Compared to the traditional breeding methods, genomic selection (GS) accelerates the genetic progress of breeding by early selection and increasing the accuracy of selection for the individuals to be retained. It is unlikely shortened the generation interval by improving the GS methods. Therefore, how to improve the accuracy of GS to obtain additional genetic progress is the core issue of GS research. With the development of omics technology, it becomes possible to obtain multi-omics biological prior information from the public available databases or the in-house research. Therefore, how to integrate this useful biological prior information into GS models to improve the accuracy of GS has become an important topic in current animal breeding research. In this paper, the types of biological prior information and the GS methods that can integrate prior information is reviewed. Then,the application and prospects of these methods applied to livestock breeding is discussed. It is expected to provide useful information for the GS research that integrates biological prior information on livestock breeding.

Super enhancers are a class of large clusters containing multiple enhancers, mainly enriched with a high density of transcription factors, cofactors and enhancer-associated epigenetic modification sites. Compared with enhancers, super enhancers have higher transcriptional activation capacity and play a key role in cell type-specific development, differentiation, and tumor development processes. Studying the mechanism of super-enhancer action in gene transcription is of great biological importance to reveal the molecular mechanism of mammalian phenotypic traits. This article outlines the concept, properties and identification methods of super enhancers, summarizes the main mechanisms of super enhancers function, and finally reviews the research progress of super enhancers in regulating important phenotypes of animals, and discusses their application prospects in mammals combined with the latest research progress of super enhancers, aiming to provide a reference for studying functions of super enhancer in important phenotypic traits in animals.

Redox balance exists in animals, which maintains a relatively stable state under normal physiological conditions. This balance will be broken when the environment or the state of animal itself changed, thus leads to the occurrence of oxidative stress. Studies have found that there are many factors can cause oxidative stress, such as changes of environment and physiological conditions, as well as pathogens and viruses infection. Oxidative stresses caused by these factors could affect animal health, production performance and reproductive function in different degrees, and even cause animal death. Therefore, it is of great practical significance to seek and add safe and effective antioxidant agents to regulate the redox state of animals. At present, the main nutritional additives for animal oxidative stress regulation including probiotics, vitamins, microelement, hormones and plant active substances. This review summarized the main causes of oxidative stress in animal and their impact on animals, and also concluded nutritional feed additives with the function of alleviating animal oxidative stress, in order to provide reference for further study of animal oxidative stress and its relieving strategies.

The study aimed to explore the influence of different reference population screening methods and size on accuracy of genotype imputation, such as maximizing the expected genetic relationship for matrix A (RELA), minimized the target population genetic variance for matrix A(MCA), the highest mean kinship coefficients (KIN), random selection (RAN), common ancestor (CA). In this study, the dwarf and yellow-feathered chicken population were used, and the chicken 600K SNP array(Affymetrix Axion HD genotyping array) was used for genotyping. The body weight of 435 offspring cocks at 45, 56, 70, 84 and 91 days of age were measured. The Beagle software was used to impute low-density SNP chips into high-density SNP chips, to compare the influence of reference population screening methods and reference population size on accuracy of imputed genotype and accuracy of imputed chips for genomic prediction. The results showed that the best imputation method was using Beagle 4.0 with pedigree information, followed by Beagle 4.0, and Beagle 5.1 was comparatively the worst. MCA method had the highest accuracy of genotype imputation, RAN method had the lowest accuracy of genotype imputation, the accuracy of MCA, RELA and CA methods for genotype imputation had small difference. Compared with other methods, MCA method has higher prediction accuracy to select key individuals as reference population and to impute from low-density SNP chips to high-density SNP chips for genome selection, which was slightly different from that of real high-density SNP chips. With the increase of reference population size, the accuracy of genotype imputation were also increased, but the growth rate were gradually decreased and finally tended towards stability. In conclusion, the accuracy of genotype imputation and genome prediction,as well as lower costs were guaranteed by selecting key individual screening methods and controlling the size of reference population. This study provides technical reference for the application of genotype imputation in livestock genetic breeding.

The study aimed to explore the expression characteristics of ZP3 gene in Hailan laying hens, which could lay the theoretical foundations for elucidating the function of ZP3 gene in follicle development in chicken. Chicken ZP3 gene was cloned with rapid amplification of cDNA ends (RACE) using the follicle tissue from 50-week-old (50 W) Hailan laying hens. The sequence of CDS region of ZP3 was analyzed by bioinformatics tools. The expression profile of ZP3 gene was constructed using heart, liver, lung, kidney, ovary, chest muscle, leg muscle tissues and follicles of different grades from 6 healthy Hailan brown laying hens with similar body weight. Total RNA was obtained from the ovarian granulosa cells with the treatment of follicle-stimulating hormone (FSH) in different concentrations (PBS, 5 ng·mL^-1,10 ng·mL^-1,20 ng·mL^-1). The expression levels of ZP3 gene in different tissues were detected by real-time PCR. The total length of chicken ZP3 gene was 1 415 bp containing 9 exons, in which the coding region was 1 341 bp, encoding 446 amino acids, located at chromosome 10. Its genetic relationship was the farthest from swine. The prediction of transmembrane structure indicated that it contained a transmembrane domain and a signal peptide; By analyzing its hydrophobicity, it was predicted that the protein was a weak hydrophobic protein, and the protein structure was mainly composed of random curl. According to the results of gene expression profile, the expression of ZP3 gene was the highest in chicken ovary. Besides, the expression of ZP3 gene was the highest in granulosa cells of mature follicles (F1) relative to follicles of other degrades. After treating granulosa cells of different grades with FSH, the expression level of ZP3 gene was significantly increased (P<0.05), suggesting that ZP3 gene was regulated by FSH in granulosa cells. This stuay analyzed the structure of ZP3 gene and predicted that it contained the transmembrane structure and signal peptide region. Expression profiles revealed that the expression level of ZP3 gene increased gradually during follicular develop ment and was mainly expressed in granulosa cells, which may be regulated by FSH. Therefore, it was predicted that ZP3 gene might be involved in reproductive function of Hailan brown laying hens.

The aim of this study was to investigate the relationship between DNA methylation of TCF21 gene promoter region and its expression in chicken adipose tissue. The mRNA expression levels of TCF21 in abdominal adipose tissues of chickens which were at 7 weeks of age and from 24^th generation of Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF) were detected by RT-qPCR. The structure and function of TCF21 promoter were analyzed by bioinformatics and luciferase reporter system. The methylation level of CpG sites in the promoter region of TCF21 in the abdominal adipose tissues of the fat and lean broilers was detected by Sequenom MassARRAY platform. The TCF21 promoter reporter gene plasmid was treated with CpG methyltransferase to analyze the effect of DNA methylation on the activity of TCF21 promoter. The results showed that the mRNA expression levels of TCF21 in the abdominal adipose tissue of fat broilers were extremely significantly higher than those of lean broilers (P<0.001). There were 40 CpG sites in the promoter region, which were distributed in the proximal and distal parts of the promoter, but there was no CpG island in the promoter region of chicken TCF21 gene. The promoter of TCF21 gene was divided into 5 functional regions, namely R1 region (-2 000~-1 500 bp), R2 region (-1 500~-1 000 bp), R3 region (-1 000~-500 bp), R4 region (-500~-200 bp) and Core region (-200~-100 bp),respectively. The DNA methylation levels in the R2, R3 and R2+R3 regions of TCF21 promoter in the fat broilers were significantly or extremely significantly higher than those of the lean broilers (P<0.05 or P<0.001). The DNA methylation levels in the R2, R3 and R2+R3 regions of TCF21 promoter were significantly positively correlated with TCF21 mRNA expression levels (R2 region:r=0.438, P<0.05; R3 region:r=0.371, P<0.05; R2+R3 region:r=0.489, P<0.05). DNA methylation significantly inhibited the transcriptional activity of R2 region of TCF21 promoter (P<0.05). In summary, the expression level of TCF21 gene in abdominal adipose tissue of fat and lean broilers was mainly related to the DNA methylation level of R2 region of TCF21 promoter.

