Imprinted genes only express the genetic information from male parent or female parent. A large number of researches showed that imprinted genes have important roles in the fetus growth, fetus development, function of the placenta and postnatal behavior. Imprinted genes have been shown to determine the transport capacity of the placenta by regulating its growth, morphology and transporter capacity. When the environment of placenta or the placenta phenotype changes, the imprinted genes can adapt to the prevailing nutritional environment, by varying their placental epigenetic status. Imprinted genes may act as nutrient sensors and optimize the fetal acquisition of nutrients for growth. These genes have a major role in the epigenetic regulation of placental phenotype.

Since 2010, veterinary science became an independent discipline from a branch of integrative animal husbandry and aquatics discipline in Natural Science system of China. Accordingly, financing scope of veterinary science was adjusted. Due to multiple reasons such as the enhancing intensity of finance, the increasing attention to basic research etc., application number of National Natural Science Foundation of China (NSFC) increased significantly. In this paper, we summarized and analyzed the application and finance situation of general program, young program and region program in veterinary science section in fiscal years 2010-2012 to provide reference data for researchers in veterinary or related fields in application of NSFC.

In order to find the different expression genes in longissimus muscle of Min pig and Yorkshire pig. In this study, the digital gene expression(DGE) profile in longissimus dorsi muscle of Min pig (fat) and Yorkshire pig (lean) were built by high-throughout sequencing technology, the different expression genes were screened and annotation. Significant enrichment analysis of GO and Pathway were done. The results showed that more than 5 000 000 clean tags were obtained from Min pig and Yorkshire pig, the tags whose copy number were greater than 100 occupied the most percentage, Min pig and Yorkshire pig were 88.62% and 84.62%, respectively. Completely blast to the sense chain and one tag only blast to one gene, Min pig and Yorkshire pig were 2 351 788 and 2 388 175, respectively. Min pig compared to Yorkshire pig, different multiples more than two times were 1 098, among them, 44 were up-regulated genes and 1 054 were down-regulated genes, 11 genes expressed only in Min pig, 256 genes expressed only in Yorkshire pig. Different expression genes included some genes and transcripts related to intramuscular fat content, lipid metabolism, meat color, tenderness and muscle growth. 23 853 and 34 731 novel transcripts located at the different sites of the pig genome were predicted in Min pig and Yorkshire pig, respectively, which including one base mismatch tags. Terms from the component ontology were 18, including cytoplasm, membrane-bounded organelle, pigment granule, lytic vacuole, extracellular region part, etc. Terms from the function ontology were 2, including protein binding and oxidoreductase activity. Terms from the process ontology were 14, including response to stress, response to reactive oxygen species and carboxylic acid metabolic process, etc. Terms from pathway were 11, according to the P value from small to large, lysosome, PPAR signaling pathway and primary bile acid biosynthesis were the most significantly different pathways. The results of this study could provide a theoretical basis for studying the novel genes or finding genetic factors affecting pork quality.

The aim of this study was to analyze the effects of single and combined genotypes of MyoG and c-fos genes on muscle fiber traits in pig. Large White(180) and Beijing Black pigs（170）were used as experimental animals. The frozen section, H-E and SDH staining technology were used to detect the partial meat quality and muscle fiber traits. PCR-SSCP was used to detect the SNPs of MyoG and c-fos genes, the genotype frequency of SNPs were calculated, and the association of SNPs with partial meat quality and muscle fiber traits were analyzed. The results showed that: (1) A polymorphism site was detected in exon1 of MyoG gene, and three kinds of genotypes(AA, AB, BB) were detected; Two polymorphism sites were detected in exon4 of c-fos gene, and three kinds of genotypes(AA, AB, BB) were detected, respectively; (2) In the two pig breeds, the muscle fiber area and diameter decreased, and muscle fiber density increased with the increase of the allele A frequency at A2512G site. The muscle fiber density and percentage of red muscle fiber enhanced with the increase of the allele B frequency at G2650A site. And the allele A and B had significant effects on muscle fiber area and density at A2910G site; (3) The phenotype values of different traits(except for backfat thickness) had significant difference between individuals with favorable and unfavorable combined genotypes in Beijing Black pigs(P＜0.05 or P＜0.01), the phenotype values of different traits(except for backfat thickness and percentage of middle muscle fiber) had significant difference between individuals with favorable and unfavorable combined genotypes in Large White pigs(P＜0.05 or P＜0.01). The results indicate that the single nucleotide polymorphisms and combined genotypes of MyoG and c-fos genes have effect on the muscle fiber traits in pig, they could be considered as the candidate genes for muscle fiber traits.

