Non-thermal plasma (NTP) is the slightly ionized plasma. The temperature of its ions and neutral particles is far lower than that of electrons, making the whole discharge system in a low temperature state. At present, NTP is widely used in the sterilization, vaccine production, oral treatment, wound healing, cancer treatment and other biomedical fields. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) produced by NTP have dose-dependent regulatrory effects on the cell proliferation and apoptosis. NTP treatments at a low intensity and short time can generate low levels of ROS and RNS, which will induce the cell proliferation and angiogenesis and thus to accelerate the wound healing. NTP treatments at a high intensity and long time can inhibit the cell proliferation and even induce the cell apoptosis. However, the mechanism of NTP regulating the cell proliferation and apoptosis and its potential application in the animal husbandry are still unclear. Therefore, this paper reviews the effects and possible mechanisms of NTP on endothelial cells, keratinocytes, fibroblasts, stem cells and malignant tumor cells, so as to provide a theoretical basis for the application of NTP in the wound healing and cancer treatment. This paper also summarizes the potential applications of NTP in the breeding environment and health, the growth and reproductive performance of livestock and poultry, as well as the animal food processing and preservation, so as to lay a foundation for promoting the animal husbandry production and ensuring the animal food safety in China.

ADP-ribosylation is a kind of post-translational modification of proteins catalyzed by ADP-ribosyl transferases (ARTs). It is divided into poly-ADP-ribosylation and mono-ADP-ribosylation, which plays an important role in regulating many biological events during spermatogenesis, such as assembly of centromere and spindle, DNA damage repair and chromatin condensation. Part of the ARTs gene deletion leads to abnormal spermatogenesis, resulting in reduced semen quality and even male infertility. However, the detailed mechanism of spermatogenesis regulated by ADP-ribosylation is still unclear, and the understand on the role of mono-ADP-ribosylation within it is even less. Therefore, this paper reviews the research progress of ADP-ribosylation in spermatogenesis, hoping to provide new ideas for further revealing the regulatory mechanism of spermatogenesis, and promoting breakthrough in the prevention and control of male reproductive disorders and the treatment of male infertility.

Viruses are ubiquitous to the gastrointestinal tracts of various types of animals, including the rumen of ruminants. The rumen is a dense and taxonomically diverse consortium of bacteria, fungi, protozoa, archaea and virus. Nowadays, most of the researches by multi-omics and bioinformatics have focused on rumen bacteria. Studies have shown that diet, environment and host genotype have great impact on the diversity and community structure of rumen bacteria; meanwhile, rumen bacteria also affect the host’s metabolism, growth performance and health status in turn. Compared with rumen bacteria, rumen phages have received relatively fewer attention. With the development of molecular biology technology, high-throughput sequencing has paved way for the study of virome. Many novel viruses in rumen have been discovered by viral metagenomics, which largely enriches the understanding of the diversity of rumen viruses. Starting from the concept of rumen phage, this review focused on the analytical methods including virus DNA extraction, sequencing and bioinformatics. Then discussed the virome analytical methods both in the rumen and other ecosystems, and finally summarized the application prospects of rumen virome research.

Newcastle disease virus (NDV) is a kind of single-stranded negative-sense non-segmented RNA virus, which contains the genome size of about 15.2 kb in length that encodes six structural proteins and two non-structural proteins. Since the NDV genome is smaller, it cannot encode all the proteins needed for viral replication, thus host proteins must be employed to complete the life cycle of NDV. The NDV M protein is a non-glycosylated membrane-associated protein, which plays essential roles in inhibiting host cell gene transcription, regulating the replication and transcription of the NDV genome, and promoting the assembly and budding of progeny virions. At present, great research progresses have been made in the relationship between M protein and NDV virulence and replication, and the formation and utilization of M protein-based virus-like particles. However, there has little progress about the roles of NDV M protein interaction with host proteins in regulating NDV replication. Due to the crucial roles of M protein in NDV replication, this summary mainly reviews the structural characteristics and intracellular localization characteristics of M protein, and also the functional study on the interactions between M protein and host proteins, which will provide theoretical references for better understanding and studying the role of M protein in the replication and pathogenicity of NDV.

The aim of this study was to screen and analyze differentially expressed genes in ovarian tissues of Yunnan Dulong chicken and White Leghorn chicken using RNA-seq technology, and to lay the foundation for subsequent studies on gene function, breed improvement, and germplasm characteristics of Dulong chicken. In this study, the ovarian tissues of three 43-week-old Dulong chickens in good health were collected for transcriptome sequencing and compared with the ovarian tissue of White Leghorn Chicken in similar rearing conditions and age. The results showed that there were 1 273 differentially expressed genes in Dulong chickens compared with White Leghorn chicken, among which 522 genes were up-regulated and 751 genes were down-regulated. GO and KEGG enrichment analysis showed that differentially expressed genes were mainly involved in proteolysis, translation, exosomes, membrane components, binding, and catalysis, and played an important role in protein synthesis, ECM-receptor interaction, oxidative phosphorylation, adhesion plaque, and other pathways. Significant enrichment pathway analysis showed that Dulong chickens had certain advantages in egg quality and immune resistance, but there was still a certain gap between Dulong chickens and White Leghorn chickens in laying performance. RPL22, ATP5I, COX5A, JAKMIP1, CATH2 screened in this study were mainly enriched in the pathways related to resistance, egg-laying performance, and egg quality, which could be used as important candidate genes for further study.

This study aimed to identify the long non-coding RNAs (lncRNAs) and genes related to pork quality traits of the longissimus dorsi in Tibetan pigs and Yorkshire pigs (180 days old), and to further explore their molecular regulatory role, which could provide theoretical basis for the improvement breeding of pork quality traits. According to previously published data, 180-day-old healthy Tibetan boars (n=3) and Yorkshire boars (n=3) raised under the same conditions were used as the research objects to collect phenotype data of the quality of the longissimus dorsi and the transcriptome data of the longissimus dorsi of the same individual were used to screen differentially expressed lncRNAs and differentially expressed genes (DEGs). Cis- and trans-target DEGs were predicted based on the positional relationship and expression correlation of differentially expressed lncRNAs and DEGs, then trans-target DEGs were enriched by GO biological processes and KEGG pathway analysis. Subsequently, lncRNAs-DEGs-biological process regulatory network related to muscle growth and fat deposition was constructed. Then, association events between candidate target genes and meat quality traits were analyzed, which comprised of drip loss, pH, meat color, marbling, intramuscular fat content, muscle fiber cross-sectional area, loin muscle area and vitamin B₁ content. A total of 1 546 lncRNAs and 20 219 mRNAs were identified in the longissimus dorsi of Tibetan pigs and Yorkshire pigs,there were 49 differentially expressed lncRNAs and 154 DEGs of them based on statistical analysis. Of these, 7 cis and 138 trans regulatory DEGs targeted by differentially expressed lncRNAs were predicted. Functional enrichment analysis showed that a large number of candidate DEGs were significantly enriched in biological processes of muscle growth and fat deposition, including positive regulation of phosphatidylinositol 3-kinase signaling, muscle structure development, fat cell differentiation, positive regulation of JAK-STAT cascade, and positive regulation of MAPK cascade, etc. Evidence from regulatory network showed that MSTRG.34836.10 and MSTRG.151.1 had the most target DEGs, followed by MSTRG.21406.1, MSTRG.26709.1. Association analysis indicated that 11 candidate target DEGs were significantly related to the meat quality traits. Taken together, some differentially expressed lncRNAs identified in this study were related to muscle growth and fat deposition. It was also suggested that these differentially expressed lncRNAs had an effect on pork quality by regulating the expression of HES1, LEP, TGFB2, and PER2. This study laid the foundation for further analysis of the potential molecular mechanism of pork quality traits.

