection.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00284-014-0602-zDOI Listing
October 2014

Ray-tracing method for creeping waves on arbitrarily shaped nonuniform rational B-splines surfaces.

J Opt Soc Am A Opt Image Sci Vis 2013 Apr;30(4):663-70

School of Electronic Information, Wuhan University, Wuhan 430072, China.

An accurate creeping ray-tracing algorithm is presented in this paper to determine the tracks of creeping waves (or creeping rays) on arbitrarily shaped free-form parametric surfaces [nonuniform rational B-splines (NURBS) surfaces]. The main challenge in calculating the surface diffracted fields on NURBS surfaces is due to the difficulty in determining the geodesic paths along which the creeping rays propagate. On one single parametric surface patch, the geodesic paths need to be computed by solving the geodesic equations numerically. Furthermore, realistic objects are generally modeled as the union of several connected NURBS patches. Due to the discontinuity of the parameter between the patches, it is more complicated to compute geodesic paths on several connected patches than on one single patch. Thus, a creeping ray-tracing algorithm is presented in this paper to compute the geodesic paths of creeping rays on the complex objects that are modeled as the combination of several NURBS surface patches. In the algorithm, the creeping ray tracing on each surface patch is performed by solving the geodesic equations with a Runge-Kutta method. When the creeping ray propagates from one patch to another, a transition method is developed to handle the transition of the creeping ray tracing across the border between the patches. This creeping ray-tracing algorithm can meet practical requirements because it can be applied to the objects with complex shapes. The algorithm can also extend the applicability of NURBS for electromagnetic and optical applications. The validity and usefulness of the algorithm can be verified from the numerical results.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1364/JOSAA.30.000663DOI Listing
April 2013

The molecular adjuvant mC3d enhances the immunogenicity of FimA from type I fimbriae of Salmonella enterica serovar Enteritidis.

J Microbiol Immunol Infect 2014 Feb 24;47(1):57-62. Epub 2013 Jan 24.

College of Veterinary Medicine, Yangzhou University, Yangzhou, China. Electronic address:

Background: The fimbriae of Salmonella enterica serovar Enteritidis are used for colonization and invasion into host cells, and have drawn considerable interest because fimbriae can serve as potential immunogens against many pathogenic bacteria that colonize on epithelial surfaces. The purpose of the study is to use a molecular adjuvant, C3d, to enhance the immunogenicity of FimA proteins against Salmonella enterica serovar Enteritidis.

Methods: FimA of type I fimbriae from Salmonella enteritidis and FimA with one copy of mC3d, two copies of mC3d2 and three copies of mC3d3 were cloned into the expression vector pCold-TF. Soluble fusion proteins of FimA with different copy of mC3d were induced by IPTG and expressed into Escherichia coli BL21 (DE3). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the recombinant proteins from pCold-TF-fimA, TF-fimA-mC3d, TF-fimA-mC3d2, TF-fimA-mC3d3 were 70 kDa, 100 kDa, 130 kDa and 160 kDa, respectively. The fusion protein was recognized by rabbit anti-fimbriae polyclonal antibodies, and then visualized by goat anti-rabbit polyclonal antibodies with a chrome appearance by enzyme-subtract interaction. The recombinant proteins were purified by Ni-TED (tris-carboxymethyl ethylene diamine), immobilized metal ion affinity chromatography (IMAC). Balb/c mice were subcutaneously immunized with the purified proteins and the immune response was monitored by an enzyme-linked immunosorbent assay (ELISA) for FimA-specific antibody. The immunized mice were challenged with a 10-fold LD50 dose (i.e., 100 CFU) of Salmonella enterica serovar Enteritidis standard strain (SD-2) 1 week after the second immunization.

Results: The immunized mice with the fusion proteins FimA-mC3d2 and FimA-mC3d3 had increased levels of ELISA titer of antibody that were 2 and 4 logs, respectively, more immunogenic than the TF-FimA protein alone. The challenge results showed that immune protection rate in the mice immunized with 10 μg of FimA, FimA-mC3d2, and FimA-mC3d3 were 50%, 75% and 100%, respectively.

Conclusion: We conclude that mC3d can be expressed in a prokaryotic vector and enhance the immune response of the recombinant protein. FimA-mC3d3 is potentially a subunit vaccine against S. enterica serovar Enteritidis infection.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jmii.2012.11.004DOI Listing
February 2014

Identification and genetic characterization of new bovine viral diarrhea virus genotype 2 strains in pigs isolated in China.

Virus Genes 2013 Feb 21;46(1):81-7. Epub 2012 Oct 21.

College of Veterinary Medicine, Yangzhou University, Yangzhou, China.

Classical swine fever (CSF)-like symptoms in pigs regarded as free from CSF has been reported previously. From sick pigs with CSF-like symptoms, and who had been inoculated with the hog cholera vaccine, samples were collected and subjected to RT-PCR using specific primers. Twelve bovine viral diarrhea virus 2 (BVDV-2) strains were screened and isolated. Homology comparison showed that the E2 genes of the twelve isolates were highly conserved. The genome of the one of the BVDV-2 isolates (named as SH-28) from the sick pigs, which showed a noncytopathic effect in MDBK cell cultures and strong reactivity with monoclonal antibody (MAb) Bz-53 raised against BVDV-2, was sequenced. The genome of SH-28 comprises 12,279 nucleotides and contains a large open reading frame beginning at nucleotide 386 and ending at nucleotide 12,073. Genomic comparison and phylogenetic analyzes showed that SH-28 fall into BVDV-2 subtype and was most similar to XJ-04 (nucleotide and amino acid homologies were 89.9-93.8 % and 91.1-96.9 %, respectively), but was genetically divergent from ZM-95 (pig BVDV-1).
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s11262-012-0837-3DOI Listing
February 2013

Study on the age-dependent tissue expression of FUT1 gene in porcine and its relationship to E. coli F18 receptor.

