As one of the important livestock, the goat not only provide abundant materials for human beings, such as milk, meat and wool, but also have good adaptability to live in extreme environment such as heat, cold and drought areas. However, the molecular mechanism of environmental adaptability has not been fully explained in goats. With the development of sequencing technology and the rise of landscape genomics, genetic environment association analysis has been carried out in many studies, and a series of candidate genes related to environmental adaptability have been reported, which provided an important basis for analyzing the genetic mechanism of environmental adaptability in goats. This review introduces the methods of genetic environment association analysis from the perspective of landscape genomics, including classification test, logistic regression, general linear model and mixed-effect model. In view of the current situation of environmental adaptability in goats, the research progress in recent years was summarized from 5 aspects:high altitude adaptability, heat adaptability, cold adaptability, drought adaptability and comprehensive climate adaptability. Finally, this paper pointed out the existing problems in the study of environmental adaptability in goats, and prospected the future research trends, in order to provide a theoretical basis for collecting, protecting and utilizing the goat genetic resources.

High reproductive rate is an important guarantee for enhancing the economic benefits of dairy farms, and early pregnancy diagnosis is one of the important means in improving the reproductive rate of dairy cows. It is difficult for traditional pregnancy diagnosis methods to determine the pregnancy status of dairy cows within 21 days after mating. However, the new pregnancy diagnosis technologies developed in recent years can effectively diagnose early pregnancy in dairy cows. In this paper, color Doppler ultrasound, mid-infrared spectroscopy technology, and pregnancy-related protein, circulating nucleotides, interferon tau-induced gene detection and other technologies about the latest diagnosis techniques of early pregnancy in dairy cows were reviewed. Meanwhile, the practical application effects, the advantages and disadvantages of these technologies were compared. And the new diagnostic techniques for early pregnancy were predicted in dairy cows, which would provide some ideas and references for exploitation and application of advanced and high-efficiency early pregnancy diagnosis techniques in dairy cows.

Bone development is mainly accomplished by endochondral ossification in broilers. In the process of endochondral ossification, growth plate chondrocytes undergo proliferation, hypertrophy, transdifferentiation and cartilage matrix mineralization, and eventually osteoblasts gradually replace cartilage primordia to achieve linear bone elongation. Endochondral osteogenesis is a complex and precise process, regulated by multiple signaling and transcription factors such as SOX9, RUNX2, MEF2C, OSX, TGF-β, BMP2, FGFs, IHH and PTHrP, etc. These regulators are expressed by chondrocytes in different zones of the growth plate or specifically regulate chondrocyte proliferation and differentiation, vascular invasion and so on. In poultry breeding, leg disease often occurs in broilers and is difficult to cure, but there are few studies on the pathogenesis of broiler leg disease. This article reviewed the process of bone formation and its specific molecular regulation mechanism, providing reference for understanding the pathogenesis and reasonable treatment of broiler leg disease.

Mucosa is the frontline of body immune defense. Protective mucosal immune response can be effectively induced by oral administration, nose drop, ocular routes and other mucosal routes. However, most mucosal immune vaccines have not been developed, and there is a lack of safe and effective immunopotentiator. Chinese herbal polysaccharides have been proved as biologically-active macromolecules to promote mucosal immune response. This paper reviews different mucosal immunopotentiator of Chinese herbal polysaccharides and its effect on intestinal mucosa in recent years, in order to provide new ideas for the development and utilization of Chinese herbal polysaccharides as mucosal immune immunopotentiator.

African swine fever virus (ASFV) infection causes an acute, intense, and highly contagious disease known as African swine fever (ASF). ASF outbreaks cause significant economic losses since there is no safe and effective vaccine has been developed. During the long-term interaction with the host, ASFV evolves various routes, such as inhibition of interferon and inflammation responses, regulation of apoptosis, autophagy and cellular immunity, to escape host immune surveillance to promote its replication, but the specific mechanism is still not completely clear. The complex immune escape mechanisms of ASFV may be the key factors that impede the development of effective ASF vaccine. Bioinformatics method can be used to analyze the genome and proteome of ASFV to Screen the key genes for immune regulation and protective antigen epitopes of the virus, which will be helpful for the study of molecular mechanism of ASFV immune escape and ASF vaccine development. In this review, research progresses on ASFV infection induced immune responses and the potential immune escape mechanisms of ASFV are summarized, which provides ideas for ASF vaccine development and comprehensive prevention and control of the disease.

The aim of this study was to investigate the polymorphism of DKK3 and CCR1 genes and their effects on backfat thickness of Beijing black pigs, so as to screen out molecular markers that can be used to improve backfat thickness traits and provide basis for directional breeding of excellent traits in Beijing black pigs. DNA extraction, PCR amplification and gene sequencing were performed on 413 healthy Beijing black pigs to screen SNPs of candidate genes. At the same time, SAS 9.2 software was used to analyze the association between different genotypes of mutation sites and backfat thickness of 6-7th ribs, backfat thickness of lumbosacral region. The results showed that a total of 10 SNPs were detected in the exons of DKK3 gene in Beijing black pig, including 1 splice region variant, 1 missense variant and 8 3'UTR variants. Among them, 4 SNPs of 3' UTR variants were significantly associated with the backfat thickness of lumbosacral region (P<0.05), which were g.47443779 T > C, g.47444258 C > T, g.47444912 G > T and g.47445304 T > C, respectively, and other SNPs had no significant correlation with backfat thickness of lumbosacral region or backfat thickness of 6-7th ribs (P>0.05). A total of 3 SNPs were detected in the exon of CCR1 gene, including 2 synonymous variant and 1 missense variant. One synonymous variant (g.29229037 G > A) was significantly associated with the backfat thickness of 6-7th ribs (P<0.05). The linkage disequilibrium analysis of 10 SNPs in DKK3 showed that g. 47443779 T > C and g. 47443783 C > T were in a completely linked state (D'=1), g. 47443783 C > T and g. 47443858 C > T were in a completely linked state (D'=1), g. 47444258 C > T and g. 47444275 C > T were in a highly linked state (D'=0.98). In summary, SNPs of DKK3 and CCR1 genes were respectively significantly associated with the backfat thickness of lumbosacral region and backfat thickness of 6-7th ribs of Beijing black pig, which could provide reference for the early molecular breeding of backfat thickness in Beijing black pig.

The purpose of this study was to investigate the developmental expression pattern of NR1H3 gene in subcutaneous adipose tissues of pigs at different ages and the effects of NR1H3 gene on adipogenic differentiation of porcine preadipocytes, and determine its primary function in fat deposition. qRT-PCR was used to detect the expression of NR1H3 in subcutaneous adipose tissues of Mashen pigs aged 30, 90 and 240 days. Porcine preadipocytes were isolated from dorsal subcutaneous adipose tissues of 5-day-old DLY (Duroc×(Landrace×Yorksire)) piglets. Cellular immunofluorescence technique was used to detect the Adiponectin protein content in the cells to identify the purity of preadipocytes. The over-expression vector of porcine NR1H3 gene was constructed and the NR1H3 siRNA sequences were designed, and then both of them were transfected into porcine preadipocytes. The overexpression and interference efficiency and their effects on expression of key genes related to adipogenic differentiation were investigated by qRT-PCR, Western blot and Oil red O staining methods. The results showed that the expression of NR1H3 in subcutaneous adipose tissues increased as the age increased, the expression level was the highest at 240 days of age and the lowest at 30 days of age, with extremely significant difference between them (P<0.01). Compared with negative control (NC), over-expression (OE) of NR1H3 significantly increased the number of lipid droplets in adipocytes, the expression levels of downstream targets SREBP-1c and ChREBP extremely significantly increased (P<0.01), and the mRNA expression levels of key adipogenic genes FAS, C/EBPβ, PPARγ and FABP4 extremely significantly increased (P<0.01). On the contrary, interfering with NR1H3 significantly reduced the number of lipid droplets. The mRNA expression of downstream target genes and key adipogenic genes extremely significantly reduced (P<0.01), which inhibited the adipogenesis process. The results showed that NR1H3 gene was a positive regulator of the adipogenic differen-tiation of porcine preadipocytes, and affected the differentiation of porcine preadipocytes by regulating the expression of its downstream targets SREBP-1c and ChREBP. These results are of great significance to elucidate the molecular mechanism of fat deposition and improve meat quality.

