In recent years, the frequent outbreaks of domestic cattle diseases, the increase of potential risk of infection and the decline of natural disease resistance have received much attention from multidisciplinary fields. It has been observed that there are significant differences in disease resistance among different domestic cattle breeds. A large number of research has been conducted on dissecting the molecular genetic basis of disease resistance differences among domestic cattle breeds. To better understand the advances of this field and provide the literature basis of cattle breeding for disease resistance, in this review, we have summarized the current status of domestic cattle diseases, evidence of disease resistance differences among domestic cattle breeds and the molecular genetic basis leading to such differences.

Fascioliasis is a sort of zoonotic disease caused by Fasciola spp., which can cause a large number of deaths in animals. It is considered to be a major problem for effecting on global public health and husbandry economic development. The method of resistance of host immune response that can survive in the host during the process of long-term mutual struggle between Fasciola spp. and its host. However, the excretory-secretory products (ESP) in different developmental stages of Fasciola spp. play a significant role in this process. In fact, ESP can be divided into oxidative stress related proteins, proteins related to energy metabolism and the proteasome associated proteins according to the functions. The ESP play important roles in the immune evasion of the Fasciola spp. Therefore, this dissertation expounded its functions with the purpose of providing implications for the research of new vaccines or drugs.

High conserved Wnt signal transduction pathway regulate multiple aspects of animal development, including growth and development, disease, aging, death etc. In this review, we summarize the role of Wnt signaling pathway in flatworm (planarian, tapeworm and trematode). Wnt signaling pathway accurately guide regeneration, regulate antero-posterior (AP) axis patterning and maintain genes gradient expression along AP axis in muscle cells and stem cells in planarian. Wnt signaling pathway regulate the development of proglottid in tapeworm, academics studied the function of wnt1, wnt2, wnt4 and wnt5 genes in the trematode. There are still many questions to be settled and prospects in Wnt signaling pathway in flatworms.

The objectives of the present study were to clone the genes in MOGAT and DGAT families and characterize their expression and regulation properties in chicken. The coding sequences of genes including MOGAT1, MOGAT2, LPGAT1, DGAT2 and SOAT1 which encoded proteins with enzymatic activities of MOGAT and DGAT families were cloned and sequenced. The tissue distribution of the genes was investigated by RT-PCR, and the expression patterns of genes in liver between pre-laying and peak-laying hens were detected by real-time PCR. In addition, the expression and regulation mechanism of the genes were explored using both in vivo and in vitro models treated with estrogen. The results showed that the genes in MOGAT and DGAT families expressed extensively in chicken various tissues, and had relatively higher expression levels in the lipid metabolism organs such as liver, kidney, small intestine (duodenum, jejunum, ileum), the DGAT2 gene had lower expression in liver and the MOGAT2 gene was mainly expressed in small intestine (duodenum, jejunum, ileum). MOGAT1 was expressed in the most of tissues except pancreas. The expression levels of MOGAT1, LPGAT1 and SOAT1 genes in liver of peak-laying hens were significantly lower than that in pre-laying hens. The expression levels of the genes in liver, duodenum, kidney and primary hepatocyte maintained no change or significantly decreased after the chickens and primary hepatocytes were treated with estrogen, respectively. In conclusion, the known genes in MOGAT and DGAT families, which play important roles in mammalian TG metabolism, are not the key genes in TG synthesis in liver of chicken, and the monoacylglycerol pathway don't play a leading role in chicken TG synthesis metabolism induced by estrogen.