The aim of this study was to investigate the feedback loop between oar-miR-127 and target gene FOXO4 and its effect on sheep follicular granulosa cells. The miR-127 promoter region of sheep was predicted through Promoter 2.0. The binding sites of transcription factor FOXO4 in oar-miR-127 promoter region were predicted with JASPAR database. The pGL3-basic-miR-127 luciferase reporter vector containing the predicted binding site of FOXO4 and pcDNA-FOXO4 overexpression vector were constructed. The binding sites of oar-miR-127 located in 3'-UTR region of predicted target gene FOXO4 were analyzed with TargetScan software. Recombinant luciferase reporter plasmids of pmir-GLO-FOXO4 with wild-type, mutant-type and deleted fragment containing predicted binding sites were constructed. The pGL3-basic-miR-127 luciferase reporter vector and pcDNA-FOXO4 overexpression vector were co-transfected (or separately transfected) into sheep follicular granulosa cells cultured in vitro. The wild-type, mutant-type, and deleted FOXO4 recombinant plasmids were co-transfected with miR-127 mimic/mimic NC into 293T cells cultured in vitro, and luciferase activity was detected after 48 h. The expression levels of miR-127 (FOXO4) and apoptosis genes (Casp3, Bax and BCL2) in granulosa cells were detected by qRT-PCR after the pcDNA-FOXO4 overexpression vector (miR-127 mimic/mimic NC) was transfected into sheep follicular granulosa cells for 48 h. Overexpression of transcription factor FOXO4 significantly decreased the luciferase activity of miR-127 promoter (P<0.05) and the expression level of miR-127 (P<0.05); The luciferase activity of granulosa cells co-transfected with miR-127 mimic and FOXO4 wild-type recombinant plasmid was significantly lower than that of miR-127 mimic and FOXO4 deletion/mutant recombinant plasmid (P<0.05). The expression of FOXO4 in granulosa cells transfected with miR-127 mimic was significantly decreased (P<0.05), while the expression of Casp3 and Bax genes was extremely significantly increased (P<0.001). There was no significant change in the expression of Casp3 and Bax after overexpression of transcription factor FOXO4 in granulosa cells(P>0.05). The relative expression of BCL2was significantly decreased with overexpression of FOXO4 (P<0.05). This study confirmed a feedback loop between oar-miR-127 and target gene FOXO4, and both could promote the apoptosis of granulosa cells. These results laid a foundation for the study of its mechanism in the development of sheep ovarian follicles.

The aim of the study was to clone the sequence of DGAT1 gene, clarify its expression pattern in different tissues of goat, and further reveal the role of DGAT1 gene in regulating lipid metabolism in goat intramuscular preadipocytes. In this study, seven healthy 10-month-old Jianzhou male goats were used as experimental animals. The sequence of DGAT1 gene was cloned by RT-PCR and analyzed using bioinformatics. The relative expression level of DGAT1 in different tissues of goat were determined by real-time quantitative PCR (RT-qPCR). The eukaryotic expression vector pcDNA3.1-DGAT1 was constructed by double enzyme digestion and transfected into goat intramuscular preadipocytes. RT-qPCR was used for detecting the expression of DGAT1 and lipid metabolism-related genes. The effect of overexpression of DGAT1 on lipid droplet formation was observed by Oil red O staining method, and the GPO-Trinder enzyme reaction was used to measure triglyceride content. A length of 1 651 bp of DGAT1 gene was cloned (GenBank accession number： MT221183), including 125 bp 5'UTR, 1 470 bp CDS, and 56 bp 3'UTR, encoding 489 amino acid residues. The goat DGAT1 gene had the highest expression in small intestine and the lowest in spleen. RT-qPCR results showed that DGAT1 was extremely significantly overexpressed in cells (P<0.01) by pcDNA3.1-DGAT1 treatment, as a results, the expression of GPAM (P<0.05), ADRP (P<0.01) and ACOX1 (P<0.01) were significantly up-regulated, while the expression of AGPAT6 (P<0.05), MLYCD (P<0.01) and HSL (P<0.01) were significantly down-regulated. Oil red O staining results showed that the lipid droplet accumulation after overexpression of DGAT1 was significantly (P<0.01) increased compared with that in the control group, as well as the results of triglyceride determination (P<0.01) in intramuscular adipocytes in goats. This study successfully obtained the goat DGAT1 gene CDS region sequence and constructed the pcDNA3.1-DGAT1 eukaryotic expression vector, overexpression of DGAT1 could significantly promote lipid deposition of intramuscular preadipocytes in goats and significantly affect the expression of lipid metabolism-related genes. These results provide important data for further clarifying the mechanism of DGAT1 in regulating intramuscular lipid metabolism in goats.

The study aimed to explore the mechanism of muscle fiber type difference between fetal and adult Mongolian horses. In this study, three 4-month-old fetuses (two females and one male) and three 5-year-old healthy adult female horses were selected as a whole with 4 body-distributed and representative muscle tissues (triceps brachii, splenius, longissimus dorsi and gluteus medius). Fetal Mongolian horse muscle fiber was one group, and adult Mongolian horse muscle fiber was one group, the experiment carried out 3 biological repetitions.Firstly, the skeletal muscle samples of Mongolian horses were observed by immunohistochemical staining. Then, the skeletal muscle samples were subjected to RNA-seq to construct the mRNA expression profiles of skeletal muscle fibers in different periods of fetal and adult Mongolian horses. Differentially expressed genes were screened for GO and KEGG functional enrichment analysis. Histological results showed that the proportion of fast muscle fibers in the fetal period was significantly higher than that in the adult period. RNA-seq was used to screen candidate genes that might affect the transformation related to muscle fiber types. The gene expression profiles related to muscle fiber types in fetal and adult Mongolian horses were compared. A total of 250 differentially expressed genes were screened out. There were 27 up-regulated genes in adulthood, including TNNT1, MYOZ2 and ATP2A2, and 17 up-regulated genes in the fetal period, including MYL1, TNNT3 and ENO3. ATP2A2 and TNNT1 were mainly related to fast and slow muscle fiber type transformation, and these differentially expressed genes were also closely related to skeletal muscle growth and development. The results showed that the proportion of fast muscle fibers in fetal Mongolian horses was significantly higher than that in adult Mongolian horses. ATP2A2, MYOZ2 were mainly expressed in muscle fibers of adult Mongolian horse, ENO3, TNNI2 were mainly expressed in muscle fibers of fetal Mongolian horse. Calcium and AMPK signaling pathways are related to muscle fiber type conversion.The results of this experiment are of great significance to the study of Mongolian horse muscle fiber type transformation.

