Spermatozoa that produced in the testis still need to undergo chemical modification, morphological and functional changes during epididymal transition to gain the motility and fertility capacity, which is named sperm maturation. Compared with mammals, the epididymis of poultry is somewhat degenerated. In recent years, more studies have proven that the epididymis has a certain regulatory effect on sperm maturation. In the livestock and poultry breeding and propagation system, male semen quality directly affects the mating ratio and related with the propagation efficiency and benefit.Therefore, exploring the regulatory mechanism underlying sperm function and semen quality has important application value for improving male sterility, establishing breeding methods and technique for improved reproduction performance. This article reviews the research progress of epididymis function and the mechanism of post-testicular sperm maturation in poultry, aiming at inspiring future research on this topic.

The yak is widely distributed in the plateau area with an altitude of 3000-5000 m, and is an important animal species for the development of the local animal husbandry economy. After a long-term adaptive evolution, the yak showed high-altitude adaptation characteristics in terms of physiological structure and genetic molecules. With the wide application of omics technology, the plateau adaptation mechanism of the yak has been further revealed. In this paper, the physiological structure and molecular mechanism of lung and heart in high altitude hypoxia adaptation, the periodic regulation of coat and the molecular mechanism of fat deposition and metabolism in cold adaptation, the molecular basis of disease resistance, the physiological basis and molecular mechanism of reproduction in the yak were reviewed and outlook based on relevant reports at home and abroad.

Deoxynivalenol (DON), is one of the common mycotoxins that contaminate grain, feed and food, which seriously affects the health of humans and livestock. Pigs are very sensitive to DON, which can cause toxic effects such as antifeeding, growth inhibition, vomiting, organ damage, cytotoxicity, and immunotoxicity to pigs. The toxic effects, toxicological mechanisms and antidote development of DON on pigs were reviewed in this paper. Specially, the genetic regulation of the toxicological effects of DON was concerned, which aimed to provide more comprehensive references for improving the resistance of pigs to deoxynivalenol from genetic nature.

Meat color is an important organoleptic indicator of meat quality and directly affects consumers’ purchase intention. Plant polyphenols are natural antioxidants and their addition to diets can improve meat color and its stability by affecting the redox status of myoglobin, the degree of lipid oxidation and the type of muscle fibres. To further clarify the mechanism of dietary plant polyphenols addition in regulating meat color, this paper reviews the signaling pathways mediated by plant polyphenols for meat color stability, with a view to providing a theoretical basis for the future regulation of the body’s antioxidant capacity by supplementing plant polyphenols to the diets and promoting the transformation of muscle fibre type to improve meat color.

Mitochondria are the primary energy-generating sites in animal cells, where they can participate in the production of triphosphate and the maintenance of a constant state of mitochondrial Ca²⁺, which is crucial for animal health and energy metabolism. Studies have found that gut microflora and their metabolites can mediate the level and function of mitochondrial metabolism, which then affects the growth and development of the turnover of nutrient metabolism, feed conversion efficiency, etc. Based on the summary of the biological function of mitochondria, this review focuses on the influence of gut microflora and their metabolites on mitochondrial activity and the influencing factors, aiming to provide a theoretical reference for regulating growth and intestinal health of animals by the dietary nutrition-mediated intestinal microbial-host mitochondrial pathway.

In cattle, delayed postpartum uterine involution prolongs the reproductive interval and reduces reproductive rates. Uterine involution involves postpartum uterine tissue degeneration, repair, and remodeling of the myometrium. Stem cells, cytokines, non-coding RNAs, estrogen, and progesterone all participate in controlling postpartum uterine tissue repair and regeneration. At the same time, green and non-polluting prevention and treatment techniques such as vitamins and glucose-containing compounds, traditional Chinese medicine preparations, and immunomodulatory chemicals can successfully encourage postpartum uterine involution in cattle. In this article, the latest research progress on the regulatory mechanism of postpartum uterine involution and methods for promoting uterine involution is summarized, in the hope of providing some references for studying the molecular regulatory mechanism of animal postpartum uterine involution and creating new techniques for promoting uterine involution in cattle.

Protozoa have complex life cycle and most of them can not complete the life cycle just in vitro. As a result, most of them lack an effective gene editing system. The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (endonuclease) system facilitates the precise, efficient, and rapid genetic editing through homologous recombination, and is widely used in genetic modification of eukaryotic organisms. The CRISPR/Cas9 system opens up a new avenue for targeted investigation of gene functions in protozoans. This article introduces the principle of the CRISPR/Cas9 system and its major applications in gene tagging and depletion and genome-wide screening of functional gene families in parasitic protozoan.

Hypertrophic cardiomyopathy is one of the most common primary heart diseases in cats, which is characterized by left ventricular hypertrophy. Myocardial fibrosis is a cardinal pathological change in cats with hypertrophic cardiomyopathy, which can lead to cardiac dysfunction and rhythm disturbance, and it is an important factor contributing to poor prognosis of cats with cardiomyopathy. There is an urgent need to develop novel drugs targeting cat hypertrophic cardiomyopathy and cardiac fibrosis. This review summarizes the pathological characteristics of cat hypertrophic cardiomyopathy as well as most updated research progress on the pathogenesis of cat myocardial fibrosis, aiming to find a breakthrough point for new therapeutic drug development for cat hypertrophic cardiomyopathy.

The purpose of this study was to identify the genetic loci that affected the body conformation at birth and teat number traits, develop molecular markers that can be used for assisted breeding, and provide a theoretical basis for the continuous selection of growth and reproductive performance in Sushan pig population. The phenotype included body conformation at birth and teat number were measured in 269 Sushan newborn piglets (within 24 h after birth), and the ear tissue samples were collected at the same time. Based on whole-genome resequencing and genotype imputation strategy, GWAS and Fst methods were used to identify the significant loci associated with the target traits. GO and KEGG analyses were performed on the positional and functional candidate genes to identify the most likely major genes. A total of 4 candidate loci were identified in this study, located in the 13, 14 and X chromosomes, of which 2 were significantly associated with body conformation at birth traits, and 2 were significantly associated with the teat number traits. This study screened out 1 candidate gene ACTA2 related to the body length, 1 candidate gene COL4A6 related to the chest circumference, and 3 candidate genes ACTA2, CRTAP, and SEPTIN6 related to the teat number. This research provides important molecular markers for the genetic improvement of body conformation at birth and teat number traits in Sushan pig population.

The aim of this study was to identify genetic variant loci and genes which associated with reproductive traits at first farrowing in Rongchang pigs, and provide important molecular markers and genes for improving pig reproductive traits in Rongchang pigs. In this study, a total of 429 Rongchang sows were genotyped using porcine 50K SNP chip. After quality control, 35 046 SNPs were generated for subsequent genome-wide association study (GWAS). Then, the population structure was analyzed using principal component analysis, and GWAS was performed for total number born (TNB), total number born alive (NBA), number of stillborn (NS) and litter weight born alive (LWB) using a mixed-linear model (MLM). The birth year and birth month were used as fixed effects, and principal component values were used as a covariate in this model. The results showed that a total of 2 and 1 genome-wide significant SNPs were detected for LWB and NS, respectively. Furthermore, a total of 5, 3 and 10 suggestive SNPs were found for TNB, NBA and NS, respectively. Of these, one locus located on SSC17 (position: 57 315 180 bp) were evaluated to affect TNB, NBA and LWB and one locus located on SSC1 (position: 279 214 647 bp) were evaluated to affect both NBA and TNB, indicating the pleiotropism of genes in different reproductive traits. Considering the function of candidate genes, BMP7 gene was suggested as the promising candidate genes for TNB, NBA and LWB, and two genes(MSH3 and CBLB) were suggested as the important candidate genes for NS. These results provided novel variant loci and potential candidate genes for reproductive traits, and provided important theoretical basis for improving reproductive traits using genomic selection in Rongchang pigs.