This study aimed to study the gene structure and function of red deer β-defensin-1 (redBD-1) in order to better understand the gene's tissue expression pattern. The cDNA full-length sequence of redBD-1 gene was cloned from the red deer's lingual mucous membrane adopting the technology of PCR combining with rapid-amplification of cDNA ends (RACE), the sequence was analysed by bioinformatics, and the expression of the gene in different tissues was determined by Real-time quantitative PCR (RT-qPCR) technology. The results showed that the full-length sequence of redBD-1 gene was 455 bp, and the open reading frame (ORF) was 192 bp encoding 64 amino acids. The bioinformatics analysis indicated that theoretical molecular weight of redBD-1 protein was 6.94 ku containing 10 amino acid residues with positive charges without amino acid residue with negative charges, and its theoretical isoelectric point was 10.85. It could be predicted that redBD-1 protein had a secreting signal peptide structure, had no trans-membrane domain and mainly exereds the ectocellular physiological function; six conserved cysteine residues formed 3 intramolecular disulfide bonds with the connection of Cys1-Cys5, Cys2-Cys4 and Cys3-Cys6; the tertiary structure of maturation protein consisted of β-overlap, extension and random coils. Amino acid sequence of redBD-1 showed highest similarity with that of siBD-1 (98.4%), followed with BEBD (92.2%), HBD-2 (35.9%). RT-qPCR results indicated that redBD-1 was expressed in all detected organs, and the expression level was relatively higher in most organs of digestive system, respiratory system and reproductive system compared with a relatively lower expression level in parenchymatous organs like liver, kidney and spleen. This research could provide a theoretical basis for a better study of defensins gene functions and red deer mucosal immune system.

The objective of this study was to find the transcriptome differences of sika deer antlers at different stages, and enrich the sika deer antler transcriptome data. The sika deer antlers aged 10, 20, 28, 44 and 66 d were selected as experimental material，then the transcriptome libraries of antlers were constructed by using Illumina HiSeq^TM 2500 platform, and the transcriptome data were analyzed by bioinformatics methods, such as sequencing assess and gene annotation. The results showed that a total of 375 657 contigs with an average length of 688 bp were generated, from which 329 946 unigenes with an average length of 534 bp were defined. 90.07% of the 297 198 unigenes were annotated in public databases of Nr，Nt，Pfam，KOG/COG，Swiss-prot，KEGG and GO. Through annotation classification, a total of 39 674(12.02%) genes were divided into 41 GO function categories at level 2, and 17 732 (5.37%) genes were annotated in 26 KOG categories. Comparative analysis of differentially expressed genes in sika deer antler at 5 different stages, and 509 differentially expressed genes were screened, from which 407 genes were annotated in GO database including signal transduction, oxidation-reduction process, regulation of transcription, proteolysis, and so on. And the expression of PER1 and EGR1 (transcription regulation gene) increased with the growth of antlers, on the contrary, the expression of GAS1 decreased. It indicates that PER1, EGR1, GAS1 may have important role in regulation of antler growth. The transcriptome study of the antler tissue in the different growth period revealed the number of differentially expressed genes by using high-throughput sequencing technology, obtained the function of differentially expressed genes.

The objective of this study was to evaluate the effect of vitrification on cell apoptotic levels of porcine parthenogenetic blastocysts. The blastocoel recovery, mitochondrial membrane potential (ΔΨm), early apoptotic levels and the activities of several caspases of frozen thawed blastocysts were measured, and the mRNA expression levels of apoptosis related genes involved in different apoptotic pathways were also measured. The results showed that the rate of re-expanded blastocysts (31.03%) and total cells (31.81) of vitrified blastocysts were significantly lower than those of fresh group (100.00% and 38.17, P<0.05). The mean mitochondrial ΔΨm of vitrification group was 0.46, which was significantly lower than that of fresh group (1.02, P＜0.05). TUNEL assay revealed a significantly higher rate of DNA fragmentation in the cells of vitrified blastocysts (10.13%), which was significantly higher than that of fresh group (5.92%，P＜0.05). This study also demonstrated that vitrification caused a significant increasing of Pan-caspase, Caspase-3, Caspase-8 and Caspase-9 activities in vitrified group (20.65, 20.60, 17.58 and 19.88) comparing with fresh group (7.41, 6.46, 5.47 and 6.32, P＜0.05). qRT-PCR results showed that the relative expression levels of pro-apoptotic genes (Caspase-8, Caspase-9, TNF-α) were up-regulated and the relative expression of anti-apoptotic genes (Bcl-2 and SOD-1) were down-regulated greatly in vitrified group (P＜0.05). This study concluded that the in vitro developmental competence of blastocysts after vitrification was reduced by the decreasing of mitochondrial function, and the increasing of apoptotic levels were mediated by both death receptor and mitochondria apoptotic pathways.