The aim of this study was to analyze the genetic variation among the white-, hemp- and black-feathered breeding populations of Jingyuan chickens, and to explore the key candidate genes regulating the characteristic traits. In this study, the RAD-seq technology was used to sequence 180-day-old white-, hemp- and black-feathered Jingyuan chickens (60 individuals of each plumage color, 40 hens and 20 roosters) with distinctive features, normal development and health, and the observed heterozygosity (Ho), nucleic acid polymorphism (Pi) within the population and the average inbreeding coefficient (Fis) of the population based on SNPs markers were calculated to analyze the genetic diversity and population structure of Jingyuan chickens. The candidate genes were screened by selective sweep analysis and genome-wide association analysis (GWAS), and enriched by KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways using KOBAS. The candidate genes regulating plumage color of Jingyuan chickens were finally screened. The sequencing results showed that a total of 198.83 Gb of clean data was obtained from 180 samples of Jingyuan chickens, with Q30 reaching more than 93%. Genetic diversity analysis showed that 238 533, 233 562 and 240 820 SNPs markers were identified for the white-, hemp-, and black- feathers of Jingyuan chickens, with Ho, Pi and Fis range of 0.273 2-0.278 2, 0.304 9-0.309 6 and 0.096 1-0.109 8, respectively. The analysis of population structure showed that Jingyuan chickens were divided into different groups according to different feather colors. A total of 11 candidate genes (FZD4, WNT16, EDNRB, TYR, KRAS, CTNNB1, DDC, MC1R, CAMK2A, PRKCB, PRKCA) associated with plumage color in Jingyuan chickens were screened by selective sweep analysis and genome-wide association analysis, and the enrichment results showed that these genes were related to the pathways of melanogenesis, tyrosine metabolism, and Wnt signaling. In summary, the breeding status of the Jingyuan chicken can be comprehensively evaluated using SNPs marker information and lay a theoretical foundation for the conservation of the genetic resources of the Jingyuan chicken in this study. At the same time, the candidate genes related to the plumage color trait of Jingyuan chicken were screened to provide new genetic markers and gene targets for the breeding of different plumage colors of Jingyuan chicken.

The purpose of this study was to clarify the tissue and cell expression pattern of KLF15 in Heying Black chickens and to elucidate the effects of interfering KLF15 on the proliferation and differentiation of preadipocytes of Heying Black chickens. RT-qPCR was used to detect the expression of KLF15 in various tissues, the developmental expression pattern in abdominal fat tissues and the expression level in cells at different stages of adipogenic induction differentiation. After interfering with KLF15, cell proliferation was detected by CCK-8 and EdU, triglyceride content was analyzed, Oil red O staining was carried out, and marker genes for adipocyte differen-tiation were detected (PPARγ, C/EBPa and FAS). The effects of interfering with KLF15 on adipogenic differentiation of Heying Black chicken preadipocytes were investigated from morphological and molecular biological perspectives. The results showed that KLF15 was widely expressed in all detected tissues of Heying Black chickens, with higher expression levels in liver, leg muscle and abdominal fat tissues. KLF15 showed "L" and "U" shaped expression trend in abdominal fat of males and females, respectively, and the expression peak appeared at one day of age. KLF15 mRNA increased slowly in the early stage of cell differentiation, and its expression peak occurred in the late stage of cell differentiation. Cell proliferation detection showed that the results of CCK-8 and EdU exhibited the same trend, and interfering with KLF15 highly significantly inhibited the proliferation of preadipocytes (P<0.01), obviously reduced the accumulation of lipid droplets in preadipocytes, extremely significantly reduced triglyceride content (P<0.01). In addition, the mRNA levels of PPARγ and C/EBPa were extremely significantly down-regulated (P<0.01), and FAS mRNA levels were significantly down-regulated (P<0.05). This study revealed the expression pattern of KLF15 in Heying Black chickens and verified that interfering with KLF15 could inhibit the proliferation and differentiation of preadipocytes. It is speculated that KLF15 is a negative regulator of preadipocytes, it possibly regulate the differentiation of preadipocytes through regulating the expression of PPARγ, C/EBPa and FAS genes.

The purpose of this study was to screen candidate genes for carcass traits of slow-growing yellow-feather broiler (Tiannong partridge chickens) by transcriptome sequencing (RNA-Seq) and weighted gene co-expression network analysis (WGCNA). The total of 104 Tiannong partridge chickens were slaughtered at 126 days of age. The 7 carcass traits including breast muscle weight were measured, and breast muscle tissues were collected for RNA-Seq. After obtaining the gene expression matrix based on RNA-Seq data, combined with the phenotype, WGCNA was carried out to determine the target module, screen the hub genes of the target module. KEGG pathway enrichment, protein-protein interaction network (PPI) construction and correlation analysis were carried out, respectively. The 15 092 genes were expressed in 104 breast muscle tissues after RNA-Seq quality control. By WGCNA of carcass phenotype, 19 modules were obtained, of which 4 modules were extremely significantly correlated with carcass traits. The results of functional enrichment showed that genes in the target modules were mainly enriched in ECM-receptor interaction, focal adhesion and other pathways. The 58 hub genes were screened according to|Module membership|> 0.8,|Gene significance|> 0.2. The 5 network node genes were identified through PPI network construction. By integrating correlation analysis, COL1A2 and SPARC were identified as the most important candidate genes. This study found that ECM-receptor interaction and other pathways played a major role in the regulation of carcass traits of Tiannong partridge chickens through large-scale transcriptome analysis. COL1A2 and SPARC are selected as important candidate genes affecting the percentage of eviscerated yield. The results lay a theoretical foundation for identifying molecular markers and analyzing the molecular mechanism of carcass traits, and provide an important reference for the breeding of slow-growing yellow- feather broilers.

