Genome-wide association study and genomic selection of spike-related traits in bread wheat.

KEY MESSAGE: Identification of 337 stable MTAs for wheat spike-related traits improved model accuracy, and favorable alleles of MTA259 and MTA64 increased grain weight and yield per plant. Wheat (Triticum aestivum L.) is one of the three primary global, staple crops. Improving spike-related traits in wheat is crucial for optimizing spike and plant morphology, ultimately leading to increased grain yield. Here, we performed a genome-wide association study using a dataset of 24,889 high-quality unique single-nucleotide polymorphisms (SNPs) and phenotypic data from 314 wheat accessions across eight diverse environments. In total, 337 stable and significant marker-trait associations (MTAs) related to spike-related traits were identified. MTA259 and MTA64 were consistently detected in seven and six environments, respectively. The presence of favorable alleles associated with MTA259 and MTA64 significantly reduced wheat spike exsertion length and spike length, while enhancing thousand kernel weight and yield per plant. Combined gene expression and network analyses identified TraesCS6D03G0692300 and TraesCS6D03G0692700 as candidate genes for MTA259 and TraesCS2D03G0111700 and TraesCS2D03G0112500 for MTA64. The identified MTAs significantly improved the prediction accuracy of each model compared with using all the SNPs, and the random forest model was optimal for genome selection. Additionally, the eight stable and major MTAs, including MTA259, MTA64, MTA66, MTA94, MTA110, MTA165, MTA180, and MTA164, were converted into cost-effective and efficient detection markers. This study provided valuable genetic resources and reliable molecular markers for wheat breeding programs.

Effect of sunitinib against Echinococcus multilocularis through inhibition of VEGFA-induced angiogenesis.

BACKGROUND: Alveolar echinococcosis (AE) is a lethal zoonosis caused by the fox tapeworm Echinococcus multilocularis. The disease is difficult to treat, and an effective therapeutic drug is urgently needed. Echinococcus multilocularis-associated angiogenesis is required by the parasite for growth and metastasis; however, whether antiangiogenic therapy is effective for treating AE is unclear. METHODS: The in vivo efficacy of sunitinib malate (SU11248) was evaluated in mice by secondary infection with E. multilocularis. Enzyme-linked immunosorbent assays (ELISAs) were used to evaluate treatment effects on serum IL-4 and vascular endothelial growth factor A (VEGFA) levels after SU11248 treatment. Gross morphological observations and immunohistochemical staining were used to evaluate the impact of SU11248 on angiogenesis and the expression of pro-angiogenic factors VEGFA and VEGF receptor 2 (VEGFR2) in the metacestode tissues. Furthermore, the anthelmintic effects of SU11248 were tested on E. multilocularis metacestodes in vitro. The effect of SU11248 on the expression of VEGFA, VEGFR2, and phosphorylated VEGFR2 (p-VEGFR2) in liver cells infected with protoscoleces in vitro was detected by western blotting, reverse transcription quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA). The influence of SU11248 on endothelial progenitor cell (EPC) proliferation and migration was determined using CCK8 and transwell assays. RESULTS: In vivo, SU11248 treatment markedly reduced neovascular lesion formation and substantially inhibited E. multilocularis metacestode growth in mice. Further, it exhibited high anti-hydatid activity as efficiently as albendazole (ABZ), and the treatment resulted in reduced protoscolex development. In addition, VEGFA, VEGFR2, and p-VEGFR2 expression was significantly decreased in the metacestode tissues after SU11248 treatment. However, no effect of SU11248 on serum IL-4 levels was observed. In vitro, SU11248 exhibited some anthelmintic effects and damaged the cellular structure in the germinal layer of metacestodes at concentrations below those generally considered acceptable for treatment (0.12-0.5 µM). Western blotting, RT-qPCR, and ELISA showed that in co-cultured systems, only p-VEGFR2 levels tended to decrease with increasing SU11248 concentrations. Furthermore, SU11248 was less toxic to Reuber rat hepatoma (RH) cells and metacestodes than to EPCs, and 0.1 µM SU11248 completely inhibited EPC migration to the supernatants of liver cell and protoscolex co-cultures. CONCLUSIONS: SU11248 is a potential candidate drug for the treatment of AE, which predominantly inhibits parasite-induced angiogenesis. Host-targeted anti-angiogenesis treatment strategies constitute a new avenue for the treatment of AE.

