The maturation quality of oocytes not only is the basis of mammalian reproductive capacity, but also directly determines the quality of offspring. Premature ovarian failure usually causes a decline in the number and quality of oocytes, and how to improve its maturation quality has become a frontier topic in the field of reproductive aging. In recent years, the functions of nicotinamide adenine dinucleotide (NAD+) dependent deacetylase family Sirtuins (SIRT1-7) in reproductive aging have attracted increasing attention. Particularly, the acetylated substrates of the anti-aging factor SIRT2 are directly related to oocyte maturation events. This review focused on the recent research progress of NAD+/SIRT2 in improving the quality of aging oocytes through important physiological processes such as meiosis, energy metabolism, mitochondrial oxidative stress and mitochondrial quality control from a new perspective of acetylation regulating the aging and maturation of oocytes. It is expected to provide new ideas for improving the oocyte quality and prolonging the reproduction period in aged female livestock.

Cold stress caused by continuous low temperature in winter is an important factor that reduces the economic benefits and animal welfare of northern animal husbandry, and can induce animal inflammation and oxidative stress to endanger the health of the organism.As an important molecular chaperone protein in the organism, heat shock protein plays an important role in maintaining the homeostasis of the organism's environment and helping animals resist stress. The low temperature environment can activate the rapid production of heat shock proteins, which can play an important role in regulating the immune and antioxidant systems inside and outside the cell: outside the cell, it has the functions of protecting cells, participating in regulating immune cell functions, stimulate immune response, and improve anti-oxidation enzyme activity; inside the cell, it can inhibit the NF-κB signaling pathway to protect the organism from inflammatory damage, up-regulate the Nrf2 signaling pathway to improve the organism's antioxidant function, and alleviate the negative effects of cold stress on the organism. This article summarizes the regulatory effects and mechanisms of heat shock proteins on the immune and antioxidant functions of the organism under cold stress conditions at home and abroad and gives some methods to increase the expression level of heat shock proteins, in order to provide references for subsequent research on the cold stress theory of livestock and poultry.

Quorum sensing (QS) is a microbial cell-to-cell communication mechanism that plays a key regulatory role in bacterial virulence factor expression, bioluminescence, spore formation, and biofilm formation. In recent years, researchers have found that QS also exists in the gastro-intestinal tract of livestock and poultry. In view of the significant role of gastro-intestinal microflora in the digestion, physiological metabolisms, and immune functions of animals, research and regulation of gastro-intestinal bacterial QS is of great significance in guaranteeing health and improving production performance. In this paper, the classifications and effects of bacterial QS, and the research findings of gastro-intestinal bacterial QS in different domestic animals was reviewed, and the relevant regulation and potential mechanisms of QS were also introduced, in order to provide reference for the research and application of bacterial QS in livestock and poultry industry.

Bacterial resistance has become a key problem to be solved in clinical practice, especially the drug resistance caused by Gram-negative bacteria, which bring great challenges to clinical treatment. It is particularly important to clarify the regulatory mechanism of complex drug resistance phenotypes of important bacteria as soon as possible. Two-component regulatory systems (TCS) exist in a variety of Gram-negative bacteria and play important roles in the life activities of bacteria. TCS is one of the principal mechanisms by which bacteria perceive environmental changes and produce corresponding regulations. TCS is usually composed of two proteins, including sensor proteins (usually histidine kinases) and responsive regulatory proteins (usually transcription factors). Both of them can integrate the environmental signals of bacteria, regulate gene expression and change the physiological behavior of bacteria through phosphorylation-mediated synergy. The response mechanism studies of TCS-mediated drug resistance have become a new research hotspot recently. In this paper, the structural basis and mechanism of TCS-mediated drug resistance of clinically important Gram-negative bacteria were reviewed. It would be enhancing the comprehensive understanding of the bacterial TCS system and provide new ideas and strategies for the scientific research and development of drugs in the future.

This study aimed to identify key genes affecting fat deposition and explore the function of adipose tissue, liver and muscle in Songliao black pigs. In this study, 6 healthy Songliao black pigs with weight of about 100 kg and significant difference in backfat thickness (3 high and 3 low) were selected as experimental animals. The gene expression levels in fat, liver and longissimus dorsi muscle were detected by high-throughput transcriptome sequencing, the differentially expressed genes in pigs with different fat deposition levels and different tissues were identified, and the biological functions of the differentially expressed genes were analyzed. The results showed that 135 differentially expressed genes were found in different groups of pigs, some of which were involved in PPAR signaling pathway, AMPK signaling pathway, metabolic pathway, fatty acid metabolism and glycerol metabolism. The biological function analysis showed that EHHADH, ME1, SCD, OLR1, PHGDH, ACLY, LEP and CYP gene family were the key genes affecting pig fat deposition. Among the differentially expressed genes in different tissues, the highly expressed genes in adipose tissue were significantly enriched in insulin signaling pathway, MAPK signaling pathway, tricarboxylic acid cycle, oxidative phosphorylation pathways, and so on; The highly expressed genes in the liver were significantly enriched in the metabolism of various substances, the degradation of fatty acids and the synthesis of amino acids; The highly expressed genes in longissimus dorsi muscle were mainly involved in protein degradation, PI3K-Akt signaling pathway, oxidative phosphorylation pathway, Wnt signaling pathway and phosphatidylinositol signaling pathway. The analysis results of differentially expressed genes among different groups suggest that EHHADH, ME1, SCD, OLR1, PHGDH, ACLY, LEP and CYP gene family are the candidate genes affecting fat deposition; The differentially expressed genes between different tissues show that adipose tissue is the main site of adipogenesis, while liver and muscle tissue are mainly involved in the degradation of fatty acids. This study results has certain significance for genetic improvement and mechanism analysis of fat traits.

The aim of this study was to investigate the biological characteristics of lncRNA-6617 and its effect on the differentiation of porcine intramuscular preadipocytes, preliminarily explore its upstream regulatory mechanism, and to provide the base for further clarifying the regulation mechanism of intramuscular fat deposition in pigs. In this study, healthy Mashen boars (n=3) and Large White boars (n=3) of 1, 90, and 180 days of age were raised under the same conditions. RNA-seq technology was used to screen muscle-specific lncRNAs in fatty-type pigs (Mashen pigs) and lean-type pigs (Large White pigs). Bioinformatics analysis, RT-PCR and Sanger sequencing were used to identify lncRNA-6617 biological characteristics and expression characteristics in pig tissues. The effect of lncRNA-6617 on the differentiation ability of intramuscular adipocytes were detected after interfering with lncRNA-6617 in porcine intramuscular preadipocytes. Subsequently, the expression differences of m⁶A modification-related enzymes in different breeds of pigs were detected, and the upstream regulatory mechanism of lncRNA-6617 was studied. The results showed that lncRNA-6617 was located on pig chromosome 18, the third intron region of the RAMP3 gene antisense strand, had no protein coding ability, and mainly distributed in the nucleus. lncRNA-6617 was expressed in all detected pig tissues, with the highest expression in back subcutaneous fat, and its expression was significantly higher in MS pigs muscle tissue than that in LW pigs(P < 0.01). In the differentiation process of porcine intramuscular preadipocytes, the expression level of lncRNA-6617 showed an upward trend and reached the peak on day 5. After interfering with lncRNA-6617, the number of differentiated mature adipocytes were significantly reduced, and the expression of key adipogenic genes CEBPα, PPARγ and aP2 were significantly down-regulated (P < 0.01) on day 7 of differentiation. The expression of m⁶A-modified demethylase FTO were significantly higher in the longissimus dorsi and biceps femoris muscle of MS pigs than that of LW pigs (P < 0.01), and were significantly higher than that of LW pigs in the psoas muscle (P < 0.05). And the expression of upstream regulator ZNF217 were significantly higher in the muscle tissue of MS pigs than that in LW pigs (P < 0.01), which was consistent with the expression pattern of lncRNA-6617. After interfering with FTO, the expression of lncRNA-6617 was significantly down-regulated (P < 0.01). In conclusion, lncRNA-6617 highly expressed in the muscle tissue of Mashen pig was screened and identified. The function of lncRNA-6617 in promoting the differentiation of porcine intramuscular preadipocytes and its upstream regulation mechanism were further studied, which riched the epigenetic regulation network of adipogenesis.