To explore whether the systemic skin wrinkle of Xiang pig with wrinkled skin (XPZ) was related to the variation of HAS2(hyaluronan synthase 2) gene, all exon regions of HAS2 gene were cloned by specific PCR method with ordinary Xiang pig(XP) as control. The base variation of gene coding region was analyzed by bioinformatics method, and the distribution frequencies of variant SNP sites in population were detected. Four SNP sites were detected from HAS2 gene of Xiang pig with wrinkled skin,including T221A missense mutation in exon 1 (resulting in leucine substitution to lysine) and A228G synonymous mutation,T183C synonymous mutation in exon 2 and C537T synonymous mutation in exon 4. Bioinformatics analysis showed that 4 haplotypes were formed at T221A and A228G loci. Among them, T-A was the main haplotype (27.5%) and closely linked(r²=0.253) in Xiang pig population with wrinkled skin. Moreover, mutations in T221A and A228G could change HAS2 mRNA free energy and protein structure, and the free energy of TA, TG, AG and AA combinations were -573.29, -580.89, -572.66, -571.03 kJ·mol^-1,respectively. After leucine was replaced by lysine resulted from T221A missense mutation, the percentage of α-helix decreased from 35.52% to 32.97%, and the free crimp increased from 43.17% to 44.51%. Two SNPsites of loci T221A and A228G showed abundant polymorphism in Xiang pig population with 3 genotypes. And both frequencies of A allele in T221A(P<0.05) and G allele in A228G(P<0.01) were significantly higher in Xiang pig with wrinkled skin than that in common Xiang pig. There were no significant difference in the frequencies of C allele in T183C and T allele in C537T between Xiang pig with wrinkled skin and common Xiang pig. The results showed that the two base mutations in exon 1 of HAS2 gene might affect the stability of post-transcriptional mRNA, the three-dimensional structure of protein and HA synthesis, and have a relationship to the thickening and wrinkling of skin in Xiang pig with wrinkled skin.

The aim of this study was to explore the differentially expressed genes in the subcutaneous adipose tissue of Duolang sheep (group D) and Small Tail Han sheep (group X), study their potential effects, and to provide a basis for understanding the developmental law of sheep fat tissue and the prevention and treatment of lipid metabolism-related diseases. In the study, healthy, female adult Duolang sheep and Small Tail Han sheep with good body condition, similar in-species body weight (about 50 kg) and differences in fat deposition ability were selected as the test materials, and they were divided into group D (experimental group) and group X (control group), with 3 replicates in each group. The subcutaneous adipose tissue located in the longissimus dorsi muscle was collected. RNA-Seq technology and bioinformatics methods were used to perform the transcriptome sequencing and the results analysis. According to the criteria of|Fold change| ≥ 2 and P adjust ≤ 0.05, differentially expressed genes were screened. The differentially expressed mRNAs were analyzed by GO and KEGG enrichment to obtain the differential genes related to fat deposition and lipid metabolism. Meantime, the reliability of the sequencing data was verified by real-time fluorescent quantitative PCR for 6 randomly selected differentially expressed genes. The results showed that a total of 38 672 known mRNAs and 1 606 new mRNAs were detected in the 6 samples. There were 839 differentially expressed genes in the two groups, of which 320 were up-regulated and 519 were down-regulated in Duolang sheep. Through GO function annotation analysis, it was found that differentially expressed genes were mainly involved in lipid catabolism process, lipid biosynthesis process, lipid catabolism negative regulation process, MAPK cascade reaction regulation, and reaction to triglycerides. The enrichment results of KEGG pathways showed that PI3K-Akt, MAPK, insulin and PPAR signaling pathways were significantly enriched. The qRT-PCR results were consistent with the sequencing results, indicating that the sequencing results were reliable. Through transcriptome sequencing and bioinformatics analysis of subcutaneous fat tissue of Duolang sheep and Small Tail Han sheep, differentially expressed genes related to fat deposition and lipid metabolism were screened, these genes mainly involved in lipid biosynthesis, lipid metabolism and other processes. Among them, COL1A1, AKT2, SCD, LPL, PCK1 and PPP2R5A might play important roles in the deposition and metabolism of subcutaneous fat tissue in Duolang sheep and Small Tail Han sheep.

In order to explore the role of HSP27 gene in the follicular development of Tibetan sheep, in this study, 6 healthy (3-4 years old) Tibetan ewes were selected and slaughtered to obtain ovarian tissue for cloning Tibetan sheep HSP27 gene. The CDS region sequence of HSP27 gene was analyzed using bioinformatics methods, HSP27 gene overexpression and silencing vectors were constructed, Tibetan sheep ovarian granulosa cells were isolated and cultured, and cell transfection experiment was conducted with the constructed vectors in different groups, namely blank group, overexpression vector group, silent vector group, negative control group, each group had 3 multiple wells. Microscopic observation of the cell morphology changes and cell counts of each transfection group at 0, 24, 48, 72 h incubation time was performed. The expression of of HSP27, GDF9, BMPR-1B, Erβ genes mRNA were detected by RT-PCR. The results showed that the the Tibetan sheep HSP27 gene was successfully cloned. The CDS sequence of the Tibetan sheep HSP27 gene was 618 bp in length and encoded 203 amino acids. The Tibetan sheep HSP27 gene overexpression and silencing vectors were successfully constructed; the constructed vector was transfected into the ovary granular cells, the cell morphology of each transfection group changed with the increase of culture time, the cells of the overexpression vector group gradually changed from a long spindle to an irregular polygon, the nucleus deformed and decomposed, and the pseudopodia of the silent vector group decreased, the cells shrank and had a lot of apoptosis. The cell count results showed that, after 72 h incubation, the number of cells in the silent vector group was significantly lower than that in the other 3 groups (P<0.01), and the number of cells in the overexpression vector group was significantly lower than that in the blank group and the negative control group (P<0.05). RT-PCR results showed that the expression of HSP27 mRNA in the overexpression vector group was significantly higher than that in the blank group (P<0.01), and the expression of HSP27 mRNA in the silent vector group was significantly lower than that in the negative control group (P<0.01), the expression of GDF9, BMPR-1B, Erβ mRNA in the overexpression vector group was significantly higher than that in the blank group (P<0.01), and the expression of Erβ mRNA in the silent vector group was significantly lower than that in the negative control group (P<0.01). It can be preliminarily inferred that when HSP27 gene is overexpressed, the granulosa cell proliferation and differentiation was promoted. Silencing HSP27 gene may trigger granulosa cell apoptosis and affect follicular development. The above research results can lay the experimental foundation for the function research of HSP27 gene in the follicular development process of Tibetan sheep.