The aim of this study was to clarify the expression characteristics of yak RGS1 gene and its expression pattern in ovaries and other tissues on the estrous cycle. The primer was designed based on the GenBank published Bos taurus RGS1 gene sequences, and the expression pattern of RGS1 was detected by RT-PCR in yak various tissues which include muscle, spleen, heart, kidney, stomach, small intestine, cerebellum, liver, lung and the ovarian during in different periods. The structure and function of RGS1 were analyzed by relate bioinformatics software. The mRNA expression level of RGS1 in different periods of yak estrus cycle were detected by qRT-PCR. The results showed that the yak RGS1 gene contained a 718 bp cDNA fragment, and had high homology with other mammals in nucleotide sequence through sequence alignment and phylogenetic tree, indicating that RGS1 is relatively conservative in the evolutionary process. The length of the yak RGS1 gene was 591 bp in CDS, encoding 196 amino acids, and the molecular weight was of 22.48 ku. The protein was predicted to be hydrophilic, non-secretory, lacking cross-membrane and signal peptide. Its secondary structure and tertiary structure was mainly composed of random α-helix and coil. RGS1 gene was expressed in various tissues of yak, and high abundance in small intestine, cerebellum, heart and stomach. qRT-PCR results showed that the mRNA expression level of RGS1 was significantly higher in luteal phase ovarian of yak than that in follicular phase and red stage（P<0.01). Meanwhile, the mRNA expression level of RGS1 was higher in follicular phase than that in red stage ovarian, but no significant difference (P>0.05). The RGS1 was significant differences expression among the 3 periods of yak ovarian in estrus cycle, which indicated that RGS1 involved in the estrous cycle of ovarian reproductive endocrine activities regulation.

The objective of this study was to investigate the effect of supplement urea-molasses lick block or concentrate on morphological development of small intestine and the expression of nutrient transporter gene. Eighteen healthy, 1.5-year-old Tibetan sheep ewes ((29.4±1.79) kg) were selected and randomly assigned into control group (CON group), urea-molasses lick block supplementation group (BS group) or concentrate supplementation group (CS group). The CON group were provided with oat hay ad libitum, the BS group were provided with oat hay ad libitum and supplemented with lick block ad libitum, the CS group were provided with oat hay ad libitum and supplemented with concentrate (200 g/sheep/day), during 60 days feeding experiment period. All ewes were slaughtered after the feeding trial. Sampled each segment of the small intestine, and then, the morphological development and nutrient transporter gene expression of small intestine were measured through making sections of small intestine tissue and fluorescence quantitative PCR. The results showed as follows: (1) Supplementation with concentrate or lick block significantly increased the intake of digestible energy and crude protein of Tibetan sheep (P<0.05), and the CS group was significantly higher than the BS group (P<0.05); (2) Supplementation with concentrate or lick block significantly enhanced the villus width of duodenum, jejunum and ileum of Tibetan sheep (P<0.05), and the CS group was significantly higher than the BS group (P<0.05), and the villus height of duodenum and ileum were significantly enhanced and the crypt depth of jejunum and ileum were significantly reduced in the CS group compared with the CON group (P<0.05); (3) The relative expression of insulin-like growth factor binding protein 5 (IGFBP5) mRNA in both of the duodenum and jejunum mucosa were significantly enhanced in both of the CS and BS groups compared with the CON group (P<0.05), and the CS group was significantly higher than the BS group (P<0.05), and in the ileum mucosa the CS group was significantly higher than both of the BS and CON groups (P<0.05); (4) The CS group had the higher relative expression levels of cationic amino acid transporters 1 (CAT1), L-type amino acid transporter 1 (LAT1), peptides transporter 1 (pepT1), Na⁺-dependent glucose co-transporter 1 (SGLT1) and facilitative glucose transporter 2 (GLUT2) mRNA in duodenum mucosa compared with the BS and CON groups (P<0.05). The CAT1, pepT1 mRNA relative expression in jejunum mucosa and the CAT1, LAT1, SGLT1 and GLUT2 mRNA relative expression in the ileum mucosa in the BS group were significantly higher than both of the CS and CON groups (P<0.05). We concluded that supplementation with urea-molasses lick block or concentrate increased the intake of energy and protein of Tibetan sheep in cold season, and promoted the morphological development of small intestine, and the expression of transporter gene mRNA of amino acids, peptides and glucose were enhanced in duodenum mucosa by supplementation with concentrate and they were enhanced in both of the jejunum and ileum mucosa by supplementation with urea-molasses lick block.