The purpose of this study was to explore the associations of polymorphisms of the CRBP4 gene with egg laying traits and egg quality and to find important markers for these traits of Huainan partridge chicken. The polymorphisms of CRBP4 gene were detected by PCR-SSCP and directly sequencing in 308 Huainan partridge chicken under cage condition. The association between mutation sites of CRBP4 gene and early egg laying traits and egg quality was analyzed. The results showed that, there were three mutations, C826T in the exon2, C1240T and A1241G in the exon3, respectively, and C826T and A1241G resulted in 49 Ser→49 Leu and 97 Ile→97 Val, respectively. The three sites belonged to moderate polymorphism. At the three sites, the population was in Hardy-Weinberg equilibrium (P>0.05). The mutation site of C826T was significantly associated with the egg weight at first laying (P<0.05). The CC genotype was positive for the egg weight at first laying. The haplotype combinations consisting of C826T, C1240T and A1241G was significantly associated with egg weight at first laying (P<0.05). The results indicate that the correlation is detected between different genotypes and haplotypes combinations and egg weight at first laying at the three sites.

The experiment were conducted to analyze and compare the diversity and dynamic change of the gastrointestinal mucosa-associated microbiota (MAM) of high quality chicken line A (Pure Line♂, Group A) and Huiyang Beard chicken (Pure Line♂, Group B) during day 1-28. The V3 region of 16S rDNA of the mucosa-associated microbiota of two chicken pure lines were analyzed by Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) at day 1, 3, 7, 14, 21 and 28, respectively. DGGE profiles, based on the number of bands of the gel, indicated the MAM of ileum between the two groups were significantly different at day 28. The similarity of the MAM in craw, glandular stomach and cecum between the two groups were higher than 70%, while the similarity in duodenum, jejunum and ileum were 40%, 44% and 18% at day 28, respectively. The alterations of MAM of ileum were remarkably different between the two groups during day 21-28, with 70% change in group A and 11% change in group B. This diversity between the high quality chicken line A and Huiyang Beard chicken might be influenced by the host genetic and, at the same time, might be one of the possible cause for their different growth performance.

This study aimed to research characteristics of poultry embryo amniotic epithelial cells (AECs) which were isolated, cultured, identified and induced to differentiate to different mesodermal cells. AECs were isolated from Beijing-You chicken fetal membranes using enzymes digestion, immunofluorescence and RT-PCR were used to identify Nanog, Oct-4, Sox-2, CK19 and Vimentin. The multi-lineage differentiation potential of AECs was investigated, which were induced to differentiate into osteoblasts, adipocytes and pancreatic-islet like cells cells. The result showed that chicken AECs not only expressed the special markers of stem cells, but were induced to differentiate into osteoblasts, adipocytes and pancreatic-islet like cells successfully. To sum up, the AECs isolated from Beijing-You chicken fetal membranes have a high reproductive capacity and plasticity in vitro, and can be used as seed cells for the preservation of endangered species and for clinical treatment. The results can provide the theoretical basis for clinical treatment using AECs.

This study aimed to develop a real-time reverse-transcription polymerase chain reaction (RT-PCR) method based on SYBR Green Ι fluorescent for detection of horse Toll-like receptor mRNA. According to the gene sequence in conservative region of horse’s Toll-like receptors available in GenBank, the specific primers were designed with β-actin as an internal control to construct Real-time RT-PCR assay. The standard curves showed good linear relationships with the correlation (r²>0.990) and efficiency about 1.0 in the range of 1×10² to 1×10⁸ copies·μL^-1. The melting curve analysis showed that the product was specific to a single peak, and no primer-dimers, with high specificity and sensitivity. The good reproducibility was obtained and the coefficient of variation were less than 3.5% for the intra-assay and inter-assay. The clinical sample test indicated that the TLR1, TLR2, TLR3, TLR4, TLR6, TLR7, TLR8 and TLR9 mRNA were transcripted in medulla, and TLR4 mRNA level was highest, but TLR9 mRNA level was lowest. This assay could be successfully used for the detection of clinical samples and provide a technical platform to research horse TLRs at the mRNA levels in the quantitative analysis.