The aim of this study was to investigate the association between polymorphisms of 9 genes (HoxB1-7,9 and 13) in HoxB cluster and vertebral number and carcass traits (carcass length, straight length, diagonal length and carcass weight) in Beijing black pigs. In this study, DNA was extracted from ear tissue of 251 healthy Beijing black pigs at the age of 220 days. The SNPs of 9 HoxB genes were genotyped by primer design, polymerase chain reaction (PCR) and Sanger sequencing, and the genotype frequency and allele frequency distribution of the mutated loci were calculated. The association analysis was performed between their polymorphism and the vertebral number and carcass traits. A total of 12 non-coding region (UTR) SNPs were detected in 4 genes, including one 5'UTR mutation of HoxB2, two 3'UTR mutation of HoxB5, two 3'UTR mutation of HoxB9 and seven 3'UTR mutation of HoxB13. Statistical analysis using Duncan’s multiple test revealed that a 5'UTR mutation of HoxB2 (c.40 C>T) and two 3'UTR mutations of HoxB5(c.1766 A>G and c.1767 A>G) associated with these traits indistinctively (P>0.05). A 3'UTR mutation of HoxB9 (c.2743 C>T) were significantly associated with carcass traits(P<0.05), and two 3'UTR mutations of HoxB13 (c.1863 G>A, c.1440 A>C) significantly associated with the vertebral number (P<0.05). In conclusion, among the 9 HoxB cluster genes, only one 3'UTR mutation (c.2743 C>T) in HoxB9 gene was significantly associated with carcass traits (P<0.05), two 3'UTR mutations (c.1863 G>A, c.1440 A>C) of HoxB13 gene were significantly associated with the vertebral number (P<0.05). Above the 3 SNPs could be used as candidate functional loci for the variation of vertebral number and carcass traits of Beijing black pigs.

The purpose of the study was to explore the relationship between VGLL2-AS and VGLL2 in chicken skeletal muscle. PCR and sequencing techniques were used to verify the existence of VGLL2-AS; The tissue samples of Gushi layers at different weeks of age (1 day old, 6 weeks old, 16 weeks old, 22 weeks old and 30 weeks old, 6 for per week-old) were collected respectively, and the expression profiles of VGLL2 gene and its antisense chain transcript VGLL2-AS were analyzed by real-time quantitative PCR. Bidirectional transcription test and nuclease protection test were used to detect whether VGLL2 and VGLL2-AS could be transcribed in two directions and the relationship between them. Then, the primary myoblasts of 11 embryo-old Gushi chicken embryos were isolated and cultured in vitro, and then treated with 2 μg·mL^-1 actinomycin D (the control group was not treated), and the cells were collected at different time points (0-8 h, 3 repeats in each time point). The half-life of VGLL2-AS and VGLL2 were measured. The nucleus and cytoplasm of chicken myoblasts were isolated, and their cellular localization was determined by real-time quantitative PCR. The transcripts of VGLL2 were verified by PCR amplification and sequencing; Finally, RT-qPCR was used to detect their spatiotemporal expression and correlation in the breast muscle tissues of Gushi layers (6 birds at 1 day, 6 weeks, 16 weeks, 22 weeks and 30 weeks, respectively). The results showed that VGLL2-AS existed in the chicken transcriptome. VGLL2 and VGLL2-AS were highly expressed in muscle (P<0.05). VGLL2-AS and VGLL2 could conduct bidirectional transcription and form double stranded RNA between them. VGLL2-AS had a longer half-life than VGLL2; In myoblasts, VGLL2-AS and VGLL2 were mainly located in the cytoplasm (P<0.001). VGLL2 only contained VGLL2-mRNA, VGLL2-X2 and VGLL2-X3 transcripts. The expression trend of VGLL2-mRNA and VGLL2-X3 was consistent with that of VGLL2-AS, and the expression of VGLL2-mRNA, VGLL2-X3 and VGLL2-AS showed a strong positive correlation (r=0.943 and 0.935, respectively, P<0.01). To sum up, VGLL2-AS, as the antisense chain encoding lncRNA of VGLL2, is located in the cytoplasm, possibly through the interaction with the double stranded RNA formed by VGLL2, and then participates in regulating the expression of VGLL2 and maintaining its stability, finally playing an important role in the early muscle development of chickens. The results of this study expand the research on NATs in chickens, and lay a foundation for the study of biological function of chicken VGLL2 gene and its natural antisense chain transcript VGLL2-AS in chicken skeletal muscle development, which has certain significance for improving the growth and development of poultry.

This study aimed to clone the coding sequence (CDS) of the nuclear receptor subfamily 1 group D member 1 gene (NR1D1) in dairy cows, construct its eukaryotic expression vector, predict and analyze the biological function of the NR1D1 gene, detect the tissue expression profiles of the gene and its expression distribution in ovary. In this study, 5 healthy female dairy cows (24 months old) were used as experimental animals. The full-length fragment of the CDS region in dairy cow NR1D1 gene with homologous arms was amplified by PCR, and the target fragment was ligated to the linearized pcDNA3.1-Puro-N-3HA vector by homologous recombination to construct a recombinant plasmid. Recombinant plasmids were identified by enzyme digestion, PCR and sequencing techniques. Combined with the results of plasmid sequencing, bioinformatics softwares were used to predict and analyze the biological function of NR1D1. The correctly identified plasmid was transfected into HEK293T cells, and the expression of NR1D1 gene at mRNA and protein levels was detected by real-time quantitative PCR (qPCR) and Western blotting. In addition, qPCR was used to detect the expression profile of dairy cow NR1D1 gene in different tissues, and immunohistochemistry was used to focus on its expression distribution in ovary. The full-length fragment of the CDS (1 884 bp) region of NR1D1 gene in dairy cow with homology arms was obtained by PCR amplification. The results of restriction enzyme digestion, PCR identification and sequencing showed that the eukaryotic expression vector pcDNA3.1-3HA-cNR1D1 was successfully constructed. The results of bioinformatics analysis showed that the NR1D1 protein had no transmembrane structure and was a nucleoprotein, its nuclear localization sequence existed near the 200th amino acid, and there were 86 potential phosphorylation sites in NR1D1 protein. The tertiary structure of cow and mouse NR1D1 protein was similar and the NR1D1 protein may play an important role in the physiological process of reproduction, metabolism and immunity in dairy cow. The pcDNA3.1-3HA-cNR1D1was transfected into HEK293T cells, and the cow NR1D1 gene was successfully overexpressed at both mRNA and protein levels. The qPCR results showed that the expression of NR1D1 gene in dairy cow’s rumen, colon and liver was significantly higher than other tissues. Immunohistochemical results showed that NR1D1 was mainly expressed in granulosa cells of follicles in different developmental stages. This study successfully constructed dairy cow NR1D1 gene’s eukaryotic expression vector, and successfully overexpressed NR1D1 in HEK293T cells. The tissue expression profile of NR1D1 and its expression distribution in the ovary of dairy cows were preliminarily predicted and analyzed. This study provided the preliminary basis and key materials for in-depth exploration of NR1D1’s biological function.