This experiment was conducted to study the effects of diets with different dietary digestible energy (DE) and crude protein (CP) on the body weight (BW), nutrients apparent digestibility and metabolism, colostrum composition and fatty acid profile of milk fat of Yili mares during late pregnancy period, to determine requirements of dietary DE and CP of Yili mares during late pregnancy period. Twenty-five Yili mares aged 12-13 years old with BW (380±48) kg, parity of 5-6 in late pregnancy were selected, and were randomly divided into 5 groups (group Ⅰ, Ⅱ, Ⅲ, Ⅳ and Ⅴ), each group with 5 repeats. From group Ⅰto Ⅴ, daily supplied DE were 91.28, 96.72, 102.16, 107.61 and 113.05 MJ•d^-1, respectively; CP were 0.92, 0.99, 1.06, 1.13 and 1.20 kg•d^-1. The adaptation period was 10 d, and the trial period lasted for 20 d. BW were weighted at the end of the trial period and after parturition. Colostrums were collected, and milk composition and fatty acid profile were detected. The results showed that there were no significant differences in BW, average daily gain, nutrients apparent digestibility, metabolism of nitrogen, phosphorus and gross energy, and contents of milk fat, protein, total solids and somatic cell count among the 5 groups (P>0.05). However, with the increasing of dietary DE and CP levels, the availability of digestible Ca increased linearly and quadratically (P<0.05). Lactose content of group Ⅲ, Ⅳ and Ⅴ were significant higher than that of group Ⅱ (P<0.05). The main saturated fatty acid in mares colostrum was palmitic acid, and the main unsaturated fatty acids were oleic acid and linoleic acid. With the increasing of dietary DE and CP levels, the contents of the polyunsaturated fatty acids (PUFA) in colostrum increased linearly (P<0.01). It is concluded that DE and CP feeding levels of 91.28 MJ•d^-1 and 0.92 kg•d^-1 are able to meet the requirement of Yili mare in the late pregnancy, and further increasing the dietary DE and CP levels can improve the content of PUFA in colostrum.

The present study was conducted to investigate the effects of basal diet type on the determination of true ileal digestibility (TID) of nitrogen (N) and amino acids (AA) for growing pigs by the linear regression method and the difference method. A total of 32 barrows (initial BW, (28.9±0.7) kg) fitted with T-cannula were used in an 8 × 2 incomplete Latin square design including 8 diets and 2 digestibility tests. The 8 experimental diets consisted of 2 types of basal diet (cornstarch or corn), and 4 graded levels of soybean meals (0, 13%, 26% or 39%). Each test consisted of a 5-d adjustment period and a 2-d ileal digesta collection period. The results showed that the ileal outputs of N and the most of AA, except for Lys, Arg and Pro, were greater in pigs fed the corn basal diets than those fed the cornstarch basal diets (P<0.01). And for growing pigs, ileal outputs of N and the most of AA, except for Pro, increased linearly as soybean meal levels increased in diets (P<0.01). The result obtained from the difference method showed that, pigs fed the corn basal diets had greater TID of N and Gly compared with the pigs fed the cornstarch basal diets, whereas the TID of Met and Lys was lower for pigs fed the corn basal diets than the cornstarch basal diets (P<0.05). In addition, TID of other AA was not effected by basal diet types (P>0.05). Values of TID of the most of AA for growing pigs determined by the regression method were not affected by the basal diets types (P>0.05). In summary, the determination of TID of N and AA for growing pigs was not affected by the basal diet types.