Diets Partially Replaced With Cassava Residue Modulate Antioxidant Capacity, Lipid Metabolism, and Gut Barrier Function of Huanjiang Mini-Pigs.

Agricultural by-products have been identified as potential feed resources in animal production. The present study investigated the effects of cassava residue (CR) or fermented CR (FCR) on antioxidant capacity, immunity, gut barrier functions, and lipid metabolism in pigs. A total of 120 healthy Huanjiang mini-piglets were assigned into three groups, including control group (basal diet), CR group (basal diet + 5% CR), and FCR group (basal diet + 5% FCR). The experiment lasted for 30 days. The results showed that, dietary CR or FCR supplementation increased the jejunal catalase (CAT, P = 0.063) and glutathione peroxidase (GSH-Px, P < 0.05) levels and hepatic superoxide dismutase (SOD, P < 0.05) level while decreased (P = 0.077) ileal malondialdehyde (MDA) level, when compared with the control group. Dietary CR supplementation increased intestinal SOD and hepatic GSH-Px levels, whereas decreased jejunal and hepatic MDA levels (P < 0.05). Dietary CR supplementation increased the levels of secretory immunoglobulin A (sIgA) in the intestine and liver, as well as jejunal interleukin (IL)-10, ileal tumor necrosis factor (TNF)-α, and hepatic interferon (IFN)-γ, whereas dietary CR or FCR supplementation decreased the jejunal IL-1ß level and increased hepatic IL-10 level (P < 0.05). In the intestinal microbiota analysis, dietary CR or FCR supplementation enhanced the colonic α-diversity and ileal Actinobacteria abundance, whereas decreased ileal Verrucomicrobia and colonic Tenericutes abundances (P < 0.05). In addition, dietary FCR supplementation increased Firmicutes and decreased Bacteroidetes abundances in the ileum and colon, whereas CR supplementation increased Escherichia-Shigella and decreased Terisporobacter abundances in the ileum (P < 0.05). Moreover, dietary CR or FCR supplementation up-regulated (P < 0.05) the gene expressions related to gut barrier functions of piglets. However, dietary CR supplementation showed negative impacts on hepatic lipid metabolism by up-regulating the expression of genes associated with fatty acid synthesis and triglyceride and lipid metabolism. In conclusion, dietary CR or FCR supplementation can maintain the health of piglets by increasing antioxidant capacity, gut barrier function, and altering the intestinal microbiota composition, but CR supplementation may increase the potential risk of abnormal lipid metabolism.

Dietary Addition With Clostridium butyricum and Xylo-Oligosaccharides Improves Carcass Trait and Meat Quality of Huanjiang Mini-Pigs.

This study was conducted to investigate the effects of dietary addition with Clostridium butyricum (CB) and xylo-oligosaccharides (XOS) on growth performance, carcass trait, and meat quality of pigs. A total of 128 Huanjiang mini-pigs with an initial body weight of 9.5 ± 0.1 kg were randomly assigned to one of four groups. The pigs in control (Con) group were fed a basal diet and those in the experimental groups were fed the basal diet supplemented with 0.05% CB (CB group), 0.02% XOS (XOS group), or 0.05% CB + 0.02% XOS (CB + XOS group). Eight replicate pens were used per group with four pigs per pen. On days 28, 56, and 84 of the trial, the growth performance, carcass trait, and meat quality were evaluated. The results showed that dietary CB addition decreased (p < 0.05) the average daily gain and increased (p < 0.05) the ratio of feed intake to body weight gain at day 28 of the trial; CB, XOS, and CB + XOS addition increased (p < 0.05) the backfat thickness at day 84 of the trial compared with the Con group. Dietary CB, XOS, and CB + XOS addition increased (p < 0.05) the pH45min, while decreased (p < 0.05) the marbling score at day 28 of the trial compared with the Con group. Dietary CB + XOS addition increased (p < 0.05) the contents of Ala, Arg, Asp, Gly, His, Leu, Lys, Met, Phe, Ser, Thr, Tyr, and Val in muscle at day 56 of the trial. At day 84 of the trial, dietary CB addition increased the contents of nonessential amino acid (NEAA), total amino acid (TAA), and monounsaturated fatty acid (MUFA), while decreased (p < 0.05) the percentage of C20:1 in muscle compared with the Con group. Collectively, dietary addition with 0.05% CB and 0.02% XOS could not alter the growth performance, but increase carcass trait, meat quality, and muscular nutrient contents in Huanjiang mini-pigs.