The black shank trait favored by Chinese consumers is controlled by recessive sex-related gene or autosomal dominant gene. To explore the inheritance and molecular basis of black shank trait in a cultivated W strain, this study first conducted reciprocal crossing test between mature male and female chickens (1♂∶5♀) from the W line (black feather black shank) and the Rhode Island Red line (yellow feather yellow shank), followed by transcriptome analysis on the shank skin samples collected from the female offspring (4 chicks with black shank and 4 chicks with yellow shank) of the reciprocal crosses. The results showed that there were 293 chicks with black shank (♂172, ♀121) and 51 chicks with yellow shank (♂20, ♀31) in the cross of the W line with the Rhode Island Red line, there were 256 chicks with black shank (♂156, ♀100) and 73 chicks with yellow shank (♂29, ♀44) in the reciprocal cross, the offspring of both crosses had two shank colors, and the number of the chicks with black shank was significantly higher than that with yellow shank in each cross, indicating that the black shank trait of the W line belongs to autosomal dominant inheritance. Based on the statistics of shank color and feather color in the offspring of the breeding groups with shank color separation, it was found that the chicks with black shank had black feathers and the chicks with yellow shank had yellow feathers, the number of the chicks with black shank to that with yellow shank (121∶117) conformed to the separation ratio of 1∶1, suggesting that the black vs. yellow shank traits in the cultivated strain were controlled by a pair of alleles. The transcriptome analysis of shank skin samples showed that the differentially expressed genes (DEGs) were significantly enriched in Melanogenesis pathway (P < 0.01), the expression of Mc1r and the genes, such as Tyr and Tyrp1, which are known to be involved in the regulation of melanin synthesis was significantly different between black and yellow shank skin samples, and some other genes, such as Wnt16, Wnt3a, Fzd10, that are not previously reported to be involved in shank color formation, also showed significant difference. In addition to the Melanogenesis pathway, black shank formation was also involved in extracellular matrix and receptor signaling, signal transduction, cell cytoskeleton and migration, cell adhesion, sphingolipid and glycolipid metabolisms. In conclusion, based on genetic analysis and transcriptome analysis, this study inferred that the black shank trait in the cultivated W strain was regulated by Mc1r gene, was inherited in autosomal dominant mode, the formation of black shank trait involved multiple signaling pathways, which laid a foundation for clarifying the mechanism underlying formation of black shank trait.

This study aimed to explore the genetic diversity and population structure of Loumen duck conservation population and evaluate the conservation effect. In this study, a total of 163 healthy 90-day-old Loumen ducks (39 males and 124 females) of the same generation were randomly selected from the Loumen duck conservation population, and genomic DNA was extracted after wing-vein blood collection to detect genome-wide single nucleotide polymorphisms (SNPs) using reduced-representation genome sequencing techniques. Six genetic diversity indexes, including polymorphism information content (PIC), fixation index (Fix-index), Shannon information index (SHI), gene diversity index (Nei), effective number of alleles (Ne) and relative PIC (rPIC), were calculated by R software, and the genetic diversity indexes at different chromosome levels were compared. At the same time, admixture software was used to analyze the population genetic structure, and gcta software was used to analyze the principal component analysis (PCA) and genetic relationship, and the runs of homozygosity (ROH) and genomic inbreeding coefficient F_ROH were analyzed to evaluate the conservation effect. The results showed that 622 205 SNPs were detected in 163 Loumen ducks, and 374 455 high-quality SNPs were obtained after quality control filtration, of which 50.70% were distributed in 4 chromosomes of NC_ 040046, NC_ 040047, NC_ 040048 and NC_ 040049. The PIC, Nei, Ne, SHI and rPIC values of Loumen duck population were 0.154 1, 0.192 9, 1.336, 0.296 9 and 0.410 9, respectively. For SNPs, 38.80% of the loci in this population belonged to highly polymorphic loci and had rich genetic diversity. Fix-index value was 0.320 8, indicating that the Loumen duck population had differentiated, which was consistent with the result that the Loumen duck population was divided into 3 populations by genetic structure, PCA and kinship analysis. All the distribution patterns of PIC, Nei, Ne and SHI on different chromosomes were consistent, and the correlation coefficients of the 4 indexes was more than 0.97, while the correlation between Fix-index and the other 4 indexes was lower than 0.23, indicating that a few indexes such as Fix-index and PIC could be selected to evaluate the genetic diversity of Loumen duck population. A total of 2 966 ROH fragments were detected in 163 Loumen ducks, and the length was mainly concentrated in the range of 0-2 Mb. The genomic inbreeding coefficients F_ROH obtained based on ROH were 0.027 5 and 0.043 3 in male and female ducks, respectively, indicating that the inbreeding degree of the conserved population of Loumen duck was low. At the chromosome level, the F_ROH values of chromosome NC_040068 and NC_040074 were 0.352 4 and 0.319 3, respectively. Loumen duck conservation population has rich genetic diversity and low population genomic inbreeding coefficient, but the inbreeding coefficient of individual chromosomes is high, and the population has partial differentiation. Therefore, appropriate non-random mating between individuals of the 3 differentiated subpopulations obtained from genetic structure analysis can be used in subsequent conservation to eliminate the current population differentiation, and the genomic regions with high inbreeding coefficient shall be monitored to avoid the rapid increase of inbreeding coefficient of special chromosomes.

The study aimed to investigate the expression of miR-15a in different tissues of broilers and explore the effect of overexpression of miR-15a on chondrocytes. In this study, the chondrocytes were isolated and culture, and it was identified by inverted microscope observation, PCR, gel electrophoresis and toluidine blue staining. Real-time fluorescence quantitative PCR was used to detect the expression level of miR-15a in each tissue of broilers in the valgus-varus deformity (VVD) group and the healthy group (3 each). After that, the cell proliferation of chondrocytes in miR-15a overexpression group was detected by CCK8 and EDU assay. After chondrocytes were transfected with miR-15a mimics, the expression of marker genes of chondrocytes Collagen-2, Aggrecan, Collagen-10, mature differentiation genes Runx2, Sox9, VEGF, MMP9, inflammatory factors IL-1β, IL- 6, IL-8, IL-10, TNF-α, TGF-β3 and the apoptosis genes Fas, FasL and Bcl-2 were detected by qPCR. The wild-type vector and mutant vector of FKBP5 3'UTR were constructed, and the targeting relationship between miR-15a and FKBP5 was detected by dual-luciferase detection reporter. The results showed that the trypsin, collagenase II and hyaluronidase combined digestion method used in this study could successfully isolate chondrocytes. The qPCR results showed that miR-15a was expressed in all detected tissues. The expression of miR-15a in the liver (P < 0.01), spleen (P < 0.05) and thymus (P < 0.01) in the VVD group were significantly higher compared with the healthy group. And the expression level in the heart and pectoral muscle tissues was significantly lower in the VVD group (P < 0.01). The results of CCK8 and EDU showed that, compared with the NC group, the proliferation rate of chondrocytes in the miR-15a overexpression group was significantly decreased (P < 0.01), and the number of proliferating cells was significantly decreased (P < 0.01). The qPCR results showed that, the expression of Aggrecan, Sox9 and Runx2 genes in the miR-15a mimics group were significantly decreased (P < 0.05), and the expression of Fas gene was extremely significantly increased (P < 0.01), the expression of FasL gene and anti-apoptotic gene Bcl-2 were extremely significantly decreased (P < 0.01) compared with the NC group. The FKBP5 3'UTR wild-type and mutant vectors were successfully constructed, and the dual-luciferase detection reporter showed that the predicted target gene FKBP5 had no targeting relationship with miR-15a. In this study, chicken chondrocytes were successfully isolated and identified, and the results suggest that the overexpression of miR-15a can inhibit the proliferation, maturation, differentiation and promote apoptosis of the chicken chondrocytes.