This study aimed to explore the antiviral effect and immunomodulatory functions of porcine tumor necrosis factor receptor-associated factor 6 (TRAF6), an important adaptor molecule in innate immune signaling pathway. Firstly, porcine TRAF6 gene was cloned by RT-PCR from porcine peripheral blood mononuclear cells and gene sequence was analyzed by sequence alignment. Secondly, stably expressing porcine TRAF6 cell line was established by Piggybac transposon system. And finally, we detected the impacts of TRAF6 on Pseudorabies virus (PRV) replication and porcine IFN-β promoter activity at the cellular level. The results showed that porcine TRAF6 gene was 1 623 bp. It was connected to Piggybac transposon (PB) plasmid and successfully constructed the eukaryotic expression vector PB-TRAF6, and then the recombinant plasmid was transfected into PK-15 cells to establish stably-expressed cell line by drug treatment and fluorescence screening. The high-expression of TRAF6 was confirmed by real time PCR and Western blot in the cell line. In vitro antiviral tests showed that the replication of PRV could be significantly inhibited in the cell line. Dual luciferase reporter system showed that porcine TRAF6 significantly enhanced poly(I:C)-mediated type Ⅰ interferon responses. These results indicated that porcine TRAF6 possesses significant anti-PRV activity and immunomodulatory functions, which will provide the basis for further research on immunology functions of TRAF6.

The aim of the present study was to screen the differentially expressed immune-related genes from duck spleen by DEV infection. Ducks of 50 days old were selected to inoculate with DEV in leg muscle, and the duck spleens were collected at 66, 90 and 114 h after inoculation, and were extracted for total RNA, then sequencing by high-throughput RNA-Seq technology. The sequenced segments were compared and noted for screening of the differentially expressed immune-related genes by GO and KEGG database in NCBI. Some of the differentially expressed genes were selected to verify by real-time PCR technology. The results showed that there were 511 of the differentially expressed genes in duck spleen at 66 h after DEV infection, which 70 involved in the biological immune process, including 46 up-regulated genes and 24 down-regulated genes. At 90 h after DEV infection, there were 485 of the differentially expressed genes, which 64 involved in the biological immune process, including 49 up-regulated genes and 15 down-regulated genes. At 114 h after DEV infection, there were 531 of the differentially expressed genes, which 66 involved in the biological immune process, including 35 up-regulated genes and 31 down-regulated genes. The differentially expressed immune-related genes were mainly involved in cell adhesion, antigen processing and presentation and complement activation through GO database analysis. And these genes were mainly enriched in the signaling pathways as cell adhesion molecule, ECM receptor interaction and PPAR, etc. The expressed contents of the selected 7 genes by the FQ-PCR, were basically the same with the sequencing results by RNA-Seq technology. These results provided a theoretical basis for further understanding of the mechanism of DEV infection and duck immune respone.

The pathogenic E. coli of different serotypes is a major sort of the causative agents of the diarrhea of young animals. In the present study a microencapsulated E. coli inactivated oral vaccine was prepared and its immune effect in rats was evaluated. The prepared ovine pathogenic E. coli inactivated vaccine with propolis-adjuvant as the core material was microencapsulated by natural alginate polymer as wall material, in which 7.52×10¹¹ bacteria•g^－1 dry powder were encapsulated. A hundred Wistar rats were divided into randomly 3 groups, i.e., the control, injection immunization and oral immunization groups, in which the oral immunization group was futher-divided in to the basal dose, 10-fold dose and 20-fold dose groups. After the inoculation the specific antibody was tested by the micro-agglutination test and the indirect ELISA, and the cellular immunity and mucosal IgA were detected. The micro-agglutination test showed that the specific antibody was produced from day 7 after the inoculation and reached to the peak at day 28, then declined thereafter. The indirect ELISA test showed that the antibody was positive from day 14 to day 28 after inoculation. The antibody titers in oral vaccine groups were significantly higher than that in the control (P<0.05), but not in the injection group (P>0.05), and were not significantly between the oral vaccine groups (P>0.05). The antibody titers in the oral vaccine groups declined to negative level at day 35 after inoculation whereas the injection group remained to be antibody-positive；T lymphocyte transformation test showed that SI index in 3 oral vaccine groups was higher than that in the control or the injection group significantly (P<0.05). The sIgA levels in oral vaccine groups were also significantly higher that either in the control or injection group, reaching to the peak at day 21 after inoculation. These data suggest that microencapsulated E. coli inactivated oral vaccine could produce the humoral immunity equivalent to the injected vaccine but with better cell and mucosal immunity in rats.