In order to research the relationship between the organic ingredients in eggshell and eggshell quality, 30 eggs from each breed were collected randomly. All breeds including Beijing You chicken, Large Silky Fast chicken, Bian chicken, Chahua chicken, Tibetan chicken and Jinhu Wufeng chicken were bred under the same feed and management conditions for 38 weeks. These eggs were used to test their eggshell quality and organic ingredients in eggshell. The results showed that there was significant difference (P<0.05) on eggshell quality and the organic ingredients in eggshell among different breeds. The content of protein in eggshell was extremely significantly and positively correlated with eggshell strength (r=0.839，P<0.01) and had significant positive correlation with eggshell thickness (r=0.224，P<0.05). The concentration of uronic acid and amino acid glucan in eggshell both had extremely significant and positive correlation with eggshell strengh (P<0.01). The conclusion of this research was that the difference of the content of the organic ingredients in eggshell was one of the most important factors affecting eggshell quality.

The experiment was conducted to investigate the effect of chromium picolinate (CrPic) on Ca²⁺ concentration of lymphocyte and immune response from heat stress broiler chickens. Three hundred 1-day-old Arbor Acres male broiler chickens were randomly allotted to 5 treatments (Ⅰ-Ⅴ), each treatment represented by 5 replicates of 12 birds each. At 15-day-old，the broilers in treatmentⅠ-Ⅳ were pre-fed 0, 400，800，1 200 μg·kg^-1 Cr of diet for 2 weeks, then they were housed in 3 controlled environment chambers with daily high cyclic temperature (27 ℃—35 ℃—27 ℃) for 14 d, the broilers in treatment Ⅴ were kept in 22 ℃ as the control group. The result shows that supplemental chromium (800-1 200 μg·kg^-1 Cr) decreased serum insulin concentration, supressed the increasement of the lymphocytes ［Ca²⁺］_i and IL-2 concentration, increased the lymphocyte proliferation. The results sugget that the CrPic could alleviate the negative effect of heat stress on the cellular immune response of heat stress broiler chickens.

The aim of this study was to investigate a safe and effective universal vaccine. A consensus HA gene sequence was found by phylogenetic analysis all of the HA gene sequences of H1 subtype classical swine influenza virus and influenza A virus from GenBank and optimized according to the swine bias codon usages（optimized consensus, opti-CS）, we also selected another two HA genes and optimized (one is from A/Swine/Guangdong/1/01 strain and the other one is from rPan09 strain),then the three opti-HAs were inserted into pCAGGS vector, and constructed three recombinant plasmids named pCA-opti-CS, pCA-opti-G11 and pCA-opti-r09, respectively. The HA protein transiently expressed in 293T cell was detected in indirect immunofluorescence. Groups of six to eight week-old BALB/c were intramuscular inoculated singly with pCA-opti-CS, pCA-opti-G11, pCA-opti-r09 and PCAGGS vector, respectively. A total of immunity was three times interval of three weeks with the same dosage. Mice of each Group were divided into two parts after two weeks of the third immunity, then challenged with 10^3.87EID₅₀ of A/Swine/Guangdong/1/01(H1N1) and 10^7.45EID₅₀ of rPan09, respectively. Sera were collected, antibodies to HA and HI antibodies, level of IL-4 and IFN-γ were detected. The lungs were used for virus titer detection and histopathological observation. The result showed that HA can be expressed in 293T cell, plasmid pCA-opti-CS can induce high level of HA antibodies, high HI titer and can induce high level of IL-4 and IFN-γ. The result of determination of virus content in lungs and histopathological observation were showed that pCA-opti-CS DNA vaccine could effectively limited viral replication and attenuated histopathological damage in mice lungs. Based on our research, we conclude that recombinant plasmid pCA-opti-CS can induced high level of humoral and cellular immune response and own a good heterologous protecting capacity. Our study had made beneficial exploration in universal vaccine against influenza virus.