The objectives of the study were to obtain the sequence of yak FHL2 gene and to elucidate its sequence, protein structure and properties, mRNA expression in tissues, protein localization in the reproductive organs and granulosa cells of female yaks (Bos grunniens). Tissue samples of heart, liver, spleen, lung, kidney, ovaries, uterus and oviducts of 5 healthy non-pregnant adult female yaks and ovaries, uterus and oviducts of 5 pregnant female yaks were used as study materials. The FHL2 gene was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and its bioinformatics properties were analyzed using online softwares. mRNA expression levels in tissues were detected by a real-time quantitative PCR (RT-qPCR). The distribution of FHL2 protein in female reproductive organs and localization in granulosa cells was detected by immunohistochemistry (IHC) and immunofluorescence (IF) techniques. The results showed that the CDS region of FHL2 gene was 840 bp (ON456866) in length and encoded 279 amino acids. The FHL2 protein was a weakly basic hydrophilic unstable protein with no transmembrane structure. Based on amino acid sequence comparison and evolutionary tree results among multiple species, the coding region of the FHL2 gene was relatively conserved during evolution. RT-qPCR results showed that the FHL2 gene was expressed in all detected tissues, and the expression in the heart was significantly higher than in other tissues(P<0.01). Its expression levels in lung, ovary, oviducts and uterus were significantly higher than in liver, spleen and kidney (P<0.01). Its expression in the uterus during pregnancy was significantly higher than that in the non-pregnant period (P<0.01), but in the ovary its expression was significantly lower during pregnancy than in the non-pregnant period (P<0.01). IHC result showed that FHL2 protein was mainly expressed in follicular granulosa cells, endometrium and oviductal mucosa. IF result showed that FLH2 protein was mainly localized in nuclear of granulosa cells. In conclusion, the sequence of yak FHL2 gene is relatively conserved during animal evolution. It is highly expressed in heart, lung, ovaries, oviducts and uterus, indicating that it may be involved in the regulation of physiological functions such as hypoxia adaptation, follicular development and pregnancy maintenance in yak.

The study aimed to investigate the expression pattern and function of KRT16 during hair follicle development in Angora rabbit. In this experiment, 12 healthy 6-month-old Wan Strain Angora rabbits were selected for dorsal skin sampling during the anagen, catagen, and telogen after hair shearing. The coding sequence of the rabbit KRT16 gene was obtained by cloning, and the biological properties of the KRT16 coding sequence (CDS) were initially analyzed using bioinformatics softwares. Quantitative real-time PCR (qRT-PCR) was performed to analyze the expression of KRT16 in hair follicles at different times. Overexpression and knockdown of KRT16 in dermal papilla cells (DPC) were performed to investigate the regulatory role of KRT16 on genes related to hair follicle growth and development, and the effect on DPC proliferation. The results showed that the coding sequence of the KRT16 gene was 1 431 bp, could encode 476 amino acids, and showed high homology in different mammals. qRT-PCR results showed that KRT16 showed different expression levels during the period of hair follicle development, with high expression during the anagen phase. The changes in the expression levels of hair follicle development-related genes were examined by overexpressing and knocking down KRT16 in rabbit hair papilla cells. The mRNA expression of SFRP2 and TGFβ1 genes was highly significantly decreased (P<0.01), and the mRNA expression of BCL2, CCND1, EGF, LEF1, and CTNNB1 genes was highly significantly increased after overexpression of KRT16 (P<0.01); while the expression of SFRP2 and TGFβ1 genes was highly significantly increased (P<0.01), and the expression of BCL2, CCND1, EGF, LEF1, and CTNNB1 genes was highly significantly decreased (P<0.01) after knockdown of KRT16. Meanwhile, KRT16 was able to significantly promote the proliferation of hair papilla cells. In summary, it is showed that KRT16 is involved in the cyclic development of hair follicles and can regulate hair follicle growth-and development-related genes to promote the proliferation of hair papilla cells. This study provides a reference for the basic research of hair follicle development through the preliminary study of KRT16 gene function and also provides a basis for the functional study of hair follicle development-related genes in Angora rabbits.

The study aimed to establish a system for isolation and culture of epidermal stem cells (EpSCs) of Simmental cattle, study its biological characteristics, explore a new method for the conservation of Simmental germplasm resources, and provide seed cells for stem cell therapy research. The EpSCs were isolated from the dorsal epidermis of 3-to 4-month-old Simmental cattle by enzyme digestion method and tissue adhesion method. Simmental cattle EpSCs were then cultured to draw the growth curve and to further investigate the proliferation ability. EpSCs surface markers (ITG β1, P63 and CD71) were identified by immunofluorescence, and the expression analysis of specific genes (ITG β1, KRT19 and ITG α6) were performed by RT-PCR. Simmental cattle EpSCs were induced to differentiate into adipocytes, osteoblasts and chondrocytes in vitro for detecting their differentiation potential. The results showed that the purity of EpSCs obtained by tissue adhesion method was much lower than that obtained by enzyme digestion method, and they began to age gradually when passed to P28 generation. The growth curve of EpSCs was shaped as “S”. Immunofluorescence result showed that positive expression of surface specific markers in EpSCs such as ITG β1, P63 and CD71, RT-PCR showed that EpSCs highly expressed gene ITG β1, KRT19 and ITG α6 but CD31. When induced to adipocytes, EpSCs produced lipid droplets that could be stained by Oil red O and positively expressed specific genes of adipocyte PPAR-γ and LPL. When induced to osteoblasts, EpSCs produced calcium nodules that could be stained as orange red by Alizarin red, and positively expressedgenes such as Collagen Ⅰ and RUNX2. When induced to chondrocytes, they produced hyaline cartilage matrix that could be stained as blue by Alcian blue and positively expressed specific genes BGN and Collagen Ⅱ of chondrocytes. In this research, Simmental cattle EpSCs in good condition were successfully isolated from epidermis of fetal bovine with the potential to differentiate into osteogenesis, cartilage and adipose cells, the isolation and culture system in vitro for Simmental EpSCs was established.

The high lipid content of porcine oocytes is considered one of the most important factors for its low freezing efficiency. The purpose of this study was to evaluate the effect of chemical lipid-lowering agent forskolin on the lipid degradation and anti-freezing ability of porcine oocytes during in vitro maturation. The content and ultrastructural changes of lipid droplets, mitochondrial membrane potential, reactive oxygen species level, early apoptosis rate, survival rate and developmental potential of matured porcine oocytes with or without forskolin treatment were detected before and after vitrification. The results showed that forskolin treatment partially improve the maturation rate of oocytes, but the difference was not significant (P>0.05). The results of nile red fluorescence staining showed that the number and area of lipid droplets before and after vitrification in forskolin-treated oocytes were significantly lower than those untreated (P<0.01). Ultrastructural observation showed that the ratio of the number of heterogeneous lipid droplets to homogeneous lipid droplets in the oocytes treated with forskolin increased, and the area of heterogeneous lipid droplets was larger than that of homogeneous lipid droplets. The heterogeneous lipid droplets were irregularly distributed in oocytes after vitrification as the main lipid droplets form, of which became smaller and less in number. After forskolin treatment, the proportion of lipid droplets between heterogeneous and homogeneous further increased. Forskolin treatment also significantly increased the mitochondrial membrane potential of vitrified oocytes (1.04 vs. 0.51, P<0.01), alleviated oxidative stress and reduced early apoptosis rate (68.30% vs. 86.03%, P<0.01), which effectively improved the survival rate of vitrified oocytes (71.17% vs. 51.47%, P<0.01) and its cleavage rate after parthenogenetic activation (16.63 vs. 8.23%, P<0.05). In conclusion, the addition of forskolin during the in vitro maturation of porcine oocytes effectively reduced the content of intracellular lipids, and improved the survival rate and development ability after vitrification through alleviating oxidative damage and improving mitochondrial function.