As a major component of membrane structure, membrane proteins not only have important physiological functions in the life of pathogen, but some of them are closely related to pathogen infection and immune response. Therefore, the main aim of this study was to isolate, screen and identify unknown possible immunogenicity proteins of Mycoplasma synoviae (MS) using immunoproteomics approach, which will lay a foundation not only for the study of infection and immune mechanism of MS but also for the establishment of diagnosis and prevention methods of related diseases. Chickens and New Zealand rabbits were immunized with MS cultured to mid-to-late logarithmic growth phase, respectively. Then the antiserum against MS from the chicken and the rabbit were prepared. Based on these, the membrane proteins of MS were isolated and then separated by two-dimensional electrophoresis. Finally the immunogenicity membrane proteins were screened and identified by Western blot combined with mass spectrum. The results showed that among the eight screened protein spots, three protein spots were identified as elongation factor EF-Tu, the rest of protein spots were identified as pyruvate decarboxylase alpha subunit, beta subunit, NADH oxidase, histidyl-tRNA synthetase and dihydrolipoyl dehydrogenase, respectively. Among them, it was confirmed for the first time that the pyruvate decarboxylase alpha subunits and beta subunits were the immune-related membrane proteins of MS, whereas the histidyl-tRNA synthetase need to be confirmed by further studies because protein score and protein Score CI% were too low. Several immune-related membrane proteins of MS strain WUV1853 were identified, which will be useful for further study on the biological function of target protein and development of new vaccines and diagnostic reagents.

The purpose of this paper was to isolate and identify the pathogenic bacteria of lymphadenitis in goat of Chongqing area and establish the test method. Sterile separation operation was carried out to isolate pathogenic bacteria from fester of the goat. The phylogenetic tree was founded according to the gene sequence of 16S RNA. And the specific primers were designed to establish PCR test method. The result showed that Corynebacterium pseudotuberculosis, Arcanobacterium pyogenes and Staphylococcus aureus were isolated from the samples, and three pairs of specific oligonucleotide primers were respectively designed according to the gene sequence of Phospholipases D, 16S RNA and nuc for multiple PCR detection. The multiple PCR detection showed that a fragment of 507, 1 378 and 278 bp amplified from the mixture genome .The sensitivity test showed that the multiplex PCR could detect minimum detectable concentration of 10³ cfu•mL^-1.The detection of 80 clinical samples indicated that, the detection rates of mixed infections of Corynebacterium pseudotuberculosis and Staphylococcus aureus, Corynebacterium pseudotuberculosis and Arcanobacterium pyogenes, three kinds of bacterias were 12.5%, 3.75% and 7.5%, respectively. Three pathogenic bacterias of lymphadenitis in goat were isolated successfully and the multiplex PCR detection method was detected in this study, it laid a foundation for the research of the prevention and control and the pathogenesis for lymphadenitis in goat.

Transglutaminases (TGases) are a widely distributed group of enzymes that catalyze the formation of isopeptide bonds either through protein cross-linking via ε-(γ-glutamyl)lysine bonds or through incorporation of primary amines at selected peptide- bound glutamine residues, and involved in multiple important physiological events. This study was aimed at cloning and expression of a transglutaminase gene from a tick Haemaphysalis longicornis Shanghai strain (HlTGase, GenBank accession number: KX59300), and analysis of the molecular characterization and the enzyme activity of the recombinant HlTGase. Total RNA of the adult ticks was extracted and HlTGase gene was amplified. The open reading frame of HlTGase was inserted into a plasmid pPICZC and transformed electrically into yeast Pichia pastoris, which was then induced to express the recombinant HlTGase. The result showed that the open reading frame of the gene was 2 262 bp, which encoded a polypeptide of 756 aa. The polypeptide possessed four transglutaminase domains. The calculated molecular weight of the polypeptide was 84.6 kDa. A phylogenetic tree showed that the polypeptide had a closest relationship with that of Drosophila melanogaster among TGases from 11 typical species, which was correspondent with the traditional taxonomical status. The antibody against the recombinant HlTGase recognized the endogenous HlTGase in a Western blotting analysis. The results also showed that the recombinant HlTGase had enzyme activity to catalyze cross linking between proteins. However, the activity was lower than a commercial available TGase from guinea pigs. HlTGase was expressed in yeast successfully. More modification in the expression and purification of the recombinant HlTGase might be required to improve the enzyme activity. This study would provide basic information for further study on the function and potential application of HlTGase.