The purpose of this study was to identify the expression of circSESN1 in chickens and to further explore the expression characteristics of circSESN1 in chicken pectoral muscle. In experiment 1, the expression of circRNA of chicken SESN1(circSESN1) was identified by RT-PCR, Sanger sequencing, and qRT-PCR. Then the spatio-temporal expression characteristics of circSESN1 and its source gene SESN1 were detected by qRT-PCR with tissue samples of silky fowls at different growth stages (E10, E19, 21 d, 49 d, n=4). In experiment 2, the effects of exogenous stimuli on the expression of circSESN1 were explored with the pectoral muscle of broilers collected at different time points after intraperitoneal injection of glucose (18 d, n=6), sodium pyruvate (21 d, n=6) or insulin (24 d, n=6). In experiment 3, 21 d (n=10) broilers were conducted with 15% energy restriction and 15% protein restriction, and the effect of feeding restriction on the expression of circSESN1 was explored in the pectoral muscle tissue of broilers. In addition, the potential function of circSESN1 was predicted by the bioinformatics analysis software. The results showed as follows:1) circSESN1 was a circular RNA with high stability in chicken and can not be digested by RNase R enzyme. 2) The relative expression levels of circSESN1 and source gene SESN1 were the highest in the pectoralis of 21 d silky fowl, followed by leg muscle and pancreas (P<0.01). The expression level of circSESN1 dramatically varied in the pectoral muscle of birds at different growth stages, with the highest level at 21 d and the lowest at E10 (P<0.01). The expression of SESN1 was highest in the pectoral muscle of silky fowl at E19 and the lowest at E10 (P<0.01). 3) After insulin injection, the expression of circSESN1 in the pectoral muscle showed a trend of first increasing and then decreasing at different time points. CircSESN1 was significantly higher at 120 min after insulin injection than the basal state or PBS group (P<0.01). 4) Compared with the control group, the expression of circSESN1 decreased significantly by 15% energy restriction and 15% protein restriction in the pectoral muscle of birds (P<0.05). 5) After predicting the potential function of circSESN1, it was found that circSESN1 functioned as efficient miRNA sponges, but can not encode proteins. The above results showed that circSESN1 was expressed in chicken, and it had similar spatio-temporal expression characteristics with its source gene SESN1. Exogenous insulin and feeding restriction can regulate the expression of circSESN1 in the pectoral muscle.

This study aimed to explore the function and regulatory mechanism of EPB41L4A-AS with high expression in early lactation in bovine mammary epithelial cells (BMECs). In this study, EPB41L4A-AS siRNA and si-NC were transfected into BMECs, respectively. The effect of EPB41L4A-AS siRNA on the proliferation of BMECs was detected by EdU and MTT assays; qPCR and Western blot methods were used to detect the expression of ErbB3, PIK3R1, AKT and STAT5a genes. EdU and MTT results showed that the activity of BMECs was significantly increased with the interference of EPB41L4A-AS expression (P<0.001) and the number of EdU positive cells obviously increased. qPCR and Western blot results showed that the relative mRNA abundance of ErbB3, PIK3R1, AKT and STAT5a were significantly higher than those of the control group after interfering EPB41L4A-AS expression (P<0.05), and the relative protein expression levels of ErbB3, PIK3R1, AKT, p-AKT and STAT5a significantly increased (P<0.05). The results of this study suggested that reducing the expression of EPB41L4A-AS promoted the expression of ErbB3, and activated the PI3K/AKT and JAK-STAT signaling pathways to promote the proliferation of BMECs. This study revealed the role and regulatory pathway of EPB41L4A-As in BMECs proliferation, which provided a theoretical basis for further exploring the regulation mechanism of lncRNA on BMECs.

The aim of this study was to investigate the role of long non-coding RNA BCL2 (lncRNA BCL2) in inflammation and apoptosis of dairy cow mammary epithelial cells. The lncRNA BCL2 sponge adsorbed miRNAs and the mRNAs they targeted were analyzed using RNAhybrid, Targetscan and miRwalk softwares, and KEGG pathway enrichment analysis was performed on these target genes; lncRNA BCL2 expression in mammary tissue and mammary epithelial cell inflammation and its effect on the expression of genes involved in inflammation and apoptosis in mammary epithelial cells were studied using qPCR and lncRNA super expression technology. The results showed that, compared with control group, the lncRNA BCL2 was significantly down-regulated (P<0.05) in mammary gland tissues of clinical mastitis cow and inflammatory mammary epithelial cells induced by LPS. In the presence of lncRNA BCL2 overexpression, the expression of the inflammatory cytokines IL-1βand IL-8 and the key gene NF-κB (p65/p50) of the inflammatory signaling pathway were significantly up-regulated (P<0.05). The expression of CASP3 and CASP9 genes related to apoptosis were significantly decreased (P<0.05), BCL2 expression was significantly up-regulated (P<0.001), and the ratio of BCL2 to BAX was greater than 2. The above results were consistent with the results of bioinformatics analysis, which indicated that lncRNA BCL2 might regulate inflammation and apoptosis of bovine mammary epithelial cells through lncRNA BCL2-miRNA-BCL2 (NLRP1, CASP1, NLRP3) network. It can conclude from the above results that in the process of cow mastitis, the expression of lncRNA BCL2 is down-regulated, which weakens its effect on increasing the expression of IL-1β, IL-8, and NF-κB, subsequently alleviates mammary inflammation. At the same time, the decrease of lncRNA BCL2 expression attenuates its role in down-regulating CASP3 and CASP9 gene expression and up-regulating BCL2 gene expression, which might promote the apoptosis of cow mammary epithelial cells.

This study aimed to apply the single-step genomic best linear unbiased prediction with metafounders (MF-SSGBLUP) to joint genomic breeding and compare it with other classical genomic selection methods. QMSim software was used to simulate 3 dairy cattle populations with independent pedigrees; The generalized least squares (GLS) and naïve (NAI) methods were used to estimate the ancestral relationship matrix Γ between different populations. MF-SSGBLUP, SSGBLUP and BLUP were used respectively to joint breeding for the simulated populations, and the performance of each method in estimating genetic parameters and breeding values was evaluated. For different heritabilities, the Γ matrix obtained by GLS was slightly lower than that obtained by NAI method in diagonal elements, but there was no significant difference in non-diagonal elements. The correlation coefficient in diagonal elements between the genomic and genetic relationship matrices based on metafounders (0.750-0.775) was higher than that between genomic and traditional relationship matrices (0.508-0.572). The deviations of heritability estimates by MF-SSGBLUP (0.138, 0.140, 0.297 and 0.298) from the current population heritability (0.107 and 0.296) were smaller than those of the other two methods (0.145, 0.173, 0.273 and 0.340). Correspondingly, the accuracies of estimated breeding values by MF-SSGBLUP (0.888-0.908) were higher than that by SSGBLUP (0.863-0.876) and BLUP (0.854-0.871). The results showed that MF-SSGBLUP had less biased estimates of genetic parameters and breeding values with higher accuracies. Based on the simulation results, the MF-SSGBLUP performed better than other classical genomic selection methods in joint breeding.