The aim of this study was to analyze the expression patterns and function of the effector gene YAP1 in Hippo signaling pathway in endometrium of Hu sheep with different fertility. Based on the genealogical archives and polymorphism analysis of BMPR-1B, a total of 9 healthy pluriparous ewes were divided into 3 groups (HBB, LBB and LB+, n=3). RNA was extracted from endometrial tissues of Hu sheep with different fertility, and reversed transcript into cDNA. Real-time quantitative PCR (RT-qPCR) was used to analyze the expression of several major genes in the Hippo signaling pathway in endometrium of Hu sheep with different fertility. The amino acid homology and genetic relationship of YAP1 protein in different species were analyzed by bioinformatics methods. Hu sheep endometrial stromal cells (ESCs) were isolated in vitro, and the expression of YAP1 was downregulated by using RNAi and cell transfection techniques in vitro. The effect of YAP1 gene on the expression of uterine receptivity related genes was analyzed by RT-qPCR. Moreover, the effect of interfering YAP1 on cell apoptosis was analyzed by flow cytometry, and the expression of apoptosis and mitochondria related genes was analyzed by RT-qPCR and Western blot. The expression levels of YAP1, LATS1 and MST1 in Hippo signaling pathway were different in endometrium tissues of Hu sheep with different fertility, and the expression level of YAP1 gene in high fertility Hu sheep (HBB) endometrium was significantly higher than that in low fertility Hu sheep (LB+, LBB) endometrium. Amino acid homology and phylogenetic tree analysis showed that YAP1 protein of goat and cattle had the highest homology and the closest genetic relationship with sheep. The mRNA expression levels of receptivity related genes (FOXO1, PRL, VEGF, OPN, COX-2) in ESCs were significantly changed after interfering YAP1. Meanwhile, the apoptosis rate of ESCs was significantly increased, the expression levels of Bax, BIM, p53, caspase3, caspase8, caspase9, the ratio of Bax/Bcl-2 were upregulated, and expression levels of Bcl-2 was downregulated after YAP1 silencing. In addition, the mRNA expression levels of genes related to mitochondrial function (FIS1, DNM1L, MFN2, PPARGC1A, OPA1) were also significantly changed. YAP1 may establish good endometrial receptivity by inhibiting apoptosis and stabilizing mitochondrial function.

The purpose of this study was to obtain the sequence of the rabbit GnIH gene, detect its expression patterns in different tissues and different developmental stages in the hypothalamus, and investigate its effect on the secretion of reproductive hormones in young male rabbits. In this study, the 90-day-aged healthy male Minxinan black rabbits were used as experimental animals, the sequence of the GnIH gene was cloned by RACE (rapid-amplification of cDNA ends)and analyzed by bioinformatics methods; the relative expression levels of GnIH in different tissues of male rabbits aged 90 days (n=5) and in the hypothalamus aged 11, 30, 60, 90, 120 and 150 days (n=6) were determined by real-time quantitative PCR (qPCR); the male rabbits aged 80 days were chosen and intraperitoneally injected with 0, 0.5, 5 or 50 μg quail GnIH related peptides for 10 consecutive days, respectively (n=10). These rabbits were slaughtered and their testes were collected after ear vein blood sampling on the morning of the 11th day, and then, the concentrations of reproductive hormones (GnRH, FSH, LH, INHB, and T) in serums and the mRNA levels of testosterone synthesis related enzyme genes (StarR, 3β-HSD and p450scc) in testes were tested, respectively. A total length of 904 bp GnIH gene sequence was cloned successfully, including 41 bp of 5'UTR, 606 bp of CDS, and 227 bp of 3'UTR, encoding 201 amino acids. The GnIH gene expressed in most tissues and its mRNA level was higher in the hypothalamus than in other tissues (P<0.001); Aged from 11 to 150 days, its mRNA levels in hypothalamus reached the lowest point at 90 day-aged (P<0.01), and then significantly increased with increasing age(P<0.001). The young male rabbits were administrated at different doses of GnIH polypeptide. The results showed that the concentration of serum testosterone in group 50 μg was signicantly lower than those in control and 0.5 μg groups(P<0.001), the concentration of testosterone in group 5 μg was significantly lower than those both in control and 0.5 μg groups(P<0.05), and the concentration of LH in group 50 μg was significantly higher than that in control group (P<0.05); the mRNA level of p450scc gene in group 5 μg was significantly higher than those in the other groups(P<0.001), and there were no significant differences in the mRNA levels of other testosterone synthesis related enzyme genes among the GnIH administrating-groups and control group (P>0.05). These results indicated that the GnIH gene might be related to the synthesis and secretion of LH and testosterone, and participate in the regulation of reproductive development of male rabbits, chronic injection with a high dose of GnIH-related peptide leads to drug insensitivity of male rabbits, and the specific regulatory mechanism needs subsequent studies.

Animals acquire energy and nutrients mainly by feed intake. Exploring the effect of gut microbiota on pig feeding behaviors and investigating its potential mechanism would provide theoretical basis for improving pig feeding behaviors through regulating gut microbiota. A total of 210 commercial Duroc pigs were used in this study. Phenotypes relating to feeding behaviors, including average daily eating time (ADET), average daily eating visits (ADEV) and average daily feed intake (ADFI) were recorded by automatic feeders. Stool samples were collected from the anus at the age of 140 days, and the structure and composition of gut microbiota were obtained by sequencing V3-V4 region of 16S rRNA gene. The Two-part model and co-abundance groups (CAGs) were used to identify the gut microbial taxa associated with feeding behaviors of pigs. The results showed as follows: 1) Correlation analysis between phenotypes showed that ADET was significantly and positively correlated with ADEV (r=0.41, P<0.05) and RFI(r=0.32, P<0.05); ADFI was significantly and positively correlated with RFI (r=0.56, P<0.05) and average daily gain (ADG) (r=0.63, P<0.05). In addition, RFI was significantly positively correlated with backfat thickness (r=0.16, P<0.05). 2) In CAGs analysis, a significant negative correlation was found between CAG2 and ADFI (P<0.05). This CAG mainly included OTUs annotated to Ruminococcaceae, Bacteroidales and Oscillospira, etc. However, CAG9 containing OTUs annotated to Faecalibacterium prausnitzii, Eubacterium biforme and Asteroleplasma anaerobium was significantly and positively correlated with ADFI (P<0.05). 3) In the correlation analysis, there was no OTU that was identified to be significantly associated with feeding behaviors at FDR<0.05. However, 14, 21 and 25 OTUs were identified to showing a tendency of association with ADET, ADEV and ADFI, respectively (P<0.01). Among them, OTUs showing positive correlations with the ADFI were mainly annotated to Prevotella, Ruminococcaceae and Erysipelotrichaceae, etc, while those OTUs having negative correlation were annotated to YRC22, Burkholderiales, Lachnospiraceae and Ruminococcaceae. 4) The functional prediction analysis of the gut microbiota showed that there was a significantly positive correlation between ADFI and biological funtions including Insulin signaling pathway and Lipid metabolism. Whereas the Sphingolipid metabolism was significantly and negatively correlated with ADFI (P<0.05). The results of this study suggest that some bacteria which can produce short-chain fatty acids or lactic acid, such as Burkholderiales, Ruminococcaceae, Christensenellaceae, Lachnospiraceae, etc., may play an important role in inhibiting feed intake, while Prevotella, especially Prevotella copri has the potential effect enhancing host appetite, are the key microorganisms that regulating the host feeding behaviour.

The purpose of this experiment was to investigate the effect of xylanase (Xyl) added to wheat flour on the development of gastrointestinal tract of Tibetan lamb. Sixty plateau Tibetan lambs aged 2-3 months with good homogeneity were randomly divided into two groups with 30 lambs in each group and 5 replicates each. The control group was fed without xylanase, and the test group was fed the diet supplemented with 0.2% xylanase. The experimental period was divided into 10 days of pre-feeding period and 90 days of positive trial period. At the end of the feeding experiment, six lambs in each group were randomly selected for slaughter, the digestive tract tissues were collected and fixed in paraformaldehyde. And blood, the contents of rumen and jejunum were collected at the same time. The results showed that: 1) Compared with the control group, the thickness of rumen cuticle and granular layer, the length and width of omasum nipple, the thickness of omasum cuticle and granular layer, the thickness of abomasal mucosa, and duodenal muscle layer, the height of jejunal villi and the thickness of jejunum mucosa, the thickness of ileal muscle layer in the experimental group were significantly increased (P<0.05); 2) The contents of rumen and serum LPS in the control group were significantly higher than those in the experimental group (P<0.05); 3) Compared with the control group, the activities of rumen chymotrypsin, trypsin and lipase, the activities of jejunal chymotrypsin and α-Amylase increased significantly (P<0.05); 4) The total antioxidant capacity and the activities of glutathione peroxidase and catalase in rumen, the total antioxidant capacity of jejunum, the activities of glutathione peroxidase, superoxide dismutase and catalase of jejunum in the experimental group were significantly higher than those in the control group (P<0.05); 5) There was no significant difference in rumen and jejunum pH between the two groups (P>0.05). In conclusion, the addition of 0.2% xylanase to wheat feed can effectively reduce the concentration of LPS in rumen and serum of plateau Tibetan sheep, promote the morphological development of gastrointestinal tract, enhance digestive enzyme activity, and improve antioxidant capacity.