In this study, phenotypes and differentially expressed genes in the white and black regions of the body of Mongolian spotted horse were analyzed and verified, in an attempt to elucidate the molecular mechanism of hair color formation of Mongolian spotted horse, and lay a foundation for future protection, development and utilization of Mongolian spotted horse. First, skin tissue sections of the white and black regions of the body of Mongolian spotted horse were made and stained by HE staining method and phenotypic differences were observed under a microscope. Secondly, transcriptome sequencing was performed on the skin of white and black regions to compare the differentially expressed mRNAs. Finally, the selected typical genes were tested by immunofluorescence immunohistochemistry and qRT-PCR, the results of transcriptome sequencing were further verified. The results showed that there were significant differences in the pigment distribution between the black and white skin tissues of Mongolian spotted horses. Melanocytes were mainly distributed in the epidermis and hair follicle parts of the black skin tissues, while no melanin deposition was found in the corresponding positions of the white skin tissues. The expression levels of 740 genes were significantly different between black and white skin tissues (P<0.05), among which 109 genes were highly expressed in black skin tissue of Mongolian spotted horse, and 215 genes were highly expressed in white skin tissue of Mongolian spotted horse. Differentially expressed mRNAs were mainly enriched in melanin production pathway contained TYR, TYRP1, DCT, PAX3 and DAPL1, etc.genes. TYR and TYRP1 proteins were not expressed in the hair bulb of the hair follicle of Mongolian spotted horse. DCT was expressed in the mother material of hair follicle. The hair color formation of Mongolian spotted horse is mainly determined by the expression of genes related to melanin formation.

The purpose of this study is to evaluate the level of genetic diversity and explore the genetic structure between subspecies of Lepus tolai in Xinjiang by molecular genetics methods. In this study, the CO1 and D-LOOP gene sequences of mitochondrial DNA were obtained from 78 Tolai hares from the northern, northwestern, central and eastern regions of Xinjiang by PCR amplification and sequencing, and then the data were analyzed by bioinformatics methods. The results showed that 68 haplotypes were defined from the combined sequence of 78 samples of Xinjiang Tolai hare, and relatively high haplotype diversity (h=0.996±0.002) and nucleotide diversity (π=0.049±0.023) were detected. The phylogenetic relationship and median-joining network showed that the Tolai hare samples used in this study were divided into three branches (Clade A-C), in which Clade A and C contained the majority of hares from the northern and northwest groups of Xinjiang, belong to the Lepus tolai lehmanni, while the majority of hares in Clade B were from the central and eastern regions of Xinjiang, belong to the Lepus tolai centrasiaticus. Except for few mixed samples of Tolai hare, the clustering of the rest majority was consistent with their distribution range and geographical distance. The results of AMOVA analysis, Fst and gene flow showed that the genetic differentiation between subspecies of Tolai hare was larger and the gene flow was smaller, while the genetic differentiation within subspecies was relatively small and the gene flow was relatively large. The results of neutral test showed that the Tajima’D value of Xinjiang Tolai hare population was 1.075 (P>0.05), and Fu`s Fs was -15.100 (P<0.01). The mismatch distribution of nucleotide indicated a multi-peak phenomenon, and EBSPs analysis showed that the population expansion occurred at about 0.01 Ma. The results of comprehensive analysis based on CO1 and D-LOOP gene showed that Xinjiang Tolai hare was rich in genetic diversity and had an obvious systematic geographical distribution pattern. There was a strong gene flow and a small genetic differentiation between northern and northwest groups of L. t. lehmanni. However, there was a distinct differentiation between the central and eastern groups of L. t. centrasiaticus, which was probably due to the greater geographical distance. In addition, Tolai hare has recently experienced a population expansion.

The aim of this study was to investigate the expression pattern of DIO3 during the embryonic development in mice, assess the effects of intragastric gavage of levothyroxine (L-T₄) on DIO3 expression and embryo number, and provide basic data for the study of maternal thyroid hormone abnormalities during pregnancy. In this study, the healthy Kunming mice (70 females, 35 males) aged 6 weeks were used as experimental objects. After adaptive feeding for one week, female mice were given intraperitoneal intragastric administration of 10 U PMSG (dissolved in 0.3 mL normal saline). After normal feeding for 48 h, male and female mice were caged together at the ratio of 1∶2 overnight. Once the milky vaginal suppository was visible in the morning of the next day, it was considered as the first day of pregnancy. Pregnant mice were randomly divided into 5 groups. The mice in experimental groups were intragastric administrated with 0.25(SG, n=10), 1.25(MG, n=10), 2.50(LG, n=10) and 5.00 μg·μL^-1(HG, n=10) of L-T₄, re-spectively, and the control group was intragastric administrated with the same amount of normal saline(CG, n=10), 80 μL per day for 10 days. Placentas were collected from mice in exprmental groups on the 10th and 18th day of gestation (n=4/group) and the numbers of embryos were recorded. In addition, ovaries of non-pregnant mice and placentas of mice from the control group on the 10th, 14th and 18th day of gestation (n=3/group) were collected. The expression of DIO3 in placentas of mice was detected by RT-qPCR, in situ hybridization and immunohistochemistry. The contents of TSH, FT₃ and FT₄ in serum of pregnant mice were detected by chemiluminescence meter after intragastric administration of L-T₄. The results of RT-qPCR showed that，under normal physiological condition, the expression level of DIO3 in placenta decreased with the increase of gestation time, and the highest expression level was found on the 10th day of gestation, and the lowest expression level was found on the 18th day of gestation, which was lower than that in non-gestation (P<0.01). In situ hybridization and immunohistochemistry analysis of paraffin sections were consistent with the results of RT-qPCR. The serum levels of TSH, FT₃ and FT₄ increased with the increase of concentration of L-T₄ intragastric administration, and the differences among groups were significant (P<0.05). In addition, the expression of DIO3 increased with the increase of intragastrically administered dosage of L-T₄. On the 10th day of gestation, there was a significant difference in DIO3 expression between the control group and HG group (P<0.01), there was no significant difference between the control group and the SG, MG and LG groups. On the 18th day of gestation, the expression of DIO3 in the control group was significantly higher than that in gavage groups (P<0.01). On the 10th day of gestation, the number of embryos of pregnant mice in the gavage groups was slightly higher than that in the control group. On the 18th day of gestation, the number of embryos of pregnant female mice in the control group and the gavage groups was approximately the same. In conclusion, intragastric administration of L-T₄ could increase the level of thyroid hormone in the serum of pregnant mice and induce the expression of DIO3 in the placenta. However, intragastric administration of L-T₄ had no significant effect on the number of embryos in pregnant mice. This study provides a theoretical basis for further study on the molecular mechanism of thyroid hormones level regulation during pregnancy.