This experiment was conducted to compare the pathogenicity of ALV-J related acute fibrosarcoma extract inoculated on embryos and chicks. Acute fibrosarcoma extract was inoculated on yolk sac of 5-day-old-embryos, chorioallantoic membrane of 11-day-old-embryos and intraperitoneal inoculation of 1-day-old SPF chicks to compare pathogenicity of different inoculation methods. The results showed that the mortality of yolk sac inoculation was 14/30 at the age of 18-22 days, the incidence of visceral sarcoma was 8/14. The mortality of Chorioallantoic membrane inoculation was 17/30 at the age of 18-22 days, the incidence of visceral sarcoma was 6/17. As to the pathogenicity of the chicks, 13 hatched chicks from the chorioallantoic membrane inoculation group were all died, 11 of them developed visceral sarcoma. These results indicated that the pathogenicity of chorioallantoic membrane inoculation is higher than those inoculated on yolk sac of 5-day-old-embryos and those intraperitoneal inoculation of 1-day-old SPF chicks. Chorioallantoic membrane inoculation not only caused high incidence of visceral sarcoma, but also the sarcoma occurs earlier and faster, so it can be used as further acute fibrosarcoma model of artificial disease.

This experiment was conducted to study the immune protection effect of a Recombinant Pseudotype Baculovirus expressing the Sl protein of infectious bronchitis virus. Sixty-six 7-dayold SPF chickens were randomly divided into 6 groups, and were inoculated with Ac-V-EGFP (2×10⁹ pfu), Ac-V-S1, ^poly156S1 subunit vaccine, commercialized inactivated vaccine (M41) and PBS (control) respectively, boosted immunization was conducted at day 21. The results showed that, in the humoral immune, the inactivated vaccine group induced the highest level of IBV-specific ELISA antibody, significantly higher than other groups(P<0.01), in turn were prime-boost group and the Ac-V-S1 group; in cellular immunity, the Ac-V-S1 immunized group showed significantly higher than other groups(P<0.01), in turn were prime-boost group and the wild-type baculovirus group. Two weeks after the second immunization, the chickens were challenged with IBV M41 strain. The post-challenge results showed that the protective rate of Ac-V-S1, Ac-V-EGFP, and ^poly156S1 subunit vaccine were 55%(6/11), 27%(3/11), 37%(4/11), respectively, and the protective rate of the prime-boost immunity group was 64%(7/11), a little lower than the regular inactivated vaccine group 73%(8/11). Results demonstrated that the recombinant baculovirus Ac-V-S1 showed better effects on cellular immunity, which can make up for deficiencies in humoral immunity, and the prime-boost immunization strategy can further enhance the immune effect of recombinant baculovirus. This study laid the foundation of the research of recombinant baculovirus as genetic engineering vaccine of IBV.

The aim of this study was to construct a recombinant secreted Lactobacillus oral live vector strain co-expressing goose parvovirus (GPV) VP3 protein and goose interleukin-2 (gIL-2) protein. The VP3 gene was ligated with gIL-2 by enzyme digestion, and the signal peptide (SP) gene from Lactobacillus brevis 1.2028 was located on the N-terminal of fusion protein. The three genes (as a complete open reading frame) were subcloned into Lactobacillus integrated expression vector pMJ67, and then the recombinant plasmid pMJ-SP-GPVVP3-gIL2 was electro-transformed into Lactobacillus casei CECT5276 to construct the oral live vector vaccine strain. The recombinant Lactobacillus was identified by PCR method and immunogenicity of expression protein was tested by SDS-PAGE and Western blot methods. The specific anti-GPV antibody in serum, secretive immunoglobin A (sIgA) and spleen lymphocyte proliferation were detected to evaluate the immune effects after oral immunization goose with the recombinant Lactobacillus. The results showed that the recombinant plasmid has been constructed successfully, PCR proved that the plasmid pMJ-GPVVP3-gIL2 has been integrated into Lactobacillus genome, SDS-PAGE and Western blot could detect the GPV VP3-gIL2 protein band in supernatant and it could be recognized by the positive serum against GPV. Spleen lymphocyte proliferation responses results indicated that recombinant Lactobacillus could also induce an obvious cellular immune response, the lymphocytes proliferation level of GPV VP3-gIL2 group was obviously higher than GPV VP3 group and gIL2 group in the 4^th, 5^th and 6^th week. Neutralization antibodies could be produced in GPV VP3-gIL2 group, GPV VP3 group and gIL2 group, but the neutralization antibodies level of GPV VP3-gIL2 group was significantly higher than that of the latter two groups. Since the 2^nd week, the number of sIgA in intestinal mucosal fluid was significantly higher than GPV VP3 group and Vaccine group. Challenge experiment results showed that goose immunized with the GPV VP3-gIL2 fusion protein obtained 85% protection; the results confirmed that the recombinant Lactobacillus could enhance the protection against GPV. A recombinant oral live vaccine Lactobacillus CECT5276 (pMJ-GPVVP3-gIL2) secretive expressing fusion protein GPV VP3-gIL2 was constructed successfully, this recombinant Lactobacillus could be a basis to oral live vector vaccine for GPV infection.