The aim of this study was to clone the HABP4 gene sequence of Jianzhou goats, analyze the proteins predicted to interact with HABP4, and determine their mRNA expression, so as to identify the correlations among different genes. These data will be beneficial for exploring the role of HABP4 gene in regulating cell apoptosis in goat. Sixteen 10-month-old healthy Jianzhou goats, with the average weight of 55 kg, were randomly selected from Sichuan Jianyang DAGEDA Animal Husbandary Co. Ltd.. Nine tissue samples of heart, liver, spleen, lung, kidney, pancreas, large intestine, small intestine and rumen of each goat were quickly collected after slaughter. The total RNA was extracted by TRIzol method and reverse transcribed into cDNA. The HABP4 gene sequence was cloned by RT-PCR and sequenced for further bioinformatics analysis using online softwares. Real-time fluorescent quantitative PCR (RT-qPCR) was performed to determine the expression levels of HABP4 in different tissues (n=16), and followed by the analysis of the mRNA expression levels of 10 proteins interacted with the HABP4 protein in pancreatic tissues(n=16). Also, the correlation analysis was performed in expression among these genes. A total length of 1 496 bp HABP4 gene sequence was cloned successfully, including 61 bp of 5' UTR, 1 254 bp of CDS, and 181 bp of 3' UTR, encoding 417 amino acids. The HABP4 expressed in all detected tissues. The mRNA level of HABP4 was higher in pancreas compared with other tissues. The results of correlation analysis showed that the expression of HABP4 was significantly positively correlated with UBA52, RPL32 and RPS9. The results of functional enrichment analysis showed that HABP4 might play an important role in regulating cellular localization, metabolism, negative regulation of biological processes, translation initiation, ribosome transcript and cytoplasmic metabolism. In this study, a full length of 1 496 bp of HABP4 gene was cloned successfully, and which might be significantly correlated with UBA52, RPL32 and RPS9. This data will be beneficial for further exploring the role of HABP4 gene in regulating cell apoptosis.

The study aimed to explore the effects of myosin binding protein C1 (MyBPC1) on the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells, and provide a basis for further study of the regulatory role of MyBPC1 in cell differentiation and muscle development. The Simmental fetal bovine primary bovine skeletal muscle satellite cells to inducing myogenic differentiation model in vitro was used to simulate the growth and development process of bovine skeletal muscle. qRT-PCR and Western blot were used to detect the cellular temporal expression profile of MyBPC1. There were two groups in this experiment. At the RNA level, there were 4 replicates in each group, and 20 μL in each replicate. However, at the protein level, there were 3 replicates in each group, and 15 μg in each replicate. qRT-PCR and Western blot were used to detect the overexpression effect of MyBPC1 transfected into bovine skeletal muscle sate-llite cells, and to further detect the expression changes of cell proliferation marker factors Pax7, Ki67 and cell differentiation marker factors MyHC, MyOG. The myotube formation status of bovine skeletal muscle satellite cells was observed. The expression level of MyBPC1 was significantly different before and after the differentiation of bovine skeletal muscle satellite cells. After the differentiation of bovine skeletal muscle satellite cells, the mRNA and protein expression of MyBPC1 were significantly higher than those in the proliferation phase (P<0.01). After overexpression of MyBPC1, the number of myotubes formed by cell differentiation was significantly greater than that of the control group. There was no significant difference at the mRNA and protein expression levels of the proliferation marker factor Pax7. The mRNA and protein expression levels of the differentiation marker factor MyHC were significantly higher than that of the control group (P<0.01). Overexpression of MyBPC1 can promote the myogenic differentiation of bovine skele-tal muscle satellite cells in vitro, which lays the foundation for further studying the regulatory mechanism of MyBPC1 on bovine skeletal muscle satellite cells.

In mammals, spermatozoa acquired the ability of motility and fertilization only after being matured in the epididymis. To explain the maturation process of buffalo spermatozoa in the epididymis, sexually mature buffalo (Bubalus bubalis) epididymis were selected and the spermatozoa from the caput, corpus and cauda of the epididymis were purified by Polyvinylpyrrolidone coated silica (Percoll) gradient centrifugation. Computer-assisted sperm analysis system (CASA) was used to examine sperm motility, transmission electron microscopy to observe the ultrastructure of spermatozoa in different regions of the epididymis, and after fluorescent labeling of spermatozoa, flow cytometry and fluorescence microscopy were employed to determine the plasma membrane integrity, mitochondrial membrane potential and acrosome variations in different regions of the spermatozoa. As the result showed, the purity of spermatozoa obtained from the head, body and tail of epididymis by Percoll were 95%, whereas the motility of spermatozoa in various regions was 8.35%, 20.21% and 65.60%, respectively; All the spermatozoa in different regions of epididymis existed the same abnormality pattern, and the spermatozoa in the cauda of epididymis showed the highest ratio of mitochondrial membrane potential, the sperm plasma membrane integrity rate and and the sperm acrosome integrity rate increased gradually from the caput to the cauda region of epididymis. Overall, this study demonstrated the characterization and variation of spermatozoa in distinct regions of the buffalo epididymis which provides a theoretical basis for investigating the mechanism of sperm maturation.

The study aimed to investigate the differences in testicular development between Liao-ning cashmere goat and Ziwuling black goat at sexual maturity and to compare the reproductive performance between the two breeds. In this study, testicular tissues were collected from five healthy Liaoning cashmere goats and five healthy Ziwuling black goats at sexual maturity stage. The testicular tissue development and morphological differences between the two goat breeds were compared using anatomy and Hematoxylin-eosin (HE) stained, respectively. The concentration of androgen were detected using ELISA assay. The expression of DEAD box polypeptide 4 (DDX4) and Deleted in azoospermia-like gene (DAZL) in testis of the two breeds were detected using Real-time quantitative PCR(RT-qPCR) and Western blot technique. The study revealed that the total weight and length circumference of testicular in Liaoning cashmere goats were significantly higher than those in Ziwuling black goats (P<0.01), while the difference testicular breadth circumference, total testicular weight/pre-slaughter live weight and total testicular weight/carcass weight between the two goat breeds were not significant (P>0.05). The spermatogenic epithelial thickness of Liaoning cashmere goats was significantly higher than that of Ziwuling black goats (P<0.05), there were no significant differences in the area and diameter of the seminal tubules, number of seminal tubules per unit area between the two goat breeds (P>0.05). The difference of androgen secretion in testis between the two goat breeds was not significant (P>0.05). The expression levels of DDX4 mRNA and DDX4 protein in the testis of Liaoning cashmere goats were significantly higher than those of Ziwuling black goats (P<0.01 or P<0.05). The expression levels of DAZL mRNA in the testis of Liaoning cashmere goats was significantly higher than that of Ziwuling black goats (P<0.01), while the expression of DAZL protein between two goat breeds was not significant (P>0.05). In short, the degree of gonad development in Liaoning cashmere goats was identical with Ziwuling black goats, while the spermatogenic epithelium was thicker than Ziwuling black goats, there were relevant difference in the expression levels of reproductive marker genes between the two goat breeds, which might affect the spermatogenic ability between the two breeds.

The aim of study was to reveal the effect of ovine miR-200b on the follicular granulosa cells. The predicted target genes of miR-200b by 3 online tools (TargetScan, miRTarBase and miRDB) were analyzed with KOBAS for GO and KEGG pathway enrichment. The isolated and cultured sheep follicular granulosa cells were divided into 4 groups transfected with miR-200b mimic, mimic NC, miR-200b inhibitor and inhibitor NC,respectively, with 6 replicates in each group. CCK8 was used to detect the proliferation of granulosa cells at 24, 48 and 72 h after transfection. The expression level of miR-200b, CDK4, CDK6, CCND1, CCND2, Bax and Bcl-2 genes were measured by fluorescence quantitative PCR. A total of 25 common target genes of miR-200b were predicted by 3 online tools. These predicted target genes were enriched in cell proliferation and differentiation, cell cycle and reproductive development by GO and KEGG enrichment analysis. The survival rate of granulosa cell with time was a "V" decreased trendency and reached the lowest at 48 h (P<0.01) after transfection. There were no significant difference in miR-200b expression (P>0.05) between miR-200b inhibitor and inhibitor NC groups. Compared with mimic NC group, miR-200b mimic group significantly increased the expression of miR-200b (P<0.001), significantly down-regulated the expression of CDK4 (P<0.01), CDK6 (P<0.001), CCND1 (P<0.001), CCND2 (P<0.05) and Bcl-2 (P<0.01), while the expression of Bax did not significantly change (P>0.05), and extremely significantly decreased the ratio of Bcl-2 to Bax (P<0.001). In summary, miR-200b can inhibit the cell cycle and proliferation, and promote the apoptosis of follicular granulosa cells in sheep.