The aim of the present study was to investigate the effect of mitochondrial tRNA-Lys(T7719G) gene variation on the litter size in sheep population with Small-tailed Han sheep as female parent, and to explore the possibility of mitochondrial tRNA-Lys(T7719G) gene variation as molecular marker for assisted selection. Healthy and suitable breeding ewe with Small tail-Han sheep as female parent were selected. The polymorphism analysis of tRNA-Lys(T7719G) gene was conducted by sequencing of PCR amplificatin product, genetic analysis of the influence of tRNA-Lys (T7719G) gene, season and parity on the litter size were conducted. The results showed that the mitochondrial tRNA-Lys gene mutated into G and T alleles, with allele frequency of 57.8% and 42.2%, respectively. The average litter size of ewes with G allele was 0.08 more than that of ewes with T allele in average litter size. Litter size of population increased with the number of parities and the number of individual ewes with G allele in the same season. The formula of prediction model of litter size was established: Litter size = 1.201 + (－0.009)×dummy variable 1 + 0.043×dummy variable 2 + 0.194×dummy variable 3 + 0.061×parity + 0.123×ewes number carrying G allele, which indicated that adding an individual with G allele in population, the litter size was expected to increase 0.123. tRNA-Lys(T7719G) gene can be used as a molecular marker for litter size traits of population with Small-tailed Han sheep as female parent.

This experiment aimed to clone goat SRSF10 gene sequence, reveal its biological characteristics, and clarify the effect of SRSF10 on goat intramuscular adipocyte differentiation through overexpression and interference techniques. Taking Jianzhou big ear goat(Capra hircus)as the experimental object, the goat SRSF10 gene sequence was cloned by RT-PCR technique, along with bioinformatics analysis. The real-time quantitative PCR(qPCR)technique was used to investigate its expression level in adipogenic differentiation of cells at different stages. pcDNA3.1-SRSF10 overexpression vector and SRSF10-siRNA were transfected into goat intramuscular preadipocytes, followed by inducing differentiation. Oil red O and Bodipy staining were used to clarify the effect of SRSF10 on lipid droplet accumulation, and the expression changes of adipocyte differentiation marker genes were detected by qPCR technique. The results showed that the obtained goat SRSF10 gene sequence was 1 026 bp in length, the CDS region was 552 bp, encoding a total of 183 amino acids. The highest expression level of SRSF10 in goat was detected at 96 h in intramuscular adipocytes after inducing differentiation. Overexpression and interference of goat SRSF10 promoted and inhibited the accumulation of lipid droplets in intramuscular adipocytes, respectively. After overexpression of SRSF10, the relative expression of SREBP1, PPARγ and C/EBPα extremely significantly up-regulated(P < 0.01), and the relative expression of differentiation marker genes SREBP1, PPARγ and C/EBPα extremely significantly down-regu-lated after interference(P < 0.01). The results show that goat SRSF10 is a positive regulatory factor for the differentiation of intramuscular adipocytes, and this regulation may be achieved mainly through regulating the expression of SREBP1, PPARγ and C/EBPα.

This study aimed to clone the retinoic acid-related orphan receptor alpha (RORα) gene and construct its eukaryotic expression vector, and systematically analyze the biological characteristics of RORα by bioinformatics tools. In addition, the regulatory role of RORα in goat circadian clock system was investigated. One healthy male Saanen dairy goat (12 months old) was used as experimental animal. Total RNA extracted from goat liver tissue was reverse transcribed into cDNA by reverse transcription PCR, and the coding sequence (CDS) fragment of goat RORα gene was amplified by PCR using the cDNA template. Then, the PCR product was ligated to pcDNA3.1-Puro-N-3HA vector by homologous recombination. The recombinant plasmid was identified by PCR, restriction enzyme digestion and sequencing. The positive plasmid was named as pcDNA3.1-3HA-gRORα. The pcDNA3.1-Puro-N-3HA and pcDNA3.1-3HA-gRORα plasmids were transfected into HEK293T cells, respectively. The expression of RORα was examined by real-time quantitative PCR (qPCR) and Western blotting (WB) analysis. Meanwhile, the goat RORα gene was systematically analyzed by bioinformatics softwares including ExPASy and ProtScale. In addition, the luciferase reporter plasmids of pGL4.10-BMAL1-Promoter-Luc and pGL4.10-NR1D1-Promoter-Luc containing the promoter fragments of goat circadian clock genes BMAL1 and NR1D1 were constructed by homologous recombination methods. At last, the dual luciferase reporter assay was used to explore the function of goat RORα protein in regulating BMAL1 and NR1D1 gene promoter activity. PCR, enzyme digestion and sequencing results showed that the eukaryotic expression vector of pcDNA3.1-3HA-gRORα was successfully constructed. The qPCR and WB results showed that the mRNA and protein expression of goat RORα gene in HEK293T cells with the transfection of pcDNA3.1-3HA-gRORα were significantly higher than those in the group of pcDNA3.1-Puro-N-3HA transfection(P < 0.01). Bioinformatics analysis showed that the similarities of the CDS region of goat RORα gene were 97.5% with sheep, 97.1% with cattle, and 95.2% with pig, respectively. Goat RORα protein was a hydrophilic protein, and its secondary structure mainly consisted of α-helix, extended chain, β-turn, and irregular coiled structures. It had a small probability of having a signal peptide, and it had no transmembrane region. Meanwhile, the tertiary structure of goat RORα protein was highly similar to the corresponding protein of mouse and human. PCR, enzyme digestion and sequencing results showed that pGL4.10-BMAL1-Promoter-Luc and pGL4.10-NR1D1-Promoter-Luc recombinant plasmids were successfully constructed. The results of dual luciferase reporter assay showed that goat RORα protein could significantly upregulate the transcriptional activity of goat BMAL1 and NR1D1 gene promoters. In this study, the eukaryotic expression vector of goat RORα was successfully constructed. Meanwhile, it is proved that goat RORα protein can positively regulate the activity of goat BMAL1 and NR1D1 gene promoters. These results provide a preliminary basis for further investigate the biological role of goat nuclear receptor RORα and the transcriptional regulation mechanism of goat circadian clock system.

Mannose binding lectin C(MBL-C)is a member of the C-type (Ca²⁺dependent) lectin superfamily. As an acute phase protein, MBL-C plays an important role in the innate immune response. The aim of this study was to identify the key transcriptional factors in the promoter region (1 009 bp) of MBL2 gene, and explore the expression regulation mechanism of MBL2 gene. The promoter sequence 1 009 bp of MBL2 gene was selected from Hainan black goat, and a serial of 5'-flanking deletion regions of the goat MBL2 promoter were amplified and subcloned into pGL3-Basic vector. The recombined plasmids were transfected into 293T cells and detected with the dual-luciferase activity detection system to screen the core promoter region of MBL2 gene. The transcription factor binding site of the core promoter region of Hainan black goat MBL2 gene were predicted by online bioinformatics software. The site mutation technology and dual-luciferase activity detection system were utilized to identify the transcription factor binding sites. The results showed that the core promoter region of Hainan black goat MBL2 gene located in the range of -304 to -45 bp of the transcription initiation site. Online software analysis indicated that this region existed three transcription factors binding sites, including RELA, NF-κB2 and MZF1. Dual-luciferase report analysis results showed that the deletion of the binding sites of RELA and NF-κB2 extremely significantly reduced the transcriptional activity of goat MBL2 gene (P < 0.01). These results indicated that RELA and NF-κB2 might positively activate transcriptional regulation of goat MBL2 gene. This study can provide a theoretical basis for further exploring the function of MBL2 gene in Hainan black goat.