This study was conducted to explore the effects of dietary resveratrol supplementation on liver anti-oxidant capacity and anti-apoptosis of Shanma ducks under acute heat stress, and to determine the effects of resveratrol supplementation on SIRT1 signaling pathway, antioxidant enzyme system and apoptosis related genes in the liver of Shanma ducks. A total of 120 healthy female Shanma ducks with similar weight at 60 days old were randomly divided into two groups, 60 in each group. The control (C) group was fed a basic diet, and the resveratrol (RES) group was fed a basic diet supplemented with 400 mg·kg^-1 resveratrol. Animals in two groups were kept at a temperature of (24±2) ℃. After 15 days, there was no significant difference in the weight of the animals between the two groups. Then the animals in each group were randomly divided into 3 groups,20 in each group, and placed in an artificial climate room at 39 ℃ for 0, 30, and 60 min, respectively. After heat stress treatment, duck serum samples and liver tissues were collected, and the mRNA, protein level and apoptosis number of SIRT1 signal pathway and apoptosis related genes were detected using qRT-PCR, Western blot and TUNEL techniques. Biochemical method was adopted to detect the oxidation index in serum and liver. The results showed that the mRNA expression of HSP70and HSP90 in RES group was higher than those in control group. At the same time, resveratrol activated the SIRT1 pathway and increased the mRNA expressions of downstream PGC-1α and TFAM, and significantly increased the nuclear proteins expression of Nrf1 and Nrf2 under heat stress for 60 min. Resveratrol increased the levels of T-AOC in serum. Resveratrol significantly increased the levels of SOD, CAT, AIHR and decreased the content of MDA under heat stress for 60 min，and under the same heat stress condition， the T-AOC, CAT and GSH-Px in liver of RES group were higher than those of C group, while MDA content in RES group was lower than that in C group. Under the condition of acute heat stress, the number of hepatocyte apoptosis in RES group was significantly lower than that in C group, and resveratrol decreased the mRNA expression of Bax, Bak1, Bax/Bcl-2, increased the protein expression of Bcl-2, and decreased the protein expression of Bax, Caspase3, Bax/Bcl-2. In conclusion, the addition of 400 mg·kg^-1 resveratrol to the diet can activate the SIRT1 signaling pathway in duck liver, reduce the expression of pro-apoptotic genes and increase the expression of anti-apoptotic genes. Resveratrol can improve the anti-oxidant capacity and anti-apoptosis ability of duck liver, and can improve the oxidative damage of duck liver caused by acute heat stress.

To study the effects of recombinant phosphatidylinositol transfer protein (HCPITP) from Haemonchus contortus on goat immunity, prokaryotic expression recombinant plasmid pET-28a-HCPITP was constructed and the recombinant protein was obtained. The differences of transcription level of HCPITP gene in egg, the third-stage larvae, female and male adult worm were analyzed. Goat peripheral blood mononuclear cells (PBMCs) were isolated and incubated with recombinant HCPITP, the effects of recombinant protein on the proliferation of goat PBMCs, the effects of recombinant protein on the transcription levels of pattern recognition receptors (PRRs) and cytokines in goat PBMCs were analyzed. Results showed that the relative transcription levels of HCPITP mRNA in female, male, the third-stage larvae, and egg were 1, 10.2, 0.47 and 0.58, respectively. The recombinant protein showed reactogenicity by Western blot analysis. It was found that recombinant HCPITP significantly inhibited the proliferation of ConA on PBMCs. Recombinant HCPITP promoted the transcription of IL-2, IL-4, IL-6, IL-17, TLR1 and TLR10 in goat PBMCs. In conclusion, the transcription level of HCPITP gene shows obvious stage differences, recombinant HCPITP has a certain effect on the proliferation and mRNA transcription of PRRs and cytokines of goat PBMCs.

To investigate the effect of Echinococcus multilocularis infection on the lipid metabolism in the liver of mice, 60 clean-grade BALB/c mice (6-week-old, male) were randomly divided into two groups with 30 mice in each group. The mice in the experimental group were intraperitoneally injected with 600 protoscoleces, while mice in the control group were treated with an equal volume of PBS. Hepatocytes were obtained by separation and purification. High-throughput sequencing was used to analyze the differentially expressed mRNAs of hepatocytes from mice liver at 3 months post infection (Mpi) and KEGG,GO analysis and qRT-PCR were used for verification. Liver tissue sections of mice at 2, 3 and 6 Mpi were stained with Oil Red O. The expression levels of lipid metabolism-related genes in hepatocytes at different infection stages were detected by qRT-PCR. The results showed that the fat deposition in the liver increased significantly compared with the control group at 3 and 6 Mpi. Lipid synthesis-related genes Scd1, Fasn and Srebf1, and lipid oxidation-related genes Acox1 and Cpt1α were significantly up-regulated in hepatocytes at 2 Mpi of E. multilocularis; the expression levels of Scd1, Fasn, Srebf1 and Cd36 were significantly increased, however, expression levels of Acox1, Cpt1α, Pparγ, and Pparα were significantly decreased in hepatocytes at 3 Mpi of E. multilocularis. The above results suggest that E. multilocularis infection may induce hepatic lipid deposition by promoting lipid synthesis in hepatocytes and inhibiting lipid oxidative.

H3N2 canine influenza virus (H3N2 CIV) circulate in dogs from multiple area in China, which is a novel branch formed by trans-host infection of avian influenza. Many research showed that the PA-X gene may play roles in the adaptation of influenza A viruses to new hosts, and can impact the replication efficient and virulence of influenza A virus by its length changing. To better understand the function of PA-X gene in H3N2 CIV, eight-plasmid operating systems were used to rescue three recombinant viruses, the parental virus CIV_PA-X_232 strain (232 aa), the CIV_PA-X_Knock strain, and the CIV_PA-X_252 strain (252 aa). To explore the effect of the length change of PA-X gene on H3N2 CIV, we compared the viral polymerase activity, replication efficiency in MDCK cells, and pathogenicity in mice of the three recombinant viruses. In vitro, the polymerase activity of CIV_PA-X_252 and CIV_PA-X_Knock were significantly (P<0.05) higher than that of CIV_PA-X_232, and that of CIV_PA-X_252 was higher than that of CIV_PA-X_Knock; the replication ability of CIV_PA-X_252 and CIV_PA-X_Knock were significant (P<0.05) higher than that of CIV_PA-X_232 in MDCK cells. In vivo, three recombinant viruses had no significant difference in the body weight of mice and were not lethal; TCID₅₀ of three recombinant viruses could be only detected in the lung of infected mice on 1 dpi, and the titer of CIV_PA-X_252 and CIV_PA-X_Knock was slightly higher than CIV_PA-X_232 with no significant difference; three recombinant viruses all caused lung pathological damage in mice, in terms of pathogenicity, CIV_PA-X_252> CIV_PA-X_232> CIV_PA-X_Knock. These results showed that the expression of PA-X protein with different size can affected the H3N2 CIV in transcription, replication, and the pathogenicity of mice. This study preliminarily explored the effects of PA-X gene within different length by H3N2 CIV, providing a reference for subsequent studies on the pathogenesis of H3N2 CIV.