The purpose of this study was to analyze the differential expression proteins(DEPs) of yak pulmonary arterial smooth muscle cells (PASMCs) under different culture conditions of hypoxia and normoxia by using TMT technology, so as to understand DEPs for PASMCs of plateau yak. In this study, PASMCs were cultured by sticking culture method and cultured for 72 h under hypoxia and normoxia conditions, respectively. Then cells were collected and the total protein of cells was extracted. DEPs were screened for analysis using TMT labeled protein quantitative technology, and GO functional annotation and KEGG pathway analysis were performed. A total of 6 859 proteins were obtained by comparing the samples in 2 groups of PASMCs under hypoxia and normoxia. With the standard of FC(fold change) ≥ 1.3 or ≤ 0.78 and P<0.05, a total of 531 DEPs were obtained, among them, 186 DEPs were up-regulated and 345 DEPs were down-regulated. The results of enrichment analysis showed DEPs were mainly enriched in the hypoxia adaptation related pathways such as central carbon metabolism, glycolysis and glucose metabolism synthesis, HIF-1 signaling pathway, and biosynthesis of secondary metabolites. Seven important DEPs (PGK, HK, LDH, PGAM, pfkA, IL6ST and PDK1) were found in these pathways. Subcellular localization of plasma membrane (28.9%) accounted for the highest proportion. Epidermal growth factor-like domain was the main domain in domain analysis. The results indicated that DEPs were enriched in metabolism, enzyme activity, hypoxia adaptation and other related categories and pathways. Seven important DEPs (PGK, HK, LDH, PGAM, pfkA, IL6ST and PDK1) may be related to the adaptation to hypoxia environment. These results provide the theoretical support for further study on adaptation of yak PASMCs to hypoxia.

The study aimed to analyze the protein expression of sperm in epididymis at 3 different regions (caput, corpus and cauda) and obtain differentially expressed proteins using proteomics in sheep. The functional enrichment analysis on the data was performed to explore proteins regulating spermatogenesis and maturation. In this study, 3 healthy male Hu sheep around 12 months of age were selected as experimental animals, the epididymis were isolated and sperms were collected by regions. There were 3 groups of samples (caput group, corpus group and cauda group), 3 biological replicates in each group, totaling 9 samples of sheep sperm. Based on TMT-calibrated quantitative protein technology and R software, GO and KEGG enrichment analysis were performed on differentially expressed proteins, and the reliability of the results was verified by Western blot, immunofluorescence and flow cytometry. A total of 616 differential proteins were identified from 22 841 unique peptides. Among them, cauda vs. caput group had 309 differentially expressed proteins(up:213, down:96), cauda vs corpus group had 167 differentially expressed proteins(up:107, down:60), corpus vs caput group had 140 differentially expressed proteins(up:88, down:52). According to the fold change and protein function, the key protein KPNA4, which may be related to sperm maturation and nuclear material transport, was screened out. This study reveals the characteristics and differences of sperm from different regions of the sheep epididymis. These data provide a rich resources for studying the reproductive mechanism and sperm maturation in male sheep.

The study aimed to investigate the role of RBM3 in the reproductive-related regulation in yak (Bos grunniens). Three healthy female yaks aged 4-6 years old in different reproductive periods (follicular, luteal and pregnancy phases) were used in this study. Nine groups of experimental samples were collected from ovaries, uterus and fallopian tubes, and 3 biological replicates were set in each group. The RBM3 gene of yak was cloned by gene cloning technology and analyzed by bioinformatics. RBM3 relative expression levels of gene and protein in 9 groups of experimental samples were detected by quantitative real-time PCR(qRT-PCR) and Western blot(WB). The distribution of RBM3 protein in samples was detected by immunohistochemistry (IHC). RBM3 gene(GenBank No.:MF142258.1) of yak was cloned in this experiment, it had the closest relationship with wild yak(Bos mutus). Bioinformatics analysis showed that nucleotide No.51 and No.98 were different from several common mammals, and its secondary and tertiary structure was constructed. After prediction, the encoded protein was stable non-transmembrane protein. WB results showed that RBM3 was expressed in fallopian tube, ovary and uterus of yak, but the expression level at luteal phase was significantly higher than that at follicular and pregnancy phases (P<0.05). The expression in fallopian tube at follicular phase was significantly lower than that at luteal and pregnancy phases (P<0.05). The expression in uterus at follicular phase was significantly higher than that at luteal and pregnancy phases(P<0.05). Immunohistochemical results showed that the main expression sites of RBM3 in the ovary were theca follicle, granular layer and luteal cells, while the expression sites in the fallopian tube were epithelium mucosae. For the uterus, the expression sites was mainly in the endometrium and uterine glands. The results of this study suggest that RBM3 may be involved in the regulation of individual estrous cycle and pregnancy process in yak, as well as pregnancy recognition. It provides a reference for the research of RBM3, a cold stress regulatory factor, in the physiological mechanism of yak in cold and hypoxic environment at high altitude, and other related aspects.

This experiment was conducted to evaluate the effects of dietary metabolizable energy (ME) levels during rearing on growth performance and the development of Rugao yellow chicken at the initiation of the laying period. Three hundred and sixty 8-week-old Rugao yellow chicken with similar body weight were assigned to 3 groups with 8 replicates per group and 15 hens per replicate. The pullets were fed one of three diets that were nutritionally equal with the exception of ME levels of 10.88, 11.30, and 11.72 MJ·kg^-1 from 8 to 18 weeks of age. From 18 to 20 weeks of age, all pullets were fed a diet with the same nutritional level. The trial period lasted for 12 weeks (from 8 to 20 weeks of age). The results showed as follows:1) The body weight of hens at 18 weeks of age, the average daily weight gain (ADG) and the average daily ME intake (MEi) of hens aged from 8 to 18 weeks increased linearly (P<0.05), while the ratio of feed to gain (F/G) and the ratio of CPi to gain (CPCR) of hens aged from 8 to 18 and 8 to 20 weeks decreased linearly (P<0.05) with the increase of dietary ME levels from 8 to 18 weeks of age.2) The development of carcass, internal organ and digestive tract at 18 and 20 weeks of age were not significantly affected by dietary ME level from 8 to 18 weeks of age (P>0.05). 3) The tibia weight and tibia index showed a decreasing and then increasing response to increasing dietary ME level from 8 to 18 weeks of age (P<0.05), while there was no significant difference in the development of comb and femur at 18 weeks of age and the development of comb, femur and tibia at 20 weeks of age. 4) The ovary stroma index and small yellow follicle index at 18 weeks of age increased linearly with the increase of dietary ME levels from 8 to 18 weeks of age, while there was no significant difference in the other reproductive organs at 18 weeks of age and all the reproductive organs at 20 weeks of age. It was indicated that the body weight of hens at 18 weeks of age and the ADG of hens aged from 8 to 18 weeks were increased, the F/G and CPCR of hens aged from 8 to 18 weeks were improved, and the development of ovary stroma and small yellow follicle was promoted when the dietary ME levels was 11.72 MJ·kg^-1 from 8 to 18 weeks of age. But no significant difference was observed in the indexes of growth performance and development among the treatments at the initiation of the laying period (20 weeks of age). The results of the present study showed that it has an important practical importance to study the combination of growing period and preparatory laying period of laying hens.