The aim of this study was to investigate the effects of prostaglandin F2α (PGF2α) injection at different phases of luteal on reproductive hormones and related cytokines in growing ewes. A total of 60 healthy ewes in good condition with similiar weight and normal estrous cycle were used in this study. Forty eight ewes with normal estrus were randomly divided into 6 groups after synchronization of estrus cycle with "MAP +PMSG" method. The day of oestrus was recorded as day 0. Ewes in the experiment groups of early-, mid-, and late-luteal phase were injected with 1 mL PGF2α (0.1 mg) on day 6 (early-luteal phase), 11 (mid-luteal phase) and 16 (late-luteal phase) of luteal phase, respectively. Ewes in the control groups of different luteal phases were injected with 1 mL normal saline on day 6, 11 and 16 of luteal phase, respectively. Blood was collected at 0.5, 1, 2 and 3 hours after each injection for the determination of blood indexes. The results showed as follows: There were no significant differences in levels of FSH, LH, PRL, P₄, E₂, TNF-α, IL-1β, IL-6 and IFN-β between experimental groups and control groups at 0.5, 1, 2, 3 h after injection (P > 0.05). Injection of PGF2α at different phases of luteal had no significant effect on serum FSH, LH, PRL, IL-1β and IFN-β levels at 0.5, 1, 2 and 3 h (P > 0.05). At 3 h after injection, P₄ level was significantly lower, E₂ and IL-6 levels were significantly higher in the early-luteal experiment group than that in control group (P < 0.05). The level of TNF-α in the early-luteal experiment group was significantly higher than that in control group at 2 and 3 h after injection (P < 0.05), and the level of P₄ in the mid-luteal experiment group was significantly lower than that in the control group at 1 h after injection (P < 0.05). After injection of PGF2α in the early-luteal phase, the overall levels of FSH, E₂, TNF-α and IL-6 in the early-luteal experiment group were significantly higher than those in control group within 3 h (P < 0.05), and the overall level of P₄ was significantly lower than that in control group (P < 0.05), there were no significant effects on LH, PRL, IL-1β and IFN-β (P>0.05). After injection of PGF2α in the mid-luteal phase, the overall level of E₂ in the mid-luteal experiment group was significantly higher than that in control group within 3 h(P < 0.05), and P₄ was significantly lower than that in control group (P < 0.05), there were no significant effects on FSH, LH, PRL, TNF-α, IL-1β, IL-6 and IFN-β (P>0.05). Injection of PGF2α at the late-luteal had no significant effect on reproductive hormones and related cytokines (P>0.05). In conclusion, there are stage differences of PGF2α in dissolving corpus luteum. The short-term response of ewes to PGF2α was stronger in early-luteal phase than in mid- and late-luteal phases. The ovaries could respond to PGF2α in early-luteal phase to create a better environment for follicular development.

Early embryonic death and abortion in dairy cows impair the reproductive performance and cause great economic losses. In this study, endometrial cells were treated with different concentrations of hydrogen peroxide (H₂O₂) for 6 h. The levels of intracellular reactive oxygen species (ROS) were measured by flow cytometry. The activities of superoxide dismutase (SOD) and catalase (CAT), and the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) were measured by microplate reader, which were to establish an oxidative stress model in endometrial cells. Then apoptosis and cell cycle were detected by flow cytometry, mRNA and protein expression of key genes in Fas/FasL signaling pathway were detected by fluorescent quantitative RT-PCR and Western blot, and lentivirus-mediated RNA interference (RNAi) was employed to inhibit Fas expression in endometrial cells to further verify whether the oxidative stress-induced apoptosis is mediated through Fas/FasL signaling pathway. All experiments were repeated more than three times. Results are as follows: Treatment of endometrial cells with 200 μmol·L^-1 H₂O₂ for 6 h resulted in an increase in the amount of intracellular ROS (P < 0.05), a significant decrease in the activities of SOD and CAT (P < 0.05), and the ratio of GSH to GSSG was significantly reduced (P < 0.05), which indicated that H₂O₂ caused oxidative stress damage in endometrial cells. Compared with control group, the proportion of S phase in cell cycle was significantly reduced (P < 0.05), and the proportion of apoptosis was significantly increased (P < 0.05). The levels of mRNA expressions of Fas, Caspase 8 and Caspase 3 in Fas/FasL signaling pathway significantly increased (P < 0.05); The concentration of FasL in the cultured medium significantly increased (P < 0.05) and the levels of protein expression of Fas, Caspase 8 and Caspase 3 significantly increased (P < 0.05). Further studies have found that interfering with Fas gene expression partly reduced the proportion of apoptosis and the levels of protein expression of key genes in Fas/FasL signaling pathway induced by oxidative stress. In conclusion, these results demonstrated that oxidative stress-induced apoptosis in bovine endometrial epithelial cells is mediated through Fas/FasL signaling pathway, which may provide new insights into the adverse effects of endometrial cells in response to oxidative stress.

Sheep are suffering from severe heat stress in summer recently, and present study was conducted to investigate the effect of Chinese herbal formulation (Huopopuling Powder) in diet on growth performance, digestibility, rumen fermentation parameters, and serum biochemical indexes in fattening lambs in summer. A total of 200 fattening lambs with similar body weight and the same age were selected and randomly divided into 4 groups, which were fed with 0% (control group), 0.5%, 1.0%, and 1.5% of Huopopuling Powder. The experimental period were lasted for 30 days, and the temperature and humidity index in experimental shed averaged 79.11. The growth performance of fattening lambs, apparent nutrient digestibility, rumen fermentation parameters, and serum biochemical indexes were detected at the end of the experiment. The results were showed as follows: 1) The daily weight gain of lambs fed 0.5% Huopopuling Powder was higher than that of the control (P < 0.01). There was no significant difference in daily water intake and daily feed intake among all groups (P>0.05). For physiological parameter, the rectal temperature and respiratory rate of lambs fed with Huopopuling Powder showed no significant difference compared with the control (P>0.05). 2) The apparent digestibility of nutrients involving dry matter (DM), neutral detergent fiber, acid detergent fiber, crude protein, calcium and phosphorus was higher in 0.5% group than that in the control (P < 0.05). The DM digestibility increased by 7.20% in 0.5% group compared with the control (P < 0.01), while the digestibility in 1.0% or 1.5% group exhibited no significant difference compared with the control (P>0.05). 3) For serum biochemical indexes, growth hormone content was higher in the three Huopopuling-fed groups than that in the control (P < 0.05); Among all groups, the content was the highest in 0.5% group, showing 16.90% higher than that in the control. While thyroxine content in 0.5% group significantly increased compared with other groups (P < 0.05). 4) From analysis of rumen fermentation, fermentation parameters in three Huopopuling supplementation groups demonstrated an increase ranged from 1.19 to 1.30 times in total volatile fatty acid, 1.18 to 1.24 times in acetic acid, and 1.28 to 1.43 times in butyric acid than those in the control (P < 0.05). The results indicate that under the condition of this study, Huopopuling Powder could effectively alleviate heat stress of fattening lambs and its appropriate dose is suggested for 0.5% in diet.

Listeria monocytogenes is an important food-borne pathogen. So it's important and urgent to establish a sensitive and specific method for its rapid detection. The monoclonal antibody of Listeriolysin O (LLO) was screened after prokaryotic expression of recombinant soluble protein His-LLO as antigen, followed by detection of the subtypes, titers and affinity of the antibody. Colloidal gold-based test strip for Listeria monocytogenes LLO was developed via sodium citrate method. Subsequently, the specificity, sensibility, repeatability and stability as well as the primary application of the strip in simulated samples and seafood samples were performed. The results showed that the monoclonal antibody of LLO was specific, stable, repeatable, relatively sensitive and had high affinity, with its affinity constant reaching to 4.25×10⁸ L·mol^-1. The sensibility of the LLO strip in the simulated samples was 3.0×10⁵CFU·mL^-1, which was consistant with that in the pure bacterial cultures. The positive detection of L.monocytogenes from 500 seafood samples was 2.20% (11/500) via LLO colloidal gold-based test strip, which was a bit lower than using standard method (2.40%, 12/500) and PCR detection method (2.40%, 12/500). The coincidence rate of L. monocytogenes positive detection via the aforementioned three methods was 91.67%. Besides, the detection time was deceased to 15 minutes, thereby the efficiency was increased to a greater extent. Therefore, the developed LLO colloidal gold-based test strip could be further used for the rapid detection of L. monocytogenes.