The aim of this paper was to prepare specific monoclonal antibody (mAb) against African swine fever virus (ASFV) p54 protein. The p54 protein was expressed in Escherichia coli expression system and used as the antigen in mAb production. The spleen cells from the immunized BALB/c mice were fused with myeloma cells SP2/0. To screen the positive hybridoma cells, the purified p54 protein was used as envelope antigen for indirect ELISA. After four times’ subcloning, the supernatant of hybridoma cells were used to identify mAb subtype, ascites were prepared via in vivo induction method in mice and then the mAb was purified. The titer of the mAb was detected by indirect ELISA, and the specificity of the mAb was identified by cross reactivity assay, IFA and Western blot. According to the predicted secondary structure of p54 protein, using the stepwise truncation method identified the epitope region of mAbs, and labeled the region in tertiary structure of p54 protein. Results were as follows: six hybridoma cells secreting p54 monoclonal antibody were successfully screened and named 28G12-1, 31G7-1, 31G7-2, 35F10-1, 35F10-2, 38D3-1, respectively. The heavy chains of 28G12-1, 31G7-1, and 31G7-2 were IgG2a type, the heavy chains of 35F10-1, 35F10-2, 38D3-1 were IgG1 type, light chains were all κ chains. The lowest titer of mAb was 1∶25 600, and having no cross reaction with PRRSV, PRV, PEDV, PPV, SADS-CoV, PCV2, the specificity was strong. All six monoclonal antibodies could recognize the 127-146 aa on carboxyl end. In this study, ASFV p54 protein and p54 monoclonal antibody were successfully obtained, and the epitopes of six mAbs were identified, these experimental data laid a foundation for the functional research of p54 protein and the study of ASFV epitope vaccine.

The present study aimed to screen and identify host proteins that interact with the nucleocapsid protein (N protein) of pathogenic porcine reproductive and respiratory syndrome virus (PRRSV). In this experiment, we screened host proteins that interact with N protein and investigated their effects on PRRSV replication by constructing a porcine lung yeast cDNA library, yeast two-hybrid, immunoprecipitation, laser confocal, overexpression and interference assays. The results showed that N protein and ribosomal protein S20 (RPS20) interacted with each other in yeast cells. The overexpression and interference assays also provided preliminary evidence that RPS20 can affect the proliferation of PRRSV in cells. This study enriches the network of PRRSV-host factor interactions and lays the foundation for further studies on the biological functions of RPS20 protein during PRRSV infection.

To investigate the mechanism of systemic infection caused by Glaesserella parasuis (GPS) breaking through the epithelial barrier of swine respiratory tract, outer membrane vesicles (OMVs) were extracted by hypercentrifugation and density gradient centrifugation in this study.The protein bands of OMVs were detected by SDS-PAGE and distributed between 55 to 100 ku. The particle size of OMVs was found to be 100-200 nm observed by transmission electron microscope (TEM) and nanoparticle diameter analysis(NTA).The prepared HbpA and OmpP2 polyclonal antibodies were used to perform Western blot verification on OMVs and bacterial supernatants without OMVs. The results proved that the extracted samples were outer membrane vesicles, and further results proved that the cytolethal distending toxin (CDT) mainly exists in the form of OMVs in G. parasuis. Then, swine tracheal epithelial cells (STEC) were treated with OMVs or CdtB for 36 h, and the protein expression levels of cleaved-caspase3, ZO-1 and Occludin in STEC were detected, and FITC-dextran (FD-4) was used to detect the paracellular permeability. The results showed that after treated with OMVs or CdtB, the expression level of apoptosis-related protein cleaved-caspase3 was increased and the expression levels of ZO-1 and Occludin were decreased compared with the control group, and the permeability of FD-4 was increased. At the same time, the lactate dehydrogenase (LDH)cytotoxicity assay showed that the cell viability was significantly decreased after 24h of CdtB co-culture and 36h of OMVs co-culture. STEC pretreated with inhibitor of the apoptosis p53 pathway (pifithrin-α) could inhibit the cell apoptosis and barrier permeability increase induced by OMVs or CdtB. In addition, when CdtB in OMVs was removed by immunomagnetic bead capture (OMVs-△CdtB), the expression levels of cleaved-caspase3, ZO-1, and Occludin were not different after OMVs-△CdtB treated STEC for 36h compared with the control group. In conclusion, GPS OMVs were successfully extracted and purified, and the experiment proved that OMVs induced p53-dependent apoptosis and disrupted tight junctions in STEC by transporting CdtB, and finally increased the permeability of the tracheal epithelial barrier, providing a new idea for the mechanism of G. parasuis breaking through the respiratory epithelial barrier.

To investigate the role of the Nrf2/HO-1 pathway in neuronal damage caused by Japanese encephalitis virus (JEV) infection, an in vitro infection model was developed in which Nrf2 agonist tert-butylhydroquinone (TBHQ) was used to treat JEV-infected mouse neuroblastoma (N2a) cells. N2a cells were divided into following four groups: DMEM control group (Control), JEV-infected group (JEV), TBHQ-treated group (TBHQ), and JEV-infected +TBHQ -treated group (JEV+TBHQ). The cytopathic lesions were observed after different treatments and the changes in ROS levels and MDA, SOD and CAT contents were detected. The mRNA and protein expression of Nrf2 and HO-1 were detected by qPCR and WB techniques, and Nrf2 protein expression in each group of cells was detected by immunofluorescence, and inflammatory factors and viral content were measured. The results showed that the level of ROS gradually increased with time after JEV infection of N2a cells and was reached the highest at 36 h. The mRNA of Nrf2 and HO-1 increased at 36 h after JEV infection, and the level of inflammatory factors increased. JEV infection of N2a cells resulted in cell crumpling, cell membrane rupture, increased ROS and MDA levels and decreased SOD and CAT levels; treatment with 40 μmol·L^-1 TBHQ for 6 h reduced cell damage, decreased ROS and MDA levels and increased SOD and CAT activities. Both JEV and TBHQ activated the mRNA and protein expression of Nrf2 and HO-1, and the expression levels of proteins related to the Nrf2/HO-1 pathway were significantly increased by combined treatment with JEV+TBHQ, and the nucleation of Nrf2 in the cells by immunofluorescence was evident, and the mRNA levels of TNF-α and IL-6 and the levels of viruses decreased. The above results suggest that TBHQ is able to reduce cellular oxidative stress and inflammation levels induced by JEV through the Nrf2/HO-1 pathway.

This study investigated the virulence genes, drug resistance genes, drug resistance spectrum and genetic diversity of Klebsiella pneumoniae from sheep in Guangdong province. One hundred and fifty sheep nose swab samples were collected from sheep farms with different scales in 3 cities of Guangdong province. The pathogenic bacteria were isolated and identified by means of culture, purification, staining and microscopic examination. The biological characteristics of the isolated strains, such as virulence genes and drug resistance etc., were studied by means of molecular biology and drug sensitivity test according to CLSI manual. The results showed that 42 strains of K. pneumoniae were isolated from the samples. The drug susceptibility test results showed that the drug resistance rate of amoxicillin and ofloxacin was up to 100%, and there were different degrees of resistance to other drugs; the virulence gene test results showed that rmpA, kfuBC and ureA were 88.0%, 85.7% and 83.3%, respectively. The detection results of drug resistance genes showed that the detection rates of drug resistance genes aphA, qnrA, aacC4, aacC2 were 42.9%, 40.5%, 28.6%, 23.8%, and the detection rates of other drug resistance genes were low. Eric-pcr typing results showed that the homology was more than 80.0%, which could be divided into seven groups, among which population Ⅰ was the dominant population, accounting for 66.6%.This study was conducted to analyze the virulence genes, epidemic status and drug resistance of sheep K. pneumoniae in Guangdong area, which has certain guiding significance for the prevention and control of sheep K. pneumoniae.