This study aimed to construct the overexpression vectors encoding two isoforms of goat CREBZF protein (long isoform SMILE and short isoform Zhangfei), and to preliminarily explore the apoptotic effect of CREBZF on goat endometrial epithelial cells (gEECs). In this study, pairs of primers were designed for the CDS region encoding two isoforms of goat CREBZF protein， and the gene fragments were amplified by PCR; Target gene fragments were connected to linear pcDNA3.1(+) by seamless cloning to construct the recombinant overexpression vectors of pcDNA3.1(+)-SMILE and pcDNA3.1(+)-Zhangfei which were identified by double enzyme digestion and sequencing; The overexpression vectors were transfected into gEECs, then total RNA and protein were extracted. Overexpression efficiency was determined by qRT-PCR and Western blot; Flow cytometry was used to investigate the effect of SMILE and Zhangfei overexpression on apoptotic rates of gEECs. PCR and double enzyme digestion results showed that the target bands were in the correct locations;Sequencing results showed that the target gene fragments were 100% consistent with the predicted CREBZF sequence in NCBI GenBank. The results of qRT-PCR and Western blot showed that two isoforms of CREBZF were overexpressed significantly in transcription and translation levels, and SMILE may be modified after translation on gEECs. The apoptotic rate of empty vector group was 8.45%; The apoptotic rate of pcDNA3.1(+)-SMILE group significantly increased to 12.41% (P<0.01); The apoptotic rate of pcDNA3.1(+)-Zhangfei group increased to 9.83%, however, there was no significant statistical difference compared with the empty vector group. It is suggested that CREBZF may regulate the apoptotic process of gEECs. This study combines the segmented cloning of target gene with seamless cloning to construct the above two overexpression vectors, and lays a foundation for further research on the role of CREBZF in ruminant embryo implantation.

The aim was to explore the effect of N-acetyl-L-cysteine (NAC) on goat endometrial stromal cells (gESCs). In this study, goat endometrial stromal cells were used as the object. Three concentration gradients of 100,200,400 μmol·L^-1 NAC were added to the culture medium. The 0 μmol·L^-1 NAC was regarded as the blank group and CCK -8 method was used to screen the optimal concentration of NAC and judge its effect on cell proliferation.Then DCFH-DA probe and RT-qPCR were used to detect the optimal concentration of NAC on the intracellular reactive oxygen content, cell cycle and proliferation related factors, and goat reproductive performance-related genes TGF-β1 and TGF-β3 mRNA expression levels. The results showed that the concentration of 200 μmol·L^-1 NAC had a more significant effect on the proliferation of goat endometrial stromal cells than other gradients. In addition, 200 μmol·L^-1 NAC reduced the ROS content of cells compared with the blank group(P<0.01), thereby delaying cell senescence. RT-qPCR results showed that the addition of 200 μmol·L^-1 NAC significantly increased the expression levels of cell cycle and proliferation-related factors CyclinA1, CyclinE, PCNA mRNA (P<0.01), and reduced the mRNA expression of cell cycle factor CyclinD2(P<0.01); and extremely significantly reduced the TGF-β1 gene(P<0.01) of the transforming growth factor family and significantly reduced the mRNA expression level of the TGF-β3 gene(P<0.05). Therefore, this study speculates that 200 μmol·L^-1 NAC may promote cell proliferation by regulating the expression levels of CyclinA1, CyclinE, PCNA, and CyclinD2 in cells, and affect goat reproduction traits by regulating the expression levels of transforming growth family factors TGF-β1 and TGF-β3. This study provide a theoretical basis for further research on the mechanism of NAC’s influence on goat reproductive traits.

The study aimed to explore the effects of absent, small, or homeotic 1-like (ASH1L) gene knockdown on mRNA profiling in bovine cumulus cell. In this study, the mRNA profiling in cumulus cells with or without interference of ASH1L gene was measured using RNA sequencing. Differentially expressed genes (DEGs) were identified, and bioinformatics analyses (differential expression of genes,function,signaling pathway,alternative splicing (AS) and SNP) were carried out. Transcriptional levels were confirmed using quantitative RT-PCR,3 replicates per trial. Results showed that a total of 2 046 DEGs were screened in cells with or without interfere-nce of ASH1L gene, of which 1 078 were up-regulated and 968 were down-regulated. Those DEGs mainly enriched into 2 802 GO terms including cell division, cell junctions and histone methyltransferase activity. Fifty-three pathways were involved by those DEGs such as leukocyte transendothelial migration, PI3K-Akt and cell adhesion molecule. The results of gene set enrichment analysis showed that ASH1L gene was closely involved in developmental induction, H3K4 specific histone methyltransferase activity, histone demethylation, etc. Moreover, 117 994 AS events with 12 patterns were identified; In the meantime,522 205 SNPs were detected and the frequency of base transitions was significantly higher than that of transversions. In summary, the mRNA profiling between bovine cumulus cells with or without interference of ASH1L gene were different. It was speculated that ASH1L gene may be involved in leukocyte transendothelial migration and cell adhesion molecule to regulate cell proliferation. Also, ASH1L can affect gene expression by AS and SNP, which may provide a theoretical basis for further study on the function of ASH1L in cumulus cell proliferation and embryo development.

The objective of this study was to explore the population characteristics and influencing factors of plasma anti-Müllerian hormone (AMH) concentration in Holstein cows, and to estimate its genetic parameters. A total of 625 blood samples were collected from 398 healthy Hols-tein cattle in a large-scale dairy farm in Beijing. The plasma AMH concentration was determined by ELISA method. The MIXED procedure in SAS 9.2 was used to analyze the influencing factors of the plasma AMH concentration. A single trait animal model was used to estimate the variance components and genetic parameters of the plasma AMH concentration and the approximately genetic correlations between the plasma AMH concentration and some reproductive traits were calculated by DMU. The results from this study showed that the mean of the plasma AMH concentration in Holstein cattle was (96.30±14.25) ng·L^-1. The plasma AMH concentration was significantly (P<0.01) influenced by the sampling season, and was not significantly influenced by the parity, pregnant stage and lactation stage. The plasma AMH concentration was a moderately heritable trait with a heritability of (0.10±0.08). The plasma AMH concentration was associated with the interval from calving to the first service, interval from the first to the last inseminations and stillbirth in heifer and cow with moderate to high genetic correlations and approximate genetic correlation coefficients ranging from 0.3 to 0.7. In conclusion, this study provided a reference for further research on the genetic basis of the plasma AMH concentration in Holstein cattle, which was helpful to investigate the reproductive performance of dairy cows by AMH concentration.