This study aimed to characterize the effects of neuropsin (OPN5) on apoptosis, prolife-ration and steroid hormone production of granulosa cells of duck. The duck granulosa cells were isolated and transfected with OPN5 overexpression plasmid and OPN5 siRNA, respectively. After continuous culture for 72 h (n=6), both proliferation and apoptosis of granulosa cell were detected using EdU cell proliferation assay, flow cytometry, Annexin V-FITC. The mRNA and protein expression levels, as well as hormone secretion were investigated by RT-PCR, Western blot and ELISA, respectively. The results showed that overexpression of OPN5 promoted prolife-ration and inhibited apoptosis of follicular granulosa cells in ducks. Moreover, overexpression of OPN5 significantly promoted the expression of GnRHR, FSHR, LHR, inhibited the expression of GnIH and GnIHR (P<0.01), significantly or highly significantly upregulated the mRNA levels of StAR, CYP11A1, 3β-HSD, CYP17A1 and CYP19A1 (P<0.05 or P<0.01), significantly increased the protein expression levels of OPN5, 3β-HSD and CYP19A1 (P<0.05), highly significantly increased the secretion levels of E2 and P4 (P<0.01). However, the levels of INHβ was highly significantly decreased (P<0.01). OPN5 knockdown with siRNA could effectively downregulate OPN5 expression in transfected cells (P<0.01), inhibit proliferation and promote apoptosis of follicular granulosa cells. siRNA transfection significantly downregulated the expression levels of GnRH, GnRHR, FSHR, LHR (P<0.01), promoted the expression of GnIH and GnIHR (P<0.01), significantly inhibited the expression of StAR, CYP11A1, CYP17A1 (P<0.01). OPN5 knockdown significantly decreased the protein expression levels of OPN5, 3β-HSD and CYP19A1 (P<0.05), highly significantly decreased the secretion levels of E2 and P4 (P<0.01), and highly significantly increased the level of INHβ (P<0.01). In conclusion, OPN5 promotes the proliferation and inhibits the apoptosis of duck follicular granulosa cells, and promotes the production and secretion of steroid hormones in granulosa cells.

This experiment was conducted to study the effect of prolactin (PRL) inhibitor on hair follicle development of cashmere goats during anagen. Twenty healthy Yanshan cashmere goats with similar body weight ((23.01±4.82)kg) aged 3 months old were selected. Goats were randomly divided into two groups with 10 goats in each group, which were control group and experimental group, respectively. The goats in experimental group were fed PRL inhibitor (0.06 mg·kg^-1) for 90 days. At the end of the experiment, cashmere fiber, blood and skin samples were collected to analyze the effects of PRL inhibitor on the length and fineness of cashmere, blood indexes, hair follicle traits and the expression levels of PRL, MTNR1a and RORα mRNA in skin tissues. The results indicated that the length and diameter of fibers were not significantly influenced (P>0.05) by the PRL inhibitor. The PRL inhibitor significantly increased primary and secondary hair follicle numbers and the ratio of secondary number to primary number (S/P,P<0.05), and significantly increased the percentage of active primary and secondary hair follicles in anagen (P<0.05). The PRL inhibitor had no significant effect on the serum concentrations of PRL, melatonin (MT), insulin-like growth factor-1(IGF-1), cortisol (COR), growth hormone (GH), tetraiodothyronine (T₄) and triiodothyronine (T₃), but significantly decreased mRNA expressions of PRL and MTNR1a in skin (P<0.05), and had no significant effect on mRNA expression of RORα (P>0.05). Inhibition of PRL secretion during the anagen phase of goat hair follicle may promote hair follicle development by reducing the expression of PRL and MTNR1a genes in the skin.

This experiment was conducted to investigate the effects of dietary different whole-plant corn silage levels on growth performance, slaughter performance, meat quality and serum biochemical parameters of geese. The experimental results could provide a basis for whole-plant corn silage in the application of geese. A total of 280 healthy 28-day-old Sanhua geese with similar body weight(982-985 g) were randomly divided into 4 groups with 7 replicates per group and 10 geese per replicate. The control group was fed a basal diet, the experimental groups were fed experimental diets supplied with 8%, 16%, or 24% whole-plant corn silage, the experimental period was 42 days. Recording the body weight (BW) and feed consumption of the geese at 28, 42, 56, and 70 days, and calculating the average daily gain (ADG), average daily feed intake (ADFI) and feed/gain (F/G). At 70 days of age, two geese with average body weight were selected and slaughtered to determine the slaughter performance, meat quality and serum biochemical indexes. The results showed as follows:compared with the control group, the ADG and ADFI of geese aged 28 to 42 days in the 16% experimental group were increased significantly(P<0.05); the ADFI of geese aged 42 to 56 days in the 8% experimental group was increased significantly(P<0.05); the ADG, ADFI and F/G of geese aged 56 to 70 days in the all experimental groups had no significant difference(P>0.05); the ADG of geese aged 28 to 70 days had a quadratic curve relationship with the supplemental level(P<0.05), when the ADG achieved the highest, the supplemental level was 9.01%. Compared with the control group, the BW of 70-day-old geese in the 24% experimental group was significantly reduced(P<0.05). The dressing percentage and eviscerated percentage of geese were reduced significantly in all experimental groups at 70 days of age(P<0.05). The dressing percentage of geese had a quadratic curve relationship with the supplemental level(P<0.05), when the dressing percentage achieved the lowest, the supplemental level was 14.93%. There were no significant differences in meat quality among all experimental groups at 70 days of age(P>0.05).The triglyceride content of geese in the 8% experimental group was significantly reduced at 70 days of age(P<0.05) compared with the control group. In conclusion, under the conditions of this experiment, appropriate supplemental level of whole-plant silage corn can improve the growth performance of the geese and reduce the content of triglyceride content in the serum, there is no significant impact on meat quality, but a certain negative impact on slaughter performance. According to the ADG maximum limit and dressing percentage minimum limit,it is recommended that the dietary optimal whole-plant corn silage supplement level is in the range of 9.01% to 14.93%.