SDF-1 secreted by mesenchymal stem cells promotes the migration of endothelial progenitor cells via CXCR4/PI3K/AKT pathway.

Cell-based therapeutics bring great hope in areas of unmet medical needs. Mesenchymal stem cells (MSCs) have been suggested to facilitate neovascularization mainly by paracrine action. Endothelial progenitor cells (EPCs) can migrate to ischemic sites and participate in angiogenesis. The combination cell therapy that includes MSCs and EPCs has a favorable effect on ischemic limbs. However, the mechanism of combination cell therapy remains unclear. Herein, we investigate whether stromal cell-derived factor (SDF)-1 secreted by MSCs contributes to EPC migration to ischemic sites via CXCR4/Phosphoinositide 3-Kinases (PI3K)/protein kinase B (termed as AKT) signaling pathway. First, by a "dual-administration" approach, intramuscular MSC injections were supplemented with intravenous Qdot® 525 labeled-EPC injections in the mouse model of hind limb ischemia. Then, the mechanism of MSC effect on EPC migration was detected by the transwell system, tube-like structure formation assays, western blot assays in vitro. Results showed that the combination delivery of MSCs and EPCs enhanced the incorporation of EPCs into the vasculature and increased the capillary density in mouse ischemic hind limb. The numbers of CXCR4-positive EPCs increased after incubation with MSC-conditioned medium (CM). MSCs contributed to EPC migration and tube-like structure formation, both of which were suppressed by AMD3100 and wortmannin. Phospho-AKT induced by MSC-CM was attenuated when EPCs were pretreated with AMD3100 and wortmannin. In conclusion, we confirmed that MSCs contributes to EPC migration, which is mediated via CXCR4/PI3K/AKT signaling pathway.

Fumaric acid and succinic acid treat gestational hypertension by downregulating the expression of KCNMB1 and TET1.

The present study hypothesized that fumaric acid and succinic acid may exhibit therapeutic effects on gestational hypertension. During pregnancy, estrogen upregulates ten-eleven translocation 1 (TET1) expression, which subsequently increases calcium-activated potassium channel subunit ß1 (KCNMB1) expression. KCNMB1 is associated with hypertension. Fumaric acid and succinic acid are understood to inhibit TET. Therefore, the present study investigated whether fumaric acid and succinic acid exhibit therapeutic effects on gestational hypertension and whether these effects are mediated by TET1 and KCNMB1. Nω-Nitro-L-arginine methyl ester hydrochloride was injected into rats to establish a gestational hypertension model. Dimethyl fumarate (DMF) and succinic acid were administrated into rats to treat gestational hypertension. Rats were divided into five groups: i) Control; ii) model; iii) DMF; iv) succinic acid; and v) DMF + succinic acid. Blood pressure was monitored by a noninvasive meter and urinary protein was determined using a urinary protein kit. Placenta pathology was examined by hematoxylin-eosin staining. Compared with the control group, urinary protein and blood pressure in the model group increased significantly. The placental cells in the control group were arranged orderly. However, in the model group, decidual cellular edema of placenta and vacuolar degeneration were observed, and the intervascular membrane was markedly thicker with plenty of fibrin deposition. These results indicate successful establishment of a gestational hypertension model. However, compared with the model group, urinary protein, blood pressure, edema, vacuoles and fibrin deposition were markedly reduced in the DMF, succinic acid and DMF + succinic acid groups. mRNA and protein levels of TET1 and KCNMB1 in placenta were evaluated by immunohistochemical analysis, reverse transcription-quantitative polymerase chain reaction and western blotting. The TET1 and KCNMB1 levels in the model group were markedly increased compared with those in the control group. However, compared with the model group, the expression levels were markedly downregulated in the DMF, succinic acid and DMF + succinic acid groups. In conclusion, fumaric acid and succinic acid may treat gestational hypertension by downregulating the expression of KCNMB1 and TET1.