The purpose of this study was to investigate the effects of time-restricted feeding on the growth performance and liver transcriptional and metabolomic profiles of growing pigs. A total of 12 healthy growing Duroc×Landrace×Large White pigs with similar body weight ((56.29±1.93) kg) were selected in the present study and were randomly divided into the control group (CON) and time-restricted feeding group (TRF), each group had six replicates with one pig per replicate (pen). Pigs in the CON group had free access to feed throughout the whole experimental period. While pigs in the TRF group were fed three times per day at 7:00, 12:00, and 17:00, respectively, each feeding time lasted for an hour, and pigs were fed ad libitum during the feeding time. During the 21 d-experiment, feed intake of the pigs was recorded daily. Pigs were weighed weekly to calculate the feed conversion ratio (FCR). At the end of the experiment, all pigs were slaughtered, venous blood was collected to determine biochemical parameters, and liver tissues were collected for the determination of metabolome and transcriptome. The results showed that the average daily gain of pigs in the TRF group was significantly higher than that in the CON group (P<0.05), and the FCR in the TRF group was significantly lower than that in the CON group (P<0.05). Compared with the CON group, the concentration of total bile acid in serum of pigs in the TRF group trended to decrease (P<0.1), and concentrations of alanine aminotransferase and creatine kinase trended to increase (P<0.1). TRF significantly increased the expression of genes concerning the inhibiting protein decomposition (P<0.05), and reduced the decomposition of proteins in the liver, and thus decreased the level of amino acids in the liver. In addition, TRF also significantly increased the expression of genes related to fatty acid metabolism (P<0.05), increased liver fat synthesis and reduced liver fatty acids content. In conclusion, TRF can affect nitrogen metabolism and amino acid metabolism, change the metabolism of fatty acids and proteins in the liver, and then regulate the utilization of nutrients by growing pigs and affect the growth performance of pigs. These results collectively indicate that the growth performance and metabolism of growing pigs can be improved through regulating feeding regime.

To explore the B cell epitopes of African swine fever virus (ASFV) p30 protein, the monoclonal antibody (mAb) of p30 protein was prepared, and it was used as a tool for screening its B cell epitopes. First of all, p30 protein was obtained by prokaryotic expression system and purified by Ni affinity method. BALB/c mice were immunized with purified protein and hybridoma cells were prepared. Positive hybridoma cells were screened by indirect enzyme linked immunosorbent assay (iELISA). The mAb was then prepared by cell expression method. The specificity of mAb was identified by indirect immunofluorescence assay (IFA) and Western blot. Then, IEDB epitope prediction software was used to predict the B cell epitopes of p30 protein. According to the prediction results, CP204L gene was truncated to express in prokaryotic cells. IFA, Western blot and iELISA were used to identify the epitopes. Finally, a phage-displayed 12-mer peptide library was used to conduct four rounds of biological panning for the p30 monoclonal antibody, and identify the specific epitope of p30, which was further compared with the truncated protein mapping described above. The results of specific identification showed that the mAb could successfully identify ASFV in porcine alveolar macrophages. The results of truncated protein mapping showed that the epitopes region recognized by the mAb of p30 protein was ⁸⁴M-K¹⁴². The biopanning experiment indicated that ¹¹⁶TSSFETLFE¹²⁴ was a core domain of the B cell linear epitope of p30 protein, which further narrowed the range of the epitope distribution. Altogether, a mAb against ASFV p30 protein was prepared, and the epitopes were identified, which provided a reference for the development of serological diagnostic reagents and lay a foundation for the study of p30 protein function

This study aimed to investigate the effect of estradiol (E2) and progesterone (P4) on the replication of African swine fever virus (ASFV) in vitro. Non-pregnant swine serum (NPSS) and pregnant swine serum (PSS) were collected and treated. Porcine alveolar macrophages (PAMs) and Bone marrow-derived macrophages (BMDM) were cultivated with Fetal bovine serum (FBS), NPSS and PSS respectively and infected with ASFV. The difference of ASFV replication were detected by absolute quantitative PCR and Western blot. Using E2 and P4 ELISA kits to detect the contents of E2 and P4 in each serum. ASFV was infected with E2 and P4 alone or in combination, and the replication differences of ASFV were detected by absolute quantitative PCR and Western blot. NPSS and PSS treated BMDM for 24 hours, the transcriptional difference of beta-interferon and downstream antiviral factors was detected by relative quantification. The results showed that ASFV replication in PSS group was more effective than NPSS group. The contents of E2 and P4 in PSS were higher than NPSS, but the contents of E2 and P4 in FBS were low levels, and ASFV replication is more effective after adding E2 and P4 alone or mixed in FBS group. And PSS treatment of BMDM can inhibit the transcription of beta-interferon and downstream antiviral factors. High levels of E2 and P4 in PSS promote ASFV replication in vitro. This study provides a scientific basis for explaining the African swine fever (ASF) occurs preferentially in sows and symptoms are more serious in pregnant sows.

The C-terminal domain (CTD) of porcine deltacoronavirus S1 subunit is the main region which induces the neutralizing antibody. S1-CTD was expressed by HEK-293T eukaryotic expression system and purified, and porcine ileal epithelium cells membrane proteins were extracted to investigate porcine host proteins that interact with it. Thirty-two suspected interacting host proteins were obtained by co-inmunprecipitation (Co-IP) and mass spectrometry. Eukaryotic expression plasmid of KIF1 binding protein (KIFBP) was constructed, and the interaction between KIFBP and S1-CTD was identified by Co-IP and laser confocal microscopy. All results proved that KIFBP interacted with S1-CTD and co-located in cytoplasm. Further research indicated that overexpression of KIFBP could effectively reduce the viral mRNA level and the viral titer in which the mRNA level decreased by about 70%, and the viral titer decreased by 10^1.6TCID₅₀. In conclusion, a host protein KIFBP interacting with PDCoV S1-CTD was screened and identified in this study which provides a theoretical basis for understanding the pathogenesis of PDCoV.

Our study aimed at investigating the regulatory role of the PERK/ATF4/CHOP pathway on NLRP3 inflammasome of human monocyte macrophage THP-1 cells infected with Bacillus Calmette-Guérin (BCG). THP-1 macrophages were infected with BCG alone or in the presence of small interference to PERK for 24 h, then the expressions of NLRP3 inflammasome related molecules and PERK/ATF4/CHOP pathway molecules in THP-1 cells were detected at mRNA level by qRT-PCR and protein level by Western blot; THP-1 macrophages were infected with BCG alone or in the presence of specific inhibitors to PERK for 24 h, then the expression of NLRP3 inflammasome related molecules and PERK/ATF4/CHOP pathway molecules in THP-1 cells was detected at mRNA level by qRT-PCR and protein level by Western blot, and ELISA was used to detect the release of interleukin-1β (IL-1β) and interleukin-18(IL-18), the viability of THP-1 cells was detected by CCK-8 method, the co-localization of NLRP3 and ASC was detected by immunofluorescence. The results showed that the expression of PERK, NLRP3, ASC and Caspase-1 proteins increased with the infection time, and reached the peak at 24 h (P<0.001), and the release of IL-1β and IL-18 very significantly increased with time (P<0.001). THP-1 macrophages were infected with BCG alone or in the presence of small interference to PERK for 24 h, compared with uninfected control group, the expression of NLRP3, ASC, Caspase-1, PERK, ATF4, CHOP in siNC+BCG infected group were significantly (P<0.05) or extremely significantly up-regulated (P<0.001) both at mRNA level and protein level, while compared with siNC+BCG infected group, the expression of NLRP3, ASC, Caspase-1, PERK, ATF4, CHOP in siPERK+BCG infected group were significantly (P<0.05) or extremely significantly (P<0.01, P<0.001) down-regulated both at mRNA level and protein level; THP-1 macrophages were infected with BCG alone or in the presence of inhibitor to PERK for 24 h, compared with uninfected control group, the expression of NLRP3, ASC, Caspase-1, PERK, ATF4, CHOP in BCG infected group were significantly (P<0.05) or extremely significantly up-regulated (P<0.01) both at mRNA level and protein level, and the release of IL-1β and IL-18 very significantly increased (P<0.001). Compared with BCG infection group, the expression of those molecules were significantly (P<0.05) or extremely significantly (P<0.01) down-regulated both at mRNA level and protein level, and the release of IL-1β and IL-18 significantly (P<0.05) or extremely significantly (P<0.001) decreased, the viability of THP-1 cells was significantly up-regulated (P<0.05). The results of immunofluorescence also showed that NLRP3 and ASC proteins were co-located, and GSK2656157 could significantly inhibit the expressions of NLRP3 and ASC proteins induced by BCG infection(P<0.001). The above results show that PERK/ATF4/CHOP pathway regulates the activation of NLRP3 inflammasome in macrophages infected by BCG.