The experiment was conducted to investigate the effects of puerarin on mucosal morphology, mRNA expression of tight junction protein and antioxidant capacity of small intestine in yellow-feathered broilers fed oxidized soybean oil. The experiment was designed with 2×2 factors including oil quality (fresh soybean oil and oxidized soybean oil) and puerarin addition levels (0, 750 mg·kg^-1). A total of 360 1-day-old healthy female yellow-feathered broilers were randomly divided into 4 treatment groups with 6 replicates per group and 15 chickens per replicate, which were fresh soybean oil diet group, fresh soybean oil + puerarin (750 mg·kg^-1) diet group, oxidized soybean oil diet group and oxidized soybean oil + puerarin (750 mg·kg^-1) diet group, respectively. At 28 and 56 days of age, one chicken per replicate was randomly selected, and the morphology of small intestinal mucosa, the mRNA expression of tight junction protein and antioxidant indices of small intestine mucosa were determined. The results showed that:1) Consumption of oxidized soybean oil significantly decreased villus height and villus height/crypt depth (V/C) in duodenum of 28-day-old broilers, V/C and mRNA expression levels of claudin-1 in ileum, villus height and V/C in three intestinal segments of 56-day-old broilers (P<0.05). Consumption of oxidized soybean oil significantly increased crypt depth of three intestinal segments at 56 days of age (P<0.05). Dietary supplementation of puerarin significantly increased villus height and V/C in jejunum and Zonula occluden1 (ZO-1) mRNA expression level in ileum of broilers at 28 days of age, and V/C in duodenum, jejunum and ileum and villus height in jejunum and ileum of broilers at 56 days of age (P<0.05), and significantly decreased crypt depth in three intestinal segments (P<0.05). 2) Oxidized soybean oil significantly increased the glutathione (GSH) content in duodenum and activity of glutathion peroxidase (GSH-Px)in jejunum at 28 days of age (P<0.05), and significantly decreased the GSH content, superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC) in ileum at 28 days of age (P<0.05). Dietary supplementation of puerarin significantly increased the activity of GSH-Px in duodenum of broilers at 28 days of age (P<0.05). Dietary oxidized soybean oil significantly decreased the activity of SOD and T-AOC in ileum at 56 days of age (P<0.05), dietary supplementation of puerarin significantly increased the activities of SOD in duodenum and ileum, and GSH-Px in ileum (P<0.05), and significantly decreased the content of GSH and the activity of SOD, T-AOC in jejunum and T-AOC in ileum (P<0.05) of broilers at 56 days of age. In conclusion, consumption of oxidized soybean oil damaged intestinal mucosal morphology and decreased the mRNA expression of tight junction protein and antioxidant capacity of small intestinal. Puerarin could increase the mRNA expression of tight junction protein in small intestinal mucosa of broilers, improve the morphological structure of intestinal mucosa under oxidative injury, increase the activity of antioxidant enzymes and enhance the antioxidant capacity of broilers.

This study was conducted to investigate the potential regulatory role of circRNAs in the definitive host liver in the pathogenesis induced by Toxocara canis. The six-to-seven-week-old Beagle puppies (n=18) were averagely divided into three stages (0.5 day post infection, 1 dpi, and 36 dpi), and their liver samples were collected at the corresponding time points after infection. Each stage included infected group and control group. Total RNA was extracted from the livers of infected and uninfected puppies, and the circRNA libraries were constructed and sequenced by high-throughput RNA sequencing. The differentially expressed (DE) circRNAs were analyzed, and GO and KEGG enrichment analysis were performed using the source genes of DE circRNAs. The nine DEcircRNAs were randomly selected for quantitative real-time PCR (RT-qPCR) verification. Compared to the control groups, 94, 103, and 84 DE circRNAs of dogs' livers were detected in the livers of infected dogs at the three infection stages, respectively. Venn plot analysis showed that there were no co-expressed DE circRNAs among the three stages of infection. GO and KEGG analysis showed that some DE circRNAs were related to immunity and inflammation in the host liver. Among these DE circRNAs, novel_circ_0016108, novel_circ_0016184 and novel_circ_0027468 were related to the natural immunity of host against T. canis infection; novel_circ_0002212 was involved in liver inflammation caused by T. canis; novel_circ_0002212 and novel_circ_0019907 play a role in the host liver's resistance to T. canis invasion. RT-qPCR validation showed that the expression trends of DE circRNAs obtained by both methods were consistent. These results suggest that circRNAs play an important role in the infection of definitive host (dogs) by T. canis. This study provided new information for further understanding of the interaction between T. canis and its definitive host, and will also facilitate the development of interventions for the disease.

The Theileria annulata infected host cells obtained unlimited proliferation as tumor cells, and the phenomenon of parasite-dependent transformation is reversible. The T. annulata transformed cells provide an ideal model for better understanding the molecular mechanism of parasite-host cells interactions. In the present study, the targeted metabolomics method was applied out to investigate the changes of metabolites of the transformed cells related with energy metabolism. Aimed to investigate the difference of cells proliferation, Buparvaquone treatment group (BW720c), DMSO control group and blank control group were set. Cell samples were collected at 72 h, and the energy metabolism-related metabolites of the cell samples were detected by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) targeted metabolomics method. Then, the compound types of the metabolites were identified by the energy database. The differential metabolites were screened by the Partial Least Squares-Discriminant Analysis (PLS-DA).With the results of cells number counting, it was found that there has no significant change between the three groups within 24 h. However, the cell growth rate of the experimental group was significantly lower than the other two control groups at 36 h, and the difference between the two groups was largest at 72 h. PLS-DA analysis showed that metabolites variables could be clustered and distinguished between the drug treatment group and DMSO control, and 21 differential metabolites between two groups were screened out. In the drug treatment group, 8 metabolites were up-regulated and 13 metabolites were down-regulated. After enrichment analysis, it was found that these metabolites were mainly related to energy metabolism, such as glutamine, L-lactic, malic, pyruvic acid, and isocitrate. Meanwhile, it was found that the changes of glutamine, L-actic and pyruvic acid contents in the drug treatment group and the control group were consitent with the results of targeted metabolomics detection by using the corresponding kits. The results indicated that T. annulata infection can significantly affect the energy metabolism of the host cells, which lays a foundation to illustrate the mechanisms of cells transformation by T. annulata.

This study was conducted to explore the methods and its feasibility of identifying Canidae species and to investigate the infection of Echinococcus in wild Canidae based on non-invasive fecal sample analysis. A total of 101 DNA samples extracted from feces of wild Canidae were amplified and sequenced with a pair of universal primers designed for the mitochondrial DNA control region of mammalian species. The species origins of the fecal samples were molecularly identified by the analysis of nucleotide variation, the comparison of sequence similarity, the analysis of genetic distance and the construction of phylogenetic tree. Meanwhile, copro-antigen ELISA was used to investigate the infection of Echinococcus. The test success rate for the species identification of the samples was 73.27%. A total of 20 haplotypes were inferred from the 74 sequences obtained in the study. Each of the haplotype sequences was found to match with one single species in the GenBank database with 98.52%-100% sequence similarity. All the haplotypes can be divided into 4 haplotype sets according to sequence differences. The average number of nucleotide differences among the 4 haplotype sets is 17.83-70.10, and the number of nucleotide differences among the haplotypes within each set is 1-10. The genetic distance among the 4 haplotype sets is 0.068-0.342. The value is much larger than that of the genetic distance among the haplotypes within each set, which is 0.009-0.022. The phylogenetic analysis showed that haplotypes in the same sets clustered into same clades with high bootstrap values. Based on all the analysis results, it can be inferred that the sources of the fecal samples were Canis lupus familiaris, Canis lupus, Vulpes vulpes and Vulpes ferrilata respectively. A total of 11 positive samples were found in the copro-antigen ELISA test. The positive rate of Echinococcus antigen was 10.89%. The analysis of mitochondrial DNA control region sequence can be used for the molecular identification of the Canidae. The detection accuracy rate would be further improved if other molecular markers such as nuclear genes were aslo used. The positive rate of Echinococcus antigen is high in feces of wild Canidae in the study area.