The study aimed to investigate the function of DBI gene and reveal its role in reproductive physiology of yak. The ovaries, fallopian tubes and uterine tissues of healthy female yaks (3-6 years old) at different stages of reproductive cycle (follicular stage, luteal stage and gestation stage) were collected and divided into 9 groups with 3 biological replicates in each group. Real-time quantitative PCR (qRT-PCR) and Western blotting (WB) assays were used to detect the rela-tive expression of DBI in reproductive organs of female yaks at different stages of reproductive cycle. Immunohistochemistry (IHC) was used to determine the distribution of DBI protein in the ovaries, fallopian tubes and uterus of female yaks. The results showed as follows: 1) qRT-PCR result showed that,in ovary,the expression of DBI was significantly higher in luteal stage and pregnancy stage than in follicular stage (P<0.05); In fallopian tube, the expression of DBI in luteal stage was significantly higher than that in follicular and gestation stages (P<0.05). In utero, the expression level of DBI was highest during gestation stage, followed by luteal phase, and the lowest during follicular phase. 2) Western blotting results showed that,in ovary, the level of DBI was significantly higher in pregnancy stage than in follicular and luteal stages (P<0.05); In fallopian tube, the level of DBI in luteal stage was significantly higher than that in follicular stage and gestation stage (P<0.05); In uterus, the expression level of DBI in pregnancy stage was significantly higher than that in luteal phase and follicular phase (P<0.05). 3) IHC results showed that DBI was mainly located in reproductive epithelial cells, granulosa cells, follicular membrane cells, luteal cells, tubal epithelial cells, uterine stromal cells and uterine glands. The expression levels of DBI in the ovaries, fallopian tubes and uterus tissues of yaks at different stages of reproductive cycle are significantly different, indicating that DBI is involved in the reproductive physiological regulation of yaks, which lays a theoretical foundation for further exploring the role of DBI gene in the reproductive aspects of plateau mammals.

The study aimed to explore whether the long non-coding RNA of Tubb4b (lncRNA-Tubb4b) has protein coding capability and the regulatory effect of lncRNA-Tubb4b on the Tubb4b gene. The ATG, 0/1/2 T tails and Flag sequence were added to lncRNA-Tubb4b to analyze the coding ability of lncRNA-Tubb4b by constructing vector, liposome transfection and Western blot; Then, overexpression vector of lncRNA-Tubb4b was constructed and the siRNA of lncRNA-Tubb4b was synthesized, while the liposome transfection and real-time fluorescent quantitative PCR technology were used to detect the influence of lncRNA-Tubb4b on the Tubb4b gene.The results showed that the Flag fragments with 0, 1, and 2 T-tail, was amplified in this study, were fused with lncRNA-Tubb4b, then were successfully constructed into the pcDNA3.1(-) vector; After transfection of mouse spermatogonia, it was found that lncRNA-Tubb4b with 0, 1, and 2 T-tail had no FLAG protein bands. With pcDNA 3.1(-)-EGFP vector as a control, overexpression of lncRNA-Tubb4b significantly reduced the mRNA expression level of Tubb4b in mouse spermatogonia (P<0.01), while there was fluorescence in control cells. And interfering the expression of lncRNA-Tubb4b significantly increased the mRNA expression level of Tubb4b in mouse spermatogonia (P<0.01). It is indicated that lncRNA-Tubb4b does not encode protein, and overexpression or interference with the expression of lncRNA-Tubb4b will extremely significantly reduce or increase the expression of Tubb4b.

PICA behavior of cattle refers to an abnormal behavior characterized by licking and gnawing some foreign matters without nutritional value, which is often considered to be caused by some nutrients disorders and living environmental confinement. It has been proven that a strong association between gut microbiota and host nutritional metabolism is existed. The link between gut microbiota-gut-brain axis(MGB) and the animal behavior is being revealed. However, there is a lack of enough investigation on the relationship between the occurrence of PICA and the changes of gut microbiota in cattle. In this paper the gut microbiota was compared between PICA cows and the normal ones in order to reveal the possible correlation between gut microbiota and PICA behavior. After the clinical survey for PICA cows in an Angus cow breading farm, 10 healthy Angus pregnant cows as control group (Ctrl) and their 10 counterparts with typical PICA behavior as PICA group (PICA) were chosen.Blood and hair samples were collected to measure the minerals, fat-soluble vitamins (VA, VD, VE), and 17 free amino acids and total serum proteins. Feces samples were collected and the composition and the structure of gut microbiota were analyzed and compared. The results showed that the serum content of Iso and Phe in PICA cows were significantly higher than those in the Ctrl group(P<0.05). Though the α-diversity and β-diversity of gut microbiota in PICA cows were not significantly different from those in the Ctrl group(P>0.05), the relative abundance of Oscillospira and CF231 were significantly lowered than those in the Ctrl groups (P<0.05)。Pearson correlation analysis showed that there was a significant negative correlation between Iso content and the relative abundance of CF231, as well as Phe and Tenericutes (P<0.05). The results suggest that the changes of Iso and Phe contents in the serum and the changes of Oscillospira and CF231 relative abundance in gut microbiota could be associated with the generation of PICA behavior in Angus cows, which provides a possibility for developing a therapeutic way to treat PICA via gut microbiota manipulation.

Feline infectious peritonitis (FIP) is a highly lethal infectious disease caused by coronavirus and characterized by severe abdominal cavity inflammation. However, there is no approved standard method for the diagnosis or treatment of pathogen. Therefore, this study describes the diagnosis and treatment process from a case report of feline infectious peritonitis and explores reasonable diagnosis and treatment methods for the disease. High-throughput sequencing technology, clinical symptoms, biochemical analysis, blood routine and etc. were comprehensively used to diagnose this case, to evaluate the treatment effect of the antiviral drug GS-441524 with albumin/globulin ratio and fSAA level, and to monitore the sequelae. Results showed that high-throughput sequencing technology can be applied to the diagnosis of FIP. After high-dose drug treatment, the clinical symptoms were significantly improved, the albumin/globulin ratio and fSAA were restored to normal at 8 weeks, and the discharge standards were met at 10 weeks. No signs of disease recurrence were found during the 16-week review. The successful treatment of this case proves the potential of high-throughput sequencing technology in veterinary clinical diagnosis, and the effectiveness of GS-441524 in the treatment of critical cases. The serum albumin/globulin ratio (≥0.6) and fSAA (<5 μg·mL^-1) can be a clinical indicator to evaluate the therapeutic effect.

