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Giardia duodenalis, also known as G. intestinalis or G. lamblia, is the major cause of giardiasis leading to diarrheal disease with 280 million people infections annually worldwide. Extracellular vesicles (EVs) have emerged as a ubiquitous mechanism participating in cells communications. The aim of this study is to explore the roles of G. duodenalis EVs (GEVs) in host-pathogen interactions using primary mouse peritoneal macrophages as a model. Multiple methods of electron microscopy, nanoparticle tracking analysis, proteomic assays, flow cytometry, immunofluorescence, qPCR, western blot, ELISA, inhibition assays, were used to characterize GEVs, and explore its effects on the host cell innate immunity as well as the underlying mechanism using primary mouse peritoneal macrophages. Results showed that GEVs displayed typical cup-shaped structure with 150 nm in diameter. GEVs could be captured by macrophages and triggered immune response by increasing the production of inflammatory cytokines Il1β, Il6, Il10, Il12, Il17, Ifng, Tnf, Il18, Ccl20 and Cxcl2. Furthermore, activation of TLR2 and NLRP3 inflammasome signaling pathways involved in this process. In addition, CA-074 methyl ester (an inhibitor of cathepsin B) or zVAD-fmk (an inhibitor of pan-caspase) pretreatment entirely diminished these effects triggered by GEVs exposure. Taken together, these findings demonstrated that GEVs could be internalized into mouse peritoneal macrophages and regulate host cell innate immunity via TLR2 and NLRP3 inflammasome signaling pathways. Author summary: G. duodenalis, one of the most common cause of diarrheal diseases, is widely existed in the contaminated water and threatening the public health especially in developing countries. Along with the increasing resistance to anti-G. duodenalis drugs occurs, new targets against giardiasis are of urgently needed. The innate immune system is the first defense line of organism to resist multiple pathogens invasion through recognizing pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs), termed Toll-like receptors (TLRs) on the surface of cell membrane and nucleotide oligomerization domain (Nod)-like receptors (NLRs) inside immune cells. Recently, extracellular vesicles have emerged as a ubiquitous mechanism participating in cells communications. In this study, EVs secreted by extracellular protozoan G. duodenalis were obtained and displayed typical cup-shaped structure with 150 nm in diameter. Moreover, GEVs could enter into primary mouse peritoneal macrophages and regulate host cell innate immunity by up-regulation of various inflammatory cytokines expression. Furthermore, TLR2 and NLRP3 inflammasome signaling pathways involved in this process. This study demonstrated that GEVs could be internalized into primary mouse peritoneal macrophages, regulate host cell innate immunity via TLR2 and NLRP3 inflammasome signaling pathways, and may provide new targets against giardiasis. [ABSTRACT FROM AUTHOR]

The objective of this study was to identify the key changed subtype of T helper cells (Th cells) and their cytokines in pristane-induced arthritis (PIA) in rats. The severity of arthritis was evaluated by body weight, clinical score, the perimeter of ankle and mid-paw and histological assessment of ankle joints. Cytokines of Th1, Th2 and Th17 were determined in the spleen and inguinal lymph nodes at 28 days after pristane injection by real-time qPCR. The mRNA levels of IL-22 receptors, IL-22R1 and IL-22BP, in the spleen were quantified by real-time qPCR. Additionally, IL-22 expression in synovial membrane was detected by Western blotting, and serum IL-22 concentration was determined by ELISA. Correlation between IL-22 concentration and clinical score was analyzed. By screening the cytokines of Th1, Th2 and Th17 expression profile, we found that the mRNA levels of Th17 cytokines were significantly increased in PIA rats. Particularly, a significant increase in the protein expression of IL-22 was determined in synovial membrane and serum from PIA rats, and correlated with clinical score. We conclude that IL-22 expression level was increased and correlated with disease severity, which indicated that IL-22 may play an important role in development of PIA, and was helpful to explorer the pathogenesis of rheumatoid arthritis. [ABSTRACT FROM AUTHOR]

To solve the problems of yield reduction and low water-use efficiency (WUE) of winter wheat (Triticum aestivum L.) caused by winter and spring drought, a 2-year field experiment (2017–2019) was performed under movable shelter conditions with the large- and multispike cultivars Shannong 23 and 29, respectively, to explore the optimal supplemental irrigation regime. Three wetting layers were used for irrigation at the jointing stage: 0–10 cm (T2), 0–20 cm (T3) and 0–30 cm (T4). No irrigation at the jointing stage (T1) served as the control. Within a given cultivar, the soil water content in the 0–80 cm soil layers increased after irrigation, and the rate of tiller mortality decreased with increasing depth of the wetting layer used for irrigation at jointing. No significant differences were found between the T3 and T4 treatments in the photosynthetic rate (Pn) of the apical leaf of the main stem (O), the first primary tiller (I) and the fourth tiller (IV) after jointing. However, compared with the T3 treatment, the T4 treatment had a significantly higher transpiration rate (Tr) and lower instantaneous water-use efficiency (WUEleaf) of the apical leaf of the O and tillers I and IV. This eventually led to a decreasing WUE, although there was no significant change in the spike number or grain yield. These results indicated that moderate irrigation at jointing can effectively reduce the tiller mortality, improve the leaf Pn of the tillers, and increase the spike number and grain yield. However, excessive irrigation can significantly increase the leaf Tr of the tillers, lead to inefficient water consumption and significantly reduce the WUEleaf of the tillers and the WUE. Irrigation at the jointing stage brought the soil water content in the 0–20 cm profile to 100% of field capacity, making it the most suitable supplemental irrigation regime for both the large- and multispike cultivars in the North China Plain. [ABSTRACT FROM AUTHOR]