This study was conducted to establish a V⁵⁺-induced oxidative stress model by using primary oviduct magnum epithelial cells (OMECs) of laying hens. OMECs were isolated from Lohman layers (2-3 weeks before onset of laying), cultured and characterized subsequently. Cultured OMECs were then treated with 0, 25, 50, 100, 250, and 1 000 μmol•L^-1 vanadium for 12 h to measure cell viability, apoptosis ratio as well as cellular ROS, SOD, MDA, CAT and LDH released. The results showed that: 1) cell island formed after 24 h and showed classical flagstone morphology. Cultured cells were positive to anti-CK18, anti-OVA, anti-ESR1 and anti-PGR antibody, while were negative to anti-vimentin antibody. 2) Cell viability rate were （69.90±2.78）% and （63.60±1.57）% after 50 and 100 μmol•L^-1 vanadium treated for 12 h, respectively, which were lower (P<0.05) than that of 25 μmol•L^-1 and higher (P<0.05) than that of 250 and 1 000 μmol•L^-1 vanadium. 3) The apoptosis rate increased(P<0.05) as vanadium levels increased, with the 1 000 μmol•L^-1 vanadium treatment had the highest apoptosis rate. 4) The cellular ROS, FITC mean values in 100 and 250 μmol•L^-1 vanadium groups were significantly higher than in 0 μmol•L^-1 vanadium treatment (P<0.05), meanwhile FITC mean value in 250 μmol•L^-1 was significantly higher than in 100 μmol•L^-1 vanadium treatment (P<0.05). 5) Compared with 0 μmol•L^-1 vanadium, cells treated with 50 μmol•L^-1 vanadium had lower (P<0.05) SOD activity, while 100, 250 and 1 000 μmol•L^-1 vanadium groups significantly increased MDA contents(P<0.05). The activity of CAT in 50, 100, 250 and 1 000 μmol•L^-1 vanadium treatments were significantly lower than in 0 μmol•L^-1 vanadium treatment (P<0.05). The amount of released LDH increased as vanadium levels increased, with the 1 000 μmol•L^-1 vanadium treatment had the highest value (P<0.05), and 100 μmol•L^-1 vanadium group had higher value than in 0 μmol•L^-1 (P<0.05), up to 141.61±2.81 U•gprot^-1. These results indicated that the primary OMECs were cultured successfully and supplementation of 100 μmol•L^-1 vanadium in primary OMECs can be used to establish an oxidative stress model in OMECs.

The study was conducted to investigate the effects of folacin on quinestrol induced abberant reproduction in adult male rat. Twenty 8-week-old adult male SD rats were randomly divided into 4 groups after 1-week of adaptive feeding. Control group: olive oil + physiological saline; Quinestrol group: 1.0 mg•kg^-1 quinestrol; Quinestrol + folacin group: 1.0 mg•kg^-1 quinestrol + 1.1 mg•kg^-1 folacin (dissolved in saline); Folacin group: 1.1 mg•kg^-1 folacin. Quinestrol and folacin were given intragastric administration to male rats daily for 2 weeks. After the treatments, the sexual organs were weighed, semen quality were tested, antioxidant status of testes were investigated, testis tissue section were made, histopathology of testes were observed, the expressions of Caspase-3 and endothelial nitric oxide synthase (eNOS) were detected by immunohistochemical staining. The results showed that atrophy of sexual organs were induced, the weights of sexual organs were decreased, abnormal spermatogenesis were abberant, sperm quality were reduced, antioxidant enzyme activity were decreased, oxidative stress were happend in testes after the treatment of quinestrol alone. By contrast, the expressions of Caspase-3 and eNOS in the testes were increased. In the group of quinestrol and folacin treatment, oxidative stress were reduced by decreasing eNOS expression, the expression of Caspase-3 were inhibited, the sperm deformity rate and the damage of the tubule were reduced, the reproductive function of male rats were improved significantly. Compared with the control group, the testicular antioxidant enzyme activities, tissue structure and sperm quality were ameliorated to some extent in the folacin treatment group, there was no significant difference. The results clearly demonstrate for the first time that folacin effectively inhibited the damaging effects of quinestrol on reproductive function in adult male rats.