Gene 2012 Apr 28;497(2):336-9. Epub 2012 Jan 28.

College of Animal Science and Technology, Yangzhou University, Yangzhou, China.

Escherichia coli (E. coli) that produces adhesin F18 is the main pathogen responsible for porcine post-weaning diarrhea and edema disease. The receptor for E. coli F18 has not been described in pigs, however the alpha (1,2)-fucosyltransferase (FUT1) gene on chromosome 6 has been proposed as a candidate. The objective of this study, therefore, was to investigate the relationship between FUT1 gene expression and E. coli F18 receptor in Sutai pigs of different ages (8-, 18-, 30- and 35-day-old). FUT1 gene expression was detected in 11 pig tissues with the highest level in lung, and expressed consistently at the four time points. In most tissues, FUT1 gene expression levels decreased from days 8 to 18, then continually increased on days 30 and 35, with expression around weaning time higher than that on day 8. Gene ontology and pathway analysis showed that FUT1 was involved in 32 biological processes, mainly those integral to the membrane, or involved in glycosylation, as well as regulation of binding, interestingly participating in three pathways related to glycosphingolipid biosynthesis. From this analysis and the high linkage disequilibrium between the FUT1 gene and the E. coli F18 receptor locus, we can speculate that higher expression of the FUT1 gene in small intestine is beneficial to the formation of receptors to the E. coli F18 strain and is related to the sensitivity to the pathogen.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.gene.2012.01.035DOI Listing
April 2012

Relationship between the expression level of SLA-DQA and Escherichia coli F18 infection in piglets.

Gene 2012 Feb 13;494(1):140-4. Epub 2011 Dec 13.

College of Animal Science and Technology, Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, Yangzhou University, Yangzhou, China.

The expression of SLA-DQA was assayed by Real-time PCR to analyze the differential expression between ETEC F18-resistant and -sensitive post-weaning piglets, and then to compare the expression levels of SLA-DQA in 11 different tissues from 8-, 18-, 30- and 35-day-old ETEC F18-resistant piglets, which aimed at discussing the role of SLA-DQA in resistance to ETEC F18. The results showed that SLA-DQA is broadly expressed in 11 tissues with the highest expression level in lymph nodes, and a relatively higher expression level in lung, spleen, jejunum, and duodenum. In tissues of lymph node, lung, spleen, jejunum, and duodenum, the mRNA expression of SLA-DQA in resistant individuals was significantly higher than that in sensitive ones (P<0.05). In most tissues, the expression of SLA-DQA increased from 8 to 18 and 30 days (weaning day), and increased persistently to 35 days of post-weaning. Expression levels of SLA-DQA on 35 days in most tissues were significant higher than that on 8, 18 and 30 days (P<0.05). The results demonstrated that the resistance to ETEC F18 in post-weaning piglets is related to up-regulation of mRNA expression of SLA-DQA to a certain extent. The analysis suggested that SLA-DQA may be not the direct immune factor that resisted the Escherichia coli F18, but perhaps enhanced humoral immunity and cell immunity to reduce the transmembrane signal transduction of ETEC F18 bacterial LPS and then led to the resistance to ETEC F18 in piglets.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.gene.2011.11.030DOI Listing
February 2012

Beneficial genotype of swine FUT1 gene governing resistance to E. coli F18 is associated with important economic traits.

J Genet 2011 Aug;90(2):315-8

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225009, People's Republic of China.

View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s12041-011-0059-9DOI Listing
August 2011

Evaluation of M307 of FUT1 gene as a genetic marker for disease resistance breeding of Sutai pigs.

Mol Biol Rep 2012 Apr 19;39(4):4223-8. Epub 2011 Jul 19.

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, China.

Alpha (1,2) fucosyltransferase (FUT1) gene has been identified as a candidate gene for controlling the expression of the receptor for ETEC F18. The genetic variations in the position of M307 nucleotide in open reading frame of FUT1 have been proposed as a marker for selecting ETEC F18 resistant pigs. The polymorphisms of M307 in FUT1 of breeding base group for ETEC F18 resistance of Sutai pigs (Duroc × Meishan) was detected and their correlations to some immune indexes, growth and development ability, carcass traits and meat quality were also analyzed, which aimed to investigate feasibility of further breeding for diseases resistance based on M307 of FUT1 for Sutai pigs. After digested by Hin6 I, M307 of FUT1 gene could be divided into three kinds of genotypes, AA, AG, and GG. The frequencies were 0.235, 0.609, and 0.156, respectively. The results indicated that Sutai pigs with the AA genotype in M307 of FUT1 gene not only have relatively strong general disease resistance ability in piglets, but also have higher growth and development ability and stable carcass traits and meat quality. It is entirely feasible to raise the new strains of Sutai pigs resistant to Escherichia coli F18 based on genetic marker of the M307 position in FUT1gene.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s11033-011-1208-1DOI Listing
April 2012