The study aimed to analysis the prevalence, structural characteristics and transduction ability of prophages in porcine methicillin-resistant Staphylococcus aureus (MRSA) ST9, and to explore the role of prophages in the formation of porcine MRSA epidemic clones. Based on whole genome sequences, we analyzed the prevalence, typing, phylogeny, and structural characteristics of prophages in 131 MRSA ST9 strains isolated from several provinces of China in recent years. Strains with prophages of various types were selected to be induced into phage particles which were then performed transduction experiment. We determined the antimicrobial resistance and in vitro fitness cost of the transductants. The results showed that the carriage rate of prophages in porcine MRSA ST9 was 78.6% (103/131), of which 63 isolates carried prophages with complete structure. None of prophage harbored resistance gene, while only 2.9% sequences (3/103) carried virulence genes. The profiles of prophages were abundant, of which the integrase typing were mainly Sa2int and Sa4int. Different types of prophages had high structural homology, and the complete prophages could be induced into Siphoviridae. Resistance genes aadD and tet(L) of donor strains could be packaged into phage particles and transduced to recipient strain. The transductants acquired resistance phenotypes to kanamycin and tetracycline, while the growth ability of transductants had no significant difference with the recipient strains in vitro (P>0.05). These results indicated that the prophages of porcine ST9 MRSA have high carriage rate, various types, and do not carry resistance gene. Resistance genes of donor strains could be packaged and transduced into recipient strains by part of phages, resulting in little fitness costs.

This study aimed to investigate the effect of endoplasmic reticulum stress on lipopolysaccharide (LPS) induced autophagy in bovine mammary epithelial cells (BMECs). Firstly, the experiment was divided into control group (CON) and LPS group (LPS, 4 μg·mL^-1) to study the effects of LPS on endoplasmic reticulum (ER) stress and autophagy in BMECs. Then, cells were pretreated with different concentrations of PERK inhibitor GSK2606414 (GSK), and the mRNA expressions of PERK, ATF4, eIF-2α and CHOP were measured to determine the optimal inhibitory concentration of GSK. Finally, the BMECs were divided into 4 groups: control group (CON), LPS group (LPS, 4 μg·mL^-1), GLPS group (GSK+LPS) and GSK group, to study the effects of PERK inhibition on LPS-induced autophagy in BMECs were investigated. The expressions of endoplasmic reticulum stress and autophagy related genes and proteins were analyzed by RT-qPCR and Western blot. The fluorescence intensity of GRP78 and p62 was measured by immunofluorescence. The results showed as follows: 1) Compared with the control group, the mRNA and protein expressions of PERK, IRE1α, ATF6, GRP78 and CHOP in LPS group were significantly (P<0.05) or extremely significantly (P<0.01) increased. LPS significantly also increased the mRNA and protein expressions of LC3, ATG5, ATG14 and Beclin1 (P<0.01), and significantly decreased the mRNA and protein expressions of p62 (P<0.01). In addition, LPS reduced the fluorescence intensity of p62, increased the fluorescence intensity of GRP78 and lysosome formation; 2) Compared with LPS group, GSK pretreatment significantly reduced the protein expressions of PERK, ATF4, eIF-2α and CHOP (P<0.05 or P<0.01), and also significantly reduced the mRNA and protein expressions of LC3, ATG14, ATG5, Beclin1 and increased the mRNA and protein expressions of p62 (P<0.01 or P<0.05). The immunofluorescence results further indicated that GSK pretreatment could increase the fluorescence intensity of p62. Therefore, in this study, LPS induced endoplasmic reticulum stress and autophagy, and inhibition of the PERK/eIF-2α/ATF4 signaling pathway can alleviate LPS-induced autophagy in BMECs.

This study aimed to investigate the effect of N-acetyl-L-cysteine (NAC) on oxidative damage of lens epithelium in the beagle model of type 1 diabetes mellitus. Forty beagles, 2-3 years old, were randomly divided into control group (CON group), diabetes model group (DM group), insulin treatment group (INS group), NAC combined with insulin treatment group (NAC+INS group) and NAC treatment group (NAC group), 8 dogs in each group. The establishment of experimental dog model in each treatment group lasted for 120 days. Before and after modeling, insulin release test, fasting blood glucose and aqueous humor glucose were tested and cataract grade evaluation was carried out by slit lamp. The lens epithelium was histopathologically observed, and ELISA and RT-PCR detected the oxidative stress index and inflammatory factors in the lens. The results of insulin release test and fasting blood glucose showed that the model of type 1 diabetes beagle was established successfully; Compared with CON group, at the end of the experiment, the serum and aqueous glucose of DM group increased significantly (P<0.05), and the lens function decreased significantly; Slit-lamp results showed that the lens opacity of the DM group reached grade IV at the end of the experiment, while the other four groups showed no apparent signs of cortical opacity; In addition, obvious pathological changes such as nuclear pyknosis and necrotic bodies were observed in the lens epithelium of DM group; Blood glucose in NAC, INS and NAC+INS groups showed a downward trend，but was still higher than that in CON group; The blood glucose in NAC+INS group was significantly lower than that in DM group (P<0.05); and the lens opacity was also significantly improved; At the end of the experiment, compared with CON group，in the canine crystal epithelium and aqueous fluid, MDA content of DM group increased (P<0.05 or P<0.01), while GSH-Px activity and GSH/GSSG all had the noteworthy decrease (P<0.05 or P<0.01), GR activity decreased (P>0.05 in crystal epithelium; P<0.01 in aqueous fluid); After treatment of NAC combined with insulin, oxidative stress markers in crystal epithelium showed an opposite trend. The expression levels of SOD₂ and GR in DM group and all treatment groups were significantly increased (P<0.05). In conclusion, therapy of NAC combined with insulin has improvement for glycemic control and lens function in diabetic dogs, the protective effect of NAC combined with insulin on lens epithelial injury in diabetic dogs may be related to the inhibition of oxidative stress of the lens.

This article aims to evaluate the clinical outcomes and complications of distal radial and ulnar fractures in toy-breed dogs treated with conventional T-shape bone plate fixation. Medical records of toy-breed dogs with distal radius and ulna fractures repaired with open reduction and internal fixation utilizing conventional T-shape bone plate in the animal hospital of China Agricultural University were reviewed, and these cases were followed up. The inclusion criteria were: body weight of ＜7 kg; Fracture located in the distal antebrachium (distal-to-total radial length ratio ＜0.25)；The follow-up time was longer than 12 months; Case information records were complete. Results: 29 cases of radius and ulna fractures in 29 dogs were included, 26 cases (89.7%) had a successful return to normal function, and 3 cases (10.3%) had barely detectable lameness. Minor complications in 6 fractures (20.7%) were identified and there was no major complications occurred. Conventional T-shape bone plate internal fixation are effective means for stabilization of distal radius and ulna fractures in toy-breed dogs that result in good clinical outcome with no major complication.