A combination of pirfenidone and TGF-ß inhibition mitigates cystic echinococcosis-associated hepatic injury.

Cystic echinococcosis (CE) occurs in the intermediate host's liver, assuming a bladder-like structure surrounded by the host-derived collagen capsule mainly derived from activated hepatic stellate cells (HSCs). However, the effect of CE on liver natural killer (NK) cells and the potential of transforming growth factor-ß (TGF-ß) signalling inhibition on alleviating CE-related liver damage remain to be explored. Here, by using the CE-mouse model, we revealed that the inhibitory receptors on the surface of liver NK cells were up-regulated, whereas the activating receptors were down-regulated over time. TGF-ß1 secretion was elevated in liver tissues and mainly derived from macrophages. A combination of TGF-ß signalling inhibitors SB525334 and pirfenidone could reduce the expression of TGF-ß1 signalling pathway-related proteins and collagen production. Based on the secretion of TGF-ß1, only the pirfenidone group showed a depressing effect. Also, the combination of SB525334 and pirfenidone exhibited a higher potential in effectively alleviating the senescence of the hepatocytes and restoring liver function. Together, TGF-ß1 may be a potential target for the treatment of CE-associated liver fibrosis.

[Study on effect of echinococcus granulosus protoscolices on fibrosis of bone marrow mesenchymal stem cells].

OBJECTIVE: To investigate the effect of echinococcus granulosus protoscolices on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into fibroblasts. METHODS: Femur bone marrow of 4-week-old C57BL/6 mice was taken and BMSCs were isolated and cultured by adherent culture. Echinococcus granulosus protoscolices was extracted from the liver of sheep infected with echinococcus granulosus. The experiment was divided into two groups. The experimental group was co-cultured with the 3rd generation BMSCs and the echinococcus granulosus protoscolices, and the control group was the 3rd generation BMSCs. Before and after co-culture, the morphology of BMSCs and the activity of echinococcus granulosus protoscolices were observed by inverted microscope. After cultured for 1, 3, 5, and 7 days, the mRNA expressions of transforming growth factor ß 1 (TGF-ß 1), collagen type Ⅰ, and collagen type Ⅲ were detected by real-time fluorescent quantitative PCR, the protein expressions of TGF-ß 1, collagen type Ⅰ, collagen type Ⅲ, Smad7, and phosphorylated Smad2/3 were detected by Western blot, and the contents of collagen type Ⅰ and collagen type Ⅲ in the supernatant of the two groups were detected by ELISA. RESULTS: After 7 days of co-culture, the morphology of BMSCs changed into fusiform and irregular triangle, which was closer to the mouse fibroblasts. The relative mRNA expressions of TGF-ß 1, collagen type Ⅰ, and collagen type Ⅲ in the experimental group were significantly higher than those in the control group; the relative protein expressions of TGF-ß 1, collagen type Ⅰ, collagen type Ⅲ, and phosphorylated Smad2/3 in the experimental group were significantly higher than those in the control group, and the relative protein expression of Smad7 in the experimental group was significantly lower than that in the control group; the contents of collagen type Ⅰ and collagen type Ⅲ in the supernatant of the experimental group were significantly higher than those in the control group. The differences between the two groups were significant ( P<0.05). CONCLUSION: Echinococcus granulosus protoscolices may promote the secretion of collagen type Ⅰ, collagen type Ⅲ, and TGF-ß 1 by TGF-ß 1/Smad signal pathway, which can promote the fibrosis of BMSCs that related to the formation of fibrocystic wall by echinococcosis.