This study aimed to study the function of sapC in ABC transporters of SalmonellaTyphimurium in its pathogenic mechanism. The sapC gene deletion mutant SM△sapC of S. Typhimurium was constructed by λ-Red mediated mutagenesis procedures, and the growth characteristics, acid stress test, polymyxin B sensitivity test, biofilm detection, intracellular survival and toxicity test in mice were performed. The results showed that the growth rate of the deletion strain SMΔsapC was not significantly different from that of the parent strain and the supplemented strain. Under acid stress condition, the survival rate of SMΔsapCsapC was significantly lower than that of the parent strain; the deletion of sapC gene enhanced the drug resistance of polymyxin B and decreased the ability of biofilm formation of S. Typhimurium. The proliferation ability of SMΔsapC strain in mouse macrophages and the virulence in mice were significantly lower than that of the parent strain. This study showed sapC gene decreased the acid resistance and biofilm formation ability of S. Typhimurium, thus reducing the virulence of Salmonella in vivo and in vitro. This study laid a foundation for further explaining the pathogenic mechanism of S.Typhimurium.

The aim of this study was to develop monoclonal antibodies (MAbs) against Mycoplasma mycoides subsp. Mycoides (Mmm), and to provide a specific monoclonal antibody (MAb) for immunological methods for detection of Mmm, the causative agent of contagious bovine pleuropneumonia (CBPP). In this study, the protein M0071 was selected based on bioinformatics analysis of the whole genomes of five representative generations of Mmm Ben-1 isolates. A soluble fusion protein M0071 was expressed in prokaryotic expression system, the purified recombinant protein M0071 (rM0071) was used as an immunogen to immunize BALB/c mice. The hybridoma 3C4A1 stably secreting MAbs against rM0071 protein, was produced after screening by limited dilution method and an indirect ELISA method. MAb 3C4A1 were purified from mouse ascites. The specificity of the MAbs was identified using Western blot method, the titer and isotype of the purified MAbs were also determined. Further, indirect immunofluorescence (IFA) assay was used to evaluate the potency of the MAbs as primary antibodies for the detection of Mmm infection. One hybridoma 3C4A1 was successfully obtained, and its secreted antibody was named MAb 3C4A1. Western blot results showed that MAb 3C4A1 specifically reacted with Mmm isolates and type strain, while no reaction occurred with Mycoplasma capricolumsubsp.capripneumoniae, Mycoplasma mycoides subsp.capri, Mycoplasma bovirhinis, Mycoplasma agalactiae, Mycoplasma bovis, Mycoplasma leachii and bovine Pasteurella type A. Isotype analysis showed that MAb 3C4A1 belonged to IgG1, and the light chains belonged to κ. The ascites titer was 1:256 000. IFA results showed that MAb 3C4A1 only reacted with EBL cells infected with Mmm, while did not react with Mycoplasma bovirhinis, Mycoplasma agalactiae and Mycoplasma bovis, showing excellent specificity. MAb 3C4A1 showed excellent specificity and immunoreactivity, and could be used as a valuable tool for immunological diagnosis of CBPP pathogens, which provides basic material for further development of differential diagnostic techniques of CBPP pathogens.

Toxoplasma gondii and Neospora caninum are closely related Apicomplexa parasites. The identified cross-reacting antigens may account for the cross-protection between them. This study aimed to express and identify the cross-reacting antigen TgMIC17A to evaluate the cross immune-protective effect against T. gondii and N. caninum infection in mice. First, TgMIC17A was cloned and expressed in E. coli(DE3), and its cross reactivity and immunogenicity were analyzed by Western blot and ELISA, respectively. Then, mice were immunized with recombinant protein, and challenged with 1×10³ tachyzoites of T. gondii Pru or 1.5×10⁷ N. caninum Nc1 tachyzoites 10 days post the second immunization. The body weight change and survival rate of mice were recorded, and the parasite load in the brain was measured 30 days post-infection to assess the cross-protective efficacies of rTgMIC17A against T. gondii and N. caninum. The results indicated that recombinant TgMIC17A could be recognized by both positive sera of T. gondii and N. caninum. Compared with the unimmunized group, the immunized mice produced high level of specific IgG antibodies (P<0.01), and the amount of T. gondii or N. caninum load in brain significantly decreased as well (P<0.01). In summary, TgMIC17A was identified as a cross-reacting antigen between T. gondii and N. caninum, which can induce robust immune response to protect mice against T. gondii and N. caninum. Taken together, TgMIC17A is a potent cross-protective gene for use as recombinant vaccine against T. gondii or N. caninum infection.

The present study aimed to explore the effect of histone deacetylase(HDAC) inhibitors on peste des petits ruminants virus (PPRV) replication, and to determine the role of HDACs involved in the PPRV replication.The mRNA expression of HDACs (HDAC1-11) was examined by RT-qPCR assay in PPRV-infected Vero cells. To screen the inhibitors that affect PPRV replication, the expression level of N protein was evaluated in infected Vero cells by Western blot following treatment with the various inhibitors for different types of HDACs for 48 h. The selected inhibitors were used to treat PPRV-infected Vero cells, and the effects of inhibitors on PPRV N RNA, N protein and virus titer were further analyzed by RT-qPCR, Western blot and TCID₅₀, respectively. The results showed that PPRV infection in Vero cells resulted in a significant increase of the mRNA expression level of HDAC2 (P<0.05). After treatment with SAHA, TMP269 and MGCD0103, the expression level of PPRV N protein was significantly reduced and negatively correlated with the doses of these inhibitors. SAHA, TMP269 and MGCD0103 also significantly reduced the expression level of PPRV N RNA (P<0.05, P<0.05, P<0.01) and virus titer (P<0.05, P<0.05, P<0.01), respectively. These results indicated that class IIa HDACs inhibitors and class I HDACs inhibitors significantly inhibited PPRV replication in Vero cells, suggesting that class I HDACs (HDAC1-3) and class IIa HDACs (HDAC4-5, HDAC7, HDAC9) may be involved in the regulation of PPRV infection.