This study aimed to evaluate the characteristics of mannosylated chitosan polylactic acid-glycolide copolymer[poly(D,L-lactide-co-glycolide), PLGA] nanospheres as a delivery system of FMDV nucleic acid vaccine. A certain degree of substitution of mannose modified chitosan (MCS) was prepared by schiff base reaction and elemental analysis. Then, Mannose modified chitosan PLGA nanospheres (MCS-PLGA-NPs) were prepared by double emulsification and volatilization. The particle size distribution and surface potential (zeta) of MCS-PLGA-NPS were measured by nanometer particle size analyzer; The morphology of MCS-PLGA-NPS was investigated by scanning electron microscopy; The ability of plasmids adsorbed on MCS-PLGA-NPs to resist nuclease degradation was observed by agarose gel electrophoresis; The cytotoxicity of MCS-PLGA-NPs was detected by CCK-8 method; The MCS-PLGA-NPs-DNA complex uptake by macrophages was observed by confocal laser scanning; The expression of MCS-PLGA-NPs loaded plasmid DNA in cells was verified by fluorescence microscope and Western blot. The results of elemental analysis showed that MCS with a substitution degree of 5% to 10% was successfully prepared; The results of nanometer particle size measurement and scanning electron microscopy showed that the zeta of MCS-PLGA-NPs was positive, the particle size distribution was uniform and the morphology was spherical; The results of agarose gel electrophoresis showed that the ability of MCS-PLGA-NPs to adsorb plasmids increased with the increase of their quality and could resist nuclease degradation of plasmid DNA to a certain extent; In the cytotoxicity test, the survival rate of RAW264.7 cells was still more than 85% after 24 h co-incubation with different concentrations of MCS-PLGA-NPs; In the cell uptake experiment, confocal laser microscopy could clearly observe the plasmid DNA binding to the surface of the nanomicrosphere and uptake by RAW264.7 cells; Fluorescence microscopy and Western blot experiments showed that MCS-PLGA-NPS-loaded plasmid DNA could be expressed in cells. In summary, MCS and MCS-PLGA-NPs with the ability to deliver DNA vaccine of FMDV were successfully prepared, which provided new directions and insights for the delivery of FMDV nucleic acid vaccine, and also laid a foundation for further research on the carrier carrying specific antigen targeting mannose receptor on APCs surface and its application in animal immunity.

H9 subtype avian influenza is widely prevalent in poultry in China, which has caused huge economic losses to the poultry industry and also seriously threatened the public health safety. The high genetic variability of H9 subtype avian influenza virus (AIV) leads to poor antigenic matching between the circulating and vaccine strains, thus affecting the clinical protective effect of the vaccine. Therefore, it is urgent to develop a highly effective cross-protective vaccine against H9 subtype AIV. Mosaic vaccine is designed for the genetic diversity of pathogens by integrating all antigenic sequences to obtain a mosaic protein with the most extensive epitopes coverage, and to generate vaccine. In order to develop an efficient and universal H9 subtype avian influenza vaccine, a mosaic HA sequence of H9 AIV HAm/H9 was designed, optimized and synthesized according to the principles of mosaic vaccine. Utilizing the reverse genetic technology, a recombinant virus rPR8-HAm/H9 was constructed and rescued by replacing the HA segment of influenza virus PR8 strain (H1N1) with the HAm/H9 gene. The SPF chickens were immunized with the inactivated rPR8-HAm/H9 virus, and the effect of cross-protection was evaluated by detecting the antibody titer, the protection rate and viral shedding. The recombinant virus rPR8-HAm/H9 could induce high level of HI antibody and neutralizing antibody, as well as reduce virus shedding, and the protection rate against H9N2 AIV JM0305 strain was 80%. Our results showed that rPR8-HAm/H9 inactivated vaccine could provide efficient cross-protection against H9N2 AIV strain JM0305, and provided a preliminary basis for the research and development of universal vaccine for avian influenza based on mosaic technology.

This study was conducted to evaluate the immune enhancement effect and mechanism of pig spleen transfer factor(TF) on Newcastle disease virus(NDV) attenuated vaccine LaSota strain. Specific pathogen free(SPF) chickens were inoculated with different doses(10^-2, 10^-3,10^-4, 10^-5) of NDV strain LaSota(vaccination group without TF), NDV strain LaSota combined with TF(vaccination group with TF), respectively, and challenged with virulent NDV strain F₄₈E₉ on 14 days post vaccination(dpv). The control group(non-immunized but challenged) and blank group(non-immunized and non-challenged) were set up simultaneously. In addition, the change of IL-4, IFN-γ and IL-12 P40 in peripheral blood were analyzed by ELISA and protein chip technology. The protection rate of different doses and median protective dose(PD₅₀) were 100%, 55%, 0%, 0%, 0.000 8 doses in the plume in vaccination group without TF, respectively, and 100%, 75%, 0%, 0%, 0.000 5 doses in the plume in vaccination group with TF. In contrast, the mortality of control group and blank group were 100% and 0%. After immunization with 10^-3 dose of vaccine, the levels of IL-4 and IFN-γ in vaccination group with TF were higher than in other groups and the difference was extremely significant on 7 and 14 dpv(P<0.01). After challenge, the levels of IL-4, IFN-γ and IL-12 P40 in vaccination group with TF were higher than in other groups. The differences of IL-4 on 1 and 14 days post challenge (dpc), IFN-γ on 1 and 3 dpc, IL-12 P40 on 1 dpc were extremely significant(P<0.01). These findings indicated that TF could reinforce cellular immunity by IFN-γ, IL-12 and humoral immunity by IL-4, improve the immune protection rate of NDV attenuated vaccine LaSota strain and reduce the PD₅₀. Vaccination group with TF was more effective than vaccination group without TF against virulent NDV strain F₄₈E₉.

This study aimed to investigate the binding domain and intracellular localization of chicken invariant chain (Ii) to the endosome transporters Rab5a and Rab7b. First, the cytoplasmic region and transmembrane region (Ii_(Cyt-Tra)), Ii CLIP (class Ⅱ-associated invariant chain peptide) with trimer region (Ii_(CLIP-TRIM)) and Ii mutants (Ii_(M81-87aa),Ii_(M91-99aa)and Ii_(M81-99aa)) obtained by PCR and gene mutation techniques were inserted into pET-32a and pEGFP-C1 to consturct prokaryotic and eukaryotic recombinant plasmids, respectively, the corresponding eukaryotic and prokaryotic recombinant expression plasmids were constructed. Secondly, the constructed recombinant plasmid containing green fluorescent protein and the recombinant plasmids containing red fluorescent Rab5a and Rab7b were co-transfected into human embryonic kidney cell line 293 T, and their co localization was observed. Finally, the binding domain of Ii with Rab5a and Rab7b was detected by pull-down method and Western blot. The results showed that the recombinant plasmids of Ii domain and mutant were successfully constructed. Both Ii _(cyt-tra) and Ii mutants could co localize with Rab5a and Rab7b, but Ii _(CLIP-TRIM) and empty vector could not. The cytoplasmic and transmembrane domains of Ii are the main domains binding Rab5a and Rab7b, not CLIP and trimer domains. In conclusion, the colocalization and binding regions of chicken Ii with Rab5a and Rab7b are cytoplasmic and transmembrane regions, not endoplasmic reticulum cavity regions. These results further suggest that Rab molecule is involved in the transport mechanism of Ii in intracellular organelles, and provide a new way for further study of the transport mechanism and function of Ii and its carrier in cells.