In order to develop the antioxidant polysaccharide preparations, based on our previous researches, 18 compounds composed with Lycium barbarum polysaccharide (LBP), Atractylodes macrocephala polysaccharide (AMP), Chinese angelica polysaccharide (CAP), Codonopsis pilosula polysaccharide (CPP), Garlic polysaccharide (GP), Lily polysaccharide (LP), Schisandra chinensis polysaccharide (SCP), selenizing LBP (sLBP), selenizing AMP (sAMP), and selenizing CAP (sCAP) with stronger antioxidant activities according to carbohydrate content at 9:1 ratio of polysaccharide to selenizing polysaccharide were chose. Their antioxidant activities were compared. In vitro tests, the scavenging abilities of 18 compounds were compared for 3 kinds of free radicals. In vivo tests, the chicks were injected respectively with 3 compounds each at high and low dosage when they were vaccinated with Newcastle disease vaccine, and the dynamic changes of serum contents of 3 antioxidases and MDA were determined. The results of in vitro tests showed that 18 compounds each at 5 concentrations had different extent scavenging abilities for DPPH radical, hydroxyl radical and ABTS radicals, and the abilities of LBP-sAMP, CAP-sLBP, and CAP-sAMP were stronger. The results of in vivo test showed that in 6 compound groups at 4 time points, the activities of CAT, SOD and GSH-Px were higher or significantly higher than those in control group, and MDA contents were lower or significantly lower than those in control group. In CAP-sLBP group, 3 antioxidases activities were the highest and MDA contents were the lowest. These results indicate that 18 compounds possess differen extent antioxidant activities, the activity of CAP-sLBP is the strongest among 18 compounds and can be considered as the candidate of the antioxidant polysaccharide preparation.

In order to study the effect of compound Kuqin on T cell subsets and cytokines of blood in canines with canine parvovirus, we established the canine parvovirus model, and separate 120 canines into 6 groups, like blank control group, model group, positive medicine group, compound Kuqin high dose group, middle dose group, low dose group, with 20 canines in each group. We set the time point that 6 hours after the virus was inoculated as 0 d, and collect the blood sampling from the canine saphenous vein at the day 0, day 7, day 14, day 21, day 28, and day 35. T lymphocyte subsets were detected by flow cytometry, and the cytokines IL-2, IL-4, IFN-γ were detected by kit. The result showed that, compared with the model group, compound Kuqin can promote the secretion of IL-2, IL-4, IFN-γ, and increase the percentage of CD3⁺, CD3⁺CD4⁺T cells in peripheral blood, and decrease the percentage of CD3⁺CD8⁺T cells. In conclusion, compound Kuqin can regulate canines’ immune function and improve the ability of antiviral after they were infected by canine parvovirus.

To investigate the roles of Ccna1 gene in duck skeletal muscle development, its promoter sequence was amplified by RT-PCR, then was analyzed using bioinformatics methods. qRT-PCR was used to detect the expression profiles of Ccna1 during duck embryonic muscle development, as well as the expression profiles of myogenic transcription factors predicted to take part in regulation of Ccna1 transcription. We obtained 2 213 bp promoter region of duck Ccna1 gene. Bioinformatics analysis showed there were typical TATA-box, CAAT-box and binding sites for transcription factors MyoD, MRF4, MyoG, Sp1, PITX1 and MEF2 in the Ccna1 gene promoter region. The qRT-PCR data showed that MyoD, MRF4, MyoG and Ccna1 expressed in duck muscle tissues at embryonic development stages. In the breast muscle, the expression of MRF4 was the highest in E23 and was significantly higher than that in D8 (P<0.05). In the leg muscle, the expression of MRF4 and MyoG were the highest in E17, and was significantly higher than that in D2, D8 (P<0.05). Clustering results showed that the expression patterns of Ccna1 was consistent with MyoD in breast muscle, while in the leg muscle, Ccna1 was clustered together with MRF4 and MyoG. It was preliminary concluded that Ccna1 was involved in the duck skeletal muscle development, and Ccna1 might be associated with the MRF4 and MyoG during the leg muscle development, whereas in the breast muscle development, its expression might be regulated by MyoD.