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Weighted gene co-expression network analysis identifies potential regulators in response to Salmonella Enteritidis challenge in the reproductive tract of laying ducks
ZHANG Yu, LUO Shu-wen, HOU Li-e, GU Tian-tian, ZHU Guo-qiang, Wanwipa VONGSANGNAK, XU Qi, CHEN Guo-hong
2022, 21 (8): 2384-2398.   DOI: 10.1016/S2095-3119(21)63888-1
Abstract222)      PDF in ScienceDirect      

Salmonella Enteritidis (SE) is a zoonotic and vertically transmitted pathogen, often colonized in the reproductive tract of adult poultry, which can result in direct contamination of eggs and threaten human health.  Previous studies have revealed that some pattern recognition receptors and resistance genes were involved in regulating immune responses to SE invasion in birds.  However, the role of these immune response genes was not independent, and the interactions among the genes remained to be further investigated.  In this study, SE burden and colonization were determined in reproductive tissue after the ducks were SE-infected, and RNA-sequencing was performed to construct co-expression networks by weighted gene co-expression network analysis (WGCNA).  The result showed that SE could be isolated from 22% of infected-birds in any segment of the reproductive tract and the SE was readily colonized in the stroma, small follicle, isthmus, and vagina of the reproductive tracts in morbid ducks.  The top central, highly connected genes were subsequently identified three specific modules in the above four tissues at the defined cut-offs (P<0.01), including 60 new candidate regulators and 125 transcription factors.  Moreover, those 185 differentially expressed genes (DEGs) in these modules were co-expressed.  Moreover, the hub genes (TRAF3, CXCR4 and IL13RA1) were identified to act with many other genes through immune response pathways including NF-kappaB, Toll-like receptor, steroid biosynthesis, and p53 signaling pathways.  These data provide references that will understand the immune regulatory relationships during SE infection, but also assist in the breeding of SE-resistant lines through potential biomarkers.

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The tissue expression level of BPI gene in piglets from newborn to weaning and its relationship with Gram-negative bacterial infection
DAI Chao-hui, CAO Yue, GAO Zhong-cheng, ZHU Guo-qiang, WU Sheng-long, BAO Wen-bin
2020, 19 (12): 3065-3073.   DOI: 10.1016/S2095-3119(20)63369-X
Abstract92)      PDF in ScienceDirect      
The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria.  In this study, qPCR was used to detect the expression of the BPI gene in twelve tissues of Meishan piglets from birth to weaning.  BPI gene overexpression, bacterial adhesions count and indirect immunofluorescence were applied to analyze the relationship between BPI gene expression and the infectivity of Escherichia coli and Salmonella.  The results showed that the BPI gene was expressed highly in duodenum, jejunum and ileum (fold changes of relative expression levels were more than 10 000, 500 and 200, respectively).  The expression of the BPI gene at 35 days of age was significantly higher (P<0.01) than that at all other days.  Transcription of the BPI gene was up-regulated 2 401-fold in porcine intestinal epithelial (IPEC-J2) cells that were transfected with the BPI gene overexpression lentivirus (IPEC-J2-BPI), and significantly higher (P<0.01) than that in negative control cells (IPEC-J2-NC).  Protein expression levels in IPEC-J2-BPI cells were also increased.  When IPEC-J2 cells were incubated with E. coli and Salmonella, respectively, for 2, 4, 6, 8, 10 and 12 h, the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.05) than that in IPEC-J2-NC cells.  The results of indirect immunofluorescence analysis showed that the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.01) than that in IPEC-J2-NC cells.  These results demonstrated that the BPI gene might play an important role in regulating weaning stress especially intestinal-mediated immune response.  Overexpression of the BPI gene at the cellular level could significantly enhance the anti-bactericidal ability against Gram-negative bacteria such as E. coli and Salmonella.  This has important biological significance in piglet resistance to bacterial diarrhea. 
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