To assess the immunoefficacy of recombinant Serpin against E. tenella infection in chickens, Serpin gene of E. tenella was cloned and expressed, subsequently, the recombinant Serpin was purified and refolded. Three experimental groups i.e. intramuscular injection, oral administration and intranasal administration groups were designed. At the same time, the challenged and unchallenged groups were also designed as positive and negative control, respectively. Seven-day old chickens were immunized with the recombinant Serpin protein,and boosted with the same method at 14 and 28 days of age. All chickens were orally challenged with sporulated oocysts at 35 days of age and killed at day 7 post challenge. Survival rate ,relative weight gain, oocyst decrease ratio and anticoccidial index were used as the main criteria for the assessment of the protective effect. The results showed the weight gains of the immunized chickens were significantly higher than that of the positive control group (P＜0.05). The oocyst outputs in immunized chickens dropped significantly when compared with the positive control. In comparison with the positive control, the ACI in immunized groups were significantly higher,and the ACI of group immunized orally with recombinant antigen was the highest, the results that the ACI of all immunized group were below 160 implied that the Serpin protein is not an ideal coccidiosis vaccines candidate antigen.

This experiment was conducted to study the role of cell signal transduction and cytoskeleton in invasion of primary chicken intestinal epithelial cell by human source S. flexneri. Primary chicken intestinal epithelial cell was treated with Genistein, sodium orthovanadate, staurosporine, Nifedipine and Cytochalasin B respectively, which are the corresponding inhibitor of protein tyrosine kinase, protein tyrosine phosphorylase, protein kinase C, Ca²⁺ channel signal transduction and microfilament polymerization. The influences of above inhibitors on invasion of primary chicken intestinal epithelial cells by ZD02 strain were detected by immunohistochemistry. F-actin distribution of primary chicken intestinal epithelial cells infected with Human S. flexneri ZD02 strain was detected by indirect immunofluorescence double labeling. Results were as follows: Genistein and Nifedipine significantly reduced internalization of ZD02 strain into primary chicken intestinal epithelial cells. In contrast, staurosporine and sodium orthovanadate have no effect on internalization of ZD02 strain. Cytochalasin B could prevent internalization of ZD02 strain. F-actin aggregated when ZD02 strain invaded into primary chicken intestinal epithelial cells. The process of human source S. flexneri invading into primary chicken intestinal epithelial cell depends on protein tyrosine kinase, Ca²⁺ channel activation and actin rearrangement.

This experiment was conducted to study the influence of Lactobacillus acidophilus (LAB1) on the focal adhesion kinase (FAK) phosphorylation and tight junction protein occludin expression. IPEC-J2 cells were infected by three concentrations of pathogenic Escherichia coli (E. coli) and Salmonella choleraesuis (S. cho), and then treated with LAB1. Occludin protein level was detected by ELISA, and FAK(Y³⁹⁷) phosphorylation was detected by Western blotting. Results were as follows: In LAB1 treated S. cho test, Occludin protein and FAK phosphorylation levels were positive correlated with the LAB1 concentration, but were negative correlated with S. cho concentration, and the differences were significant among groups that treated by mid and low concentrations of S. cho (P<0.05); In LAB1 treated E. coli test, Occludin protein and the level of FAK phosphorylation were positively correlated with LAB1 concentration, and the differences were significant among groups that treated by low concentrations of E. coli (P<0.05). Conclusion: LAB1 can activate the phosphorylation of FAK which inhibited by E. coli and S. cho, and raise regulate the cells occludin expression.