To clarify the role of the 43 ku outer membrane protein (43K OMP) in the adhesion of Fusobacterium necrophorum to cells, the mouse mammary epithelial cells and the recombinant protein or the natural protein were co-incubated in this study, and the adhesion of the 43K OMP of F.necrophorum was observed under a laser confocal microscope. Meanwhile, the adhesion inhibition assay with antibodies against the recombinant 43K OMP or the natural protein was used to determine the effect of the 43K OMP on the adhesion of F. necrophorum to host cells, and the effect of 43K OMP gene deletion on the adhesion of F. necrophorum to host cells was investigated using deletion mutant strain. Immunofluorescence results showed that the natural 43K OMP and recombinant 43K OMP could bind to the cell membrane of mouse mammary epithelial cells. When mouse mammary epithelial cells or mouse liver cells were preincubated with the natural 43K OMP, the adhesion of F. necrophorum significantly decreased (P < 0.05). When F. necrophorum was preincubated with polyclonal antibody or monoclonal antibody against the recombinant 43K OMP, the adhesion of F. necrophorum to mouse mammary epithelial cells or mouse liver cells significantly decreased compared with the negative serum control and the no antibody control (P < 0.05). The number of bacterial cells attached to cells was higher with F. necrophorum A25 strains than that of the mutant strain A25Δ43K OMP(P < 0.01), The 43K OMP gene deletion reduced the binding of F. necrophorum to mouse mammary epithelial cells or mouse liver cells by 94.4% and 90.4%. Therefore, the 43K OMP plays a key role in the adhesion of F. necrophorum to host cells, and further study of the adhesion mechanism will provide a theoretical basis for revealing the pathogenic mechanism of F. necrophorum.

This study aimed to investigate the pathogenicity of Pasteurella multocida CS strain by analyzing virulence genes of its complete genome. The bacteria strain was isolated from swine pasteurellosis by conventional methods and its virulence was tested by mice. The complete genome of P. multocida CS strain was sequenced based on the second generation sequencing platform, and analyzed its differences with others submitted to GenBank from different regions and hosts by comparative genomics methods. The LD₅₀ of P. multocida CS strain was 5×10² CFU·mL^-1, which showed strongly toxic to mice. The genomic information of CS strain was submitted to NCBI, and the accession number was SUB11119617. The analysis results of the complete genome sequence of P. multocida CS strain showed that the total genome size of CS strain was 2 599 048 bp, the content of G+C was 39.932%, the CDs encoding gene was 2 580, accounting for 69.6% of the whole genome length. The average length of CDs encoding gene was 952 bp. Besides, there were 53 tRNA and 3 rRNA genes. 87.95% of the coding genes could be assigned as COGs, and the functions of 29 genes were unknown. Phylogenetic analysis using maximum likelihood method showed that CS strain was closely related to 1 pig origin serotype A strain and 2 cattle origin serotype A strain and 1 pig origin serotype D strain. The P. multocida CS strain contained 254 virulence-related genes which were divided into 4 major classes and 11 subclasses. The distribution of several genes related to iron uptake system, adhesion system, secretion system and two-component regulatory system was different among different serotypes. By analyzing the frequency of these gene occurrence of different P. multocida serotypes, it was found that tbpA and iron complex outer membrane receptor protein gene (irp) related with iron uptake system, and ppdD and ompA genes related with adhesion appeared to be associated with bacterial toxic differention among different strains. The secretory system of P. multocida CS strain consisted of three types: type I (hly gene), Tat type and Sec-SRP type. The frequency of hlyD gene among different serotypes was different. P. multocida CS strain's two-component signal transduction system (TCSs) consisted of 16 regulatory genes, 12 induction-related genes and 2 hybrid genes. The relationship between the polymorphism of TCSS genes in different serotypes and virulence of different strains needs to be further studied. To sum up, the gene frequency of iron uptake system, adhesion system, secretion system and two-component regulatory system had polymorphism among different serotypes and different isolates of the same serotypes. These may be one of the reasons for the virulence differences among different P. multocida strains.

The present study aimed to investigate the effect of signal peptidase complex subunit 1 (SPCS1) on the replication of bovine viral diarrhea virus (BVDV). The sgRNA targeting SPCS1 gene was designed by using bioinformatics platforms such as Benchling and CHOPCHOP, following with cloning into lentivirus vector lentiCRISPR v2, and transfected into human embryonic kidney cell HEK-293T plus packaging plasmids pSPAX2 and pMD2.G. After lentivirus generation, lentivirus was used to infect MDBK cells, and puromycin was used for continuous screening for 5 generations. The knockdown (KD) of SPCS1 protein was determined by Western blot. At different time intervals after BVDV infected SPCS1 gene knockdown cells, the mRNA level of 5' untranslated region (UTR) and the accumulation level of double-stranded RNA (dsRNA) of BVDV were detected by real-time quantitative PCR and immunofluorescence assay. The cytopathic effect (CPE) was observed under a microscope. The progeny virus suspension was collected and its titer was determined by Reed-Muench method. We observed that the expression of SPCS1 protein decreased significantly, and SPCS1 KD cells were successfully constructed. Compared with the scramble cells, after BVDV infected SPCS1 KD cells, BVDV 5'UTR mRNA level and dsRNA accumulation decreased significantly (P < 0.01), CPE phenomenon was significantly delayed and weakened, and the virus titer of progeny virus decreased significantly (P < 0.01), up to 95.8% reduce. Our findings suggest that SPCS1 KD significantly inhibits BVDV replication, which provides an important target for the establishment of new technology for BVDV prevention and control.

To study the function of D1133L gene encoded by African swine fever virus (ASFV), we constructed MA-104/D1133L cell line for overexpression of D1133L using lentivirus expression vector, and compared ASFV replication ability in MA-104/D1133L and MA-104 cells. The transcription and protein expression levels of p30 and p72 in ASFV infected MA-104/D1133L cells were evaluated by real-time qPCR and Western blot respectively, and the proliferation of ASFV was evaluated according to virus titer and virus copies number. The transcriptional and protein levels of ASFV p30 and p72 proteins in MA-104/D1133L cell line were higher than those in wild MA-104 cell, and the viral proliferation capacity of ASFV in MA-104/D1133L cell line was higher than that in wild MA-104 cell line. The results showed that overexpressing ASFV D1133L gene of MA-104 cell line was successfully constructed in this study, that compared with wild type cells, MA-104/D1133L cell line was more conducive to ASFV replication, and D1133L promoted ASFV proliferation. These results provide biomaterials for further study of the function of ASFV D1133L gene.

This study aimed to understand the infection situation of suspected rabbit hemorrhagic virus type 2 (RHDV2) in Henan province, and preliminarily analyze the pathogenicity of RHDV2. The liver tissues of dead rabbits were collected, then the microblood coagulation assays, RT-PCR amplification and sequencing, VP60 gene phylogenetic tree analysis and animal regression test were conducted to identify the pathogen. The results of the microblood coagulation assay showed that the tissue sample suspension were able to produce a lect reaction with the human type "O" red blood cells; RT-PCR showed the detection of RHDV2-specific bands, the fragment size was 829 bp; Results of the phylogenetic tree analysis showed that the VP60 gene similarity was as high as 98.2% between the isolated virus and the first RHDV2 strain SC2020/04 found in Sichuan; The autopsy of clinical cases showed that the bleeding of thymus, trachea, lung, liver, spleen, kidney and other substantial organs was more serious; Animal regression test showed that the mortality rate of rabbits was 100% and the average death time was 65.8 h after challenge test, and the specific bands of RHDV2 were detected by RT-PCR amplification. In conclusion, the RHDV2 virus was detected for the first time in the rabbit field in Henan Province, which provided a scientific reference for the scientific and reasonable prevention and control of RHDV2.