Microencapsulated SP4 phage was prepared by using cationic etherified starch/sodium alginate/xanthan gum/nano TiO₂/chitosan oligosaccharide as raw materials, to provide biological agents for controlling Salmonella infection. Salmonella SP4 phage was used to prepare microencapsulated SP4 phage by droplet method, and its titer, cationic etherified starch concentration, sodium alginate concentration, xanthan gum concentration, chitosan oligosaccharide concentration, pH stability, thermal stability, simulated gastric juice stability, simulated intestinal juice release behavior and preservation stability were analyzed. The results showed that when the concentration of cationic etherified starch was 2.4%, the concentration of sodium alginate was 2%, the concentration of xanthan gum was 1%, and the concentration of chitosan oligosaccharide was 0.6%, the shape of microencapsulated SP4 phage microspheres was regular, and the encapsulation efficiency was 97.5%. The phage maintained high biological activity in the range of pH 5.0-8.0, temperature 10-30℃, simulated gastric juice pH 2.0 for 0-30 min and pH 3.0 for 0-150 min. The phage was completely released after 4 h in simulated intestinal juice, and the phage titer did not decrease significantly after 6 weeks at 4℃. The results show that microencapsulated Salmonella SP4 phage significantly improves the stability and sustained-release performance of phage, its acid resistance is conducive to resisting the decomposition of gastric acid, and has the potential to be used in biological control, which lays a foundation for the further identification of microencapsulated Salmonella SP4 phage by multi factor test.

Duck tembusu virus (DTMUV) infection is a disease that causes a decline in laying ducks and brings great economic losses to the poultry breeding industry. In this study, the virus location and pathological damage of DTMUV-infected duck ovaries were observed to understand the effect of DTMUV on ovarian damage. Pathological necropsy, routine paraffin sections, HE staining, and immunohistochemistry were used to study diseased ducks naturally infected with DTMUV. The clinical manifestations of laying ducks were mainly characterized by a sudden drop in egg production. In severe cases, they could not stand steady, fell to the ground and convulsed, and finally died of exhaustion. Necropsy showed that follicles were hemorrhage and rupture. Histopathological changes were mainly the proliferation, apoptosis or necrosis of granulosa cells in growing follicles and mature follicles; there were a large number of vacuolar cells and small arteries around the follicles. Arterial smooth muscle cell necrosis and a large number of inflammatory cells infiltrated; immunohistochemical examination results showed that the proliferating vacuolar cells are epithelial cells, and DTMUV is mainly located in the granular cells of the follicles. According to the observation results, DTMUV is mainly distributed in the granulosa cells. The granulosa cells proliferate, apoptosis or necrosis, which leads to follicle atresia, and causes the egg production of laying ducks to decrease. It provided a theoretical basis for elucidating the pathogenic mechanism of DTMUV infection.

The aim of this study was to explore the protective effect and possible mechanism of aqueous extract of Yinshanlian (YSL) on acute liver injury induced by carbon tetrachloride (CCl₄) in mice, so as to provide theoretical guidance for further research and clinical transformation of YSL. Sixty ICR mice were randomly divided into 6 groups (n=10):control group, model group, low (1.17 mg·g^-1), medium (2.34 mg·g^-1) and high (4.68 mg·g^-1) YSL dosage groups. The drugs were administered by gavage once a day for 7 days. One hour after the last administration, except the control group, the mice in other groups were injected intraperitoneally with 0.2% CCl₄ to establish the model of acute liver injury. After 12 hours, blood was collected from mouse eyeballs, serum was isolated, and liver tissue was collected. The serum total protein (TP) and activity of glutathione S transferase (GST), as well as the activity of GST, succinate dehydrogenase (SDH), catalase (CAT) and total superoxide dismutase (T-SOD) in liver tissue were detected by biochemical analysis. The liver pathology and liver fibrosis were observed by HE staining and Masson staining. Expression and localization of IL-1β, tumor necrosis factor alpha (TNF-α), angiogenesis factor A (VEGFA), matrix metalloproteinase 9 (MMP9) and transcription factor NF-κBp65, protein kinase IKK, and p38MAPK in liver tissue were detected by Western blot and immunohistochemistry. The results showed that 2.34 mg·g^-1 YSL aqueous extract could significantly reduce the level of serum GST and significantly increase the level of intrahepatic CAT compared with the model group. There was no significant difference in T-SOD among these groups. There was no significant difference in the levels of liver GST, SDH and serum TP between 2.34 mg·g^-1 YSL aqueous extract group and control group or model group. Besides, 2.34 mg·g^-1 YSL aqueous extract could alleviate liver pathological injury and fibrosis induced by CCl₄, and decrease the expression of liver IL-1β, TNF-α, VEGFA, NF-κBp65 and IKK. It is suggested that the aqueous extract of YSL probably inhibits liver oxidative stress and inflammation through NF-κB and p38MAPK pathways to alleviate acute liver injury in mice.

The aim of this study was to investigate the antioxidant effect of artesunate (ART) on acute kidney injury induced by gentamicin (GM) in dogs and its mechanism. Twenty dogs were randomly divided into 4 groups:control group (Control), gentamicin model group (GM), artesunate group (GM+ART), artesunate + ML385 group (GM+ART + ML385). In addition to the control group, other groups were injected with GM to establish AKI model. After successful modeling, Dogs in GM+ART group were given ART, and GM+ART+ML385 group were given ART and ML385 at the same time. The control group and GM group were given the same volume of normal saline. The test period is 12 days. Different concentrations of GM were co-cultured with MDCK cells, and the optimal concentration was 4.0 mmol·L^-1. The optimum concentration of ART was determined to be 50.0 μ mol·L^-1 by the same method. The MDCK cells, MDCK cells overexpressing kelch like ECH related protein 1 (Keap1) (M-K) and MDCK cells knockdown nuclear factor E2 related factor 2 (Nrf2) (M-SiNrf2) were divided into three groups respectively:Healthy control group, GM control group and GM + ART intervention group, co-culturing for 24 hours. The results show as follows:1) In animal experiments, the levels of Cr, UN and KIM-1 in GM + ART group were significantly lower than those in GM Group, and the levels of Cr, UN and KIM-1 in GM + ART + ML385 group were significantly higher than those in GM + ART group. 2) In animal experiments, compared with GM Group, the expression of Nrf2 and glutathione cysteine ligase catalytic subunit (GCLC) protein was up-regulated, Keap1 protein was down-regulated, the levels of total superoxide dismutase (T-SOD) and glutathione (GSH) in renal homogenate were increased, and the level of malondialdehyde (MDA) was decreased in GM + ARTgroup. Compared with GM + ART group, the expression of Nrf2 and GCLC decreased, and the levels of T-SOD and GSH decreased after intervening by ML385. 3) In the cell test, compared with MDCK cells co cultured with 4.0 mmol·L^-1 GM, the dose of 50.0 μmol·L^-1 ART added could significantly increase the cell proliferation rate, reduce the level of ROS, down regulate the expression of Keap1 and up regulate the expression of Nrf2, heme oxygenase 1 (HO1) and GCLC. 4) In the cell test, compared with MDCK cells, the expression of Keap1 was up-regulated, the expression of Nrf2, HO1 and GCLC were down regulated, the cell proliferation rate was decreased, and the content of ROS was increased both in M-K cells and M-SiNrf2 cells under the same treatment of GM + ART(P<0.05 or P<0.01). In conclusion, artesunate has a protective effect on acute renal injury induced by gentamicin in dogs, and its mechanism is related to inhibit oxidative stress via Keap1/Nrf2 signaling pathway.