Bovine group A rotavirus (BRVA), bovine coronavirus (BCoV), bovine viral diarrhea virus (BVDV), bovine norovirus (BNoV) and bovine nebovirus (BNeV) are common viruses that cause calf diarrhea, and there are mixed infections in clinic. The purpose of this experiment was to establish a multiplex PCR method that can simultaneously detect the above five viruses. The multiplex PCR method for simultaneous detection of BRVA, BCoV, BVDV, BNoV and BNeV was successfully established by designing primers, optimizing reaction system and conditions. This method only specifically detected the target virus, but did not amplify the diarrhea pathogens such as bovine astrovirus, bovine kobuvirus, bovine parvovirus, enterotoxigenic Escherichia coli, Salmonella and Campylobacter jejuni, which had good specificity and stability. The lowest detection limits for each virus were as follows:BRVA 1.63×10⁵copies·μL^-1, BCoV 2.0×10⁶copies·μL^-1, BVDV 1.9×10⁵copies·μL^-1, BNoV 1.96×10⁵copies·μL^-1 and BNeV 2.0×10⁵copies·μL^-1. The comparison experiment showed that the coincidence rate between the multiplex RT-PCR method and the RT-PCR method for detecting single virus was 100%. The detection results of 220 yak diarrhea samples collected from northwest Sichuan grassland from 2019 to 2020 showed that the detection rates of BRVA, BNoV and BNeV were 40.5%, 5.9% and 4.5%, respectively, while BVDV and BCoV were not detected. The multiplex RT-PCR method established in this study can simultaneously detect five common diarrhea viruses in fecal samples of calf diarrhea, providing technical support for the rapid diagnosis of bovine diarrhea.

This study aimed to establish an antibiotic-resistance-free gene knockout method for Glaesserella parasuis based on the Flp-FRT site-specific recombination system, thus providing a genetic manipulation tool for the research on virulence factors, pathogenic mechanism, genetic engineering vaccines of G. parasuis. Firstly, the cI857/P_RM/P_R expression regulation system from λ bacteriophage was used to control the expression of Flp recombinase to realize resistance gene excision from the resistance-marker mutants of G. parasuis. Then, the Flp expression plasmid was introduced into the G. parasuis CF7066 by electroporation, and the obtained recombinant strain was naturally transformed with the suicide plasmid containing the homologous DNA sequence flanked with FRT sites to generate the marker mutants. Following the regulation expression of Flp recombinase, the markerless mutants were screened by colony PCR and the Flp expression plasmid was cured by improving the culturing temperature to 42 ℃. Furthermore, this knockout system was improved by mutation of unilateral FRT site. Initially，the Flp recombinase carried by temperature-sensitive shuttle plasmid pSHG5C-Flp was verified to be able to express under the control of λ repressor and promotor of cI857/P_RM/P_R and to excise the resistance gene in the mutant of ΔnanH::Kan^R. Subsequently, the G. parasuis CF7066 was electrotransformed with pSHG5C-Flp in advance, then it was naturally transformed with the suicide plasmids of pKF-ΔnanH and pKF-Δapd2 successively. Correspondingly, the markerless single gene and double gene deletion mutants of ΔnanH and ΔnanHΔapd were generated from the primary transformants of the primary generation, and the Flp recombinase expression plasmid pSHG5C-Flp also was cured. Although we achieved successive two gene knockout, the knockout efficiency for the second gene was obviously declined, which was probably due to the accumulating Flp recombinase that probably interfered the homologous recombination mediated by the plasmid DNA from natural transformation. Thus, we screened the mutated one FRT site and introduced it into the plasmid of pKFM-Δapd2 to conduct natural transformation. As a result, the introduction of mutated FRT site into the suicide plasmid guaranteed the occurrence of homologous recombination and deletion mutation, which increased the effectiveness and stability of this method. This study provided a novel markerless and double gene knockout system for G. parasuis and paved a way for the research on molecular pathogenetic mechanism and genetic engineering vaccines.

Three isolates of Streptococcus subsp. equi HTP133, HTP123 and HTP232 were obtained from a donkey farm in Xinjiang. To identify the genotypingand understand the biological and epidemic characteristics of these three isolates,the biochemical characteristics analysis,drug sensitivity test and 16S rRNA sequence comparison analysis and phylogenetic tree construction were performed. The SeM alleles were amplified by PCR and sequenced for genotyping. The results showed that these three isolates were Streptococcus equi subsp.equi. These three isolates were sensitive to 18 drugs, including amoxicillin, ampicillin and gentamicin etc, whereas they are resistant to penicillin and cefuroxime. Form the 16S rRNA gene analysis and the phylogentic tree, we found that these isolated strains belonged to group I and there is an obvious variation in their variable regions of V1-V5. The homology of these three isolates are closely related with S. equi JQ726493. In addition,two new SeM genotypes, SeM 210 (HTP232) and SeM 211 (HTP133, HTP123) were identified by SeM allele analysis and they were closely related to SeM 136, SeM 138, SeM 142 and SeM 146 genotype.This work enriches the molecular biological foundation of prevalence and transmission characteristics of donkey S. equi in China and provides the theoreitical basis for the prevention and control of donkey strangles as well.

Heparin could inhibited the adhesion of Trueperella pyogenes to host cells in a dose-dependent manner. To understand the heparin binding proteins from T. pyogenes and their adhesion characteristics, the proteins were extracted from the lysate of T. pyogenes by heparin aga-rose gel, and were identified by protein mass spectrometry and Western blotting. The reactivity of the extracted proteins with the serum from a goat naturally infected with T. pyogenes was detected by Western blotting. The recombinant proteins were prepared. The heparin binding activity of recombinant proteins and the inhibitory effect of heparin or the antiserum on the binding of recombinant proteins to heparin were analyzed. The cell adhesion activity of the recombinant proteins and the inhibitory effect of the antiserum on the adhesion of T. pyogenes to host cells were analyzed. The results showed that multiple substrate binding proteins of ATP binding cassette transporters including iron-binding protein (IBP) were extracted. The IgG antibody against IBP was detected in the serum of naturally infected goat with T. pyogenes, which indicated that IBP was expressed in the process of T. pyogenes infection. The result of SDS-PAGE showed that rIBP could bind heparin agarose gel, and the physiological concentration of NaCl could partially dissociate the rIBP bound to heparin agarose gel. The result of Western blotting showed that heparin could inhibit rIBP binding heparin agarose gel in a dose-dependent manner, indicating that rIBP specifically binds heparin. Western blot showed that rIBP could adhere to MDBK cells. Western blotting results showed that anti-rIBP serum could not inhibit the binding of rIBP to heparin, but the results of viable bacteria counting showed that anti-rIBP serum could inhibit the adhesion of T. pyogenes to MDBK cells. The result of this study laid a foundation for further understanding heparin binding proteins of T. pyogenes and their role in the adhesion of T. pyogenes to host cells.