This study established a rat model of damp-heat diarrhea (DHD) to explore the efficacy of Echinacea purpurea extract (EPE) on DHD, so as to provide scientific basis for EPE in the prevention and treatment of DHD. Rats with DHD were established by “internal and external Dampness-heat+Escherichia coli”, and were treated with 1 and 3 g·kg^-1 doses of EPE, respectively. The DHD model was evaluated by clinical symptom score, the blood routine test, blood biochemistry, serum inflammatory factors, organ index and colon related gene expression level of rats in each group were measured, and the histopathology of rat organs was observed. The results showed that the DHD rats were fluffy, depressed, piled up, lazy, secretions appeared in the eyes, loose stool with yellow and smelly, and perianal dirt was visible. After 6 days of administration, the spirit, diet and appearance of rats returned to normal in the high and low dose EPE groups and the positive drug group. Compared with the control group, the indexes of heart, spleen and lung in the model group increased significantly (P<0.05), and the numbers of red blood cell (RBC), hemoglobin (HGB) and hematocrit (HCT) in blood decreased significantly (P<0.05); the levels of urea and alanine aminotransferase (ALT) in serum decreased significantly (P<0.05), the levels of IL-6, IL-17 and IL-23 increased significantly (P<0.05). The levels of TGF-β, IL-2 and IL-10 decreased significantly (P<0.05); the expression of colon TGF-β1 and Foxp3 mRNA decreased significantly (P<0.05), the expression of RORγt mRNA increased significantly (P<0.05); Massive bleeding and stasis in the heart, edema in the liver, edema and old bleeding in the spleen, inflammatory cell proliferation and deposition of proteinuria in the kidneys. Compared with the model group, the heart, spleen and lung of the high and low dose EPE groups were significantly lower (P<0.05), the level of HCT in blood was significantly higher (P<0.05), the levels of urea, total cholesterol (TC), total protein (TP) and albumin (ALB) in serum were significantly higher (P<0.05), the levels of IL-17 and IL-23 in serum were significantly lower (P<0.05), the level of TGF-β increased significantly (P<0.05); The expression of colon TGF-β1 and Foxp3 mRNA increased significantly (P<0.05), the expression of RORγt mRNA decreased significantly (P<0.05). The organ cell damage in high and low dose EPE groups recovered. The data may be extrapolated to suggest that Echinacea purpurea extract can alleviate DHD in rats by improving organ function, reducing blood lipid and regulating the level of serum inflammatory factors.

The aim of this experiment was to investigate whether kushen-cangzhu (Sophora flavescens Ait. and Atractylodes Lancea) granules could improve the abnormal cholesterol synthesis induced by Escherichia coli (E. coli) through regulating liver cholesterol metabolism in broilers. Forty 21 days old white-feathered broilers were randomly divided into 5 groups (8 in each group): Blank control group, model control group, low-, medium-and high-dose of kushen-cangzhu groups. The model control group and different doses of kushen-cangzhu groups were intramuscular thoracic injection treated with pathogenic E. coli bacterial solution for 0.7 mL (8.37×10⁸ CFU·mL^-1). After inoculating pathogenic E. coli bacterial solution for 24 h, different doses of kushen-cangzhu granules (low-dose group: 3.6 g·L^-1, medium-dose group: 5.5 g·L^-1, high-dose group: 7.3 g·L^-1) were supplemented in drinking water for chickens in low-, medium-and high-dose groups, respectively, and the trail lasted for 7 days. At the end of the experiment, all the broilers were slaughtered, body and organs were weighed, blood and tissue samples were collected. The morphological changes of liver were observed by hematoxylin-eosin (HE) staining. The levels of serum and liver total cholesterol (TCH), hepatic triglyceride (TG), high-density lipoprotein-cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were detected, RT-qPCR was used to detect the expression of cholesterol metabolism-related genes in liver. The results showed that, compared with the blank control group, the broilers in the model control group had significantly lower body weight, higher liver index and serum TG content (P＜0.05), while infiltrated inflammatory cells in the central vein and the hepatic sinusoid and disordered hepatocyte was observed in model control group. TG and SCAP mRNA expression was significantly lower (P＜0.05) in liver of model control group compared with blank control group. Compared with the model control group, the body weight and liver LDL-C content was significantly increased in the low-dose group (P＜0.05), while the liver index and serum TG content were significantly decreased (P＜0.05), and the hepatic morphological structure was at a better state. The HDL-C, LDL-C and the LDL-C/HDL-C ratio in liver were significantly increased in the medium-dose group compared with model control group. Meanwhile, the mRNA expression of HMGCR, SREBP2 and SCAP was significantly enhanced (P＜0.05) in medium-dose group than model control group. In the high-dose group, the serum TG concentration was significantly reduced (P＜0.05) but hepatic LDL-C was significantly increased (P＜0.05), and the ratio of LDL-C/HDL-C was significantly raised (P＜0.05) compared with the model control group. In conclusion, the liver abnormal synthesis of cholesterol in broilers induced by E. coli was alleviated by kushen-cangzhu granules through regulating cholesterol metabolic pathway.

The aim of this study was to explore the mechanism of immune-enhancement effects of Eucommia ulmoides leaf extract (ELE) based on network pharmacology and immunosuppressive mouse model. Three active components of ELE were screened out from the TCMSP database. Their 306 potential corresponding targets were predicted by Uniprot and Swiss Target. After comparison with GeneCards and Online Mendelian Inheritance in Man(OMIM), 105 common genes of immune dysregulation and ELE were obtained. The “active ingredient-gene” network diagram was constructed by Cytoscape 3.8.2 software. STRING was used to construct a protein-protein interaction(PPI) network of the overlapping genes, network topology analysis was employed by CytoNCA to screen the 21 core genes, which were then subjected to enrichment analysis with gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG). The results showed that the main compounds of ELE, such as kaempferol, quercetin and chlorogenic, participated in the IL-17 and THF signaling pathways by regulating tumor necrosis factor (TNF), interleukin-6 (IL-6), vascular endothelial growth factor A (VEGFA) and interleukin 1β (IL-1β) to exert immunoregulation. The immunoregulation of ELE were investigated by in vitro experiments of peritoneal macrophages and in vivo experiments of cyclophosphamide immunosuppression. The results showed that, 1 000~5 000 μg·mL^-1 ELE could promote the proliferation and phagocytosis of peritoneal macrophages. Compared with the model group, ELE could increase immune organ indices, phagocytic index, peripheral blood leukocytes and lymphocytes contents of mice, enhance the earlap swelling of the delayed type hypersensitivity (DTH). These findings indicated that the immunomodulation of ELE was realized through multiple components and targets, so as to provide a scientific basis for the application of immune regulation of ELE in healthy breeding of livestock and poultry.

This study was conducted to examine the distribution of phylogenetic clustering, virulence genes, serotypes and drug resistance of Escherichia coli (E. coli) from diarrhea with piglets in Xinjiang. In this study, 154 fecal samples were collected from diarrhea with piglets, and isolation, identification of E. coli was conducted. Phylogenetic clustering, virulence genes, serotype were tested by multiple PCR method. Drug sensitivity of isolates was determined by using a Kirby-Bauer disk diffusion method and drug resistance gene was detected by PCR. The results showed that 154 E. coli were isolated, including ETEC (n=24), STEC (n=21), EPEC (n=1), EPEC/STEC (n=2), ETEC/STEC (n=1), ETEC/EPEC (n=1), and other 104 strains. Phylotyping assays showed that most strains largely belong to group A (37%) and B1 (31%). O serogroups were identified for 44 E. coli isolates, of which O154, O12, O8, O141 and O175 were dominant serogroups. One hundred and fifty-one strains were multiple drug resistance (MDR), and the drug resistance rates of 154 E. coli isolates to cotrimoxazole, tetracycline, ampicillin, streptomycin and chloramphenicol ranged from 81% to 100%, that to amoxicillin/clavulanate, cefotaxime, gentamicin, ceftriaxone, ciprofloxacin and amikacin ranged from 31% to 66%, that to levofloxacin, polymyxins B, ceftazidime, cefepime, ampicillin -sulbactam, piperacillin -tazobactam and imipenem ranged from 1% to 19%. The prevalence of drug resistance genes tetA (88%), tetG (60%) and cmlA (45%) was higher, while bla_CTX-M-2G, bla_TEM, bla_SHVand tetE were all lower than 30%. bla_CTX-M-2G, bla_TEM, bla_SHV and tetE were not detected. The results indicated that the types of E. coli from diarrhea with piglets in Xinjiang were complex and the situation of MDR was severe, the prevalence of drug resistance genes was diversified, and important antibiotic resistance phenotypes were detected in human and clinical, so the monitoring of drug resistance of E. coli in pig farms should be strengthened.