In order to study the effect of restraint stress on the neuroplasticity of pregnant sows, 6 Bama miniature sows aged 8-9 months were randomly divided into two groups:stress group (n=3) and control group (n=3). The remaining conditions were consistent.The pregnant pigs were sacrificed on the 18th day of gestation. The hippocampal and blood samples were collected and the hormone levels were detected by ELISA and radioimmunoassay.After the paraffin-embedded hippocampal sections were prepared and stained by Nishner staining and silver staining, the neuron loss rate, dendritic complexity and the number of dendritic spines were observed, and the expression level of brain-derived neurotrophic factor (BDNF) in the hippocampus was detected by Western blot. The results showed that the plasma levels of corticotropin-releasing hormone (CRH), adrenocorticotrophic (ACTH) and cortisol (COR) increased significantly in the stress group (P ≤ 0.05), in the hippocampus dentate gyrus (DG) and CA3 area neurons leakage phenomenon, CA3 area down cone cells dendritic complexity, mature dendritic spines of neurons and the quantity of immature dendritic spines are significantly reduced, DG mature dendritic spines significantly reduced, and the amount of BDNF protein expression in hippocampus was reduced by 70.6% (P ≤ 0.01). Therefore, restraint stress attenuates the neuronal plasticity of CA3 and DG in the hippocampus of pregnant pigs.

The purpose of this study is to observe the effects of different durations of thermal stimulation on the neurobehavior of mice at 34℃ and the specific changes in the blood physiological indicators of mice under this condition. Eight weeks old mice were selected and divide them into 4 groups with 10 mice in each group. After grouping, the mice were placed in a 34℃ environment for thermal stimulation for 0, 2, 4 and 6 hours. Immediately after the stimulation, the mice were subjected to open-field and elevated plus maze tests to observe and record the movement of the mice within 5 minutes, and take blood samples from the mice to detect their blood physiological indicators. Results were as follows:Compared with the control group, the exploratory behavior of the mice in the 2 h heat stimulation group was significantly increased, while the anxiety-like behavior of the mice in the heat stimulation 4 h group and the heat stimulation 6 h group was significantly increased, and the exploratory behavior was reduced; the levels of EPI and CORT of the mice in heat stimulation 2 h group increased in a time-dependent manner and the difference were significant; the levels of norepinephrine (NE) in the 2 h, 4 h, and 6 h heat stimulation groups increased extremely significantly, and it reached a peak at 4 h of heat stimulation and slightly decreased at 6 h; the level of corticotropin-releasing hormone (CRH) in mice in the heat-stimulated 4 h group increased significantly, and decreased slightly at 6 h. The results suggest that heat stimulation at 34℃ for 2 hours promotes spontaneous movement and exploratory behavior in mice. As the stimulation duration increases, the spontaneous movement and exploratory behavior of mice are significantly inhibited, and anxiety-like behaviors increase significantly, showing a time dependence, and suggest that the emotional and behavioral abnormalities of mice after thermal stimulation may be regulated by the central nervous system.

In order to explore the mechanism of Aspergillus terreus causing liver injury of mice, 20 Kunming mice were divided into a control group and a test group. The test group was intraperitoneally injected with 0.3 mL (5×10⁷CFU·mL^-1) spore suspension. The trail was 7 days. The indicators of oxidative damage in mouse liver tissue were evaluated by malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-PX) and superoxide dismutase (T-SOD) kits. The iron content of liver tissue was determined by iron kit. Prepare liver paraffin sections to observed the pathological changes and iron accumulation by HE and Perls stain. The ultrastructure of hepatocytes was observed by transmission electron microscope. The mRNA relative transcription level of ferroptosis-related genes in liver tissue were detected by qPCR. The results were as follows:compared with control group, the content of MDA in test group was significantly increased, but the content of GSH,the activities of GSH-PX and T-SOD in test group were significantly decreased (P<0.01). Meanwhile, the content of iron ions in test group was significantly increased (P<0.01). Some hepatocytes were swollen and necrotic. A few inflammatory cells were infiltrated. Iron ions accumulated in the center of hepatic loules. The mitochondria atrophied, cristae reduced, and membrane density increased in hepatocytes. The relative mRNA transcription level of transferrin receptor protein 1 (TFR1), iron uptake-related proteins divalent metal transporter-1 (DMT1), ferritin heavy chain 1 (FTH1), and voltage-dependent anion channel 3 (VDAC3) genes were significantly up-regulated (P<0.05), while, the relative mRNA transcription level of glutathione peroxidase 4 (GPX4) and solute carrier family-7 member-11 (SLC7A11) genes were significantly down-regulated (P<0.05). These results indicated that Aspergillus terreus MSF2 could cause ferroptosis in the liver of mice. It benefits for further study of the pathogenic mechanism of Aspergillus terreus.

The purpose of this experiment was to prepare Phyllanthus emblica polysaccharide (PEP) and analyze its structural characterization and investigate the effect of PEP on immune effect of Newcastle disease (ND) vaccine. The structural characterization of PEP was analyzed by ultraviolet spectroscopy (UV), Fourier transform infrared spectroscopy (FT-IR), high performance liquid chromatography (HPLC) and scanning electron microscope (SEM). At the same time, 175 1-day-old Hetian chickens were randomly divided into 5 groups with 35 chickens in each group, including blank control group, vaccine control group, PEP low-dose group, PEP medium-dose group and PEP high-dose group. All chickens were immunized with ND vaccine at 7 days old except the blank control group, and immunized second times at 28 days old. At the same time of immunization, the low-, medium- and high-dose groups of PEP were given 0.5 mL of PEP solution according to the doses of 10, 20 and 30 g·L^-1, respectively, while vaccine control group and blank control group with physiological saline, once a day for 3 successive days. On 7, 14, 21, 28, 35, 42, and 49 days old, the levels of Newcastle disease virus (NDV) antibody, INF-γ and IL-4 were tested. On 28 and 49 days old, the proliferation of splenic lymphocytes and CD4⁺/CD8⁺ratios were measured. Chickens were slaughtered and the thymus, spleen, bursa and liver were take at 49 days old, then the organs index were calculated. The results showed that PEP was white cotton flocculent, the total carbohydrate content of PEP was (86.70±1.05)%. It was a kind of homogenous polysaccharide containing α-type glycosidic bond without nucleic acid and protein. Also, it was determined to be composed of glucuronic acid (GlcA), glucose (Glu), xylose (Xyl) and fucose (Fuc) in the molar ratio of 0.5:8.3:1.8:0.3. It was distributed in sheet shape, and its overall structure was regular and dense. The results of animal experiment found that PEP significantly increased the level of NDV specific antibody, IFN-γ and IL-4 (P<0.05), the proliferation of splenic lymphocytes, the immune organ index and CD4⁺/CD8⁺ratios (P<0.05). Also activated immune function of spleen, bursa, thymus and liver. The total carbohydrate content of PEP was high and its structure was stable. Also, PEP could improve the immune activity of the body, possess good adjuvant activity and control effect on the ND for chicken.