This experiment was conducted to detect existence of Gonadotropin-releasing hormone receptor (GnRHR) in the celiac-superior mesenteric ganglia (CSMG) of goats, and determine the possible influence of GnRH on CSMG. Five CSMG were taken from mature female and male goats， respectively. The distribution characteristics of GnRHR were observed after immunohistochemical SP staining. Image-Pro Plus 6.0 (IPP 6.0) was used as the method of semi-quantitative image analysis to evaluate expression difference of GnRHR between neurons and non-neuron cells. The results showed that strong GnRHR immunoreactivity (GnRHR-ir) products mainly distributed on the membrane and whole cytoplasm of neurons and weak GnRHR-ir products on non-neuron cells. Expression difference of GnRHR between these two sorts of cells was highly significant (P<0.01). The wide distribution of GnRHR in CSMG suggests that GnRH can exert potential effects on CSMG and neurons are the major targets, which implies GnRH may regulate the functional activities of the gastrointestinal system by sympathetic nerve from celiac superior mesenteric ganglion.

This study focused on the effects of co-administration of Bacillus subtilis RJGP16 and porcine Lactobacillus salivarius B1 on intestinal villus of piglets, as well as the antagonism between the probiotics and Escherichia coli K88 in vivo. Eight litters newborn piglets were orally administrated with B. subtilis (viable count 1.0×10⁹ CFU·mL^-1), procine L. salivarius (viable count 1.0×10⁹ CFU·mL^-1), these two kinds of probiotics (volume ratio 1:1) respectively, at 0, 7, 11 and 26 days after birth. After co-administration of the two probiotics, the piglets were inoculated with E. coli K88. The results showed that the co-administration of the two probiotics could significantly promote the villus height (VH) of duodenum (P<0.05) and ileum (P<0.01), increase the villus number of duodenum (P<0.05) and jejunum (P<0.01), lower the crypt depth (CD) of jejunum (P<0.05) and ileum (P<0.01), and increase the villus height/crypt depth (VH/CD) of duodenum (P<0.05), jejunum (P<0.05) and ileum (P<0.01). In addition, the co-administration of the two probiotics could effectively antagonize the injury caused by E. coli K88 to intestinal epithelium of piglets. These results suggested that the B. subtilis and L. salivarius could promote the development of the intestinal villus and increase the resistance to infection.

The research was conducted to prepare the polyclonal antibody against goat estrogen receptor β (ERβ) and build the specific immunohistochemitry assay to detect ERβ in the goat’s ovary and uterus. The ERβ gene was amplified from Jining grey goat ovary by RT-PCR. The cloning sequence was identified by enzyme digestion and sequencing, then the gene was cloned into a prokaryotic expression vector pET32a(+) to construct recombinant plasmid named as pET32a(+)-ERβ. The expression of fusion protein was induced by IPTG in E. coli BL21( DE3) system and then identified by SDS-PAGE and Western blotting. After purification with Ni-NTA under denaturing condition, the fusion protein was applied as antigen to immunize New Zealand White rabbits for preparing specific polyclonal antibody. The titer of the antibody was detected by ELISA, and the specificity of the antibody was detected by Western blotting; the tissue sections of goat’s ovary and uterus were assayed with immunohistochemistry method mediated by the prepared antibody. The results showed the protein of ERβ was expressed as fusion protein, 53 kDa. Western blotting result indicated that ERβ fusion protein could specially reacted with polyclonal antibody against human ERβ. The titer of the antibody detected with ELISA was 1:2¹³, and the prepared antibody combined to ERβ was specific.the results of immunohistochemistry assay indicate ERβ mainly exists in the granule cells of ovary follicles and the epithelial cells and smooth muscle cells of the uterus endometrial layer. In conclusion, the polyclonal antibody against goat ERβ are successfully obtained in the present study and the ERβ distribution in the ovary and uterus of Jining Grey goat was firstly reported with immunohistochemistry assay mediated by the antibody against goat estrogen receptor β.