This study aimed to analyze the differential expression of Th1 and Th2 type immune response-related genes in macrophages RAW264.7 in response to Echinococcus granulosus protoscoleces stimulation, this study lays a theoretical foundation for further revealing the immune regulation mechanism of macrophages against protoscoleces.The protoscoleces and macrophage RAW264.7 were cultured for 6, 24 and 72 hours, total RNA was extracted from RAW264.7 cells, and cDNA library was constructed. The results showed that when the protoscoleces(PSC)were cultured with macrophages RAW264.7 for 6， 24， and 72 hours, compared with the PBS control group, there were 848, 3 745 and 7 009 genes in PSC treatment group showed significant difference, in which 415, 1 159 and 2 237 genes were up-regulated and 433, 2 586 and 4 772 genes were down-regulated respectively. The results of differential expression analysis of Th1 and Th2 immune response-related genes showed that when macrophages RAW264.7 were co-cultured with protoscoleces for 6 hours, a total of 31 Th1 and Th2 type immune response-related genes showed significant changes in expression, including 15 genes significantly up-regulated, 16 significantly down-regulated. When macrophages RAW264.7 were cultured with protoscoleces for 24 hours, there were 112 of Th1 and Th2 type immune response-related genes were significantly changed, of which 34 were up-regulated and 78 down-regulated. When macrophages RAW264.7 were cultured with protoscoleces for 72 hours, the expression of 212 Th1 and Th2 type immune response related genes changed significantly, among which 50 genes were up-regulated and 162 genes were down-regulated. These differentially expressed genes include the Leucine-rich repeat protein LRRCs, the G protein coupled receptor GPCRs, the C type Lectin receptor CLRs, the Scavenger receptor SRS, and some downstream molecules of PRR. At the same time, the results showed that the expression of Th1-related genes such as Tnfrsf1a, Ifnar1 Tnfrsf12a and so on was significantly up-regulated when the protoscoleces were cultured with macrophages RAW264.7 for 6 and 24 hours. However, when the protoscoleces were cultured with macrophages RAW264.7 for 72 hours, the expression of Th2-related genes such as IL4 and IL6 was significantly up-regulated. At the same time, some differentially expressed genes were selected at random and verified by qRT-PCR. The results showed that the expression trend was consistent with that of RNA-seq. To sum up, this study systematically analyzed the different expression profiles of genes related to macrophages RAW264.7 responding to protoscoleces stimulation in different time periods, the genes associated with macrophage RAW264.7 response to protoscoleces stimulation and innate immunity were screened, this study provides a basis for further understanding the role of these differentially expressed genes in response to protoscoleces stimulation by macrophages RAW264.7 and their regulatory mechanisms.

This study aimed to investigate the effect of STAT6 mediated macrophage polarization on intracellular survival of Brucella. Brucella smooth strain S2308(S2308) and rough vaccine strain RB51(RB51) were used to infect macrophages.M1-type macrophage marker factors p65, NOS2 and IL-1β were detected by qRT-PCR. mRNA expression levels of M2-type macrophage marker factors STAT6, ARG1 and IL-10 were detected by qRT-PCR. The expression of M1-type marker CD86 and M2-type marker CD206 was detected by flow cytometry. The inhibitory effect of p-STAT6 protein and inhibitor AS on the protein was detected by Western Blot.The expression levels of M1-type cytokines TNF-α and IL-12 and M2-type cytokines IL-4 and IL-10 were detected by ELISA. Finally, CFU count of intracellular colonies was carried out. The qRT-PCR results showed that the mRNA expression of M1 type factor was significantly induced at 8 h and 12 h after infection, the expression of M2 type factor was low at 72 h, and the expression of M2 type factor was high at 72 h. Flow cytometry showed that S2308 could significantly induce CD86 expression at 12 h after infection, and CD206 expression at 72 h after infection, but RB51 had no effect on both.Western Blot results showed that strain S2308 activated STAT6 signaling pathway 72 h after infection, while RB51 hardly activated STAT6 signaling pathway. The inhibitory effect of inhibitor AS was the best at 2 μmol·L^-1 concentration. ELISA results showed that AS inhibitors could significantly inhibit the release of IL-4 and IL-10, and promote the release of TNF-α and IL-12.CFU count results showed that intracellular bacteria in S2308 group were firstly decreased and then significantly increased, and S2308+IL-4 group was significantly higher than S2308 group, S2308+AS group was significantly lower than S2308 group, and there was no significant difference in RB51 group. Brucella S2308 can induce the transformation of M1 type macrophages to M2 type macrophages through STAT6 in the late stage of infection, and promote the release of Th2 type cytokines, which is conducive to the intracellular survival of Brucella. RB51 does not activate this pathway and does not affect intracellular survival.

The objective of this study was to analyze the differential expression profile of lncRNA in pig kidney epithelial cells PK15 infected by Japanese encephalitis virus (JEV), and to explore the potential function of host lncRNA in JEV infection. In this study, JEV was used to infect PK15 cells for 36 hours, and the cells were collected. Meanwhile, an uninfected control group was established. There were 3 biological replicates in each group. The total RNA was extracted, and the cDNA libraries were constructed and then subjected to high-throughput sequencing to identify differentially expressed lncRNA. The cis- and trans-acting target genes of lncRNA were obtained by calculating the positional relationship and sequence similarity between differentially expressed lncRNA and mRNA. Next, the predicted target genes were analyzed by GO and KEGG. Finally, 9 differentially expressed lncRNAs were randomly selected for expression verification using real-time fluorescent quantitative PCR (quantitative reverse transcription PCR, qRT-PCR) technology. The results showed that a total of 856 differentially expressed lncRNAs were identified, of which 452 lncRNAs were significantly up-regulated and 404 lncRNAs were significantly down-regulated. GO analysis results showed that the target genes of lncRNAs were mainly enriched in innate immune response; KEGG pathway analysis results showed that the target genes of lncRNA were significantly enriched in signal pathways such as tumor necrosis factor, Toll-like receptor and NF-κB. The quantitative results are the same as the sequencing results through qRT-PCR verification. This study lays a theoretical foundation for further exploring the roles of lncRNA in JEV infection.