[Prokaryotic expression and immunogenicity of IgG-binding protein of Streptococcus equi subspecies equi].

To analyze the immunogenicity and protective ability of recombinant IgG-binding protein (EAG) of Streptococcus equi subspecies equi and to evaluate its value when used as equine vaccine antigen, EAG gene was amplified by PCR and inserted into pET-28a vector. The EAG recombinant proteins were expressed and purified to immune mice. The serum antibody and challenge protection were tested. The purified recombinant protein of EAG was 26 kDa, and the protein reacted specifically with positive serum of Streptococcus equi subspecies equi. The mice antibody level for EAG immunization group was 1∶8 100. The immunological protection result showed that the protection rate of the EAG recombinant protein was 90%. The results suggested that the EAG protein has good immunogenicity and immunological protection, and it can effectively increase the humoral immune response and immunological protection of mice.

[Characterization of antiserum after co-immunized with adhesins expressing ClfA, FnBPA-A and FnBPA-BCD of Staphylococus aureus].

OBJECTIVE: To compare the antigenicity, adhesion inhibition activity and immune protection of the serum immunized with recombinant adhesins Clumping factor A (CfA), Fibronection binding protein A-A ( FnBPA-A) and Fibronection binding protein A-BCD (FnBPA-BCD) of Staphylococcus aureus. METHODS: ClfA, FnBPA-A and FnBPA-BCD genes were expressed and the resulting recombinant proteins were induced and purified. After immunizing mice with these purified proteins in combination or alone, the serum antibodies of mice were analyzed and compared for their antigenicity, adhesion inhibition activity and immune protection. RESULTS: The recombinant FnBPA-BCD protein had better fibronection (Fn) binding ability than that of fibrinogen (Fg) binding and ClfA. FnBPA-A had better binding capacity to Fg than that of FnBPA-BCD. The antibody titer induced by ClfA and FnBPA-A were better than that of FnBPA-BCD, and three proteins co-immunization group had better antibody titer and adhesion inhibition to Staphylococcus aureus than those of single recombinant protein immunization group (P < 0.05). The immune protection rate for both ClfA and three proteins co-immunization group were 100% and for FnBPA-A and FnBPA-BCD immunization group the immune protection rate were 80% and 50%, respectively. CONCLUSION: Our data suggest that coimmunization with these 3 recombinant proteins helps to achieve better immune effect.

[Immunological comparison of Efb and ClfA of Staphylococcus aureus isolated from bovine].

To compare immunological characteristics of Extracellular fibrinogen-binding protein (Efb) and Clumping factor A (CfA) of Staphylococcus aureus, we constructed two prokaryotic expression vector pET28a-Efb and pET28a-ClfA. After prokaryotical expression and purification, Efb and ClfA were used to immunize experimental animal. After the second immunization the antisera were collected and the antibody titers, the bacteria binding activity and adhesion inhibition activity of these antisera were detected by enzyme linked immunosorbent assay, adhesion inhibition assay and challenge. Both Efb and ClfA had Fibrinogen binding activity whereas the former had better Fibronectin binding activity. The bacteria binding capability of antisera of rabbits immunized with ClfA was better than that with Efb (P < 0.01). Both antisera of Efb and ClfA could inhibit adherence activity of Staphylococcus aureus to Fibrinogen and Fibronectin adherence compare to the control group (P < 0.01), and Efb had better adhesion inhibition activity than ClfA. The antibody titer of immunized group could reach 1:40 500. After the second immunization, both Efb and ClfA had good protective efficacy. This result constitutes a good foundation for Staphylococcus aureus subunit vaccine development.