To reveal the effects of interferon τ (IFN-τ) and progesterone on the expression level of extracellular matrix (ECM) and their ligands genes in bovine endometrial cells (bECs). In the study, bECs were cultured in medium containing bovine IFN-τ (bIFN-τ), progesterone and combination of bIFN-τ and progesterone, respectively, and then expression level of ECM related genes (Collagen, laminin, Fibronectin, THBS and Tenascin) and their ligand genes (Syndecan, CD44, CD47 and RHAMM) in bECs were detected by quantitative PCR after in vitro cultured bECs were treated for 24 h with bIFN-τ, progesterone, combination of bIFN-τ and progesterone, respectively. The results showed that, bIFN-τ significantly induce the expression of Tenascin, Collagen, RHAMM and Laminin, and progesterone significantly induced the expression of CD44, Fibronectin, RHAMM, and Laminin, but under co-action of bIFN-τ and progesterone, the expression level of CD44 and THBS was inhibited, and the expression of Tenascin, Fibonectin and Collagen returned to the expression level in the control group. The results suggest that bIFN-τ and progesterone might regulate the expression of ECM and their ligands in the process of embryo implantation.

In order to isolate the ES cells from Kunming mouse efficiently, the feeder cells, developmental stage of embryos and culture conditions were optimized in the study. Mouse embryonic fibroblast (MEF) were inoculated at concentrations of 1×10⁴, 1×10⁵ and 1×10⁶·mL^-1 as the feeder layers of ES cells after they were treated with Mitomycin C within 3 passages, and cultured in H-DMEM+15%KSR+LIF, the growth of the ES cells were observed. The effects of developmental stages of embryos and added SCF, SCF+insulin in culture medium on growth of Kunming mouse ES cells were investigated. The results showed that the formation rates of the 1^st and 2^nd passage ES cells in 1×10⁵·mL^-1 MEF were higher than those cultured on the other 2 groups(P<0.05). The 2^nd passage ES cells colonies formation rate of blastocysts were higher than that of compacted morula(P<0.05). It had improved the attachment rate was significantly enhanced(P<0.05) when the culture medium added with SCF, and the highest attachment rate, 1^st and 2^nd passage ES cells colonies formation rate were got when the culture medium added with SCF and insulin (P<0.05). The isolated ES cells, which were positive for AKP staining and immunofluorescence against antigens of Oct-4 and SSEA-1, had a series of characters of mice ES cells. These results suggest that the blastocysts on the MEF with a density of 1×10⁵·mL^-1 and addition with SCF and insulin in the culture medium is more suitable for the isolation and passage of Kunming mouse ES cells.

This experiment was conducted to investigate the effects of Lycium barbarum polysaccharide (LBP) on cell proliferation of chicken Lymphocytes in vitro. Different concentrations of LBP were added into cultured with plants hemagg-lutinin (PHA) stimulated chicken thymus, spleen and peripheral blood lymphocyte, and cultured with bacteria lipopolysaccharide (LPS) stimulated chicken bursa of Fabricius, peripheral blood lymphocyte, spleen, MTT method and ELISA method were used to determine the lymphocyte proliferation and IL-2 excretion respectively. The results showed that: (1) LBP could stimulate T, B lymphocyte proliferation and IL-2 excretion in certain concentration range and with the increase of the concentration of polysaccharide increase; (2) The OD value of T, B lymphocyte with the source of the same organ were significantly growth trend with chickens dayage growth in same concentration of LBP. (3) The OD value of thymus T lymphocyte and bursa of Fabricius B lymphocyte were higher than in peripheral blood and the spleen T, B lymphocytes with the same concentration LBP and the same day age of chicken. These results indicated that LBP could stimulate T, B lymphocyte proliferation and IL-2 excretion，and the promoting function of lymphocyte proliferation have certain concentration-response, time relationship and organizational correlation.

This experiment was conducted to investigate the distribution and the dominant serotype’s mutation of avian Salmonella in Eastern China. Using the methods of enrichment, purification, Polymerase Chain Reaction (PCR), serotype identification and biochemical identification, 1 730 samples were detected and seven isolates with mutated serotypes were sequenced to analyze serotypes of avian Salmonella. The results showed that 87 isolates were identified as Salmonella and belong to five subtypes, including S. typhimurium, S. derby, S. saintpaul, S. pullorum and S. abaetetuba. Furthermore, Hd and Hg were mutated in seven S. pullorum isolates. These results indicated that poultry were mainly infected by S. pullorum in Eastern China. The serotype of the seven S. pullorum isolates mutated and the mutated rate was increasing. The research provided the experimental evidence of serotypes’ distribution and mutated serotypes in Salmonella isolates.