The aim of this study was to compare the differences of pathogenicity, innate immune factors and oncogenic factors expressions of chickens induced by two different avian leukosis virus subgroup J (ALV-J) strains. Seventy-five 1-day-old chickens were inoculated with 10³ TCID50 acute fibrosarcoma ALV-J strain (ALV-J-SD1005), 10³ TCID50 myeloma ALV-J strain (ALV-J-NX0101), and same dose sterile PBS saline. The body weight, tissue hyperplasia lesions, mortality, viral loads in blood and subcutaneous fibrous tissues, innate immune factors (MDA5, IRF7, IFN-α/β) and cell proliferation-related factors (TRIM25, P53, 14-3-3σ, STAT1) of the infected chickens were detected at 7 days post inoculation (dpi), 14 dpi, 21 dpi. The results showed that the fibrosarcoma in ALV-J-SD1005 group was firstly observed at 10 dpi, the fibrosarcoma incidence was 100% (18/18) and mortality was 5.2% (1/19) at 14 dpi, and the sarcoma index and the mortality increased continuously from 14 dpi to 21 dpi, and respectively reached 34.4%(74.5/209.5) and 58.3% (7/12) at 21 dpi. Viral loads in blood and subcutaneous fibrous tissues were also significantly increased. Meanwhile, the mRNA expressions of MDA5, IRF7, P53 in the livers were significantly up-regulated, and the mRNA expressions of IFN-α/β and 14-3-3σ genes were significantly down-regulated, while chicken TRIM25 was significantly down-regulated at the early stage of virus infection (at 7 dpi), and significantly up-regulated at the late stage (14~21 dpi). In ALV-J-NX0101 group, the tissue hyperplasia lesions and death were not observed within 21 days after infection, but the viral loads increased significantly, the mRNA expressions of TRIM25, MDA5, IRF7 and IFN-α/β genes were decreased significantly, and the expression of STAT1 gene was up-regulated significantly. It can be concluded that ALV-J-SD1005 strain and ALV-J-NX0101 strain can cause significantly different antiviral immune responses and antitumor responses during their infections, which result into dramatically different virus proliferation and pathogenicity. This study provides scientific basis for further undestanding the tumorigenic mechanism of two ALV-J viruses and exploring new diagnostic markers.

Based on the structure-activity relationship of the antimicrobial peptide tachyplesin Ⅰ(TP Ⅰ), a novel antimicrobial peptide TP Ⅰ-Y4 was designed out. TP Ⅰ-Y4 was obtained by replacing four cysteines with tyrosines to delete two disulfide bonds of TP Ⅰ, keeping the chain length and charge number of the parent peptide. The results of bioinformatics software analysis had discovered that compared with TP Ⅰ, TP Ⅰ-Y4 had better thermal stability, stronger hydrophilicity, and similar structure and antibacterial activity to propeptide. After chemical synthesis, the results of circular dichroism spectroscopy had showed that TP Ⅰ-Y4 also adopted β-sheet structure in aqueous phase and 50% trifluoroethanol solution which simulated the hydrophobic environment of cell membrane. The β-sheet contents of TP Ⅰ-Y4 in hydrophobic environment were higher than that in aqueous phase. TP Ⅰ-Y4 exhibited higher β-sheet contents than TP Ⅰ in different environments. TP Ⅰ-Y4 owned potent antimicrobial activities against bacteria and fungi, and TP Ⅰ-Y4 displayed stronger antimicrobial activities against bacteria tested than TP Ⅰ. TP Ⅰ-Y4 reduced the hemolysis of mouse red blood cells and retained a strong endotoxin neutralization ability, the neutralization rate reached more than 50% when the concentration was 40 μg·mL^-1. The results suggest that the increase of TP I-Y4 antibacterial activity and the decrease of hemolytic activity may be related to the substitution of tyrosine, as β-sheet strong-former which further leads to the enhancement of β-sheet structure in antimicrobial peptide.

To investigate the effect of double-expression DNA vaccine of SS and CST (hereinafter referred to as double-expression DNA vaccine) immunized calves through different routes, 27 dairy buffalo calves aged 2-6 months were randomly divided into four groups. The first three groups were orally (group O), nasally (group N), and orally plus nasally (group ON) vaccinated with dual-expression DNA vaccine at a dose of 3×10⁷ CFU·mL^-1, respectively. The control group (C) was orally plus nasally immunized with PBS, and boosted immunization once after 2 weeks. The results showed that all three immunization methods could induce calves to produce antibody of SS and CST. After 2 weeks of primary immunization, the positive rates in N and ON groups were the highest, both at 85.71%. At the 6^th week, the positive rates in each group decreased, the levels of antibody also decreased in O and N groups, but the levels of antibody increased in ON group. At the 10^th week, the positive rates in ON group decreased to 14.29%, and no antibodies were detected in the O and N groups. With respect to weight gain, at the 2^nd week, the average daily gain in the ON group was significantly higher (P < 0.05) than that in the O, N, and C groups. During the entire experimental period (0-10 weeks), compared with the C group, the O, N and ON groups were respectively increased by 5.26%, 10.53%, and 15.79%. After 10 weeks of primary immunization, the chest girth and leg-hip circumference in the ON group were significantly higher (P < 0.05) than those in the C group. Hormone measurement results showed that the concentration of GH, IGF-1, and T₃ in each group were as ON>N>O>C group. The IGF-1 concentration of O, N and ON group was significantly higher (P < 0.05) than that of C group at the 2^nd week. At the 6^th week, the concentrations of T₃ and T₄ in the ON group were significantly higher (P < 0.05) than those in the O and C groups. The cytokine test results showed that the three immune methods could stimulate the body to produce humoral and cellular immunity. At the 6^th week, the concentration of INF-γ in the ON group was significantly higher (P < 0.05) than that in the C group, and there was no significant difference (P>0.05) between the other groups. The detection of blood biochemical indexes showed that there was no significant difference (P>0.05) in the contents of total protein, glucose, non-esterified fatty and urea nitrogen in the serum of calves between the groups. In summary, oral, nasal and oral plus nasal could induce good immune effect in calves, especially oral + nasal spray combined immunization had the best effect.

This paper aimed to study the distribution characteristics of immune cells in the epididymis and vas deferens of healthy yaks. The location of CD68⁺ macrophages, CD3⁺ T lymphocytes, CD79α⁺ B lymphocytes, IgA⁺ and IgG⁺ plasmocytes, and the expression of surface markers of immune cells in the epididymis (head, body and tail) and vas deferens of juvenile (5-6 months old) and adult (3-4 years old) yaks were detected by immunohistochemistry and real-time fluorescence quantitative (qRT-PCR). The results showed that CD68⁺ macrophages, CD3⁺ T lymphocytes, CD79α⁺ B lymphocytes, IgA⁺ and IgG⁺ plasmocytes were mainly distributed in the epithelium and interstitium of the epididymis and vas deferens. Additionally the CD68 and CD3 mRNA and protein level in the caput and corpus of yaks were significantly higher than that of the cauda and vas deferens (P < 0.05), while in the cauda and vas deferens CD79α, IgA and IgG mRNA and protein level were significantly higher than those of the caput and corpus epididymis (P < 0.05). Moreover the CD68, CD3, CD79α, IgA and IgG mRNA and protein level in the epididymis and vas deferens of the adult yaks were significantly higher than those of the juvenile yaks (P < 0.05). The above results suggest that the caput may be the main site of cellular immunity, while the caudal epididymis and vas deferens are the main area of humoral immune response. Moreover the local immunity of the epididymis and vas deferens of the yak may mature and expand to a fully functional state in adult yaks. The above data would provide morphological data for further research on local reproductive immunoregulation and pathology in plateau yaks.

The present study was aimed to investigate the regulatory effect of Leonurus artemisia decoction on the expression of coagulation and anticoagulation cytokines in human dermal microvascular endothelial cells (HDMECs) by proteomics. The safe concentration of Leonurus artemisia decoction to HDMECs was evaluated by MTT method. The changes of protein expression of HDMECs treated with 50 and 100 μg·mL^-1 Leonurus artemisia decoction for 24 h were analyzed by isobaric tags for relative and absolute quantification (iTRAQ) technology. By comparing the changes of protein profiles in each group, the significantly different related pathways were screened out, and the credible differentially expressed proteins (DEPs) screened in the corresponding pathway were analyzed. The antagonistic factors related to coagulation and anticoagulation in credible DEPs were selected for RT-PCR and ELISA validation. Results showed that Leonurus artemisia decoction less than 1 mg·mL^-1 had no toxic effect on HDMECs. Leonurus artemisia decoction could regulate both platelet activation related to coagulation, and heparin binding related to anticoagulation. The analysis of five credible DEPs in the screened complement and coagulation cascade pathways showed that compared with the control group, the levels of prothrombin (F2), antithrombin-Ⅲ (AT-Ⅲ), tissue plasminogen activator (t-PA), coagulation factor Ⅴ (F5) and kininogen (KNG) were significantly decreased in 50 μg·mL^-1 Leonurus artemisia decoction group. The levels of F2, AT-Ⅲ, t-PA, KNG were significantly decreased in 100 μg·mL^-1 Leonurus artemisia decoction group. Compared with 50 μg·mL^-1 Leonurus artemisia decoction group, the levels of F2, AT-Ⅲ, t-PA, KNG and F5 were significantly increased in 100 μg·mL^-1 Leonurus artemisia decoction group. This study indicate that Leonurus artemisia decoction incubation could significantly change the protein expression profile of HDMECs, and bidirectionally regulate antagonistic factors related to coagulation and anticoagulation by regulating the expression of complement and coagulation cascade-related factors F2, AT-Ⅲ, t-PA, F5 and KNG.