Echinacea is an important immune enhancer, which is often used as a substitute for antibiotic as feed additive, however, its immune mechanism needs further study. The purpose of this study was to investigate the effects of Echinacea on enhancing immune function of immunosuppressed chicken by regulating TLR4-NF-κB pathway. A total of 120 7-day-old chicken were randomly divided into 4 groups:the control group, cyclophosphamide group, Echinacea group and Echinacea + cyclophosphamide group (combined group), respectively. The chickens of cyclophosphamide group and combined group were chest muscles injected with saline compound cyclophosphamide solution (80 mg·kg^-1) for 3 consecutive days,while the control group and Echinacea group were injected with the same amount of normal saline for 3 days. The chickens of Echinacea and combined groups were fed with basal diet which containing Echinacea herb powder (1%) daily, and the rest of the chickens were fed with the basal diet. All the chikens were weighed and recorded every day. The experiment lasted for 21 days. At the end of the experiment, the changes of body weight and spleen index were analyzed, and the mRNA and protein expression levels of inflammatory factors (NF-κB, IκB, IL-2, IL-4, IL-6, TNF-α, IFN-γ) and TLR family (TLR2, TLR4, TLR7, MyD88) were detected. The results showed that Echinacea improved the growth performance of chickens, while cyclophosphamide inhibited the growth of chickens. The spleen coefficient of chickens in cyclophosphamide group was significantly lower than that in other groups (P<0.05), while the pathological changes of spleen tissue sections in the cyclophosphamide group were obvious, and the pathological changes in the combined group were relieved. Compared with the control group, the mRNA levels of NF-κB, IL-2, IL-4, IL-6, IFN-γ, TLR2, TLR4, TLR7 and MyD88 in cyclophosphamide group were significantly decreased to varying degrees (P<0.05, P<0.01 or P<0.001). Compared with the cyclophosphamide group, the above genes(except TLR7) significantly increased in the combined group(P<0.05, P<0.01 or P<0.001). In addition, the supplementation of Echinacea significantly increased the expression levels of inflammatory factors and TLR family-related proteins. The results indicated that Echinacea enhanced the immune function of immunosuppressed chickens by regulating TLR4-NF-κB pathway.

The differences in the feces and plasma metabolites of Kunming mice before and after matrine administration were analyzed using the untargeted metabolomics method. The potential mechanisms of matrine on pharmacological actions were explored through the combined analysis of metabolomics and microbiome. Twenty Kunming mice were randomly divided into two groups, named matrine treatment group (MT) and normal saline treatment group (NC). In the MT, 40 mg·kg^-1 of matrine was injected intraperitoneally to mice twice a day for 5 days. In the NC, saline was injected intraperitoneally with the same volume and method. On the 6th day of administration, the feces and plasma of each group of mice were collected for the untargeted metabolomics analysis and the 16S rDNA sequencing results obtained in the preliminary study were co-analyzed. The significant differential metabolites were obtained both in feces and plasma in matrine treatment group, and 97 metabolites in feces. while 44 in plasma. Cluster analysis showed that in matrine treatment group, 35 metabolites were up-regulated and 104 metabolites were down-regulated in feces, while 20 metabolites were up-regulated and 35 metabolites were down-regulated in plasma. KEGG analysis showed that the differential metabolites were mapped to metabolic pathways such as protein digestion and absorption. Moreover, there was a correlation between fecal and plasma different metabolites and Lactobacillus acidophilus obtained by 16S rDNA sequencing. Matrine regulated the metabolism in Kunming mice, and differential metabolites were obtained both in feces and plasma. These differential metabolites and their interactions with the microflora might be the key to the pharmacological action.

The study aimed to explore the effects of chaetocin on histone methylation modification in adipose-derived stem cells (gADSCs) of cashmere goat. In this study, gADSCs were treated with different concentrations of chaetocin for different durations, and the concentration and duration of chaetocin treatment that had the least effects on cell viability and the greatest inhibition on histone methylation in gADSCs were screened. The optimal concentration and duration of chaetocin treatment was used as the experimental group, and the chaetocin-free treatment was used as the control group. The expression of H3K9 me2 and H3K9 me3 methylation-related enzymes and embryonic development pluripotency genes were detected by RT-qPCR, and the effects of chaetocin on histone H3K9 me2 and H3K9 me3 proteins expression were observed as well. The results showed that there was little effect on cell viability when gADSCs were treated with 20 nmol·L^-1 chaetin for 48 h, and the expression of histone methyltransferase G9A significantly reduced (P<0.05), the optimal chaetin treatment concentration and duration was confirmed. RT-qPCR results showed that the expression of H3K9 me2 and H3K9 me3 methyltransferase EHMT1, EHMT2, SUV39H1 and SUV39H2 significantly reduced(P<0.05), and the expression of H3K9 me2 and H3K9 me3 demethylase KDM3A, KDM3B, KDM4B and KDM 4Dsignificantly increased(P<0.05),the expression of multipotency-related genes SOX2, OCT4, NANOG increased in experimental group. The results of immunofluorescence and WB experiments showed that the H3K9 me2 protein level did not significantly change, while the H3K9 me3 protein level significantly reduced(P<0.05) in the experimental group. The modification of histone H3K9 me2 and H3K9 me3 in gADSCs treated with 20 nmol·L^-1 chaetocin for 48 h could be reduced significantly, and the expression of multipotency-related genes could be increased.

The aim of this study was to construct of Felis catus C11orf96 (fC11orf96) gene, prepare polyclonal antibody with good reactivity to the protein and in order to analyze the localization of the C11orf96 gene in cells. Firstly, the fC11orf96 gene cloned by using Crandell Reese Feline Kidney cells (CRFK) cDNA as the template, the recombinant plasmid pET-32a (+)-fC11orf96-Fe was successfully constructed by seamless recombination. Then, the recombinant expression vector was transformed into BL21(DE3), and the recombinant protein fC11orf96-Fe was successfully expressed after IPTG induction. The fC11orf96-Fe protein was used to immunize rabbits to prepare polyclonal antibody. Finally, Western blot and indirect immunofluorescence assay (IFA) were used to analyze the validity and subcellular localization of polyclonal antibody. The results showed that the CDS sequence of the fC11orf96 gene was 372 bp, encoding 124 amino acids. The recombinant protein fC11orf96-Fe exists mainly in inclusion bodies, and the protein size was 49 ku. The polyclonal antibody prepared from the recombinant protein could identify endogenous fC11orf96 protein and eukaryotic expressed foreign protein. And we found that fC11orf96 protein was localized in the cytoplasm. In this study, the fC11orf96 gene was successfully cloned and localized in the cytoplasm, which laid a foundation for further research on the biological functions of the C11orf96 protein.

This research aimed to clarify the effect of ubiquitin-specific protease (USP7) on the resistance of chickens to Salmonella Typhimurium infection. Forty 3-day-old White Leghorns were randomly divided into two groups, and respectively intraperitoneal injected with the USP7 inhibitor P5091 and control solvent every other day for 5 consecutive injections (n=20 in each group). Four days after the last injection, they were artificially orally infected with S.Typhimurium. Samples were collected 24 hours after infection. The mRNA abundances of IL-1β, IL-8, TNF-α and NF-κB were examined by RT-qPCR. The bacteria load in the liver of chickens was measured by the plate coating method. Liver and lung inflammation was checked by H&E staining. Inhibition of the USP7 activity by P5091 treatment significantly downregulated the mRNA level of NF-κB, and its downstream cytokines IL-1β, IL-8 and TNF-α. The bacterial load in the liver of chickens decreased. H&E staining showed aggravated inflammation in the liver and lung in USP7 inhibited chickens. USP7 might have critical role in chicken to resist Salmonella infection.