This study aimed to determine and characterize the biological and infection characteristics of an Escherichia coli phage C6 which was previous isolated from feacal samples of pre-weaned piglet. Bacteriophage classification was confirmed by TEM observation and sequencing. Host range was determined by spot assay and double-layer agar plates method. Infecting characteristics including Optimal multiplicity of infection (MOI), efficiency of plating (EOP), adsorption rate, one-step growth curve as well as bacteriostatic curves of phage C6 were determined. Results were as follows: Escherichia coli phage C6 was a T4-like phage. Phage C6 infected a Escherichia coli O157 strain (E. coli W1), two Shiga toxin-producing Escherichia coli strains (STEC W3, STEC W5) and a enteropathogenic Escherichia coli strain (EPEC 143). The results of each infection index were shown in descending order as follows. Plaque size and EOP: E. coli W1>EPEC 143>STEC W3>STEC W5; optimal MOI: EPEC 143, STEC W3, STEC W5>E. coli W1; EOP and adsorption rate: E. coli W1>EPEC 143, STEC W3, STEC W5; phage proliferation and bacteriostatic efficacy in LB medium: E. coli W1>STEC W3>EPEC 143, STEC W5. E. coli phage C6 was a T4-like phage that infected different pathogenic E. coli strains and had significant bacteriostatic effects on pathogenic E. coli strains despite of varied infection characteristics.

To understand and master the relationship between prophage and virulence, environmental adaptability, drug resistance and metabolic activity of Streptococcus suis (SS), in this study, the pathogenicity test, LD₅₀ test, the number of bacteria loaded in tissue, histopathological observation, biofilm formation(BF) ability test, drug sensitivity test and transcriptome sequencing were carried out for the prophage positive strains (positive bacteria for short) and prophage negative strain(negative bacteria for short). The results showed that among 39 strains of positive bacteria and 7 strains of negative bacteria, 11 strains and 2 strains of bacteria had a lethal rate of more than 80.0%, The LD₅₀ of 11 positive bacteria ranged from 8.4×10⁸ to 2.36×10⁹CFU·piece^-1, and that of 2 negative bacteria were 1.88×10⁹ and 2.72×10⁹ CFU·piece^-1, respectively. There was no significant difference in LD₅₀ between positive and negative bacteria (P>0.05). After being challenged with positive bacteria with LD₅₀ of 8.4×10⁸CFU·piece^-1, the number of bacteria in the organs (lung, liver, spleen, kidney, heart and brain) of mice was the most, and the pathological changes were the most obvious, after being challenged with the negative bacteria with LD₅₀ of 2.72×10⁹CFU·piece^-1, the number of bacteria in each organ tissue of mice was the least, and the pathological changes were the least; The positive rate of BF formation was 97.4% for positive bacteria and 100.0% for negative bacteria; Both positive and negative bacteria showed multiple drug resistance to tetracycline, erythromycin, clindamycin, azithromycin, ceftriaxone and doxycycline, and there was no significant difference in drug resistance between them (P>0.05); Compared with positive bacteria, virulence related genes, BF formation related genes, drug resistance genes, fatty acid biosynthesis pathway and arginine and proline metabolism pathway related genes were up-regulated in negative bacteria. the results showed that there was no significant correlation between the presence or absence of prophage and the increase or decrease of SS virulence, high or low drug resistance and BF formation ability; However, the presence of prophage will lead to the down-regulation of genes associated with SS fatty acid biosynthesis and arginine and proline metabolism, resulting in the reduction of fatty acid and amino acid metabolism.

Oxytropis glabra is a poisonous plant widely distributed in grassland and desert area of Inner Mongolia, its main toxic compound is an alkaloid (swainsonine, SW). Livestock is poisoned or even dead after feeding, resulting in a major loss of grassland animal husbandry in serious cases. This research explored the relationship between SW from Oxytropis glabra in diffe-rent populations and endophytic fungi in Inner Mongolia. One hundred and twenty plants of Oxytropis glabra from 8 populations in Inner Mongolia were collected, and the SW from indivi-dual plant and mycelia were isolated and purified by extraction, centrifugation and ion exchange chromatography. The SW levels were determined by GC-MS. The endophytic fungi were isolated from stems and leaves, and the total DNA of plants and fungi was extracted. The fungal specific sequences were amplified, and the fungus was identified both by microbiological characteristics and DNA sequence comparison. Results showed that SW was detected in 111 plants from 8 popula-tions of Oxytropis glabra with the highest level 369.05 μg·g^-1 and the average level 32.78 μg·g^-1. Endophytic fungi were isolated from 38 planets. Under pure culture condition, mycelia showed loose white villous, and the colonies were round, upheaval with neat edge and radial growth. The color of colonies gradually showed gray, dark gray or brown to dark brown. SW was detected in all endophytic fungi samples with the level range of 0.83-2 573.24 μg·g^-1. The endophytic fungus was identified as Alternaria by microbiological study and 5.8S rDNA/ITS sequence comparison. The plants with Alternaria endophytic fungi contained SW, while the plants without Alternaria endophytic fungi did not contain SW. The cultured Alternaria endophytic fungi synthesized SW.

The purpose of this study was to evaluate the adjuvant effects of cultivated Artemisia rupestris L. crude polysaccharides (CARCP) on inactivated foot-and-mouth disease vaccine (FMDV). ICR mice were subcutaneously administrated with different doses of CARCP combined with FMDV. ELISA was used for the detection of FMDV-specific antibodies. T cell subsets in spleen and lymph nodes, and the expression of dendritic cell surface molecules as well as regulatory T cells (Tregs) in spleen were detected by flow cytometry. Mice were weighed every week, and observed the growth condition of mice after immunization. Compared with FMDV alone, higher levels of FMDV-specific IgG, IgG1 and IgG2a were induced by both medium-dose and high-dose of CARCP (P<0.01). Medium-dose of CARCP not only significantly increased the percentage of CD4, CD8, and CD44 T cells in the spleen and lymph nodes of mice (P<0.05), but also promoted the expression of CD86 and MHC-II in DC (P<0.05), whereas the CD4⁺CD25⁺Foxp3⁺ T cells were significantly down-regulated (P<0.05). There was no noticeable influence on the growth and body weight of immunized mice (P>0.05). The results suggested that CARCP as an adjuvant for FMDV could augment the specific humoral and cellular immune responses via inducing DC maturation, inhibiting Tregs-level, which provided a basis for the development of polysaccharide adjuvant of FMDV.

In order to understand the pathogenic characteristics of a Gram-negative strain isolated from cows bedding litters.In this study, the isolates bacterium were identified by morphological observation, biochemical test, staining microscopy, 16S rDNA homology analysis. Drug sensitivity test was used to determine drug sensitivity, mice attack test was used to determine the virulence of the isolates. The results showed that the isolates were Gram-negative bacteria. The nucleotide sequence of 16S rDNA was 1 283 bp, the nucleotide homology with Acinetobacter johnsonii is greater than 99%. The isolated bacterium had strong pathagenicity to mice, the mortality was 80% in test group. The isolates were sensitive to 6 drugs including streptomycin, compound sulfamethoxazole, moderately sensitive to cefotaxime and cefuroxime, resistant to 4 drugs including penicillin and erythromycin. In this study, a strain of Acinetobacter johnsonii was isolated from dairy cow bedding litters, which provides a reference for the prevention and treatment of Acinetobacter johnsonii.