The aim of this study was to compare the immunoprotective effects of Codonopsis pilosula polysaccharide (CP) and sulphated Codonopsis pilosula polysaccharide (SCP) on acute injury induced by alcohol in mice and H₂O₂ in RAW264.7 cells. RAW264.7 cells was induced by H₂O₂ to build a cellular model of acute injury, and the effects of different concentrations of CP and SCP on the growth, phagocytosis, and IL-2, IL-4, IL-10, IFN-γ levels of RAW264.7 cells were detected. In addition, a mouse model of acute injury induced by alcohol was established in vivo. A total of 90 4-week-old Kunming male mice were randomly divided into 9 groups: blank group (B), model control group (MO), Vitamin C control group (VC), 3 doses of CP (CPL, CPM, CPH) and SCP (SCPL, SCPM, SCPH) groups. The effects of each group on the growth, blood physiological indexes, organ index, serum IL-2, IL-4, IL-10 and IFN-γ levels of acute injury mice were detected. The results showed that in the H₂O₂-induced cell inflammation model, compared with the model group, the number of adherent cells and the cell density in CP and SCP groups remarkably increased, and the phagocytosis ability of cells was significantly improved after 30, 60 and 90 min treatment at 37 ℃ (P < 0.05). Moreover, SCP groups showed better effects of promoting cell proliferation and phagocytosis. Both CP and SCP could significantly promote the secretion of IL-2 and IFN-γ (P < 0.05), and inhibit the secretion of IL-4 (P < 0.05). SCP groups had better inhibition effect on IL-4 than CP groups, and SCPL group had the best inhibition effect on IL-4. In acute injury model induced by alcohol, compared with model group, both CP and SCP could significantly improve the blood physiological indexes of mice (P < 0.05), and restore the WBC and HGB to normal levels, among which SCPL group had the best effect. CP and SCP could also improve serum biochemical indexes, significantly increase the contents of TC and LDH in serum (P < 0.05), and reduce the content of TG (P < 0.05). The serum levels of IL-2 and IFN-γ in CP and SCP groups were significantly increased (P < 0.05) compared with MO group, and the level of IL-4 was significantly decreased (P < 0.05), in which SCP groups had the best inhibition effect on IL-4. In conclusion, SCP significantly increased the phagocytosis ability of RAW264.7 cells after acute injury induced by H₂O₂, promoted the secretion of IL-2 and IFN-γ, and inhibited the secretion of IL-4. SCP, which had protection effect, can improve blood physiological indexes, serum biochemical indices and immune factors of acute injury mice induced by alcohol. Under certain conditions, the protective effect of SCP was better than CP.

This study aims to analyze the mechanism of Mori Folium enhancing antioxidative function of chicken. The chemical components and targets of Mori Folium were collected by the traditional Chinese medicine database and analysis platform (TCMSP) and herb database. The compound-target network and the protein-protein interaction (PPI) network of Mori Folium were constructed and analyzed by Cytoscape3.7.2 software, NCBI database and STRING database. In addition, GO enrichment and KEGG pathway of these targets in PPI network were analyzed. The results showed that a total of 30 active compounds were screened, including quercetin, rutin, kaempferol, β-sitosterol, β-carotene and arachidonic acid, which acted on 221 targets. Among them, IL-6, VEGFA, EGF, INS, CAT, CASP3, CCND1, PTGS2, IL-1B and MMP9 were the core targets. GO enrichment and KEGG pathway analysis showed that the role of the targets involved in molecular function, biological process and cell composition, and involved in a number of metabolic pathways such as NOD-like receptor signaling pathway, PPAR signaling pathway, Toll-like receptor signaling pathway and apoptosis. This study theoretically shows that Mori Folium can regulate the antioxidation of chicken based on targets including IL-6, VEGFA, EGF, INS, CAT, CASP3, CCND1, PTGS2, IL-1B and MMP9, which provides enlightenment for further experimental verification and the development of Mori Folium as antioxidation feed additives.

In order to investigate the changes in energy metabolism during the course of acute laminitis in dairy cows, 12 healthy Chinese Holstein cows were used in this study, they were randomly devided into two groups (n=6): The experimental group (OF group) was treated with 17 g·kg^-1 body weight of oligofructose dissolved in 2 L·100 kg^-1 body weight of water, and the control group (CON group) was given the same amount of water, and 72 hours later cows were euthanized to collect liver and muscle tissues, and Western blot experiments and real-time fluorescent quantitative PCR experiments were performed to detect energy changes and glucose transport in the liver and muscle tissues. Indicators: glucose transporter 1 and 4 (GLUT-1, GLUT- 4), 5'-adenosine monophosphate-activated protein kinase, AMPK and related factors (PPAR-γ, PGC1-α, PEPCK). Results were as follows: In liver tissues, the expression of AMPK gene and AMPK protein of OF group increased significantly, but the ratio of P-AMPK/AMPK decreased extremely significantly, while the protein and gene of GLUT-1, PPAR-γ gene, PGC1α gene and PEPCK gene no significantly change in expression; in muscle tissue, the protein expression of AMPK gene and AMPK in the OF group did not change significantly, but the ratio of P-AMPK/AMPK decreased significantly, the expression of GLUT-4 gene and protein decreased significantly, and the expression levels of PPAR-γ and PEPCK genes increased significantly, but there was no significant change in the expression levels of PGC1-α genes. Acute laminitis in dairy cows may inhibition the energy metabolism and glucose transport ability of muscle tissue, but has little effect on the liver.

To improve the egg production of aging laying hens, we administered intraperitoneal injections of 10 mg·kg^-1 aloe-emodin (AE) to laying hens once a day for 7 days. The effects of natural AE on ovarian weight, follicle number and body weight of 580 day old laying hens were studied. Then, detect the effects of AE on the expression level of Nrf2/HO-1 pathway related proteins and genes in D-gal-induced aging ovary, and the changes of AE in alleviating D-gal-induced aging ovary after adding Nrf2 activator and inhibitor, so as to determine whether the alleviating effect of AE on D-gal-induced aging ovary depends on Nrf2/HO-1 pathway. The results showed that AE treatment increased the number of follicles at all levels (P < 0.05), including 32.00%, 34.62%, 50.00% and 54.21% for large, small and large follicles, respectively. Besides, with the treatment of AE, the estradiol (E₂) and progesterone (P₄) levels in the serum of aging laying hens were elevated by 34.14% and 680.00%, respectively. Moreover, the AE treatment can also elevated estrogen receptors (ERs, ERα and ERβ)in follicles and TG content in liver (P < 0.05). Although the transcript levels of fatty acid synthesis related genes were significantly downregulated in D580 laying hens liver tissue, transcription levels of these genes can be elevated by AE treatment. Firstly, the putative attenuating effects, cell proliferation and apoptosis of AE (10 μg·mL^-1) was evaluate through the establishment of a 2.5 mg·mL^-1 D-gal-induced aging follicular model in vitro. The results showed that AE alleviated the D-gal-induced aging ovary and natural aging ovary by increasing the activity of antioxidant enzymes and raising the expression level of Nrf2/HO-1 pathway related proteins and genes. At the same time, AE enhanced the proliferation of senescent follicle cells and inhibited cell apoptosis. In summary, AE alleviated oxidavtive stress in aging hens by activating Nrf2/HO-1 pathway. Meanwhile, AE improved ERs in follicles and vitellogenesis related genes in liver, E₂ and P₄ levels in serum, follicle antioxidative ability and liver TC synthetic ability, so as to alleviate follicular aging.