The aim of this study was to investigate whether mitochondrial calcium uniporter(MCU) mediated mitochondrial Ca²⁺ transport was involved in hypoxia-induced mitochondrial damage in broiler cardiomyocytes. In this experiment, the primary cardiomyocytes of chicken embryos of white feathered broilers were isolated and cultured for 24, 48 and 72 h under hypoxic conditions (3%O₂, 5%CO₂, 92%N₂). In order to further determine the role of MCU in mitochondrial injury induced by hypoxia, RU360 was pretreated in the hypoxia group for 72 h to inhibit the expression of MCU. The intracellular Ca²⁺ concentration, mitochondrial Ca²⁺ concentration, mitochondrial reactive oxygen species level and mitochondrial membrane potential were detected by flow cytometry. Gene and protein expressions of MCU and its regulatory factors were detected. The results showed that the purity of cardiomyocytes cultured by differential velocity adherent method could reach over 90%. After 24 h of hypoxic culture, the expression of MCU and MICU1 genes were increased (P<0.05), the concentrations of Ca²⁺ in cytoplasm and mitochondria were significantly increased (P<0.05), and the mitochondrial membrane potential was increased (P<0.05). The expression levels of MCUR1 and MICU1 mRNA were decreased (P<0.05) and intracellular Ca²⁺ concentration was increased (P<0.05) after 48 h hypoxia culture. After 72 h of hypoxic culture, MCU mRNA expression, intracellular and mitochondrial Ca²⁺ were increased (P<0.01), and reactive oxygen species were increased (P<0.01), while mitochondial membrane potential decreased (P<0.01). Compared with the 72 h hypoxic group, the Ca²⁺ in cells and mitochondria decreased, the mitochondrial membrane potential increased and the reactive oxygen species decreased in the RU360 preconditioning group (P<0.01), the expression of MCU mRNA was decreased (P<0.01). The results showed that hypoxia-induced MCU upregulation leads to mitochondrial calcium overload, causing mitochondrial dysfunction and cardiomyocyte injury. Inhibition of MCU expression can reduce hypoxia-induced mitochondrial calcium overload and protect mitochondrial function.

The aim of this study was to explore the efficacy of adipose-derived mesenchymal stem cells condition medium (ADSCs-CM) on the inflammatory response of liver ischemia reperfusion combined with partial hepatectomy in miniature pigs. Based on the laparoscopic technique, a model of liver ischemia reperfusion (IR) combined with partial hepatectomy was established in 24 miniature pigs. Miniature pigs were divided into four groups:model group (IRI), DMEM group (DMEM), ADSCs-CM group (CM) and ADSCs group (ADSCs), with six pigs in each group, according to the four different substances namely physiological saline, concentrated basal medium, concentrated ADSCs-CM, ADSCs transplanted to the liver after surgery. The blood and liver tissue samples were harvested pre-operation, 1 day, 3 days, and 7 days after operation. The effect of ADSCs-CM on inflammation were comprehensively evaluated through histopathological inflammatory cell observation, blood indicators detection, serum cortisol (COR), C-reactive protein (CRP) and hyaluronic acid (HA) detection of ELISA, liver tissue inflammation-related IL-1β, IL-6, TNF-α, IL-10 mRNA detection of qRT-PCR. The results found that 1 and 3 days after the operation:there were more inflammatory cells in the pathological tissue sections of the IRI group and DMEM group. The results of blood routine and inflammation-related genes also showed that there was obvious inflammation after the operation. The ADSCs-CM and ADSCs treatment groups significantly reduced the infiltration of inflammatory cells in the tissues and the number of white blood cells (WBC), neutrophils (NE) and lymphocytes (LY) in the blood, and decreased the expression levels of pro-inflammatory factor IL-1β, IL-6, TNF-α mRNA in tissues, and increased the expression level of anti-inflammatory factor IL-10 mRNA. After 7 days, each group basically recovered to the pre-operation. The results showed that liver ischemia-reperfusion combined with partial hepatectomy can cause inflammation, and both ADSCs-CM and ADSCs improved the inflammatory response of liver ischemia-reperfusion combined with partial hepatectomy. Transplantation of ADSCs-CM has the potential to become a cell-free therapy for the treatment of inflammation in the future.

This study was conducted to examine the distribution of phylogenetic clustering, serotype, virulence genes, drug resistance and genetic diversity of Shiga toxin-producing Escherichia coli (STEC) from cattle, sheep and camel in Xinjiang. Phylogenetic clustering, serotype and virulence genes stx₁, stx₂ (including subtypes), eaeA and hlyA of STEC isolates were tested by PCR method from cattle, sheep and camel, drug sensitivity of isolates was determined by using a Kirby-Bauer disk diffusion method and genotypical analysis was performed by enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR). The results showed that 94 of non-O157 STEC isolates which were mainly B1 group and contained 9 serum groups, including O146 (n=14), O22 (n=7), O3 (n=4), O168 (n=4), O8 (n=3), O167 (n=2), O88 (n=1), O112ab (n=1) and O147 (n=1). The virulence gene test of showed that 46.8% (44/94) carried stx₁ only, 6.4% (6/94) for stx₂ only, and 46.8% (44/94) carried both stx₁ and stx₂. STEC isolated from sheep and camel mainly carried stx₁+hlyA (68.0% and 25.0%, respectively), while STEC isolated from cattle mainly carried stx₁+stx₂+hlyA (57.9%). stx_1awas mainly distributed in bovine STEC and stx_1cwas mainly distributed in STEC of sheep. Fourteen strains (14.9%) were drug-resistant, and the drug resistance rates of STEC isolates to ceftazidime, tetracycline, cefotaxime, ampicillin and amtronam ranged from 3.2% to 5.3%, to cotrimoxazole, cefepime, piperacillin-tazobactam, ampicillin-sulbactam, amoxicillin-clavulanate and polymyxin B ranged from 1.1% to 2.1%. ERIC-PCR results showed that STEC from cattle, sheep and camel were closely related. Cattle, sheep and camels carry a variety of known serotypes of STEC and store abundant virulence genes, which may potentially infect human beings. Therefore, it is necessary to prevent and control the contamination of meat during slaughtering and processing.

Metastasis of adrenal cortical carcinoma to the spleen in cats is very rare in veterinary clinical cases. In this report, a 10-year-old farm cat with abdominal pain, frequent vomiting, wasting, a history of chronic adrenal impairment, and splenomegaly on radiographic examination was described. Spleneectomy was performed on the cat and histopathological examination of the removed tissue was performed. Histopathological results showed that the tumor cells had abundant lipid-like vacuoles, large cell volume, and many mitotic images. The tumor cells had vascular invasion, and the nucleus was heavily stained. Immunohistochemical results showed positive expression of synaptophysin, S-100 and GATA4. Based on histopathology and immunohistochemistry, history and laboratory examination, the final diagnosis was splenic metastasis of adrenal cortical carcinoma. The clinical case and pathological diagnosis of splenic metastasis of adrenal cortical carcinoma in cats were reported for the first time, which has certain reference value for the clinical diagnosis and treatment of related diseases.

This study was conducted to explore the reason why traditional Chinese medicine extractives and swine urines from pigs fed with some herbal were tested positive by CGIA. A LC-MS/MS method was established to determine synephrine in pigs' urine and traditional Chinese medicine extractives. Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium's extractives were determined, along with urine after herbal feed. In vitro mouse liver microsomes model was used to incubate the extractives of Citri Reticulatae Pericarpium Viride, Citri Reticulatae Pericarpium, Magnoliae Officinalis Cortex, and Chaenomelis Fructus. Synephrine was detected in pig urines after feeding with Citri Reticulatae Pericarpium and Citri Reticulatae Pericarpium Viride, and the concentration was 1.36 and 1.65 μg·mL^-1 respectively. The concentration of synephrine in Citri Reticulatae Pericarpium and Citri Reticulatae Pericarpium Viride's extractive was 132.6 and 312.7 μg·mL^-1 respectively. Extractives of Magnoliae Officinalis Cortex and Chaenomelis Fructus were tested gradually from positive to negative during 2-5 h incubation. Extractives of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium were tested positive during 0-6 h incubation. The experiment group of negative herbs were tested negative during incubation, and control group of synephrine matched were tested positive during 0-6 h incubation. Great dosing of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium in swine can cause false positive results of β-agonists'CGIA test in swine urine, and this phenomenon is related to synephrine. Synephrine is hard to be metabolized by mouse liver microsomes in vitro.