Stillbirths, weak fetuses and diarrhea of weaned piglets in sows are urgent matters to be solved in the swine industry of veterinary clinics. In response to the national policy of "reducing resistance and replacing resistance" implemented in the breeding industry, this article discusses the effects of Chinese herbal compounds on the reproductive performance of sows and the growth performance of weaned piglets based on the dialectical treatment of traditional Chinese medicine. Feeding the sows (35 heads) and piglets (140 heads) that were in estrus during the same period were fed with pre-mixed feeds of traditional Chinese medicine, and continued to feed them until the end of farrowing and 30 days respectively. At the same time, the farrowing data of the sows including the number of piglets born in the litter were recorded. Litter average weight, number of stillbirths per litter, stillbirth rate, etc. and piglet growth data including initial weight, final weight, feed to meat ratio and diarrhea rate, detected by enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative PCR (RT-PCR) The secretion of IL-6, IL-1β, TNF-α, IgG, IgA, IgM in the serum of sows and IgA in the serum of weaned piglets, as well as sow circovirus, pseudorabies virus, pseudorabies virus, porcine reproductive and respiratory syndrome virus and their antibody to determine the immunity of sows. The results showed that Qiying Decoction can significantly reduce the secretion of inflammatory factors IL-6 and TNF-α in pregnant sows, reduce the stillbirth rate, weak birth rate and mortality rate of newborn piglets produced by low parity sows, respectively decreased by 1.96%, 5.83%, 8.25%, and improve the reproductive performance of sows. The combined use of Zigan Decoction and Qiying Decoction can significantly increase the IgA level of piglets in the late weaning period, reduce the rate of diarrhea and the ratio of feed to meat to improve growth performance, reduced white blood cell levels in the blood of weaned piglets and returned red blood cells to normal levels.In summary, the Chinese herbal compound Qiying Decoction and Zigan Decoction are helpful to improve the reproductive performance of sows and restore the growth performance of weaned piglets.

The aim of this study was to investigate the association between polymorphisms of NR6A1, VSX2, VRTN and LTBP2 genes 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 410 healthy Beijing black pigs at the age of (210±7) days. The exon regions of 4 genes were genotyped by PCR and Sanger sequencing. The distribution of genotype frequency and allele frequency of the mutated loci were calculated, and the correlation between polymorphisms of mutated loci and vertebral number and carcass traits was analyzed. A total of 15 SNPs were detected in VSX2, VRTN and LTBP2 genes, including one synonymous mutation in CDS region and 14 3'UTR mutations. Statistical analysis using Duncan's multiple test revealed that a mutation of VSX2 c.536 G > A was significantly associated with vertebral number (P < 0.05). Four mutations in VRTN 3'UTR were significantly associated with traits (P < 0.05). Two mutations, c.2598 A > G and c.2607 G > T, were significantly related with vertebral number, carcass length, carcass straight length and carcass diagonal length(P < 0.05). A mutation, c.3087 G > C, was significantly associated with carcass diagonal length(P < 0.05). A mutation, c.3094 A > G, was significantly related with vertebral number and carcass diagonal length(P < 0.05). Six mutants in LTBP2 3'UTR, c.7158 A > G, c.6869 C > T, c.6319 G > C, c.6246 G > T, c.6170 A > G, c.6079 G > T, were significantly associated with vertebral number (P < 0.05), but no significant association with carcass traits. In conclusion, 10 SNPs in VSX2, VRTN and LTBP2 genes were significantly associated with vertebral number (P < 0.05), 4 SNPs in VRTN were significantly associated with carcass traits (P < 0.05). Above all SNPs could be used as candidate functional loci for the variation of vertebral number and carcass traits of Beijing black pigs.

The study aimed to explore the effect of dCas9-SunTag-DNMT3A editing system on the methylation level of IGF2R gene and embryonic developmental ability in IVF blastocysts from vitrified bovine oocytes, and to lay the foundation for precise control of DNA methylation at specific sites of vitrified oocytes/embryos. In this study, the in vitro matured bovine oocytes were vitrified, and followed by in vitro fertilization, and the prokaryotic embryos obtained by fertilization were injected with dCas9-SunTag-DNMT3A editing system, and the developmental ability of oocytes were analyzed; the methylation level of the IGF2R gene promoter was detected by sulfite sequencing, and the expression levels of IGF2R and related genes were detected by fluorescence quantitative PCR. Compared with the vitrified group, after injection of different concentrations of dCas9-SunTag-DNMT3A editing system, only the 40 ng·μL^-1 group significantly improved the developmental ability of vitrified oocytes after IVF (P < 0.05). There was no significant difference between the 20 and the 60 ng·μL^-1 groups (P > 0.05), but the developmental effect of the 40 ng·μL^-1 group was still significantly lower than that of the fresh control group (P < 0.05). The methylation level of IGF2R gene promoter in the 40 ng·μL^-1 group was found to be similar to that of the fresh group and significantly higher than that of the vitrified group (P < 0.05). The mRNA level of IGF2R gene in the 40 ng·μL^-1 group was significantly lower than that in the vitrified group (P < 0.05), which was similar to the fresh group. Injection of 40 ng·μL^-1 dCas9-SunTag-DNMT3A methylation editing system can effectively increase the methylation level of IGF2R gene promoter (P < 0.05) and significantly reduce its mRNA expression level (P < 0.05), to positively regulate the development of IVF embryos derived from vitrified oocyte, so that the cleavage rate and blastocyst rate were significantly increased (P < 0.05), and the expression of embryonic development-related genes was promoted.

This study aimed to analyze the proliferation characteristics of porcine deltacoronavirus (PDCoV) in suspension cultured porcine kidney cells LLC-PK1, so as to provide Candidate cell for large-scale production of PDCoV inactivated vaccine. LLC-PK1 cells were suspended by gradually decreasing serum method. PDCoV adaptive monoclonal cell lines were screened by limited dilution method. Indirect immunofluorescence method was used to identify the infectivity of PDCoV. The initial cell density, MOI, time of receiving virus collection and TPCK pancreatin concentration were screened to determine the best suspension culture conditions. The suspension cell strain LLC-PK1Sa which can proliferate PDCoV efficiently was screened out; PDCoV can specifically infect LLC-PK1 cells; PDCoV inoculated LLC-PK1Sa cells with a density of 2×10⁶ cells·mL^-1 according to the MOI of 10^-3, When the final concentration of TPCK pancreatin reached 7.5 μg·mL^-1, the titer of virus solution harvested 48 h after inoculation was the highest. In this study, the efficient proliferation of PDCoV in LLC-PK1Sa suspension cells was realized for the first time, and the suspension culture conditions were preliminarily optimized, which could provide theoretical reference for large-scale production of PDCoV inactivated vaccine.

The purpose of this study was to establish an indirect ELISA method for the detection of gC protein antibody of new pseudorabies virus FJ-2012 strain, and then the method was used to detect the level of gC antibody in different pig farms. The gC of FJ-2012 strain was connected to pCzn1 vector, then the recombinant vector was transfected into Arctic express (DE3) competent cells. The expression product induced by IPTG was verified by SDS-PAGE and Western blot. The indirect ELISA method for detecting PRV gC antibody was established by matrix test, determination of critical value, specificity test, repeatability, and sensitivity test. two hundred and eighty sera samples from different pig farms were detected for PRV gC antibody. The results showed that the pCzn1 gC recombinant protein of FJ-2012 strain was successfully expressed in Arctic express (DE3). The optimal antigen coating concentration of the indirect ELISA method was 10 μg·mL^-1, and the optimal serum dilution ratio of sample was 1∶50. The critical value of OD_{650 nm} for negative and positive judgment were 0.406 and 0.438, and the critical value between 0.406 - 0.438 were judged as suspicious. Clinical test results showed that, the positive rates of PRV gC antibody and PRV gB antibody in 120 blind pig serum were 91.67% and 95.00% respectively. The general coincidence rate of these two antibodies testing results were 95.83%. The positive rate of PRV gC antibody in blood samples from pig farms with unstable PR was 96.25% (77/80), however the positive rate of PRV gC antibody in blood samples from pig farms with purified PRV was 78.75% (63/80). Therefore, the indirect ELISA method established in this study not only provides a specific, sensitive and stable tool for the detection of porcine serum antibody, but also lays a foundation for the serological investigation of variant PRV.